Immpact Report

MCGOVERN MEDICAL SCHOOL’S BROWN FOUNDATION INSTITUTE of MOLECULAR MEDICINE FOR THE PREVENTION of HUMAN DISEASES IMMpact Report

FISCAL YEAR 2015 About the cover

McGovern Medical School’s Brown Foundation Institute of Molecular Medicine for the Prevention of Human Diseases is led by Dr. John Hancock, center.

IMMpact Report is published by McGovern Medical School. All correspondence should be addressed to: Office of Communications 6431 Fannin, B.340 Houston, TX 77030 E-mail: [email protected]

Articles and photos may be reprinted with permission.

Editor Darla Brown, Director, Office of Communications

Contributors: Darla Brown Rob Cahill John Evans IMM Faculty

Design: Roy Prichard

For information on supporting programs at the McGovern Medical School and the IMM, contact Lise Cameron, 713.500.3602, [email protected]

OC-AbsoluteColor-1450-1/16 IMMPACT REPORT 1 • • • •

8 14 12 creates a creates “rising star” “rising IMM pioneer CPRIT funds against cancer legacy of support physics in the war stem cell biologist Brandishing math, Brandishing

6 11 behind aging Metabolic and Metabolic join Center for The basic science Two investigators Two Degenerative Diseases Degenerative Features Director's Message Director's Locations Our Genetics Center for Cardiovascular Genetics Center for Human Diseases and Autoimmune Center for Immunology Diseases and Degenerative Center for Metabolic Imaging Center for Molecular Biology and Systems Center for Proteomics Medicine Cell and Regenerative Center for Stem and Aging Research Engineering Tissue Center for Institute Therapeutics Texas IMM Service Centers the Numbers By Report Gift Contents 2 3 Mission 4 16 20 25 32 41 49 58 73 80 89 92 93 • 2 IMMPACT REPORT 2 1 Director’s Message objectives: two major The IMMhas benefit from discovery. this goalofpatient specifically toachieve Therapeutics Institute talent intheTexas IMM hasorganized medical solutions.The IMM focusesonthese benefit ofpatients.The of discoveryandtothe therapies arederivative New diagnosticsand to newsolutions. defects. Discoverieslead caused bysinglegene problems thatarenot are unsolvedmedical cardiovascular diseases cancer, Alzheimer’s,and since diabetes,obesity, is amajorchallenge, the IMMfaculty.This highest priorityfor Discovery isthe I John S.Dunn Distinguished University Chair inPhysiology andMedicine Executive Director, Institute ofMolecular Medicine John Hancock, M.A.,M.B., B.Chir., Ph.D., Sc.D. science anditsimplicationsfor thefuture ofmedicineandhealthcare. tomeetwiththemand discusstheir our facultyandhave theopportunity your calendarbecauseyou willhearexciting research storiesdirectly from symposium, whichwillbeheldonApril 20,2016.Please mark thisdatein at theIMM.Alsowe wouldbedelightedtoseeyou attheannualIMM delighted totalkwithyou personally, sopleasefeelfree tocontactushere growth anddevelopment oftheIMM. leadership ofMr. Dudley Oldham, whichplaysakeyrole inthecontinued council,underthe the strong work anddedicationoftheIMMadvisory therapies forhumandisease.In thisregard we are deeplyappreciative of the aspirationofourmissiontotranslatemoleculardiscoveries intonew develop ourrelationshipsfurther withallinourcommunitywhovalue friends anddonors. collaborations, and,mostimportantly, thecontinuinggenerosity ofour from alternativesupport sources, includingresearch charities,industry implementation ofourmissionremains heavilydependentonattracting research fundsfrom whatisanever-diminishing pool.Thatsaid,full of ourscientiststhattheIMMfacultyremains sosuccessfulinattracting the preceding year. It isaremarkable testamenttothequalityandcreativity grantsandcontractswerejust ended,ournew substantiallyupagainover IMM facultycontinuetobeextremely successful.Over thefinancial year environment forscientific research funding, especiallyfrom theNIH, details ofwhichare describedinthereport. createdthe newly IMMCenterfor Tissue Engineering andAgingResearch, research teamfrom theUniversity ofPittsburg. Dr. Huard willbeleading This includesthe of recruitment Dr. Johnny Huard andhisnearcomplete who bringwiththemexciting andinnovative research programs. excellent examplesofthesecollaborative teams. flagship programs in development,regenerative provide medicineanddrug metabolic anddegenerative diseases,molecularimaging,andtwoofour who collaboratecloselywithourclinicalcolleagues.Thecentersfor this endwe have teamsofoutstandingbasicandtranslationalscientists outcomes from research inmolecularmedicinethatbenefitspatients. To McGovern Medical School.Our missionistodeliver translational you willfind the interestingreport andinformative. describing theinnovative research inwhichtheyare that engaged.Itrust on recent developments IMMfacultymember andanaccountfrom every Human Diseases (IMM).Inside you willfindin-depthfeature articles Foundation Institute ofMolecular Medicine forthePrevention of am pleasedtointroduce ourannualIMMpactreport forThe Brown If you wouldliketoinvestigate how you canbeinvolved, we wouldbe In additiontoadvancing scienceandmedicine,we therefore wishto I amalsopleasedtonotethatdespiteapersistentchallenging This facultytoIMM, year hasseenthesuccessful ofnew recruitment The IMMisastand-alone research institutethatisembeddedwithin IMMPACT REPORT 3 • • • • As an institute of McGovern As an institute of McGovern to medical practice are of to medical practice are major importance to product in biotechnology development and the pharmaceutical the IMM has industry, to the potential and desire form important and links its collaborations between activities and research own industries to apply its various and intellectual discoveries properties to pharmaceutical opportunities. School, the Brown Medical of Institute Foundation for the Medicine Molecular Diseases of Human Prevention to set the example strives and excellence for research nationally, collaboration locally, and internationally. However, it is clear that it is clear However, the applications Because research and the future practice and the future research novel of medicine with more therapies. These approaches been most successfully have used to determine the causes of and genetic infectious disorders diseases. molecular and cell biology in will play a major role clarifying the causes of many of modern problems unsolved medicine, such as heart disease, hypertension, vascular major mental disorders, illnesses, and inflammatory and immunologic diseases. of the institute’s research The is inspiring and investigators to fulfill the mission promises of the IMM. of molecular and cell biology

he Brown Foundation Foundation he Brown of Molecular Institute for the Medicine

Advances in molecular and Advances Mission

Prevention of Human Diseases Diseases of Human Prevention institute (IMM) is a research the that seeks to investigate causes of human diseases at the cellular and molecular using DNA and protein levels, technologies to elucidate disease mechanisms. This development of particular are and progress planning for future interest important in the increasingly The of clinical research. area to design institute endeavors methods of rational therapy possible, and, wherever of strategies for the prevention human diseases. enormous cell biology have medical potential for innovative T • 4 IMMPACT REPORT at St. Luke’s Episcopal Hospital The Denton A.Cooley Building – Texas Heart Institute South CampusResearch Building –3(SCRB3) Fayez S.Sarofim Research Building Our Locations • • • • • • • • Located inthe Texas Medical Center. tech laboratory. The IMMoccupiesa31,000square-foot high- Cyclotron, offices. wet labs,andsupport Imaging, Optical Imaging Tracers, a Emission Tomography, Magnetic Resonance Opened in 2009,thisfacilityhousesPositron South Campusofthe Texas Medical Center. Six-stories, 315,000square-feet locatedonthe GE Healthcare andthe Texas Enterprise Fund. Center andUTHealth, incooperationwith University of Texas MDAndersonCancer SCRB3 isacollaborationbetween The 255,748 gross square feet. Opened in2006,thebuildingencompasses the Texas Medical Center. (UTHealth) University Center Tower within Texas Health ScienceCenteratHouston Located adjacenttotheThe University of staff. administration, andsupport Primary homeoftheIMM’s faculty, IMMPACT REPORT 5 • • • • Johnny Huard, Ph.D. Huard, Johnny Vice & Professor Wallace Distinguished Chair for Research Engineering Tissue for Center Director, Research and Aging Reversing Aging Reversing Wednesday 20, 2016 April 4-6 p.m. Building Research S. Saroﬁm Fayez Street 1825 Pressler e Institute of e Institute for the Medicine Molecular Diseases of Human Prevention IMMpact Symposium ATE D THE

Eva Zsigmond, Ph.D. Eva and Director, Assistant Professor Facility Core Cells and Stem Transgenic Treating Macular Treating Degeneration AVE S Assistant Professor, Center for Metabolic for Metabolic Center Assistant Professor, Diseases and Degenerative Stress and Alzheimer’s Ph.D. Justice, Nicholas • 6

IMMPACT REPORT Profiles in research “ A Research Institute. of theSteadman Philippon and isthechiefscientificofficer Inc., abiotechnologycompany, co-founder ofCookMyoSite, Regenerative Medicine. He is at theMcGowan Institute for Cellular Therapeutic Research and wasdeputydirector for atPittsburgh,Orthopaedics, of Therapeutics, Department for Musculoskeletal Cellular Huard as thevicechair served rehabilitation. Additionally, Dr. physical medicineand pathology, pediatrics,and genetics, bioengineering, microbiology andmolecular surgery,in orthopaedic professor appointments scientist, Dr. Huard held research. center dedicatedtothisarea of as thedirector IMM ofanew the University ofPittsburgh, the IMMMay 1,2015,from strength aswe grow older. us tomaintainourhealthand age nottostoptimebuthelp seeking answers abouthow we the clock,IMMscientistsare make claimstoturnback vials, displayed underglass. of youth packagedintojarsand store toseethevarious promises — Dr. JohnnyHuard machine with youryoungstem cells,puttingthe youthfulstuffback in. We see afuturewheretoimproveagingyou wouldbehookeduptoa An internationallyrenowned Johnny Huard, Ph.D., joined While theseproducts may The basicsciencebehindaging favorite department favorite department Just walkintoyour nti-aging isbigbusiness. which causes them to die at 21 which causesthemtodieat 21 modified micehave progeria, prematurely. Thesegenetically his teamusemicewhoage aging process, Dr. Huard and stem cellschangeduringthe noon?” When we are 50,istheclockat born, isourclocksetat12a.m.? internal clocks. When we are understand theimpactofour Surgery. “We alsowantto ofOrthopaedic the Department and thevicechairforresearch in Huard, whoalsoisaprofessor and notasgood,” explainsDr. become tired anddefective, do todelayaging? How dowe age? What canwe answer basic questions,suchas: center couldprovide.” work, which issomethingthis not have a lotofbasicscience Medicine) whotellsmetheydo of Geriatric andPalliative Dyer (director oftheDivision says. “I’ve talkedtoDr. Carmel resources together,” Dr. Huard combine allofouraging field attheuniversity. those already working inthe establish collaborationswith Research, Dr. Huard says,isto Engineering andAging goal oftheCenterfor Tissue School, themostimmediate To understandwhyour “As we age,ourstemcells The center’s scientistsseekto “It wouldbegreat to At McGovern Medical are heading.” gene therapy–thisiswhere we says. “Cell,molecular, tissue, sharper mentalcapabilities. mass, stronger bones,and could result inretained muscle science behindaging,which University to uncover thebasic Rice University, andA&M Baylor CollegeofMedicine, team work withcolleaguesat UTHealth, Dr. Huard andhis back in.” cells, puttingtheyouthful stuff machine withyour young stem you would behookeduptoa future where toimprove aging blood, thenreplaces it. We seea to amachinethatcleansyour dialysis, you are hookedup kidney problems andare on you have he explains.“When opposite ofkidneydialysis,” or whenwe age. later inlife,whenwe are injured thatcanbe used cells atbirth Huard says,istocollectstem Dr. Huard says. their lifespanby three times,” aging mice,we canincrease cells intotheseprematurely compared toayoung mouse. in numberanddifferentiation not proliferate andare limited The progeria mice’s stemcellsdo days insteadofin2to3years. “Science is a team effort, he he “Science isateameffort, Beyond collaboratorsat “Think ofthisasthe The goalforhumans, Dr. “By injectingyoung stem ” IMMPACT REPORT 77 • • • •• • • • Dr. Johnny Huard is working to help us maintain health and strength as we age. as we to help us maintain health and strength is working Huard Johnny Dr.

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IMMPACT REPORT Profiles in research “ W help us recruit some of the most help usrecruit someof themost been tremendously to important chronic diseases. discovery for stem cellsfordrug platform ofpatient-derived Therapeutics Inc., anintegrated co-founder ofMultiClonal Cell Institute. She isalsothe she wasamemberofitsStem School ofMedicine, where the University ofConnecticut McGovern Medical Schoolfrom human cancers. stem cellsfrom themostlethal these technologiesforcloning tract, andhasnow adapted as thoseofthegastrointestinal normal, regenerative tissuessuch the cloningofstemcellsfrom transformative technologiesfor Dr. Xian hasdeveloped investigator Wa Xian,Ph.D. torecruit stemcell (CPRIT) & Research Institute of Texas from theCancer Prevention was awarded $3.7million at themolecularlevel. researcher whoistacklingcancer recently recruited a new Prevention ofHuman Diseases of Molecular Medicine forthe Brown Foundation Institute in theLoneStar State, the of disease. the understanding that canbeusedto developcelltherapiesandfurther Dr. Xianbrings cutting-edgeapproachestotissue-specific stemcells “The CPRITprogram has Dr. Xian comesto McGovern Medical School cancer research program toadvance ith theaidofa CPRIT funds“rising star” ” stem cellbiologist — Dr. BrianDavis newly diagnosed with cancer diagnosedwithcancer newly 109,053 peoplein Texas willbe estimatesthat Cancer Registry College ofMedicine. a postdoctoralfellow atBaylor Harvard Medical Schoolandas training asaresearch associateat She completedpostdoctoral Anderson CancerCenter. The University of Texas MD in moleculargeneticsfrom Regenerative Medicine. Center forStem Cell& Ph.D., director oftheIMM’s of disease,” saysBrian Davis, theunderstanding and further used todevelop celltherapies specific stemcellsthatcanbe edge approaches totissue- them toUTHealth,” headds. tohelpusrecruit important iscritically CPRIT support leaders inthisarea andthe into ourfuture institutional investigators whowilldevelop Distinguished Professorship. the Roger J.Bulger, M.D., research affairsandholderof president foracademicand Stancel, Ph.D., executive vice areas inthefuture,” saysGeorge in thisandrelated research to continuebuildexcellence cancer research whowillhelpus outstanding young scientistsin This calendar year, the Texas Dr. Xian received herPh.D. “Dr. Xian bringscutting- “These are thetypeof young development research programs. prevention andproduct through itsacademicresearch, institutions andorganizations. in grantsto Texas researchers, to dateawarded $1.35billion operations in2009,CPRIThas in services Texas. Beginning and prevention programs and groundbreaking cancerresearch $3 billioninbondstofund authorizing thestatetoissue by Texas voters in2007, cancer patients. personalized treatments for assist oncologiststooptimize will bestandard-of-care to cancer stemcelllibraries time inwhichpatient-specific at McGovern Medical School. says Dr. Xian,assistantprofessor comes backasarepeat disease,” chemotherapyand that survives nasty subset target aparticularly cancer stemcellstoidentifyand patient-specific librariesof recurrence, Dr. Xiansays. to treatment andcouldcausea a tinynumbermayberesistant cancer cellswithchemotherapy, the vast majorityofovarian to thedisease. and that42,255willsuccumb CPRIT provides funding CPRIT wasestablished Dr. Xian anticipatesa “In mylab, we are generating While oncologistscankill IMMPACT REPORT 99 • • • •• • • • Dr. Wa Xian is using stem cells to fight cancer. Wa Dr.

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IMMPACT REPORT Degenerative Diseases. Dr. KristinEckel-Mahan andDr. KaiSun are thenewest members oftheCenter forMetabolic and IMMPACT REPORT • • • • 11 Profiles in research in Profiles “I study the clock from from “I study the clock scientific studies point Many goal of an organism, “One started Eckel-Mahan Dr. when “I saw it mattered of her studies is area One says the Kolonin Dr. “Both the established and axis. Other cues, such as food, cues, such axis. Other body’s influence the strongly peripheral clocks. that a stricter the perspective to our internal adherence clock 24-hour biological is important for metabolic she explains. health,” from to health issues resulting disruption of the circadian clock – insulin resistance, diabetes, and even obesity, cancer. metabolically speaking, is for its she clocks to be in synchrony,” says. clocks in her focus on circadian graduate school. testing animals – were we We what time of day it was. that feeding also discovered an animal a high fat diet the dramatically reprograms she adds. clock,” liver which is night eating syndrome, when someone eats 25 percent of their calories during or more period, after their a typical rest meal. evening addition of these new recruits will help bridge the center’s efforts. research excited are the new investigators about the clear cohesion of our efforts in the that will synergize of everystrides individual group important discoveries,” toward he says. Dr. Sun’s research focuses focuses research Sun’s Dr. and fibrosis “Inflammation of A graduate of University to want my lab we “In the IMM in May Joining clocks everywhere, have We responds The brain clock research equipment helps him research delay efficiently – without work to another of sending samples results. and waiting on source fat between on the relationship remodeling tissue pathological tries He and type 2 diabetes. to identify critical factors with clinical significance in the patholophysiology of human obesity and insulin resistance. of unhealthy fat,” hallmarks are he explains, adding that white focus. fat is his research Sun Dr. California, Riverside, molecule called a has discovered accelerates the that endotrophin and inflammation in fibrosis obese fat tissue. this when and how discover – could it be molecule is created obesity? Could for a biomarker of it be targeted for treatment obesity and type 2 diabetes?” asks. Sun Dr. Eckel-Mahan 2015, Dr. the from her Ph.D. received and Washington of University continued her training at the of California, Irvine. University of focus is the role research Her clock in the 24-hour circadian health and disease. explains. Eckel-Mahan Dr. brain clock and “master” The the peripheral tissue clocks are the body. located throughout primarily to the 24-hour induced cycle light and dark on its rotation the earth’s by

harged with understanding metabolic the health and disease,

Two investigators join Center for Center join investigators Two “It was an interesting was an interesting “It The new talented faculty are they been only have “Not who joined Sun, Dr. critical received “I have Metabolic and Degenerative Diseases Degenerative and Metabolic

IMM’s Center for Metabolic for Metabolic Center IMM’s Diseases and Degenerative two new faculty welcomes members. These investigators qualified stood out as uniquely nearly 100 candidates. from challenge identifying those applicants whose portfolio with everywould resonate says principal investigator,” and Center for Metabolic Director Diseases Degenerative Ph.D. Kolonin, Mikhail whose M.D., Ph.D., Kai Sun, in obesity are interests research and diabetes, and Kristin Eckel- who studies Ph.D., Mahan, circadian of the body’s the role clock on health. laboratories and top trained by published in first-tier journals, they also engaged every current faculty member as potential collaborators that could bridge the distinct scientific interests composing of individual groups explains Dr. the center,” Jr. the Harry E. Bovay, Kolonin, Chair University Distinguished Research. Disease in Metabolic 2015 the IMM in March Texas of the University from as an in Dallas Southwestern says the assistant professor, has atmosphere collaborative been key. input and support for hiring he and lab organization,” says, adding that the available C 12 •

IMMPACT REPORT Profiles in research W by chanceIbecamefriends University ofMinnesota when my engineeringfieldatthe complex materialsandfluids. hiscareerwho started studying trained chemicalengineer, Reuters, Dr. Cristini isa Yale- Minds in2014by Thomson Most Influential Scientific cancerous tissues. with mathematicalmodelsto applies thelawsofphysics field of research, which pioneers inthisinnovative Systems Biology, isoneofthe the CenterforProteomics and physics, Isuggested.” cancer indeedobeysthelawsof falls totheground, ofcourse.’ and thenreleases it.He said,‘It takes thattumorinhishand operation, whathappensifhe from apatient duringan removing acancerous tumor recalls withasmile. nothing todowithcancer,” he I wascrazy. Thatphysicshas and thisguystoodupsaid It earlyinmycareer, wasvery institute. topic ataprestigious cancer he delivered alecture onthe Vittorio Cristini, Ph.D., after audience memberonceasked “I wasdoingresearch in Named asoneofthe World’s Dr. Cristini, professor in “Well, then,itseemsthat “So Iaskhim–whenheis “I wassothrilledtobethere. That’s whatoneoutraged physics? have todowith hat docancertumors Brandishing math, physics in thewar against cancer of Imaging Physics at The a professor oftheDepartment and Biomedical Engineering, ofNanomedicineDepartment the outcomesare poor.” are properties transport poor, biomarkers: usually, whenthe novel physics-derived cancer we are findingaresays. “What and retrospective trials,” he patients –bothprospective mathematical modelsincancer outcomes by developing how theycorrelate withclinical andfinding properties transport pharmaceuticals. tissue andhow theyrespond to tumors are through transported colleagues studyhow cancerous problem.” physics problem asabiological says. “Cancerisasmuchofa physics oftissueingeneral,” he physics ofcancerandthe applying physicstocancer. down research anew path– moment thattookDr. Cristini crystals.” as governing thegrowth of materials andfluids,such ofthecomplex properties were similartothephysical governing how tumorsinfiltrate studying. Thephysicallaws similarities withwhatIwas tumors inmice,andInoticed the morphologyofcancerous Cristini recalls. “He showed me with apharmacologist,” Dr. Vice chairofthe “We are lookingatthe Dr. Cristini andhis “I aminterested inthe That wasthelightbulb are universal.” the biology. Thelawsofphysics we canchangetheoutcomeand can directly managethephysics, Butphysical properties. ifwe underlying causeofdifferent explains. “Biology isthe properties,” Dr. Cristini correspond totransport medication. Thesedifferences respond differently tothesame regions inthesametumorthat a specificpatient. treatment thatisoptimized for role,” Dr. Cristini explains. playsanimportant perfusion of thatpatient,andblood on theuniquetissueproperties formulas. on tissue-specificmathematical respond based tocancerdrugs, on how individualpatients now onconfirmingtheirtheory Houston,” headds. researchers are righthere in this field,andmanyofthe network ofcollaborationin biology. the director ofcomputational endowed professorship andwas Mexico, where heheldan from theUniversity ofNew to McGovern Medical School Sciences, Dr. Cristini came Graduate SchoolofBiomedical and onthefacultyof Anderson CancerCenter, University of Texas MD “We have identifieddifferent The goalistocreate acancer “The mathmodelsare based The investigators are focused “We have anextensive IMMPACT REPORT • • • •• • • • 1313 Dr. Vittorio Cristini applies the laws of physics to cancerous tissues. applies the laws of physics to cancerous Cristini Vittorio Dr.

14 • IMMPACT REPORT Dr. Irma Gigli, secondfrom left,helpscutstheribbon duringtheIMM’s dedicationceremony in 2006. IMMPACT REPORT • • • • 15 Donor spotlight Donor Dr. Gigli also recently made also recently Gigli Dr. is funded The endowment firm father was a “My “I wanted to do something will make a generosity Her is so meaningful to have “It achieved over these decades. over achieved the extraordinary decision to an estate commitment leave Theof $2 million to establish MD Gigli, and Irma Gigli Luis at in Immunology Endowment the IMM. and her Gigli Dr. money by their inherited from brother parents. and in philanthropy believer in education because he was an orphan at age 7 and did not the opportunityhave of higher she explains, adding education,” who that although her father, for well died at age 59, provided he did not provide the family, Gigli’s Dr. them with luxury. died suddenly Luis, brother, in 2014. a stroke from specific for students that to my has a relationship she says. “Supporting family,” scientists is extremely young important, and immunology is what both my husband and on our whole I had worked scientific life.” to the IMM. difference founding one of the IMM’s back in such a leaders give Hancock, F. John says Dr. way,” of the IMM. director executive gifts help “These generous of much put us on the level larger institutions in terms of for faculty and student resources recruitment.” It was in 1995, when she It says the IMM’s Gigli Dr. dedication to Gigli’s Dr. When her husband passed infused the recently She says her late Gigli Dr. scientists to bridge basicscientists to bridge clinicalimmunology with published she has dermatology, original manuscripts 160 over manyand book chapters, of which identify basic in host mechanisms involved defense and in the development of skin diseases. and her late husband Professor J. Muller-Eberhard Hans T. James Dr. by recruited were to start the IMM. Willerson has always been clear. direction trying were to put together “We that had connections in areas medical biology to understand she says. the basics of disease,” many ways, I am romantic “In do science about science – you science.” love because you science and the founding principles of the IMM have been unwavering. Gigli, away in 1998, Dr. emerita of the IMM director and Center for Immunology created Diseases, Autoimmune Muller-Eberhard Hans J. The at the Chair in Immunology IMM in his memory to advance to which he the research dedicated his life. Memorial Muller-Eberhard with an Series Lecture additional $50,000, which was the UTHealth matched by Changers Initiative. Game husband would be happy with the IMM has the progress IMM pioneer IMM

creates a legacy of support legacy a creates

ne of the founding leaders of the Institute Medicine for Molecular

The innovative spirit of The innovative institutions were “There up in Argentina Growing on to went Gigli Dr. of the early physician- One

is also one of its strongest is also one of its strongest supporters, who has overcome herselfbarriers to establish field ofas a pioneer in the translational medicine. Gigli, Irma Emerita Professor M.D., led to the creation in of the first program immunodermatology sponsored of Institutes National the by historic feat in a – a Health male-dominated environment. could at the time you where count the number of women on she says. one hand,” Gigli during the 1940s, Dr. was determined to become a physician. After graduating from medical school, she traveled to pursue States to the United in interests her joint research dermatology and immunology. took off, rising from career Her residency her first medical in at Cook County Hospital Hughes Chicago to the Howard and Miami, in Institute Medical then on the faculty of Harvard School. Medical of her become the director chair of the laboratory, own atDepartment of Dermatology of California at the University and elected to the Diego, San of Medicine Institute prestigious of Academy of the National Sciences and the American of Arts and Sciences.Academy O 16 • IMMPACT REPORT T CENTER FORCARDIOVASCULARGENETICS through pilotrandomized placebo-control trials. animal modelsandthenextended tohumans studiesare pursuedincelland intervention to prevent andreverse thephenotype.Theinitial pursued toblockthelinking pathwaysinorder are genetic andpharmacological interventions genetic mutationsandtheclinicalphenotypes, elucidation ofthemolecularlinksbetween the the causalvariants andthephenotype.Upon order todelineatethemolecularlinksbetween in cellculture systemsandanimal modelsin gene expression. Thefindings are thenapplied tissue todeterminechangesintheregulation of complemented withgenomicanalysisofcardiac cardiomyopathies. Genetic discoveries are cardiovascularfor hereditary diseases,primarily of theseactivitiesistoidentifythecausalgenes molecular geneticstudies.Themainobjective members, phenotypiccharacterizationand recruitment oftheprobands andfamily human moleculargeneticstudiesthrough already have developed thedisease. attenuate theevolving phenotypeinthose who toreversethe phenotypeenablesinterventions or molecular pathwaysthatlinkthemutationsto manifestations ofthedisease.Delineation ofthe the diseasepriortodevelopment ofclinical Genetic studies afford toprevent the opportunity elucidating theirmoleculargeneticcauses. cardiovascular diseasesinhumansthrough activities. is utilized forclinicalresearch also hasaResearch Clinic,which Outpatient Clinic.TheCenter at the Texas Heart Institute the Cardiovascular Genetic Clinic cardiovascular disorders through topatientswithgenetic services provides specialized clinical St. Luke’s Health, thecenter at the Texas Heart Institute at Denton A.CooleyBuilding on theninthfloorof diseases inhumans.Located pathogenesis ofcardiovascular of moleculargeneticsand I. Specific themesof research include: The research programs attheCenterentail Our mission istoprevent focuses onelucidation Cardiovascular Genetics he IMMCenterfor Human moleculargenetics/genomics Research Distinguished Professorship inCardiovascular George Josephine andMary Hamman Foundation Center Director &Professor AJ Marian, M.D. medical interventions. cardiovascular medicinetoguide appropriate the above studies are appliedtothepracticeof diseases. Genetic informationgarnered through cardiovascularhuman patientswithhereditary the beneficialeffectsofexperimentaltherapiesin bench are extendedtohumanpatientstest the proteins intheheart. approaches are usedtocorrect thedeficiencyof vitro andinvivo genetherapy studies.Likewise, genetically targetedinmyocytes through in of thephenotypeare pharmacologicallyand pathways thatare responsible fortheinduction delineation ofthemolecularmechanismsspecific genetically engineered animalmodels myocytes through genetransferstudiesandin diseases: Thesestudiesare conductedincardiac variants identified inhumanswithcardiovascular Arrhythmogenic Cardiomyopathy. Cardiomyopathy, Dilated Cardiomyopathy, and cardiomyopathies; namelyHypertrophic focus onthree mostcommonformsofhereditary cardiovascular diseasesinhumanswithaspecific the moleculargeneticandgenomicbasisof Studies: Thesestudiesare designedtodelineate IV. III. II. Clinical Studies: Thediscoveries atthe Experimental Therapies: Upon Functional characterizationofthegenetic IMMPACT REPORT • • • • 17 Cardiovasc Cardiovasc AJ. Marian Rodriguez G, Lombardi R, PMID: 22987565 Jan 1;97(1):44-54. 2013 Res. functional Human molecular genetic and encoding Muscle TRIM63, studies identify as a novel gene for RING Finger Protein 1, Chen human hypertrophic cardiomyopathy. Jin J, Lombardi R, Y, Tan Czernuszewicz G, SN, Res. 2012 Sep Circ AJ. Marian Willerson JT, PMID: 22821932 14;111(7):907-19. Nuclear plakoglobin is essential for differentiation of cardiac progenitor cells to adipocytes in arrhythmogenic right ventricular cardiomy- da Graca Cabreira-Hansen R, Lombardi opathy. AJ. Marian Willerson JT, Fromm RR, A, Bell M, PMID: 2011 Dec 9;109(12):1342-53. Res. Circ 22021931 LAB MEMBERS Ph.D., Auguste, Post-doctoral Fellows: Gaelle Ph.D. Jennifer Karmouch, Ph.D., Auguste, Gaelle M.S. Associate: Grace Czernuszewicz, Research Gurha, Assistant Professors: Priyatansh Faculty – Ph.D. M.D., Raffaella Lombardi, Ph.D., Integrated genomic analysis in a mouse model of dilated cardiomyopathy: Genomic data in- H3K9ac, cludes genome-wide CpG methylation, and H3K27me3 histone modifica- H3K27ac, along with transcriptomic data assigned tions, to the mouse chromosomes. trial to improve symptoms and exercise tolerance in patients with hypertrophic cardiomyopathy N-Acetylcysteine in Hypertrophic Cardio- myopathy) clinical trial (ClinicalTrial.Org NCT01537926) molecular pathways that link the genetic mutations to the clinical phenotype in patients with cardiomyopathies including delineation of the mechanical signaling pathways regulated at the intercalated discs genetic and transcriptomic changes including non-coding RNAs and histone modifications in hereditary cardiomyopathies and sudden cardiac death KEY PUBLICATIONS And Is a Causal Activated The Hippo Pathway Is Mechanism For Adipogenesis in Arrhythmogenic Lombardi R, Gurha P, Chen SN, Cardiomyopathy. - Circula AJ. Marian Willerson JT, Ruggiero R, PMID: 2014;114:454-468. . tion Research 24276085 Associ- is Variant A Rare Loss-of-Function SCN5A ated With Lidocaine-induced Ventricular Fibrilla- Hong K. AJ, Marian Hu J, Cao L, Xiong Q, tion. PMID: 2014 Jan 21, , Pharmacogenomics 24445991 Pathogenesis of hypertrophic cardiomyopathy caused by myozenin 2 mutations is indepen- Chen SN, A, Ruggiero dent of calcineurin activity. LIBERTY-HCM: An industry –sponsored clinical •LIBERTY-HCM: HALT-HCM (Hypertrophic Regression with •HALT-HCM •Identification and characterization of the •Identification and characterization of epi- Molecular genetics and pathogenesis of Molecular genetics hereditary cardiomyopathies including in ge- molecular mechanistic studies, models and cultured netically modified animal that link the cells to identify the mechanisms The phenotype. causal mutations to the disease complemented with mechanistic discoveries are utilizing preventive and therapeutic approach, genetic and pharmacological approaches that These studies target the pathogenic pathways. are initially pursued in the animal models and The latter is tested humans. in subsequently, through randomized placebo-controlled pilot clinical trials to set the stage for large-scale clinical trials. RESEARCH PROJECTS •Identification of causal genes for heart failure AJ Marian, M.D. AJ Marian, Cardiovascular Genetics Director of the Center for Professor and Professorship in Foundation Distinguished Josephine Hamman George and Mary Research Cardiovascular Our long-standing research objectives have is an AC Arrhythmogenic Cardiomyopathy (AC): Hypertrophic is Cardiomyopathy (HCM): HCM Dilated Cardiomyopathy (DCM): DCM is The overall approach entails an integrated been to delineate the molecular genetics and pathogenesis of hereditary cardiomyopathies in humans and apply the discoveries to prevent the evolving and reverse the established phenotypes of heart failure and sudden cardiac death. have active research programs in three com- We mon forms of hereditary cardiomyopathies: enigmatic form of hereditary cardiomyopathies that clinically presents with cardiac arrhyth- heart sudden cardiac death, failure and mias, A unique feature of particularly in the young. this disease is a gradual replacement of cardiac There is no myocytes with fibro-adipocytes. AC. effective therapy for the most common form of hereditary cardiomy- affecting ~ 1 in every 500 individuals opathies, The affected individu- in the general population. als are typically asymptomatic and sudden cardiac death is often the first manifestation of cause of HCM is the most common this disease. While there sudden cardiac death in the young. are effective therapies to alleviate patient’s there is no effective therapy to symptoms, prevent or reverse process. the disease genetically the most heterogeneous form of hereditary cardiomyopathies and a major cause of heart failure and heart transplantation in the The affected individuals often present young. cardiac arrhyth- with symptoms of heart failure, sudden cardiac death. mias and sometimes, There are a number of effective pharmacological and non-pharmacological therapies for DCM but currently there is no cure for DCM. approach that includes human molecular genetic studies through high throughput genomic DNA sequencing to identify the causal genomic characteriza- genes and mutations, tion to define epigenetic regulation of gene and transcriptomic analysis to link expression, Genetic and the epigenetic changes to function. genomic discoveries are then pursued through CARDIOVASCULAR GENETICS FOR CARDIOVASCULAR CENTER 18 • IMMPACT REPORT CENTER FORCARDIOVASCULARGENETICS heart failure.heart long non-codingRNAsin AC andotherformsof now investigatingtheroleofothermiRNAs and AC. the roleofmiRNA-184inpathogenesis of Based ontheseandotherfindingsweestablish mitigatesadipogenesis.of miR-184partially induces adipogenesiswhileenhancedlevels ofmiR-184 demonstrate thatreducedlevels the roleofmiR-184inadipogenesis AC. We excess myocardialadipocytes, wecharacterized Given that AC represents auniquephenotypeof most down-regulatedmiRNA(~10-fold)in AC. expression dataweidentifiedmiR-184asthe by integratingRNA-seqdatawithmiRNA namely canonical Wnt and Hippoin AC. Next the pathogenicroleofgenenetworks/pathways transcriptome RNA-seqweinitiallyidentified myocytes byfibro-adipocytes. Usingwhole pathologically byagradualreplacementof arrhythmias, failure, heart suddendeathand dium thatclinicallymanifestswithcardiac (AC). AC isaprimarydiseaseofthe myocar- (miRNAs) in Arrhythmogenic Cardiomyopathy the roleofdifferentiallyexpressedmicroRNAs andfibrosis.hypertrophy pathogenic roleinstressinducedcardiac identified noveltargetsofmiR-22withpotential sion analysisandbiochemicalapproacheswe fibrosis. usingglobalgeneexpres- Furthermore and regulatorofcardiachypertrophy as akey (one ofthemostabundantmiRNAinheart) failure.and heart Recently, IidentifiedmiR-22 coding RNAsinregulationofgeneexpression Current studieshavehighlightedtheroleofnon- failure.ease symptomsandprogressionofheart dis- failureandtotargetthesealleviate heart modulate generegulatorynetworksthatleadto objective ofthisresearchistoidentifyand failure.stand themolecularbasisofheart The Using widearrayofgenomicapproachwe are Working onsimilarlineswealsocharacterized The mainfocusofmyresearchistounder- Assistant Professor Priyatansh Gurha, Ph.D. PMID: 24311620 Research, 2013Dec6;113(12): e115-20. andDisease.Cardiovascular Biology Circulation Gurha PandMarian AJ. Non-CodingRNAsin PMID: 24412363. sion., CellReports 2014Jan30;6(2): 336-45. dramatically alteringmiRNAandmRNAexpres- tissue,is disruptedinmyotonicdystrophyheart CJ, Cooper TA .TheMef2transcriptionnetwork Kalsotra A, SinghRK, GurhaP, Ward AJ, Creighton 2013 Sep27;8(9):e75882, PMID:24086656. hormone receptortranscription.PLoSOne, coordinate suppressionofPPAR/ERR-nuclear A. failurethrough microRNA-22promotesheart S, Taffet GE, Wehrens XH, EntmanML, Rodriguez Goodger C, RamirezMO, Reddy AK, Engelhardt P,Gurha Wang T,Larimore AH,SassiY, Abreu- Jan 31;114(3):454-68. PMID:24276085. contributed equally. Circulation Research , 2014 in Arrhythmogenic Cardiomyopathy. *Authors And IsaCausalMechanismFor Adipogenesis son, A.J. Marian. The HippoPathwayIs Activated Lombardi*,Ruggiero,James T. Alessandra - Willer Suet NeeChen*, PriyatanshGurha*, Raffaella 125(22): 2751-61, PMID:22570371. contractile dysfunction. Circulation, 2012Jun5; promotes stress-inducedcardiacdilationand Rodriguez A. Targeted deletionofmicroRNA-22 Taffet GE, Bradley A, Wehrens XH, EntmanML, N, Enright AJ, LeeB, KelmRJJr, Reddy AK, Drumond AL, vanDongenS, Chen Y, Bartonicek Gurha P, Abreu-Goodger C, Wang T, RamirezMO, KEY PUBLICATIONS •Identification andcharacterizationoflong •Role ofmiRNAsinthepathogenesis AC RESEARCH PROJECTS failure heart Molecular mechanismsandfunctionsofNon-codingRNAsin heart failure.heart nisms/ functionincardiomyopathiesand non-coding RNAsandmolecularmecha- IMMPACT REPORT • • • • 19 Circ Res. 2011 Dec 9; 109(12):1342-53. Circ Leung A, Bell Rodriguez G, Dong J, Lombardi R, Marian Brugada R, Willerson JT, Schwartz RJ, TK, fate mapping identifies second Genetic AJ. heart cells as a source of field progenitor adipocytes in arrhythmogenic right ventricu- Res. 2009 May 8; Circ lar cardiomyopathy. 104(9):1076-84. Noseda M, Sidhu J, V, Senthil A, Bell Lombardi R, Differential interac- AJ. Roberts R and Marian tions of thin filament proteins in two cardiac T mouse models of hypertrophic and troponin Res. Cardiovasc dilated cardiomyopathies. 2008; 79(1):109-17. Ripplinger Chen SN, Rodriguez G, Lombardi R, Wickline SA, Betocchi S, Willerson JT, W, Li CM, Resolution of Estab- AJ. Marian Efimov IR, lished Cardiac Hypertrophy and Fibrosis and Preventionof Heart Failure in the β-Myosin Rabbits with Transgenic Heavy Chain-Q403 2009 Mar 17; Circulation N-Acetylcysteine. 119(10):1398-407. LAB MEMBERS Ph.D. Assistant Professor: Priyatansh Gurha, Ph.D., Post-doctoral Fellows: Suet Nee Chen, Ph.D. Auguste Gaelle, Ph.D., Karmouch Jennifer, Czernuszewicz, Associate: Grazyna Z. Research M.S. Infiltration of fat tissue (ORO staining) in the Arrhythmogenic heart of a mouse model of In vivo labeling of adipocytes Cardiomyopathy. in red) with the fluorescent (identified by CEBPA, protein EYFP (in green) in the heart of the transgenic mouse. ment in laminopathies tion of cellular sources of fibro-adiposis in Arrhythmogenic Cardiomyopathy in the pathogenesis of primary cardiomyopathies These findings could lead to the development For my studies on cardiomyopathies I was KEY PUBLICATIONS Alessandra Lombardi R*, Gurha P*, Chen SN*, The WillersonMarian AJ. Hippo Ruggiero, J T, And Is a Causal Mechanism Activated Pathway Is For Adipogenesis in Arrhythmogenic Cardiomy- Authors (* Res. 2014; 114:454-68. Circ opathy. contributed equally to this work) Curr Genetics and Sudden Death. Lombardi R. Opin Cardiol 2013; 28 (3): 272-81. Molecular genetics and AJ. Marian Lombardi R, pathogenesis of arrhythmogenic right ventricular cardiomyopathy: a disease of cardiac stem Cardiol. 2011 Mar; Pediatr 32(3):360-5. cells. Bell da Graca Cabreira-Hansen M, Lombardi R, Nuclear AJ. Marian Willerson JT, Fromm RR, A, plakoglobin is essential for differentiation of cardiac progenitor cells to adipocytes in arrhythmogenic right ventricular cardiomyopathy. •Molecular pathogenesis of cardiac involve- •Identification and molecular characteriza- of new aimed at preventing therapies cardiac precursor cells from switching from a muscle cell fate to a fat cell fate. and “The 2008 Louis N. awarded with the American Heart from the Katz award” Arnold M. award which is the most prestigious Association, given to young investigators in the cardiovascular field. RESEARCH PROJECTS •Delineation of the signaling pathways involved Molecular genetics and pathogenesis of genetics and pathogenesis Molecular hereditary cardiomyopathies Furthermore I have AC. source of adipocytes in identified molecular key in pathways implicated progenitor cells to the differentiation of cardiac Recently I have shown that a subset adipocytes. platelet-derived of cardiac cells expressing the protein (PDGFRA+) growth factor receptor-alpha express desmosome proteins and are able to differentiate to adipocytes; hence they represent an attractive novel candidate cell type that AC (unpublished differentiate to adipocytes in data). Raffaella Lombardi, M.D., Ph.D. M.D., Lombardi, Raffaella Assistant Professor The focus of my research is the delinea- Although significant progress has been I have developed cell culture models as well Activation of the Hippo kinase cascade tion of the molecular pathogenesis of genetic which are important causes cardiomyopathies, with an of sudden cardiac death in the young, Arrhythmogenic Cardiomyopathy emphasis on in desmo- which is caused by mutations (AC), somal genes. accomplished over the past two decades in at the present time revealing causal genes, the there is no effective pharmacological or non- pharmacological therapy for this disorder. AC as genetically modified animal models of and have utilized these models to understand the molecular links between the mutant desmosome proteins and cardiac fibro-adipogenesis, which is a unique and enigmatic phenotype The mechanistic discoveries in these AC. of models have been complemented with genetic and pharmacological intervention targeting the implicated pathways in order to prevent and re- focused I have specifically verse the phenotype. on developing an independent research career on elucidating the molecular pathogenesis of the unique phenotype of fibro-fatty replacement AC and the cellular origin of cardiac myocytes in I have implicated sup- of excess adipocytes. in part Wnt signaling, pression of the canonical by nuclear relocalization of the desmosome in the junction protein plakoglobin (PG or JUP), with my colleagues Together AC. pathogenesis of I at the Center for Cardiovascular Genetics, have shown that mutations in the desmosome proteins affect the structure as well as the signaling functions of the intercalated disks (IDs), which result in activation of the Hippo cascade. Wnt signaling pathway suppresses the canonical AC. and contributes to fibro-adipogenesis in the focus of my studies has More recently, been the identification of the cellular origin AC. of adipocytes in the heart of patients with Through genetic fate-mapping experiments I have identified a subset of cardiac progenitor cells (CPC) from the second heart field as a cell CARDIOVASCULAR GENETICS FOR CARDIOVASCULAR CENTER 20 • IMMPACT REPORT H CENTER FORHUMANGENETICS competition with thepathogensinworld systems have through beenevolving inlargepart high.Ourour immuneresponses immune isvery by thefactthatextentofgeneticvariation in challenge. Thischallengeis mademore complex and how geneticvariation inthem isanew inflammation” touncover whatgenesare involved outcome. Understanding these processes of“sterile our immuneresponses maymodulate disease vessels progresses, sothegeneticfactorsshaping toblood of ourimmunesystemandasinjury vessel wallinatherosclerosis isinvaded by cells istate ofinflammation. For example,theblood system isthatthesediseasesproduce achronic element ofchronic diseaseofthecardiovascular developing inourlaboratoriesisthatanimportant modulate diseaserisk. can influences arisingthrough ourlifehistory how theseinteractwithagingandhow genetic genes thatcontributetocardiovascular diseaseand work ofour centertargetstheidentificationof these diseasesaswell asshapinghow we age.The processes of aging.Heredity impactsourriskof life andsoare intricatelylinkedtothenormal These are diseasesthatemergeinmiddleandlater of ourpopulationthananyotherdiseaseprocess. An important emergingconceptthatis An important together have alargerimpactonthehealth pressure andstroke are linkedtogetherand andkidneydisease, high blood eart Kozmetsky Family ChairinHuman Genetics Center Director &Professor Eric Ph.D. Boerwinkle, healthspan. understanding toprevent diseaseandextend the shape our”healthspan” andhow we canusesuch focused onunderstandinghow genesanddisease contained withinourgenomes.Our work is the processing andexpression ofinformation elements oflifestyleandenvironment shape symptoms. It will allow ustoseehow different disease ineachaffectedindividual,notjustthe to targettreatments totheunderlying causeof andcriticaldirection.brain diseaseinthisnew interactions between cardiovascular functionand degeneration, extendingourstudiesofthe to Alzheimer’s diseaseandage-related neuro- initiative toidentifygeneticvariation contributing high bloodpressure. We have amajornew disease, progressive kidneydisease,stroke and susceptibility toatherosclerosis, artery coronary has provided understanding of new important immune response. variation that enhancestheadaptabilityofour immune response of genetic isthepreservation rapidly andonecounterstrategyembeddedinour around us. Assimplerorganisms,thesecanevolve The advances we pursuewillallow doctors Progress inthelaboratoriesofourinvestigators IMMPACT REPORT • • • • 21 H., Hindy G., Masca N., Stirrups K., Kanoni S., Kanoni S., Stirrups K., Masca N., G., Hindy H., Goel Xue C., Kang H.M., Y., Hu Jun G., Do R., R., Asselta Merlini P.A., Duga S., M., Farrall A., Reilly W., Yin Martinelli N., Olivieri O., Girelli D., Holmen Hveem K., Fox C.S., Speliotes E., D., Fran- T.L., Assimes Farlow D.N., Nikpay M., O.L., Martin L.W., North K.E., Robinson J., ceschini N., Jansson J.H., Escher S.A., Gupta N., DePristo M., Koch N., Wareham Palmer C.N., Zuydam N., Van Ko- Erbel R., W., Lieb A., Peters T., Meitinger W., Gottesman O., Degenhardt F., Kruppa J., nig I.R., Ballan- Psaty B.M., O’Donnell C.J., Bottinger E.P., Melander Ordovas J.M., G., Abecasis tyne C.M., Ardissino D. M., Orho-Melander H., Watkins O., Erdmann J., Willer C.J., McPherson R., Loos R.J., Schunkert Deloukas P., N.J., Samani A.S., Hall Tracy Rich S.S., Kooperberg C., Wilson J.G., H., Nickerson Gabriel S., Altshuler D., Lin D.Y., R.P., Boer- A.P., Reiner Cupples L.A., Jarvik G.P., D.A., (2014) Loss-of-function Kathiresan S. winkle E., and coronary triglycerides, APOC3, mutations in N Engl J Med 371(1):22-31. disease. Cupples L.A., Kovar C., A.C., Morrison A.H., Li Muzny Metcalf G., B., Yu Polfus L.M., Brody J.A., Mosley T., Lumley Liu X., N., Veeraraghavan D., Analysis (2015) Boerwinkle E. R.A., Gibbs T.H., of loss-of-function variants and 20 risk factor phenotypes in 8,554 individuals identifies Nat Genet loci influencing chronic disease. 47(6):640-642. (2015) Boerwinkle E. Gibbs R.A., Polfus L.M., Coronary heart disease and genetic variants Med Engl J N A2 activity. with low phospholipase 372(3):295-296. - have completed the most comprehen We Developing an internationally recognized Obtaining the DNA sequence of 100,000 individuals to study the determinants of health and disease Genetic analysis of the common chronic diseases in humans, humans, the common chronic diseases in Genetic analysis of and hypertension, arteryincluding coronary disease, disease Alzheimer’s for a variety of sive genome-wide analyses including heart risk factors, disease-related These levels. blood pressure and cholesterol identification investigations have led to the As of novel susceptibility genes in both cases. sequencing the personal genomes of individuals has become more accessible and less expen- analysis of our group has focused on the sive, the vast information contained in the human recognized in are internationally We genome. Such the analysis of large-scale genomic data. by growing problems are facilitated “Big Data” “Cloud.” our research program in a team of investigators targeting the genetics of cardiovascular disease and its risk factors ensures a productive future and further discoveries. RESEARCH PROJECTS • KEY PUBLICATIONS T.D., O’Connor A.W., Bigham J.A., Tennessen Do R., McGee S, Gravel S., E.E., Kenny W., Fu Leal S.M., Jordan D, Kang H.M., Jun G., Liu X., Altshuler Abecasis G, Rieder M.J., Gabriel S., Sunyaev S., Boerwinkle E., D.A., Nickerson D., Akey J.M.; Bamshad M.J., Bustamante C.D., on behalf of the NHLBI G.O.; Seattle Broad G.O.; (2012) Evolution Exome Sequencing Project. and functional impact of rare coding variation Sci- from deep sequencing of human exomes. ence 337(6090):64-9. Abecasis Kang H.M., Jun G., T.D., O’Connor W., Fu Shendure Altshuler D., Gabriel S., Leal S.M., G., Bamshad M.J.; NHLBI Exome Nickerson D.A., J., Analysis (2013) Akey J.M. Sequencing Project, of 6,515 exomes reveals the recent origin of most human protein-coding variants Nature 493(7431):216-20. Crosslin D.R., Auer P.L., Peloso G.M., Crosby J., Zhang Z.Z., Tang Y., Lu Lange L.A., Stitziel N.O., Eric Boerwinkle, Ph.D. Eric Boerwinkle, for Human Genetics IMM Center Director, Professor and Chair in Human Genetics Kozmetsky Family of Public Health School Department of Epidemiology, and Chair, Professor Dean, I received my B.S. in Biology from the Univer- I received my B.S. My research interests encompass the in Statistics an M.A. sity of Cincinnati in 1980, in Human Genetics and Ph.D. and M.S. (1984), Ann (1985) from the University of Michigan, where I served as senior research associ- Arbor, ate in the Department of Human Genetics from Texas- of I joined the University 1985-1986. Houston Center for Demographic/ Population Genetics in 1986 as a research assistant and became assistant professor in the same year. Human I worked to move the renamed In 1991, Genetics Center to the School of Public Health, Health Science Texas-Houston University of I was In 1996, Center as associate professor. of director and in 1997, promoted to professor, I became a faculty the Human Genetics Center. member of the Institute of Molecular Medicine in 1996 and became professor and director In of the Research Center for Human Genetics. of the I became the associate director 2011, Human Genome Sequencing Center at Baylor College of Medicine. genetic analysis of common chronic diseases including coronary artery disease, in humans, This hypertension, and Alzheimer’s Disease. which contribute work includes localizing genes, identification of potentially to disease risk, test- functional mutations within these genes, ing these candidate functional mutations in defining the impact of experimental systems, gene variation on the epidemiology of disease, and determining the extent to which these genes interact with environmental factors to Activities include contribute to disease risk. A both statistical analysis and laboratory work. large part of my current research effort consists of localizing genes contributing to disease risk using modern exome (the protein encoding part of the genome) and whole genome sequencing Success depends on keeping up with methods. The the latest genomic technical advances. laboratory is set up and operating as a high through-put sequencing and genotyping facility and efficiency are accuracy, in which speed, monitored continuously. HUMAN GENETICS FOR HUMAN CENTER 22 • IMMPACT REPORT CENTER FORHUMANGENETICS Blood pressure elevation exceedstheability Blood pressureelevation role insusceptibilitytoinjury.bine toplayakey immune systemgenescom- in eachofseveral disturbed byeffectsarisinginthesegenes. the genesinvolvedandpathwaysthat are disease.and kidney We areworkingtoidentify genetic factorsinfluencebothbloodpressure in arodentgeneticmodelwhichseparate in renalfunction. We studythisdiseaseprocess risk factorthatamplifiestheage-relateddecline structures. Increasedblood pressureisamajor the glomeruliandnephrons, aremicroscopic deep withinthebodyandtheirfunctionalunits, lie are difficulttostudyinhumansbecausethey renal functionaldeclineisnotknown. Kidneys renal disease. At present, themechanismof indicates thatinheritedfactorsshaperiskof stage” ofrenaldisease. Familialaggregation degree relativewhohasreachedthis “end haveafirstorsecond dialysis iswhetherthey functiontorequire will loseenoughkidney process isamplifiedbyhighbloodpressure. progressive lossofrenalfunctionandhowthis survival. We seektounderstandwhatcauses face long-termrenaldialysisortransplantfor and ofsocietyisoutlivingtheirkidneys portion advances andwelivelongerlives, agrowing age, functiondeclines. ourkidney As medicine blood. Itisnotsosurprisingthen, thatas we products thatmustbeeliminatedfromour continuous filtrationandconcentrationofwaste their neighborhoodismadetougherbythe arealsoourexcretoryorgans,Since thekidneys the oxygen-carryingcapacityofblood. of redcellsbythebonemarrowanddetermine our blood. controltheproduction Ourkidneys we needtoincreasethenumberofredcellsin thatareusedasamonitor whether our kidneys a “feature” rather thana “bug” itis byevolution: Indeed, thischaracteristichasbeenadaptedas are constantlyonthevergeofoxygendeficiency. pumpout,fraction ofthebloodourhearts they in ourbodies. receiveasizeable Although they Our recent work reveals thatgeneticvariation Our recentworkreveals The bestpredictorofwhetheranindividual occupyaroughneighborhood Our kidneys Disease Research DistinguishedUniversityChairinMetabolicandInflammatory Mary ElizabethHoldsworth Professor &DeputyDirector Peter A. Doris, Ph.D. pressure andimmunefunction functionasweageanditsrelationshiptoblood kidney Studiestounderstandthedeclineof Out-living ourkidneys: 31:2050-2059,Hypertension 2013 RatLines. Resistance InInbredHypertensive J. RenalDisease:Susceptibility Hypertensive And Monita, R.J. Bell, S.E. Wenderfer andP.A. Doris. Braun, M.C. S.M. Herring, N. Gokul, M. Amer. Soc. Nephrol.22:881-9, 2011 andalbuminuria. ated withserumIgGlevels J. Doris. The ratimmunoglobulinlocusisassoci- Boerwinkle, S.E. Wenderfer, M.C. BraunandP.A. Herring, S.M., N. Gokul, M. Monita, R. Bell, E. Genetics). 4:223-31, 2011 between SHRlines. Circulation (Cardiovascular genetic basisforbloodpressuredifferences identity bydescentmappinguncoversthe Grove, E. Boerwinkle, P.A. Doris. Highresolution Bell, R., S.M. Herring, N. Gokul, M. Monita, M.L. KEY PUBLICATIONS •Immune cellgeneticsandsignaling blood •Genetic mechanismsofelevated •Inherited susceptibilitytorenalandcardio- RESEARCH PROJECTS system tobepreserved. the normalprotectivefunctionsofimmune functionwhileallowing that cansustainkidney and narrowlytargettherapeuticapproaches information inhandwewillbeabletoseek a self-damagingimmuneresponse. this With of theirfunctionarealteredsoastocreate expressing thesegenesandwhichaspects goal istounderstandwhichimmunecellsare shape oftheimmuneresponse. Ourimmediate genes containingvariationthatregulatesthe response. setof We nowhaveidentifiedakey the courseofdiseasebymodifyingimmune tissuedamage.tion andfurther We canmodify repair orinjuryleadingtosustainedinflamma- to determinewhethertheoutcomeisinjuryand damage tothetissue. Geneticvariationappears in whichimmuneresponsesareprovokedby tissue. This triggersaninflammatoryprocess needs foroxygenandnutrientsinthekidney tomatchbloodflowthelocal of thekidney pressure vascular endorgandisease identical. the remaining13%ofgenomesthatare not to differencesindiseasesusceptibilitywithin us topinpointthegenomicregionscontributing genetic similarity). This closenesshasallowed between fraternalandidenticaltwinsintheir 87% oftheirgenomesidentical(i.e. are they very closegeneticrelativesofoneanotherwith (injury-resistant) rats. These inbredratlinesare week oldSHR-A3(injury-prone)andSHR-B2 of30 inthekidneys at themacroscopiclevel Different susceptibilitytorenalinjuryisvisible Clinical Fellow:Xiaoyan Wu, M.D., Ph.D. Post-doctoral Fellow:IshaDhande, Ph.D. Technician: YamingZhu (Open Monoclonal Technology Inc) (Baylor CollegeofMedicine), RolandBuelow (Baylor CollegeofMedicine), M. JohnHicks (Baylor CollegeofMedicine), Scott Wenderfer (IMM), OlehPochynyuk(MMS), MichaelBraun Myriam Fornage(IMM), ManuelGonzalez-Garay Collaborating Faculty:EricBoerwinkle(IMM), LAB MEMBERS 7:903-910, 2014 signaling. Circulation (CardiovascularGenetics) associated withgenevariationaffectingimmune Zhu andP.A. Doris. renalinjuryis Hypertensive R.J. Bell, S.E. Wenderfer. M.L. Gonzalez-Garay, Y. Braun, M.C. S.M. Herring, N. Gokul, M. Monita, and Immunity, 15(8):528-533, 2014. tible andresistanttoendorganinjury, Genes ofSHRlinessuscep- immunoglobulin repertoire and P.A. Doris. Diversityinthepre-immune Gonzalez-Garay, M.L., S.M. Cranford, M.C. Braun, IMMPACT REPORT • • • • 23 Stroke 2015; 2015; 12:607-613. Carty CL, Keene KL, Cheng YC, Meschia JF, Chen JF, Meschia YC, Cheng Keene KL, Carty CL, and METASTROKE al for the COMPASS et WM, As- Meta-Analysis of Genome-Wide Consortia. Genetic Risk Factors sociation Studies Identifies for Stroke Americans. in African 46:2063-2068. Long- BB, Worrall Meschia JF, Nalls M, Auer PL, Lung, et al for the National Heart, WT Jr, streth and Blood Institute Exome Sequencing Project. Associ- Variants Genetic Region Coding and Rare The NHLBI With Stroke: Risk of Ischemic ated 2015; . Neurol JAMA Exome Sequence Project. 72:781-788. L, Yu G, Weinstein Bis JC, Adams HH, Chauhan G, disease GWAS Alzheimer’s of Association et al. Neurobiol loci with MRI markers of brain aging. Aging. 2015; 36:1765.e7-16. Mosley DS, Knopman Fornage M, Gottesman RF, TH. Brain Aging in African-Americans: The Ath- erosclerosis Risk in Communities (ARIC) Experi- Curr Alzheimer Res. ence. LAB MEMBERS Ph.D., Post-doctoral Fellows: Xueqiu Jian, Ph.D. Melissa Richard, BS Student: Li-An Lin, Ph.D. Biostatistician: Xiaopeng Liu Research Associate: Wang, Ping Ph.D. Scientific Programmer: Xiaoping Zhao Brain magnetic resonance image showing atrophy subcortical white matter hyperintensity, and enlarged ventricles. of gray matter, cognitive function and decline in middle-aged and Hispanic African, adults of European, AG048642) ancestry (HL122658 and lar traits using gene-lifestyle interactions and and HL105756, pathway analysis (HL118305, HL120393) influencing cardiovascular traits (lipids and blood pressure) in diverse ethnic groups as Architecture and part of the NHGRI Population consortiumGenomic Epidemiology (PAGE) (HG007416) influencing risk for ischemic stroke and its etiologic subtypes in well-characterized clinical samples from the NINDS Stroke Genetics Consortium (NS069208) influence risk for brain small vessel disease and its related neurocognitive outcomes (NS087541) influencing MRI-defined white matter lesions influencing MRI-defined white to brain vascular and other MRI traits related large-scale ge- disease and dementia using (AG033193, notyping and exome sequencing and NS087541) AG049506, KEY PUBLICATIONS Ikram JA, Smith Bis JC, Debette S, BF, Verhaaren Multiethnic genome-wide association et al., MK, study of cerebral white matter hyperintensi- Genet. 2015; Cardiovasc ties on MRI. Circ 8:398-409. •Discovering novel genetic variants influencing •Discovering novel genetic loci for cardiovascu- •Discovering common and rare genetic loci •Discovering common and rare genetic variants •Discovering novel epigenetic variants that Genetic basis of brain vascular disease and brain aging Genetic basis of brain RESEARCH PROJECTS genetic variants •Discovering common and rare Myriam Fornage, Ph.D. Myriam Fornage, Professor in Cardiology Professorship and Johanna Favrot Distinguished The Laurence My research interests focus on the genetic HUMAN GENETICS FOR HUMAN CENTER basis of common chronic diseases with an emphasis on vascular disease of the brain and Patients with acute stroke and brain aging. “tip dementia represent the easily recognized but the deleterious effects of of the iceberg,” aging on the brain begin well before clinical - Brain abnormali symptoms become apparent. resonance readily detectable by magnetic ties, - are common in asymptom imaging (MRI), My atic populations beginning in middle age. research program investigates the genetics and genomics of vascular and neurodegenerative disease of the brain both in its clinical and preclinical forms in well characterized populations Research from young adulthood to old age. strategies combine genetic epidemiology and functional genomic approaches using the latest use pow- I genome resources and technologies. erful genome-wide association and sequencing studies in collaboration with researchers in the United States and Europe to discover novel Alzheimer’s genes influencing the risk for stroke, These and brain MRI abnormalities. disease, discoveries may yield new insights into disease mechanisms and lead to the development of new therapeutics to prevent or slow disease progression. 24 • IMMPACT REPORT CENTER FORHUMANGENETICS •Investigating theactionofnovelRibozyme •The regulationofPCSK9miRNAsinthebrain •The roleofPCSK9(proproteinconvertase RESEARCH PROJECTS us wouldallowtoagegracefully. the interactionofgeneticsandenvironmenton of disease. Importantly, theunderstanding of therapeutic approachestocombatprogression efficient It willprovideabasistodevelop process.ological andpathologicaldevelopment provide insightintotheunderstandingofphysi- will factors modulatediseasedevelopment molecular mechanismsbywhichproatherogenic pro-inflammatory responsetopathogenicLDL. absence ofPCSK9generesultedtoreduced less lesionsthanLDbmice). Importantly, (see Figure, tripleknockoutmicehadsignificant inLTpsal ofatherosclerosisdevelopment mice Pcsk9-/-). - We demonstratedadelayandrever knockout mousemodel(LTp; Ldlr-/-Apobec1-/- (PCSK9) fromLDbmicetogenerateatriple discovered causativegeneofhyperlipidemia development. ern high-fatdietacceleratestheatherosclerosis age.atherosclerosis asthey Feedingona West- (LDb; Ldlr-/-Apobec1-/-). These micedevelop the LDLreceptorandanRNAeditingenzyme humans withhyperlipidemiabydeletingboth We havemadeamousemodelthatmimics immunity, geneticanimalmodels. wedevelop cross-regulation betweenatherosclerosisand ment ofatherosclerosisandtoelucidatethe vasculature.arterial To- understandthedevelop maladaptive immuneresponsesthataffectthe metabolism thatleadstohyperlipidemiaand age. The diseaseincludesimbalance lipid aswe thatincreasesinseverity in theaorta phospholipase A2 (Lp-PLA2)mRNAs apolipoprotine Band lipoprotein-associated molecules inregulating theproductionof mation, andatherosclerosis subtilisin/kexin type9)inautophagy, inflam- Therefore, examination ofcellularand Based onLDbmice, wedeletedarecent Atherosclerosis isaninflammatorydisease Distinguished Professorship in Heart DiseaseResearch Distinguished ProfessorshipinHeart The JerryandMauryRubensteinFoundation Professor Ba-Bie Teng, Ph.D., FAHA Summer Intern: Li,William Highschoolstudent Research Scientist:Hua Sun, Ph.D. Michael Tan Research Assistant: (Assistant professoratSchoolofNursing) Chung, Pei-Ying Ph.D.Mentor: LAB MEMBERS PMID: 22580899 ThrombVascBiol; 32:1585-1595.Arterioscler the low-densitylipoproteinreceptor. (2012) its intracellulardegradation, irrespectiveof interacts withapolipoproteinBandprevents Subtilisin/Kexin Proprotein Convertase Type 9 Zemin Yao, MomiaoXiong, andBa-Bie Teng. Hua Sun, Amin Samarghandi, Zhang, Ningyan 24043250 Circ Physiol;305:H1668-1679.Heart PMID: promotes atherosclerosis. (2013)AmJPhysiol reducesapotosisinmacrophagesand Fortilin Mahmoud Eltorky, Ba-Bie Teng, andKenFujise. Decha Pinkaew, RachelJ. Le, Yanjie Chen, e125. PMID:24084845 (2013) MolecularTherapy-Nucleic Acids;2: scAAV8.2 vectorinhibitsatherosclerosisinmice. B mRNA-specifichammerheadribozymevia Teng. Long-termexpressionofapolipoprotein David A. Ford, Cao,Ying Peng Wei, andBa-Bie Hersharan Nischal, HuaSun, Yuchun Wang, Immunity; 40:153-165. PMID:24412615 mune T helper17cellresponsesinvivo. (2014) Proatherogenic conditionspromoteautoim- Huaizhu Wu,Ba-BieTeng,and YeonseokChung. Lee, JosephM. Reynolds, ShinoHanabuchi, Hoyong Lim, Young UkKim, HuaSun, JoyceH. Biol; 35:1787-1797. PMID:26112011 lesterolemia. ThrombVasc (2015)Arterioscler nascent atherosclerosisinmicewithhypercho- Foamy monocytesformearlyandcontributeto Simon, ChristieM. Ballantyne, andHuaizhu Wu. Donglin Yang, XinhuaXiao, Ba-Bieteng, ScottI. Lu Xu, XiaoyuanDaiPerrard, JerryL. Perrad, KEY PUBLICATIONS ofviralvectorsandendothelial •Development •Identify diseasemarkersbymetabolomics atherosclerotic vasculardiseases ofgeneticandcelltherapiesforthetreatment development Pathogenesis ofatherosclerosisandimmunitythe cells fortherapeutics and miRNAsprofiling of atherosclerosis. excellent modelforstudyingthepathogenesis inDr.developed Teng’s laboratory. are They ofanLDbmouse.in theaorta LDbmiceare atheroscleroticlesionsareshown The severe LDb mice(8.8%±3.5vs. 23.7%±3.3, p=0.004). cantly lessatheroscleroticlesionsthanthatof Ldlr-/-Apobec1-/-Pcsk9-/-) micehavesignifi- aorta. As shown, triple knockoutmice(Triple KO; formation ofatheroscleroticlesionsinmouse used anen-facemeasurementtodeterminethe ofatherosclerosis.creases thedevelopment We Deletion ofPCSK9genefromLDbmicede- IMMPACT REPORT • • • • 25 The Center’s scientists also are actively actively scientists also are The Center’s macular degeneration and diabetic retinopathy. engineered pursuing the generation of genetically immune mediated stem cell lines that will avoid during transplantation procedures. graft rejection include: interests Research • Asthma and Sinusitis Retinopathy • Diabetic & Autoimmunity Immunology • Mucosal Disease Infectious • Microbial Injury and COPD Lung • Acute Deficiencies • Surfactant Degeneration • Macular Medicine Regenerative • Pulmonary Ph.D. Rick Wetsel, & Professor Director Center and Irma M.D., Ph.D. J. Muller-Eberhard, Hans Distinguished Chair in Immunology M.D. Gigli,

he investigators of the Hans J. Müller- J. Müller- of the Hans he investigators for Center Gigli and Irma Eberhard Diseases and Autoimmune Immunology These studies explore the nature, structure, structure, the nature, These studies explore concert with the molecular studies, the In efforts have these research from Results in pulmonaryAs part of its interest immunity, are examining the molecular, cellular, and genetic cellular, examining the molecular, are autoimmune, allergic, different bases of several and infectious diseases. receptors and function of specific cell membrane and and their ligands in modulating immune inflammatoryresponses. mice with engineered scientists have Center’s specific targeted gene mutations or deletions used as models for human disease. These that are facilitated the identification of animal studies have roles that play significant key gene products as the immune system, as well in regulating contributing to the pathogenesis of human disease. therapeutic targets for the identified several of asthma, septic shock, and lupus treatment erythematosus. research a robust established the Center recently of stem cell focused on the development program of acute and therapeutics for the treatment lung diseases and for genetic deficiencies chronic well as for that affect normal lung function as

CENTER FOR CENTER AUTOIMMUNE AND IMMUNOLOGY DISEASES T 26 • IMMPACT REPORT IMMUNOLOGY AND AUTOIMMUNE DISEASES IMMUNOLOGY ANDAUTOIMMUNE CENTER FOR our laboratoryhasgenerated numerous “knock- of theanaphylatoxin receptors inlungdisease, blood leukocytes. To examinetherequisiterole and directedmigrationofnumerousperipheral histamine releasefrommastcells, vasodilation, immunity, includingsmoothmusclecontraction, biological responsesininflammationand coupled receptorsthatmediatenumerous G-protein receptors areseven-transmembrane their specificreceptors(C3aRandC5aR). These complement anaphylatoxins(C3aandC5a) of thisresearchhasinvolvedstudiesthe normal andpathologicalconditions. Much and immuneresponsesinthelungduringboth responsible formediatingtheinflammatory molecules has focusedondelineatingkey lung disease. that couldbeusedfortheeffectivetreatmentof ofnoveltherapeutics slowed thedevelopment lung immunityandtissueregenerationhas of cellularandmolecularknowledgeregarding chronic lungpathologies. However, thepaucity ofirreversible thedevelopment as preventing oflungdiseaseas well controlling theseverity and cellularrepairresponsesarecriticalin and well-regulatedimmune, inflammatory, Deficiencies).(Cystic Fibrosis-Surfactant Robust allergies (Asthma), andgeneticmutations Obstructive PulmonaryDiseases-emphysema), (Pneumonia), environmentaltoxins(Chronic monly causedbyviralandbacterialinfections chronic lungdisease. Lungdiseaseiscom- is theonlyviableoptionforpatientswithsevere of cureordiseasereversal. Lungtransplantation provide symptomaticreliefbutoffernoprospect Current treatmentsforlungdiseaseatbest for approximately20percentofinfantmortality. death inbabiesunder1yearofage, accounting deaths peryear. Itisalsoamajorcauseof States, accountingforapproximately400,000 diseaseandcancer)intheUnited (behind heart lung disease, anditisthenumberthreekiller There areover35million Americans with andmorbidityworldwide.cause ofmortality Our laboratory for the past several years Our laboratoryforthepastseveral Intractable respiratorydiseasesarealeading gene minimizestheimmunogenicity ofhuman Targeted disruption oftheβ2-microblobulin Wang D, Quan Y, Yan Q, MoralesJE, Wetsel RA. KEY PUBLICATIONS •Generate embryonicstemcelllinesthatcan •Identify andcharacterizelungprogenitorcells •Evaluate thetherapeuticpotentialofgene •Determine thebiologicalroleofcomple- •Delineate themolecularmechanismsby RESEARCH PROJECTS protein Bdeficiency. diseases affectingthelungsuchassurfactant specific iPScellsforthetreatmentofgenetic therapeutic potentialofgenecorrectedpatient model inmice. Inaddition, weareexploring the lung epithelialdamageinanacuteinjury cells wererecentlydemonstratedtoabrogate thelial typeIIcellsfromhumanEScells. These of thefirstpurepopulationlungalveolarepi- COPD. This researchhasledtothegeneration as wellbychroniclungdiseasessuch lung epitheliumdestroyedbyacuteinjury stem celltherapeuticsfortheregenerationof of program hasfocusedonthedevelopment stem cellderivedprogenitorcells. ofthis Part of embryonic(ES)andinducedpluripotent(iPS) cytokine response. lung cellularinflammation, andtheCD4+ Th2 airway hyperresponsiveness, mucusproduction, hallmarksofasthma,mediator ofkey including have demonstratedthatC3aRisaimportant in whichtheC3areceptorhasbeendeleted of lungdisease. Forexample, studiesusingmice roles oftheanayphylatoxinsinpathogenesis facilitated thediscoveryofnumerousbiological methods. The generationofthesemicehas gene targetingandhomologousrecombination regulators havebeenselectivelyablatedby receptors, theirligands, andcarboxypeptidase out” miceinwhichthegenesencodingthese pulmonary regenerativemedicine, andstemcelltherapeutics Innate immunology, inflammation, infectiousdiseases, and Hans J. Muller-Eberhard, M.D., Ph.D. andIrmaGigli, M.D. DistinguishedChairinImmunology and Professor andDirectoroftheCenterforImmunology Autoimmune Diseases Rick Wetsel,Ph.D. tor cellsthatavoidgraftrejection be differentiatedintotransplantableprogeni - intissueregeneration important cells forsurfactantdeficiencies corrected iPScell-derivedlungprogenitor and tissueregeneration ment anaphylatoxinsonlungepithelialinjury tious diseases adaptive immunityduringallergicandinfec- which complementanaphylatoxinsmodulate We alsoareinvestigatingthetherapeuticuse mouse modelofasthma. inflammatory cellsand lungepithelialcellsina Expression oftheC3a receptor(greencolor)on M.D./Ph.D. GraduateStudent:DanielCalame Li Research Assistant:Yi-Dong Senior John Mazzilli Post-doctoral Fellows:Dr. Young Uk-Kim, Dr. Senior Research Associate: Dr. PoojaShivshankar Ortiz Senior ResearchScientist:Dr. Mueller- Stacey LAB MEMBERS 104:4449-4454. embryonic stemcells. ProcNatl Acad Sci.2007, epithelial typeIIcellsderivedfromhuman Wetsel RA. A purepopulationoflungalveolar Wang D, HavilandDL, Burns AR, ZsigmondE, Therapy. 2010, 18:625-634. abrogates acutelunginjuryinmice. Molecular stem cellderivedalveolarepithelialtypeIIcells Wetsel RA. Transplantation ofhumanembryonic Wang D, MoralesJE, CalameDG, Alcorn JL, cells. Annu. Rev. Med. 2011, 62:95-105. from embryonicandinducedpluripotentstem tential oflungepithelialprogenitorcellsderived Wetsel RA, Wang D, CalameDG. Therapeutic po- 2014, 193:1278-1289. monocytogenes-induced apoptosis. J. Immunol. (C3aR) provideshostprotectionagainstListeria receptor forthecomplementC3aanaphylatoxin SL,Mueller-Ortiz MoralesJE, Wetsel RA. The Immunol. 2014, Nov15;193(10):5099-5107. infection byinhibitingtypeIIFNexpression. J. protects againstListeriamonocytogenes RA. The C5aanaphylatoxinreceptor(C5aR1) Calame DG, SL, Mueller-Ortiz MoralesJE, Wetsel 2015 Oct;4(10):1234-1245.. embryonic stemcells. StemCells Transl. Med. IMMPACT REPORT • • • • 27 Karmouty-Quintana, H., Weng, T., - Garcia-Mo T., Weng, H., Karmouty-Quintana, H., Zhong, M., Pedroza, Y., N. Chen, J., L. rales, Y., Xia, A., B. Bruckner, R., Bunge, G., J. Molina, Seethama- D., Zeng, M., Loebe, A., R. Johnston, R. M. and Blackburn, L. Belardinelli, H., raju, ADORA2B and hyaluronan modulate (2013) pulmonary hypertension associated with J. Am. chronic obstructive pulmonary disease. PMID: 1038-1047. 49, Biol. Mol. Cell. Respir. 23855769 LAB MEMBERS Assistant Professor: Tingting Weng, Ph.D. Ph.D. Volcik, Senior Research Scientist: Kelly Ph.D. Post-doctoral Fellow: Frank Lou, M.D. Student: Kemly Philip, Ph.D. Chen Associate: Ning-Yuan Research Research Scientist: Jose Molina Sr. Ph.D. Graduate Student: Josh Ko, Primary type II alveolar epithelial cells isolated from genetically modified mice. Increased expression (brown color) of proteinases in pulmonary macrophages in mice with pulmonary fibrosis (BLEO). the progression of chronic lung disease IL-6 signaling in pulmonary fibrosis chronic lung disease regulation of pulmonary fibrosis and Chronic Obstructive Pulmonary Disease and chronic lung injury expression on pulmonary macrophages during expression on pulmonary macrophages fibrosis the progression of pulmonary KEY PUBLICATIONS Y., N. Chen, K., Philip, H., Karmouty-Quintana, Acero, J., Davies, F., Luo, G., J. Molina, T., Weng, Johnston, T., Le, K., Volcik, I., Bunge, Bao, Le, L., R. M. and Blackburn, K. H. Eltzschig, Y., Xia, A., R. cells ADORA2B from myeloid (2015) Deletion of dampens lung fibrosis and pulmonary hyperten- PMID: 25318478 50-60. 29, J. FASEB sion. H., Karmouty-Quintana, M., J. Poth, T., Weng, T., N. Chen, E., Melicoff, J., L. Garcia-Morales, Loebe, A., B. Bruckner, R., R. Bunge, M., C. Evans, and Blackburn, K. H. Eltzschig, A., K. Volcik, M., Hypoxia-induced deoxycytidine (2014) R. M. kinase contributes to epithelial proliferation in 190, Care Med. Crit. J. Am. pulmonary fibrosis. PMID: 25358054 1402-1412. E., Melicoff, H., Karmouty-Quintana, T., T. T. Le, Pedroza, M., Y., Chen, N. T., Weng, T., T. T. Le, R. Bunge, J., L. Garcia-Morales, T., A. George, Seethamraju, M., Loebe, A., B. Bruckner, R., (2014) R. M. and Blackburn, K. S. Agarwal, H., Blockade of interleukin-6 trans signaling attenu- 3755- Immunol. 193, ates pulmonary fibrosis. J. PMID: 25172494 3768. Garcia- H., Karmouty-Quintana, T., Weng, Bunge, M., Pedroza, G., J. Molina, J., L. Morales, H. Seethamraju, M., Loebe, A., B. Bruckner, R., R. (2013) Hypoxia-induced R. M. and Blackburn, deoxycytidine kinase expression contributes to J. 27, apoptosis in chronic lung disease. FASEB PMID: 2339349 2013-2026. •Systems Biology approaches to understand •Understanding novel mechanistic roles for •Examination of the hypoxia as an amplifier of •Novel regulation of mRNA polyA tails in the •Investigation of adenosine transport in acute Adenosine signaling and the regulation of chronic lung disease and the regulation of chronic lung Adenosine signaling RESEARCH PROJECTS A2B adenosine receptor •Examining the role of Michael R. Blackburn, Ph.D. Blackburn, Michael R. Sciences Professor of Biomedical McGovern Distinguished P. Dean and John at Houston School of Biomedical Sciences Graduate Texas of The University Vice Chairman Department Biology of Biochemistry and Molecular Professor and Chair in Pulmonary Disease Kilroy Sr., William S. UTHealth Academic Officer, Viceand Chief President Executive Inflammation and remodeling responses are A central hypothesis of my laboratory is use of genetically modi- make extensive We prominent features of chronic lung diseases pulmonary chronic obstructive such as asthma, and pulmo- pulmonary fibrosis, disease (COPD), Although signaling pathways nary hypertension. associated with the genesis of these diseases about the little is known have been described, signaling pathways that serve to regulate the The major chronic nature of these diseases. goal of my laboratory is to identify pathways that regulate the chronicity of these disorders with the intent of developing novel therapeutic strategies. that the signaling molecule adenosine is an amplifier of lung inflammation and damage. Adenosine is generated in response to cell as adenosine and it is our belief that damage, levels in the lung they increase access pathways that serve to promote airway inflammation and by engaging Adenosine signals remodeling. specific adenosine receptors on target cells airway fibroblasts, such as inflammatory cells, and smooth muscle cells. epithelial cells, Most of the projects in my laboratory focus on understanding the mechanisms by which adenosine signaling influences the activities of these cells in the context of lung inflammation and remodeling. fied mice to examine the role of adenosine This includes signaling in chronic lung disease. knockout mice of components of adenosine also conduct We metabolism and signaling. mechanistic experiments in disease-relevant cell types and work extensively with human explanted lungs obtained following lung Medical Texas transplantation here in the These translational approaches help us Center. identify novel strategies for treating chronic lung disease. CENTER FOR CENTER AUTOIMMUNE AND IMMUNOLOGY DISEASES 28 • IMMPACT REPORT IMMUNOLOGY AND AUTOIMMUNE DISEASES IMMUNOLOGY ANDAUTOIMMUNE CENTER FOR for sinonasalmucosal. Recentstudieshave first lineofdefense against theenvironment complications andblindness, respectively. orbital cavitieswhichcanresultinintracranial separating thesinusesfromintracranial and sinus cavitiesandultimatelyerosionofbone This trappedmucuscancauseexpansion of the nasalpolypsandwithinsinuscavities. with fungalhyphaeandeosinophilsbetween accumulation ofthickentrappedmucusladen subtype ofCRSwNPthatisassociatedwithan of type2cytokinessuchasIL-4, IL-5, andIL-13. phils, mastcells, andbasophilsexpression polyps (CRSwNP)havehighpresenceofeosino- type 1(Th1)cytokineswhileCRSwithnasal nance ofneutrophilsandelevated T helpercell nasal polypsarecharacterizedbypredomi- the inflamedsinustissueinwhichCRSwithout byimmunologicprofilesof has beensupported polyps (seeimage1). This clinicalclassification defined bytheabsenceorpresenceofnasal tion ofthepatientanddiseasedmucosa. - regularly inclinicwhichallowsperiodicevalua to harvestcriticaldiseasedtissueandareseen often undergosurgeryprovidinganopportunity inflammatory respiratorydiseases. CRSpatients human researchmodelforstudiesinchronic are notcurative. CRSrepresentsanideal rent surgeriesandanti-inflammatoryagents, treatment options, whichtypicallyinvolverecur- unknown aboutitspathophysiology, andcurrent Despite thishealthcareburden, muchremains chronicillnessesinthe United States.prevalent respiratory diseasesrepresentsomeofthemost asthma. Together, CRSandasthmaaschronic suffering fromCRSoftenarediagnosedwith of over3billiondollars. Inaddition, patients with anannualdirecthealthcaretreatmentcost vatively 18-22millionphysicianvisitsyearly tion. These symptomsultimatelydrive conser- and pressure, nasalcongestion, andobstruc - rhinosinusitis (CRS), whichcausesfacialpain Respiratory epithelial cellsrepresentthe Allergic fungalrhinosinusitis(AFRS)isa CRS isclinicallyclassifiedinto2groups Over 40million Americans sufferfromchronic Porter PC,Porter Yan, T, Luong A, DelclosGL, KEY PUBLICATIONS • • • RESEARCH PROJECTS it relatestoCRS. the elucidatingspecificsofthispathwayas role.important Ongoingstudiesarefocusingon shown thatfungalproteasesseemtoplayan tory responseinsomeCRSsubtypes. We have responsible forsignalingintheinflamma- signaling pathway(s)andthefungalcomponent molecular characterizationoffungi-mediated helper cellsinvariousCRSsubtypes. in thefunctionofinnatelymphoidcellsand T interested inthedistributionandultimately immune responseinCRS, wearecurrently and knowndataoftheroleadaptive release IL-33. can stimulaterespiratoryepithelialcellsto that fungalantigens, specificallyAspergillus, 13 inresponsetoIL-33. Interestingly, wefound and representthemajorcelltypeproducingIL- These ILC2expressST2, thereceptorforIL-33, in CRSwNPpatientsrelativetohealthcontrols. nate lymphoidtype2cells(ILC2)preferentially We demonstratedanincreasedpresenceofin- in thediseasedsinonasalmucosaofCRSwNP. firmed thatthereceptorofIL-33isupregulated characteristic ofCRSwithnasalpolyps. We con- orchestrating thetype2immuneresponse linked tothetype2immuneresponse. etin, interleukin(IL)-25andIL-33, havebeen derived cytokines, thymicstromallymphopoi- anti-microbials. Three identifiedepithelialcell through regulationofcytokinesandrelease shown thatepithelialcellsserveanactiverole chronic airwayinflammation Environmental triggersregulatinginnateimmuneresponsesin –HeadandNeckSurgery Otorhinolaryngology Associate Professor, and CenterforImmunology of Autoimmune DiseasesandDepartment Amber Luong, M.D., Ph.D. biomarkers forCRSsubtypes Clinical characterizationandidentification of immune response ing and/ormaintainingthecharacteristic Th2 environmental triggersresponsibleforinitiat- epithelial cellsoffungialoneandwithother Molecular signalingthroughrespiratory response cell typesinvolvedinthe Th2 immune Immunologic characterizationofimportant In addition, mylabisinterestedinthe Given theappreciationofinnateimmunity Our labhasfocusedontheroleofIL-33in 331; 2014 Aug. 331; Disease. J Allergy ClinImmunol, 134(2):325- Mycosis inChronic Th2-Associated 1 Airway admand F, CorryDB, andLuong A. Airway Surface Citardi MJ, FakhriS, ShawJL, Fothergil A, Kher- P,Porter LimDJ, MaskatiaZK, MakG, Tsai CL, Aug 15;188(4):432-9. nasal polyps. AmJRespirCritCareMed. 2013 source ofIL-13inchronicrhinosinusitiswith sponsive innatelymphoidcellsareanimportant DB, KheradmandF, Liu YJ, Luong A. IL-33-re- Shaw JL, FakhriS, CitardiMJ, PC, Porter Corry 16;341(6147):792-6. through Toll-like receptor4.2013 Aug Science. by proteinaseselicitsallergicresponses Kheradmand F, CorryDB. Cleavageoffibrinogen L, SongLZ, KnightJM, CreightonCJ, Luong A, Millien VO, Lu W, ShawJ, Yuan X, MakG, Roberts 2012 May;2(3):233-40. PMID:22344928 Atopy. InternationalForumof Allergy Rhinology, itis withNasalPolypPatientsIndependentof within SinonasalMucosaofChronicRhinosinus- Luong AL. IncreasedPercentageofMastCells Shaw JL, Ashoori F, FakhriS, CitardiMJ, and Nov;1810(11):1059-65. PMID:21712069 Airway Disease.Biophys Acta.2011 Biochim Proteinases asMolecular Adjuvants in Allergic Abramson SL, KheradmandF, andCorryDB. nasal cavityofCRSwNPpatient. Nasal polypsseenbynasalendoscopywithin IMMPACT REPORT • • • • 29 Flow cytometric plot demonstrating intracellular expression of Foxp3 transcription factor and its correlation to the cell surface IL2 receptor T cells (left alpha chain (CD25) within CD4+ Right figure shows the percentages of figure). T cells from healthy within CD4+ Tregs Foxp3+ donors. Kleiner D, Notarangelo LD, Rampertaap Y, Y, Rampertaap Notarangelo LD, Kleiner D, Pittaluga S, Hughes J, McElwee J, Olivier KN, Holland SM, TA, Fleisher Meffre E, Oliveira JB, 2014 Science. Uzel G. SG, Tangye Lenardo MJ, doi: 10.1126/ Sep 26;345(6204):1623-7. PMID: Epub 2014 Sep 11. science.1255904. 25213377 Lactobacillus reuteri DSM 17938 differentially Foxp3+ T cells and modulates effector memory of necrotiz- T cells in a mouse model regulatory Fatheree NY, DQ, Tran Y, Liu ing enterocolitis. Am J Physiol Gastrointest Liver Marc Rhoads J. doi: 2014 Jul 15;307(2):G177-86. Physiol. May Epub 2014 10.1152/ajpgi.00038.2014. PMID: 24852566 22. LAB MEMBERS I: Rachel Song and Colby Techician Research Hofferek. Ossama Clinical Hem/Onc Fellow: Dr. MDACC Maher M. Impact: This outcome will provide the neces- Impact: Optimizing fever and hyperthermia to modu- cancer and late immunity toward allergy, vaccine responses in mucosal and their role Tregs Investigating particularly with probiotics (Col- immunity, Yuying Marc Rhoads and Dr. laboration: Dr. Liu) in Tregs Deciphering the contribution of pulmonary inflammation and allergic lung diseases associated with obesity (Collabora- Richard Johnston) tion: Dr. Varying frequencies of Tregs in health and diseases. The plots show the percentages of Foxp3+ Tregs Tregs The plots show the percentages of Foxp3+ in health and diseases. Tregs frequencies of Varying T cells. within CD4+ ing in a more effective and safe product for ing in a more effective and and immunotherapy to treat autoimmune transplant-related conditions. into a clinical sary preclinical data for transition to prevent or treat GVHD Tregs trial with LAP+ and other autoimmune diseases. RESEARCH PROJECTS • • • KEY PUBLICATIONS cells and T Forkhead box protein 3(+) regulatory Helios(+) subset in perinatally acquired HIV. Contreras GA, W, Zhang Zakhour R, Degaffe G, Pérez N, Del Bianco G, Rodriguez G, Bell CS, DQ. Tran Heresi GP, Murphy JR, Benjamins LJ, Clin Exp Immunol. 2015 Apr;180(1):108-17. PMID: 25425428 doi: 10.1111/cei.12560. Immune dysregulation in human subjects with heterozygous germline mutations in CTLA4. Niemela Deenick EK, Lo B, W, Ouyang Kuehn HS, Stoddard J, DQ, Tran JN, Schickel Avery DT, JE, Scheinberg B, Dumitriu Frucht DM, Y, Zhang T, Heller Koh C, Price S, Frein CA, Folio LR, P, Su HC, VK, Rao A, Huttenlocher Seroogy CM, Immune modulation for the treatment of allergy, infectious infectious for the treatment of allergy, Immune modulation cancer and autoimmunity diseases, Dat. Q. Tran, M.D. Tran, Q. Dat. Associate Professor Allergy/Immunology & Rheumatology, Medicine Pulmonary Center, Pediatric Research Our group focuses on the novel strategies to Our primary research projects funded by an subsets and repertoire, Treg Assess the Aim 1: Hypothesis: Patients who develop chronic This study will provide critical Impact: Aim 2: Elucidate the function of IL1 receptor The unique and temporal Hypothesis: This study will provide invaluable Impact: stability and TCR diversity, Aim 3: Determine represent a highly Tregs Hypothesis: LAP+ harness the immune system for 1) induction of immune tolerance to treat autoimmunity, inflammation and allergy and 2) enhancement Our of immunity against infections and cancer. research primarily focuses on Foxp3+ regulatory T cells of CD4+ a critical subset T cells (Tregs), that play a vital role in orchestrating immune the Strategies that optimize homeostasis. shift the would Tregs number and function of balance toward immune tolerance; abrogat- On a similar would boost immunity. Tregs ing our we also concentrate branch of research, efforts of to understand the different pathways TGFbeta in order to develop therapeutic targets to modulate immune responses. R01 from NHLBI/NIH: function in patients post allogeneic stem cell transplant. de- TCR repertoire, GVHD will have restricted and decreased Tregs creased Helios+ and LAP+ Treg-specific demeth- Tregs based on bona fide ylated region (TSDR). biomarkers for predicting the development of cGVHD and a justification for novel LAP+ immunotherapy to prevent or treat this Treg debilitating condition. type 1 (CD121a) and type 2 (CD121b) on LAP+ Tregs. expression of CD121a and CD121b enhances Th17 stability and regulation of function, Treg differentiation. insights into the critical role of IL1 receptors on and a rationale for therapeutic use Tregs LAP+ to induce tolerance. Tregs of LAP+ Tregs. functionality of expanded LAP+ result- and potent population, stable, purified, CENTER FOR CENTER AUTOIMMUNE AND IMMUNOLOGY DISEASES 30 • IMMPACT REPORT search efforts arefocusedon1) toisolateand search efforts model tostudylungdisease processes. Myre- injuries andgeneticdisorders, butalsoagood can betherapeuticallyusedtotreatdistal lung are notonlyapromisingsourceofcellsthat cell deriveddistallungstem/progenitorcells disease-specific inducedpluripotentstem (iPS) diseases. The embryonicstem(ES)cell/lung use ofstemcellsinthetreatmentpulmonary inthepotentialclinical interest hasdeveloped injury. years, Duringthepastfew considerable diseases andthemechanismsofrepairfrom targets forexploringthepathogenesisoflung lung stem/progenitorcellsrepresentthekey distal lungepithelia. doubt,Without thedistal to facilitatetheregenerationorrepairofinjured noveltherapies an imperativeneedtodevelop notpositive.outcome isunfortunately There is within alimitedtimerange, andthelong-term best releasesymptomsandimprovelifequality treatments forthosepulmonarydiseasesat vivo andinvitro. Inaddition, currentlyavailable ing thebiologicalanddiseaseprocessesbothin biomedical researchmodelavailableforstudy- repair mechanisms, sincethereisnoreliable monary diseasesaswellthecorresponding known regardingthepathogenesisofthesepul- distress syndrome(RDS). However, littleis CODP, cysticfibrosis, andneonatalrespiratory diseases, includingpulmonaryfibrosis, asthma, pulmonary ofchronicorsevere development roleinthe cies isthoughttoplayanimportant tor celltypesduetoinjuriesorgeneticdeficien- normal functionsofanythesestem/progeni- point-associated neuroepithelialbodies. Lossof bronchioalveolar ductjunctionandthebranch 3) asubsetofvariantClaracellslocatedatthe transient amplifyingbronchiolarClaracells;and tions: 1)alveolarepithelialtypeIIcells;2)the maintaining distallungepithelialcellpopula- stem/progenitor celltypesresponsiblefor from theenvironment. There areatleastthree that areconstantlyexposedtoinjuriousstimuli airway andalveolarepithelialcellpopulations critical forthemaintenanceofhomeostasis IMMUNOLOGY AND AUTOIMMUNE DISEASES IMMUNOLOGY ANDAUTOIMMUNE CENTER FOR Lung epithelialstem/progenitorcellsare alveolar epithelialtype IIcells. STEMCELLS. tency andsubsequent isolationofderivedlung genetic modificationforinductionofpluripo - J.L., Wetsel R.A., and Wang D*. A site-specific Yan Q., Quan Y., SunH., PengX., ZouZ., Alcorn responding author) ENGINEERING. 2014Jun;20(6):464-72. (*cor- from mouseembryonicstemcells. TISSUE tion ofalveolarepithelialtypeIIcellsderived R.A., and Wang D*. Isolationandcharacteriza- Sun H., Quan Y., Yan Q., PengX., MaoZ., Wetsel 1326-3-15. (*corresponding author) cine. 2014, Jun17;3:15doi:10.1186/2001- lung diseases. Clinicaland Translational Medi - potentials ofhumanpluripotentstemcellsin Quan Y. and Wang D*. Clinical An invitedreview: # Co-correspondingauthor 2015 Aug. pii:sctm.2015-0049. Embryonic StemCells. StemCell Transl Med. Gene MinimizestheImmunogenicityofHuman RA., Targeted Disruption of theβ2-Microglobulin Wang D#., Quan Y., Yan Q., MoralesJE, Wetsel KEY PUBLICATIONS • • • • RESEARCH PROJECTS stem/progenitor cellsforlungtissuerepair. fortargetedactivationofendogenous strategy anovel the diseaseprocessesfordeveloping distal lungstem/progenitorcellfateduring ize factorsorregulatorypathwaysthatcontrol regeneration; and3)toidentifycharacter- forlungtissue cell-based genetherapystrategy monary diseaseprocessesandfordeveloping lung disease-specificiPScellsforstudyingpul- vitro andinvivo;2)togenerate “clinical grade” distal lungstem/progenitorcelltypesbothin characterize humanandmouseEScellderived Assistant Professor M.D.Dachun Wang, Lung stem/progenitorcellsandtissueregeneration eration deficient humanEScelllinefortissueregen- Generation andcharacterizationofHLA-1 tor cellsforthetreatmentoflungdiseases specific iPS-deriveddistallungstem/progeni- Therapeutic potentialofES/lungdisease- tor cellfate Pathways toregulardistallungstem/progeni- cells stem cellderiveddistallungstem/progenitor Isolation andcharacterizationofembryonic Senior researchassistant:Dr. Yuan Quan LAB MEMBERS 13;104(11):4449-54 (2007). human embryonicstemcells. PNAS, Mar alveolar epithelialtypeIIcellsderivedfrom E., and Wetsel R.A. A purepopulationoflung Wang D., HavilandD.L., Burns A.R., Zsigmond Molecular Therapy. 2010;18:3,625-634mar. Type IICells Abrogates Acute LungInjuryinMice. Embryonic StemCell-Derived Alveolar Epithelial J.L., and Wetsel R.A. Transplantation ofHuman Wang D., MoralesJ.E., CalameD.G., Alcorn author) 2014 Feb;32(2):402-13. (*corresponding cultures derived fromlungstem cellsin3Dselective veolar-, andupperbronchiolar-likestructures Alveolar-, terminalbronchiolar-, bronchioal- Lung stemcells Alveolar lung stem/progenitorcelltypes. Pluripotent Stemcell-derived bronchiolar Terminal Bronchioalveolar stem cells Bronchio- alveolar Alveolar epithelial progenitor typeII bronchiolar cells Upper- IMMPACT REPORT • • • • 31 Genetically-engineered mouse expressing GFP Wang. D., Haviland, D. L., Burns, A.L., Zsigmond, Zsigmond, A.L., Burns, L., D. Haviland, D., Wang. A pure population of lung R.A.: Wetsel, and E. derived from alveolar epithelial type II cells 104:4449- PNAS. human embryonic stem cells. 2007. 4454, LAB MEMBERS Manager: Aleksey Domozhirov Neural differentiation of mouse embryonic stem cells cell lines, including human ES cells approved including human ES cells cell lines, for NIH-funded research in the production of knock-out mice when using ES cells derived at the facility 23%) knock-out and knock-in animal models cells and MEF feeder layer cells cell lines preservation of mouse ES cells mice eggs eggs of knock-out and knock-in mice the production of transgenic animal models the production of transgenic KEY PUBLICATIONS Kawakami, A., Kudo, E., Zsigmond, N., Nonaka, Miyado, N., Kawano, M., Yoshida, K., Yoshida, H., : Epididymal R. Wetsel, and M. Nonaka, K., C4b-binding protein is processed and degraded during transit through the duct and is not 220(4): essential for fertility. Immunobiology , 2015. 467-75, N.G., Murray, M., M. Lazarus, R., Shegog, E. and Zsigmond, N., Sessions, M., P. Diamond, Using a molecular biology simulation to impact student academic achievement and attitudes. 2011. Educ. DO 10.11007/s11165, Sci. Res. of mouse and human Transfection E.: Zsigmond, - Trans embryonic stem cells by electroporation. 2009. Note: 5904, Tech Bio-Rad Labs. fection, •100% success rate in re-derivation of mice •Derivation of more than 20 mouse and human •100% success rate of germline transmission ••Consistently high transgenic rates (average ACCOMPLISHMENTS •Generation of more than 750 transgenic, •Availability of germline-competent mouse ES •Derivation of novel mouse ES cells and other Gene targeting, selection, expansion, cryo- expansion, selection, •Gene targeting, •CRISPR/Cas9 mediated genome editing in •Cryopreservation of fertilized mouse and rat Re-derivation of mice and rats from fertilized•Re-derivation of mice and rats Microinjection of ES cells for the production •Microinjection of ES cells for Transgenic and stem cells core facility and stem cells core Transgenic RESEARCH SERVICES clones for YAC BAC or •Microinjection of DNA, Eva M. Zsigmond, Ph.D. Zsigmond, Eva M. Assistant Professor Facility and Stem Cells Core Transgenic Director, The Transgenic and Stem Cells Core Facility and Stem Cells Transgenic The The mouse embryonic stem (ES) cell lines - cryopreserva In addition to the production, was established in 1998 and since that time, it has generated over seven hundred new animal knock-out and knock-in transgenic, as well models for investigators from UTHealth, as for scientists from numerous other academic institutions. derived in the Core Laboratory are highly effective for the generation of knock-out and knock-in mice and for studies involving cellular differentiation. tion and re-derivation of genetically-engineered also the services of the facility mice and rats, derivation of new cell include gene targeting, lines and technical support in different aspects and stem cell culture, of animal microsurgery, cells research. CENTER FOR CENTER AUTOIMMUNE AND IMMUNOLOGY DISEASES 32 • IMMPACT REPORT T CENTER FORMETABOLIC AND DEGENERATIVE DISEASES control thebody’s energybalance? • to treat musclediseases? • muscle tissueremodeling? • tissue affectinsulinsensitivity? • adipose tissuebeusedtherapeutically? • of cancersandotherdiseases? • progenitors determineadiposetissuefunction? • following: addressed by theCenter’s facultyincludethe cell fatedetermination.Key questionsbeing aspects ofenergymetabolism,cellsignalingand recruit investigators withfocusoncomplementary The guidingvisionfortheCenterhasbeento signaling pathwaysandphysiologicalprocesses. conditions involve defectsinmultiplerelated cell neurodegenerative diseases.Thesedifferent health diabetes, andcancer, aswell asmusclewastingand time: obesityandtheassociateddiseasessuchas of themostpressing healthchallengesofour How does thebrainandcircadian clocks What transcriptionalpathways canbetargeted How isangiogenesisimplicated inadiposeand How doesfibrosis andinflammation inadipose Can pharmacologicaldepletionofcellsin How doesadiposetissuepromote progression How dowhiteandbrown adipocyte an integrative approach totacklesome Diseases isadiverse group thattakes he CenterforMetabolic andDegenerative Stem Cell Biology Annie andBobGraham DistinguishedChairin Center Director Professor &Associate Mikhail Kolonin, Ph.D. degenerative diseases. for thebenefitofpatientswithmetabolicand and clinicianstotranslatetheirdiscoveries faculty alsocollaboratewithepidemiologists productivity andinnovation. TheCenter’s promotes research synergy, thereby increasing Collaboration amongtheCenter’s laboratories fly.organisms, includingthemouseandfruit employs methodsinmodel state-of-the-art pathogenesis? • cause neuronal degeneration? • neurodegenerative diseases? • To address thesequestions,theCenter How doesstress impactAlzheimer’s disease How doesabnormalprocessing ofproteins What are thefunctionsofgenesmutatedin IMMPACT REPORT • • • • 33 Adipocytes differentiating from progenitor cells (green) and stromal/vascular cells (red) in Lipid droplets are white. tissue culture. Distinct adipocyte populations genetically tracked with green and red reporter fluoro- blue. Nuclei are phores in adipose tissue. of intercellular communication ing and function in tumor microenvironment cyte progenitors in disease adipocytes in health and disease KEY PUBLICATIONS PRUNE2 is a human prostate cancer suppressor regulated by the intronic long noncoding RNA Pro- et al. Kolonin MG, et al., A, Salameh PCA3. Sciences Academy of ceedings of the National of the USA. 2015; 112(27):8403-8. Stem cells from adipose tissue and breast Petit Bertolini F, risks and hope. cancer: hype, Cancer. British Journal of JY and Kolonin MG. 2015; 112(3):419-23. Depletion of white adipocyte progenitors induces beige adipocyte differentiation and AC, Daquinag suppresses obesity development. Cell Death and and Kolonin MG. et al. Tseng C, 2015; 22(2):351-63. Differentiation. Pro-apoptotic Peptides Depleting Targeted Growth. Tumor Adipose Stromal Cells Inhibit and Kolonin MG. et al. Tseng C, AC, Daquinag In press. . Molecular Therapy Adipose Induce Proteolytic Isoforms of SPARC Tseng C Stromal Cell Mobilization in Obesity. Stem Cells. In press. and Kolonin MG. LAB MEMBERS Alexis Daquinag, Scientists: Research Sr. Zhanguo Gao Tao Post-doctoral Fellow: Zhang Tseng Candidate: Chieh Ph.D. Ali Dadbin Assistant: Research Sr. •Adipose tissue cell markers and mechanisms Mechanisms of adipose stromal cell traffick- •Mechanisms of adipose stromal Experimental therapies targeting white adipo- •Experimental therapies targeting Function and targeting of adipocyte progenitor cells in disease of adipocyte progenitor cells Function and targeting RESEARCH PROJECTS white and brown •Progenitors and lineages of Mikhail Kolonin, Ph.D. Mikhail Kolonin, Diseases Metabolic and Degenerative Center for & Director, Associate Professor Research Chair in Metabolic Disease Distinguished University Jr. Bovay, Harry E. Research in my laboratory is investigating the mechanisms linking adipose tissue expansion and dysfunction in obesity with life-threatening diseases such as type-2 diabetes and cancer. are focused on pathological we Specifically, functions of adipose stromal / progenitor cells have We and on approaches to suppress them. discovered that stromal cells from white fat tissue serve as the mechanistic link between fat tissue overgrowth and obesity pathogenesis. have shown that white adipocyte progenitors We traffic to tumors and stimulate cancer progres- we reportedThis year that mobilization sion. of these cells from fat tissue is regulated by are now elucidated the protein and the SPARC molecular mechanisms of their trafficking to have also developedWe a compound tumors. adipocyte progenitors. that targets white D-WAT This year we reported in a series of publications that depletion of white adipocyte progenitors prevents and cancer progression in obesity has been licensed for D-WAT mouse models. are also We further pre-clinical development. characterizing a distinct population of progeni- which metabolically tors giving rise to brown fat, counteracts the function of white fat in animal These studies are now further carried models. out based on patient specimens in collaboration with clinicians. METABOLIC AND DEGENERATIVE DISEASES DEGENERATIVE AND FOR METABOLIC CENTER 34 • IMMPACT REPORT action. in obesityandhowthis enzymeinhibitsinsulin investigating whySIK1 abundanceisincreased for therapeuticdevelopment. We arecurrently insulin. This makesSIK1averypromisingtarget fat miceandthatSIK1inhibitstheactions of that SIK1isturnedupinskeletalmuscle of just asobesecontrolanimals. We now know a highfatdiet, thoughthemicebecame even strongly protectedfromhyperglycemiawhenfed synthesis. Instead, SIK1-deficientmicewere gene inmicehadnoaffectonliverglucose is turnedon. Surprisingly, deletionoftheSIK1 mice thatglowinthedarkwhenpathway this pathwayinlivinganimals, weengineered glucosesynthesis.new To visuallymonitor pathwaythatstimulates thought toinhibitakey monal responses. Inliver, SIK1waspreviously infine-tuning hor- out thebodyandparticipates ible kinase1, or SIK1) thatispresentthrough- pre-diabetes. proteins toincreasethetherapeuticoptionsfor there isanurgentneedtoidentify “drug-able” approved drugsattackinsulinresistance. Thus, ofthisdisease,spite oftheprevalence FDA few in theUS, mostofwhomareundiagnosed. In pre-diabetes, whichaffects~30%ofadults resistance” occursinindividualswithclinical come resistanttothehormoneinsulin. “Insulin becauseliver,part muscle, andfattissuebe- diabetes, bloodglucosebecomestoohighin oftype2diabetes.development Intype2 circulating glucoseisdetrimentalandunderlies food isscarce. Ontheotherhand, excess when andbrainfunctioneven maintain heart Humans requireaconstantglucosesupplyto physiology. expressedgeneson impact ofthosenewly induced changesingeneregulationandthe inhibited indisease. We focusonhormone- might becomeinappropriatelyactivatedor hormonal changesandhowthosepathways how cellsrespondandadapttostress-induced CENTER FORMETABOLIC AND DEGENERATIVE DISEASES Our laboratorystudiesanenzyme(saltinduc- How doesinsulinresistancedevelop? Our laboratoryisdedicatedtounderstanding Medicine. 13(5):597-603. (Coverillustration) promotes survivalofskeletalmyocytes. Nature (2007) SIK1isaclassIIHDAC kinasethat H, Wandless T, SheltonGD, andMontminyM. Berdeaux R, GoebelN, BanaszynskiL, Takemori KEY PUBLICATIONS •Chemical-genetic methodstostimulate •Regulation ofexerciseperformancebySIK1 of andseverity •Role ofSIK1indevelopment RESEARCH PROJECTS strength inagingindividuals. be targetedtopromotemusclegrowthand pathwaysthatcould we expecttouncovernew stem cellactivityandmusclegrowth. Ultimately of genesandproteinsassociatedwithmuscle cells, weareworkingtoestablishasignature Using thesemiceandisolatedmusclestem chemicalcompound.using anotherwiseinert pathways thatstimulatemusclehypertrophy created miceinwhichwecanmimichormonal muscle massduringaging. Inaddition, we on musclestemcellfunctionandultimately cells andstudytheimpactofthesepathways tors releasedbymuscleactivatestem toolstocharacterizehow novelfac- developed and exerciseabilitywithaging. Second, wehave impact ofSIK1inhibitiononmusclestrength the brakesonmusclegrowthandtesting animals. We arenowdetermininghowSIK1puts have muchlargermusclecellsthancontrol muscle growth. We foundthatmicelackingSIK1 SIK1 respondstohormonesthatpromote the effectsofSIK1onmusclemassbecause trophy, ofexistingmuscle. First, weareanalyzing of stemcellsorpromotiongrowth, orhyper- help maintainmusclemassthroughactivation pathways thatcouldbetargetedwithdrugsto amulti-facetedapproachtoidentify dertaking of normaldailyactivities. Ourlaboratoryisun- impacts metabolichealthaswellmaintenance skeletal musclemassandstrength, which and strength?Oneaspectofagingisloss muscle growth pathwaystoimprovemusclemetabolismand Harnessing new Associate Professor Rebecca Berdeaux, Ph.D. hypertrophy that promotemuscleregenerationand pathways regeneration touncovernew muscle stemcellproliferationand type 2diabetes How dohormonesregulatemusclegrowth (left) andSIK1-knockout(right)mice. area (size)ofindividualmusclecellsincontrol cent proteintomeasurethecrosssectional Musclefiberboundariesoutlinedwithafluores- M.S., KavithaRajendran, M.S. Senior Research Assistants: MariaMendoza, Dean’s Research Award Recipient Doctoral Student:RandiFitzgibbon, B.S., 2015 Ph.D. AHA Post-doctoralFellow:Dmitry Akhmedov, LAB MEMBERS metabolic switch. Cell, 155(2):455-447. Dionne M.S. (2013)Mef2isaninvivoimmune- Walker, D.W., Berdeaux, R., Geissmann, F., and Vivancos, V., Bronda, K., Pilátová, M., Fu, J., Clark, R.I., Tan, S.W.S., Péan, C., Roostalu, U., Front. Physiol., 4:371. fects ofobesityonskeletalmuscleregeneration. Akhmedov DandBerdeauxR. (2013) The ef- PNAS, 110(1):117-22. dance andstabilityiscriticalformyogenesis. Berdeaux R. (2013)RegulationofSIK1abun- R*,Stewart Akhmedov D*, RobbC, LeiterCand lustration) Endo andMetab.303(1):E1-17. (Coveril- metabolism andregeneration. AmJPhysiol: ing inskeletalmuscleadaptation:hypertrophy, R.Berdeaux RandStewart (2012)cAMPsignal- ONE 6(9):e24714. injury andpromotesmuscleregeneration. PloS Berdeaux R. (2011)CREBisactivatedbymuscle R,Stewart FlechnerL, MontminyM, and IMMPACT REPORT • • • • 35 Knocking out a specific circadian protein- interacting partner (“KO”) in the liver dampens a gene oscillation of the expression of Cyp7A1, important for cholesterol removal from the while inducing rhythmicity of the insulin- body, Glut4 (a gene not sensitive glucose transporter, normally expressed at high levels in the liver). These livers become extremely fatty relative to a control liver. normal, Eckel-Mahan KL, Phan TH, Han S, Wang HB, HB, Wang Han S, TH, Phan Eckel-Mahan KL, Circadian Storm DR. Scheiner ZS, Chan GC-K, Activity and cAMP in the Oscillation in MAPK for Memory Per- Hippocampus: Implications Vol. 2008 Sept sistence Nature Neuroscience 11(9): 1074-1082. LAB MEMBERS Aleix Ribas Latre, Post-doctoral Fellows: Baharan Fekry Assistant: Christopher Kwok Research Robust expression of circadian clock proteins in individual nuclei of CA1 neurons of the essential for a region of the brain hippocampus, long term memory formation. initiation by CLOCK-associated tumor sup- pressors Syndrome (NES) intake of peripheral vs. central clocks of peripheral vs. The clock is as important- in what it keeps os The circadian clock in health and disease The circadian clock Kristin Eckel Mahan, Ph.D. Mahan, Kristin Eckel Assistant Professor KEY PUBLICATIONS Sassone-Corsi P Phenotyping KL, Eckel-Mahan, Circadian Rhythms in Mice Current Protocols in 2015 September Mouse Biology, Ceglia de Mateo S, VR, Patel Eckel-Mahan KL, Orozco-Solis R, Dyar KA, Dilag S, Sahar SS, NJ, Reprogramming and Sassone-Corsi P. Baldi P, of the Circadian Clock by Nutritional Challenge 1464-1478 155(7), 2013 December 19, Cell, Eckel-Mahan KL and Sassone-Corsi P Metabo- lism and Circadian Rhythmicity Converge Physi- 2013 Jan;93(1):107-35 ological Reviews, Mohney Vignola KS, VR, Patel Eckel-Mahan KL, A Map of the Baldi P and Sassone-Corsi P. RP, Circadian Metabolome Reveals the Organized Transcription Interface between Metabolism and Apr 2012 A. Acad Sci U S in the Liver Proc Natl 3;109(14):5541-6 •Regulation of hepatic cell division and tumor •Circadian mechanisms underlying Night Eating •Role of the circadian clock in rhythmic energy selves appear to function as “zeitgebers” for the for “zeitgebers” selves appear to function as This nutrient-sensing capability circadian clock. of cells is mediated in part by critical nuclear rhythmicity and receptors that control 24-hour appear to be responsible for when defective, other metabolic a fatty liver phenotype among problems such as cancer progression. cillating throughout the circadian cycle as what it preventsfrom oscillating during the circadian the appropriate levels of gene or Thus, cycle. protein activity throughout the circadian cycle is essential for optimal metabolic activity and is Understand- controlled in a tissue-specific way. ing the mechanisms underlying this control is necessary for discovering why perturbation of the clock at the environmental and behavioral including levelslead to specific diseases, may obesity and cancer. RESEARCH PROJECTS •Mechanisms underlying nutritional regulation The internal circadian (24-hour) clock is Interests of our lab center on how specific Recent work has proved that even simple METABOLIC AND DEGENERATIVE DISEASES DEGENERATIVE AND FOR METABOLIC CENTER tethered to our environment and functions as an ever-present regulator of our human physiol- Examples of circadian oscillations are the ogy. intake, food seeking and sleep/wake cycle, There is and hormone production and release. now substantial evidence that disrupting our endogenous circadian clocks (such as occurs time traveling across during night shift work, or taking certain drugs and medications) zones, has deleterious effects when it comes to me- as disparate as influencing processes tabolism, body weight regulation and cognitive efficiency. A surprising number of metabolically-related diabetes, including obesity, disturbances, cancer and dementia have all been associated with an impaired circadian clock in either the associ- Diseases brain or the peripheral organs. ated with clock disruption are disparate in their is present in as the circadian clock phenotype, all tissues of the body and is ultimately supported(such by time keeping in individual cells as individual neurons of the hippocampus) that are synchronized and coordinating within and sometimes even across tissues. “zeitgebers”) control the circadian cues (or clock in a tissue-specific way to direct cellular we are interested in In particular, metabolism. what happens to the normal function of tissues, when the clock such as the liver and the brain, Understanding the mechanisms breaks down. underlying the diseases associated with clock and obesity, including cancer, disturbance, cognitive decline will shed light onto potential behavioral or pharmacological steps that might or better synchro- restore, be taken to preserve, nize our internal circadian clocks. such as the consumption behavioral deviations, profoundly affects the circa- of a high fat diet, reprogramming it in a dian clock of the liver, manner in which oscillations in gene expression phase-shifted, or protein activity are either lost, or there is a genesis of oscillations where they nutrients them- Thus, should not take place. 36 • IMMPACT REPORT CENTER FORMETABOLIC AND DEGENERATIVE DISEASES slowing down these devastating diseases. slowing downthesedevastating stress pathwaysmight beaneffectivemeansof suggest thatpharmacological manipulationof neurodegenerative diseaseprogression, and stress responseasacriticalinfluenceon experimental andclinicalresultsidentify the dementiawithage.risk ofdeveloping Our observation thatchronicPTSDincreasesthe neurologists atUTtoinvestigatetheinteresting we haveinitiatedacollaborationwithclinical might slowprogressionofthedisease. Recently, in thehopethataddressingthesesymptoms indicate progressingneurodegenerativedisease, how late-lifeneuropsychiatricsymptomsmight our workon Alzheimer’s Diseasetodetermine drives diseaseprogression. We arecontinuing and impactsofstresscreateaviciouscyclethat cognitiveloss.overt These interactingsources pathways causinganxietyanddepressionbefore early stage Alzheimer’s stress Diseaseperturbs However, inparallelexperimentswefoundthat cause Alzheimer’s Diseasetoprogressfaster. have shownthatstressandexcessCortisol including Alzheimer’s disease. We andothers also negativelyimpactotherongoingdiseases, human patients. of thesepathwaystomodulatesymptomsin anxiety-related diseases, willallowthetargeting pathways thatareactivatedduringchronic anxiety states, andthespecificmolecules sion. Determininghowneuralcircuitsmediate return aschronicstatesofanxietyanddepres- understand howtheseemotionsandmemories to stress. Furthermore, weareattemptingto brain producesemotionsandmemoriesrelated responsive pathwaystounderstandhowthe manipulate selectcircuitsinthesestress hormone, Cortisol. Usingmousegenetics, we (HPA) axisthatcontrolsreleaseofthestress (CRF), andtheHypothalamic-Pituitary-Adrenal ReleasingFactor neuropeptide Corticotropin pathways centeraroundtheactionofstress mediate thebodies’ responsetostress. These systems thatareactivatedbystressand Chronic stress, anxiety, anddepressioncan My labstudiestheneuralandendocrine Assistant Professor Nicholas Justice, Ph.D. neurodegenerative diseaseprogression Stress-related diseaseandtheimpactofstresson M. JNeurosci.M. 2013Feb20;33(8):3284-98. man MA, ContetC, JusticeNJ, Vale W, Roberto mice.amygdala ofCRFreceptor-1 reporter Her- differential effectsofethanolinthecentral Novel subunit-specifictonicGABAcurrents and Sep 4. 10.1016/j.devcel.2014.07.001. Epub2014 BR. DevCell. 2014Sep29;30(6):645-59. doi: J,Selever DeussingJM, JusticeNJ, Arenkiel Garcia I, QuastKB, HuangL, Herman AM, and CircuitIntegrationof Adult-Born Neurons. Local CRHSignalingPromotesSynaptogenesis 26;35(34):11830-47. Justice NJ,2015 Aug ChanCS.JNeurosci. DA, MP, Fiske GlajchKE, PittJE, Huang TY, Hernández VM, HegemanDJ, CuiQ, Kelver External GlobusPallidus. Are DistinctNeuronClassesintheMouse Parvalbumin+ NeuronsandNpas1+ Aug 22. S0306-4522(15)00761-7. VM, GlassMJ, Milner TA. Neuroscience.2015 Marques-Lopes J, JusticeNJ, IadecolaC, Pickel sion. Van Kempen TA, DodosM, Woods C, - following slow-pressorangiotensinIIhyperten factortype1receptor corticotropin-releasing rostral ventrolateralmedullaneuronscontaining Sex differencesinNMDA GluN1plasticityin Feb 11;35(6):2612-23. Zhu MX, Arenkiel BR, ZhengH. JNeurosci. 2015 L, JB,Tian Cole A, PruskiM, Hunt AJ Jr, FloresR, exacerbate stressresponses. JusticeNJ, Huang factorneuronsto vates corticotropin-releasing β-amyloidlevels,elevates whichdirectlyacti- Posttraumatic stressdisorder-likeinduction KEY PUBLICATIONS • Modeling Amyotrophic LateralSclerosis(ALS) • Functionalcharacterizationofneuralcircuits • LocalneuralcircuitsthatregulateHypotha- • HowdoesPost Traumatic StressDisorder • Howdoesstressimpacttheprogressionof RESEARCH PROJECTS in thec9orf72locus discoveredexpansion in mouseusinganewly that respondtostress lamic-Pituitary-Adrenal axisoutput Disease accelerate theprogressionof Alzheimer’s Alzheimer’s Disease Zhiying Jiang Post-doctoral Fellows:ShivaRajamanickam, Graduate Student: Hunt Albert LAB MEMBERS Chem. 2012Jan20;287(4):2437-45. stable natureofsolublederivatives.JBiol neuron-specificexpression andhighly revisited: CS, ZhengH. Amyloid precursorprotein Guo Q, LiH, GaddamSS, JusticeNJ, Robertson 33(11):2678-91.(*Corresponding Author) mouse model. 2012, NeurobiolAging. Nov; inan perturbation Alzheimer’s diseaseknockin Guo Q, ZhengH, JusticeNJ*. CentralCRFsystem 2012,17;109(16):6277-82. Apr and Aggregation.Solubility of RepeatedStresson Tau Phosphorylation, Releasing FactorReceptor-DependentEffects Rice KC, Vale W, SawchenkoPE. Corticotropin- Rissman RA, StaupMALee AR , JusticeNJ, Proc Natl Acad Sci, IMMPACT REPORT • • • • 37 O2 + ENERGY DELIVERY Heart Blood, Lungs, ENERGY METABOLISM Oxidative capacity (mitochondria) Fiber-type switch ERRγ OFF ERRγ ON

AD QUADS GASTROC SOL

OFF ERR ON ERR CAPILLARIES MITOCHONDRIA CAPILLARIES γ γ (A) (B) One of the exercise mimetic pathways we have genetically switched- We (A) identified is ERRγ. on ERRγ to make high endurance and fatigue thus mimicking exercise. resistant red muscle, Red color in muscle biopsies from ERRγ ON mice is due to increased mitochondria and (B) Microscopic images of blood vessels. muscles from ERRγ ON mice showing increase in mitochondrial density (top panel) and capillaries (bottom panel). Exercise has mega-physiological effects and Are there landmark molecular health benefits. pathways that can be pharmaceutically activated to mimic exercise? Discoveries from our laboratory are answering this question. diseases KEY PUBLICATIONS (2014) VA. Narkar Lorca S, A, Matsakas V, Yadav PGC1β activates anti-angiogenic program to repress neo-angiogenesis in muscle ischemia. 8(3): 783-97. Cell Rep. Lorca V, (2013) S, Narkar V. Yadav Matsakas A, Muscle ERRγ mitigates Duchenne muscular dystrophy via metabolic and angiogenic repro- 27(10): 4004-4016. Faseb J. gramming. Narkar Evans RM, Lorca S, V, Yadav A, Matsakas VA (2012) Revascularization of ischemic skeletal muscle by estrogen-related receptor-γ. Res. 110(8): 1087-96. Circ Jonker JW, RT, Yu Downes M, W, Fan VA, Narkar Lorca Karunasiri MS, Banayo E, WA, Alaynick (2011) Exercise and PGC-1α- Evans RM. S, I Muscle Type Independent Synchronization of by ERRγ. Cell Vasculature Metabolism and Metabolism. 13(3): 283-93 Wang Embler E, RT, Yu Downes M, VA, Narkar Y, Zou Nelson MC, Mihaylova MM, Banayo E, YX, (2008) Evans RM. Shaw RJ, Kang H, Juguilon H, agonists are exercise AMPK and PPARdelta 134(3): 405-15. Cell. mimetics. LAB MEMBERS Post-doctoral fellows: Pierre-Marie Badin, Danesh Sopariwala Sabina Lorca Technician: Katha Summer Students: Laura Rangel, Korgaonkar Professor, Antonios Matsakas (Asst. Alumni: Professor, (Asst. Yadav Vikas UK), Hull University, India) Amity University, Exercise mimicry in vascular, metabolic, and metabolic, mimicry in vascular, Exercise degenerative diseases RESEARCH PROJECTS •ERRγ and diabetes disease •ERRγ and skeletal muscle ischemic •ERRγ and Duchenne Muscular Dystrophy in vascular •Nuclear receptor co-activators Vihang Narkar, Ph.D. Vihang Narkar, Assistant Professor Exercise has long been known to have a dissect the molecular circuitry of exercise, To fantastic health benefit in a range of diseases. or organ-specific molecular universal However, sensors of exercise that are responsible for Uncovering exercise benefits are poorly defined. these molecular sensors has implications for designing exercise mimetic drugs for vascular, and degenerative diseases. metabolic, we have focused on the skeletal muscle – one organ that is extensively used during exercise. we use molecular and cell Experimentally, and mouse genetic pharmacology, biology, engineering to discover exercise mimetic path- we first Using these techniques, ways in muscle. AMPK identified that serine/threonine kinase β can mimic exercise and nuclear receptor PPAR by activating genes linked to mitochondrial and slow-twitch fatty acid oxidation, biogenesis, and contractile myofibers in skeletal muscles, even in absence improve endurance in mice, we are Encouraged by this finding, of training. currently investigating the role of estrogen receptor-related receptors (ERR) - a class of orphan nuclear receptors - in skeletal muscle. and particularly ERRγ is highly expressed ERR’s in high endurance muscle fibers suggesting a role for these receptors in the regulation of have genetically tar- We aerobic metabolism. geted ERRγ in mice to investigate the effect of skeletal muscle-specific receptor modification metabolism on myocellular gene expression, we are exploring the Furthermore, and exercise. potential role of ERRγ in ameliorating obesity, as well as muscular muscle ischemia, diabetes, Our findings so far suggest that ge- dystrophy. netic ERRγ activation in the muscle can mimic exercise to increase aerobic and endurance improving It also prevents obesity, capacity. and muscle vasculature to prevent ischemic, even ameliorate pathology in orphan genetic One diseases such as muscular dystrophy. future direction is to design powerful synthetic activators (which we call exercise mimetics) for which will have pharma- the above regulators, ceutical utility in various diseases. METABOLIC AND DEGENERATIVE DISEASES DEGENERATIVE AND FOR METABOLIC CENTER 38 • IMMPACT REPORT leading ahighlyunfavorable microenvironment celerates fibrosisand inflammation, ultimately COL6α3 andproducesendotrophin whichac- pre-existing unhealthyadiposetissue, itdigests hypoxia. Ontheotherhand, inthecontextof thepathologicalconditionscausedby relieving esis incombinationwith VEGF andleptin, thus collagenous proteinsandstimulatesangiogen - at earlystagesofobesity, MT1-MMPcleaves metabolic contextdependent:Ontheonehand, the regulationofECMflexibilitybyMMPsisalso Interestingly, ourrecentresearchsuggeststhat the metabolicallydysfunctionaladiposetissue. Indeed, wefound fibrosisisthehallmarkin the pathologicalexpansionofadiposetissue. during fine-tuned regulationatotherlevels of obesity-relatedmetabolicdisorders. andtreattheprogression approach toprevent foranoveltherapeutic the greatopportunity HIF1α and VEGF-A inadiposetissuemayoffer adipocytes. Ourfindingssuggestthattargeting ments inmetabolismbyablatingdysfunctional tissue, antiangiogenicactionleadstoimprove- In contrast, inpathologicallyexpandedadipose and inducinga “browning” ofwhiteadipocytes; esis isbeneficialbyimprovingvascularization in healthilyexpandedadiposetissue, angiogen- dichotomous andmetaboliccontextdependent: in whiteadiposetissueby VEGF-A couldbe effects ofmodulationangiogenicactivity importantly, demonstratedthatthe we further microenvironment inadiposetissue. More an initialstepleadingtoultimateunhealthy are frequentlyhypoxicandHIF1αinductionis Specifically, wediscoveredthatobesefatpads dynamic processofadiposetissueexpansion. many paradigm-shiftingfindingsaboutthe obesity andinsulinresistance. ofhuman significance inthepathophysiology pinpoints themascriticalfactorswithclinical and remodeling duringobesitydevelopment factors tothedynamicsofadiposetissue essential contributionsofadipocyte-derived CENTER FORMETABOLIC AND DEGENERATIVE DISEASES In mylaboratory, exploredthe wefurther In thepastfiveyears, Ihavepublished Research inmylaboratoryexaminesthe (2013) dysfunction”. MolCellBiol. 33(5):904-17. inducible factor1αamelioratesadiposetissue Scherer PE. “Selective inhibitionofhypoxia- Sun K, HalbergN, KhanM, MagalangUJ, Metab. 18(4):470-7. (2013). andadiposetissuedysfunction”.“Fibrosis Cell Sun K, Tordjman J, ClémentK, andSchererPE 5:3485. (2014). and metabolicdysfunction”. NatureCommun. “Endotrophin triggersadiposetissuefibrosis MP, Varga J, Ploug T, An ZQandSchererPE. PL, ZhangN, MarangoniRG, NicoloroSM, Czech Sun K, ParkJ, GuptaO, Holland WL, Auerbach KEY PUBLICATIONS ofadiposetissuefibrosis bynovel •Reversibility •The mechanismsbywhichendotrophin •The effectsofantiangiogenicactionbyblock- •VEGF-A stimulatedmetabolicbenefitsduring •Hypoxia inducedfibrosisandlocalinflamma- RESEARCH PROJECTS obesity andtype2diabetes. highlight endotrophinasanattractivetargetfor diet. All theseexcitingobservations inourlab metabolic dysfunctioninducedbyhigh-fat effects inadiposetissueandeffectivelyreverses neutralizing antibodyamelioratestheadverse demonstrate thatblockingendotrophinwitha ing toenhancedinsulinresistance. We further fibrosis andinflammationultimatelylead- unhealthy adiposetissuemilieu, triggering pathwayswithinthe of pathologicallyrelevant endotrophin servesasapowerfulco-stimulator overexpression modelwedemonstratethat using adoxycycline-inducibleendotrophin and mousemodelstostudyendotrophin. By to sustainmetabolicflexibility. diabetes Targeting adiposetissueremodelingforobesityand Type 2 Assistant Professor Kai Sun, M.D., Ph.D. anti-fibrotic therapies adipose tissue shapes unhealthymicroenvironmentinobese expansion) of preexistingadiposetissue(unhealthy ing VEGF-A and/oritsreceptorsinthecontext adipose tissuehealthyexpansion tion inadiposetissue More recently, weusemoleculartools Research Assistant: XueGu Post-doctoral Fellows:LiliangJin, Yueshui Zhao LAB MEMBERS (6): 2094-101. (2011). sue remodelingandobesity”.JClinInvest.121 Sun K, KusminskiCM, SchererPE. “Adipose tis- U S A. 109(15):5874-9. (2012). adipose tissuedysfunction”. ProcNatl Acad Sci Scherer PE. “Dichotomous effectsof VEGF-A on Bueno AC, Wang ZV, PollardJW, BrekkenRA, Sun K, Wernstedt Asterholm I, KusminskiCM, Clin Invest.121(6):2094-101). associated withsystemicinsulinresistance. (J Unhealthy adiposetissueexpansionisstrongly areas). (JClinInvest . 121(6):2094-101). enhanced extracellularmatrix(rightpanel, blue ofimmunecellinfiltrationand a highlevel tissue fromahigh-fatdietfedmousecontains tissue: Trichrome stainofanunhealthyadipose Healthy (left)andunhealthy(right)adipose IMMPACT REPORT • • • • 39 Pdx1-Cre neurons (red) are neither POMC (green neurons (green in B). AgRP A) nor in A diagram showing an optogenetic approach for monosynaptic circuit mapping from lateral hypothalamic (LH) GABAergic neurons to para- A)) with ventricular hypothalamic neurons (PVH, an actual recording setup (B) and inhibitory postsynaptic current traces (IPSCs) elicited by laser (C). Pictures from control (A) and knockout (B) brains showing specific deletion of vesicular monoamine transporter 2 (green) in leptin receptor neurons (red). 2013, 2013, Cell Metabolism. tial diet-induced obesity and associated behaviors restoring blood glucose in type 1 diabetes KEY PUBLICATIONS Y and Xu Mangieri LR, Y, Xu Sun H, Z, Wu Kim ER, Non-POMC GA- Hypothalamic Non-AgRP, Q. Tong Are Required for Post-weaning BAergic Neurons Neurosci. J. Feeding and NPY Hyperphagia. PMID:26203139. 35(29): 10440-10450. 2015, Corresponding author. Li DP, LR, Mangieri Y, Xu Sun H, Kim ER, Wu Z, GABAergic Q. Tong Arenkiel BR and Y, Xu Pan HL, Projections from Lateral Hypothalamus to Paraventricular Hypothalamic Nucleus Promote 35(8): 3312-3318. Neurosci. 2015, J. Feeding. Corresponding author. PMID: 25716832. Xu P, Saito K, Y, Yang C, Wang Y, Xu Y, He C., Yan Tong Wu Tso Yan X, Shun Yu L, Q, P, G, Hinton AO, A-IV inhibits food Apolipoprotein Y*. Q* and Xu intake via the central melanocortin pathway. * Co- in press. 2015, Neuroendocrinology, corresponding author. Q. Tong Y and Xu Huang C, Fan S. Kim ER, Y, Xu - Tem Profound and Rapid Reduction in Body Agonist. perature by the Melancortin Receptor Corresponding 451(2):184-189. 2014, BBRC, author. Arenkiel BR, Kim ER, Y, Zhu Sun H, Wu Z, Y, Xu Glutamate Mediates Q. Tong Y and Xu Lowell BB, the Function of MC4Rs on Sim1 Neurons in Body Weight Regulation. •Identification of factors that control differen- Corresponding author. 18 (6): 860-870. LAB MEMBERS Instructor: Yuanzhong Xu Shengjie Post-doctoral Fellows: Eun Ran Kim, Yungang Lu Fan, Graduate student: Leandra Mangieri •Novel neural pathways responsible for feeding Mechanisms underlying brain control of body weight Mechanisms underlying and glucose homeostasis rationale functions and provide a scientific against the for effective therapeutic strategies epidemic. current obesity and diabetes RESEARCH PROJECTS •Brain mechanisms underlying leptin action in Qingchun Tong, Ph.D. Tong, Qingchun Associate Professor in Molecular Medicine Cullen Chair Obesity and diabetes are imposing a huge those especially Specific groups of neurons, One current project in this regard is to de- Another ongoing project is to understand the Ultimately we try to delineate specific neural METABOLIC AND DEGENERATIVE DISEASES DEGENERATIVE AND FOR METABOLIC CENTER while the effective burden to our society, obesity The current treatment is still lacking. epidemic is due to a combination of genetic susceptibility and high-fat high caloric (HFD) we aim to understand the Thus, environment. mechanisms underlying HFD-induced obesity and its interaction with important gene functions. receive and integrate in the hypothalamus, food and then adjust nutritional status signals, intake and energy expenditure accordingly to link neuron func- To maintain energy balance. we specifically activate or tion with behavior, inhibit a distinct group of neurons with various channelrhodopsins (ChRs) by light (optogenet- ics) or with designer receptors exclusively acti- These new vated by designer drugs (DREADD). techniques in conjunction with our novel mouse genetic models will reveal important neurons and circuits in the brain for feeding and glucose hemostasis. lineate the neural pathways from lateral hypothalamus (LH) to paraventricular hypothalamus (PVH) in feeding and self-grooming behavior – a typical behavior trait in mice for obsessive The compulsive diseases (OCD) in humans. optogenetic and DREADD approaches are being used to test the hypothesis that GABAergic projections from LH to PVH promote feeding while glutamatergic projections promote The results will lead to important self-grooming. discoveries of novel neurocircuits for feeding such as and its relations with other behaviors, self-grooming. neural pathway underlying leptin in restoring glucose to normal levels in type 1 diabetes. Identification of this pathway will offer opportunities to treat type 1 diabetes without insulin, thus avoiding hypoglycemic and lipogenic risks associated with insulin treatments. pathways underlying specific physiologic 40 • IMMPACT REPORT CENTER FORMETABOLIC AND DEGENERATIVE DISEASES expansion inHuntingtin) caninterferewiththis the HD-causingmutation (i.e., polyglutamine ,autophagy raisinganintriguingpossibilitythat cellular clearancemechanismcalledselective HD gene, playerina isitselfanimportant proteins andfightthesediseases. protective mechanismstoremovemisfolded that wecanbetteremploytheseinnateself- become compromisedinaffectedneurons, so normally operateandwhytheirfunctionsoften (“self-eating”),as chaperonesandautophagy how thecell’s self-clearancemachineries, such neurodegenerative diseases. We arestudying proteins inthebrainisahallmarkofalmostall plaques andtangles)derivedfrommisfolded nisms and cellularclearancemecha- Protein misfolding, aggregation projects: animal modelforstudyinghumandiseases. mental manipulations, itprovidesanexcellent Drosophila allowsfastandcomplicatedexperi- for mosthumandiseasegenes. Because ing theexistenceinitsgenomeofcounterparts many remarkablesimilaritiestohumans, includ- simpler thanmammaliansystems, theflybears known asthefruitfly. Althoughsmallerand a classicalgeneticmodelorganismcommonly Drosophila, malian systemsandtheinvertebrate these braindiseasesbyemployingbothmam- levels. atmolecular understanding oftheiretiology ing diseases, duetoalackofclear partially approachesagainstthese debilitat- preventive (HD). Currentlytherearenoeffectivecuresor lateral sclerosis(ALS)andHuntington’s disease (AD), Parkinson’s disease(PD), amyotrophic erative disorders, suchas Alzheimer’s disease pressing threatfromaging-relatedneurodegen- ed longerlifeexpectancy, itisalsofacinga Recently, weshowedthatHuntingtin, the The presenceofabnormalaggregates(e.g., Our currentresearchfocusesonthefollowing We arestudyingtheunderlyingcausesof While oursocietyisenjoyinganunprecedent- Becker FamilyFoundationProfessorinDiabetesResearch Assistant Professor Sheng Zhang, Ph.D. 11(5):812-832). Autophagy inMammalianCells”.Induced Macroautophagy Mechanism UnderlyingProteasomeInhibition- “The GST-BHMT Assay Reveals A Distinct Rui YN, XuZ, ChenZH, and ZhangS(2015). and Viewsinthesameissue. ,Biology 17(3):262–275. CommentedinNews as aScaffoldforMacroautophagy”. NatureCell AM andZhangS. (2015). “Huntingtin functions David G, Sun YM, StimmingEF, BellenH, Cuervo Rui YN, XuZ, PatelB, ChenZH, ChenDS, Tito A, expneurol.2015.08.002 Experimental Neurology. doi:10.1016/j. ing polyglutaminediseasesinDrosophila”. Xu Z, Tito A, Rui YN, andZhangS. (2015) “Study- 10.1080/15384101.2015.1039365. disease”. CellCycle;14(11):1617-8. doi: andHuntingtin:learningfrom autophagy Cuervo AM andZhangS. (2015) “Selective KEY PUBLICATIONS •Intracellular handlingofdopamineinParkin- •Biogenesis oflysosome-relatedorganelles •Protein misfoldingandtheirclearanceby •Huntington’s diseaseandtheroleofHunting- RESEARCH PROJECTS schizophrenia. a spectrumofdisorders, suchas AD, PD, and organelles, astheirdisruptioncontributesto and regulationofthesespecializedcellular dopamine. We arestudyingthebiogenesis proper storageandregulatedreleaseof enclosed vesicles, for whichareimportant is packagedinsidethespecializedmembrane- dopamine, achemicallylabileneurotransmitter, functions andneuronalactivities. Forexample, (LROs), controldiverseaspectsofcellular aptic vesiclesandlysosome-relatedorganelles organelles Biogenesis ofspecializedcellular esis. protective mechanismandaffectHDpathogen- Molecular mechanismsofneurodegenerativediseases son’s disease chaperones andautophagy tin inselectivemacroautophagy In cells, specializedorganelles, suchassyn- p62/Ref(2)P (red)andubiquitin(blue). markers co-localizewithautophagy partially (green puncta)intheflybrain. Theseaggregates misfolded mutantHuntingtinprotein(green) autophagy. Prominent aggregatesformedby Protein misfoldingandtheirclearanceby Technician: Lili YeTechnician: Student: AntonioTito Graduate Post-doctoral Fellow:Dr. Yan-Ning Rui Dr.Instructor: ZhenXu LAB MEMBERS processes”. Cell, 108(1):45-56. tional co-repressorinmultipledevelopmental Atrophin homologfunctionsasatranscrip- Zhang S, XuL, LeeJ, Xu T. (2002). “Drosophila side twoDrosophilacells(markedinred color). (green color)ofdifferentsizesaredetected in- presence ofmanylysosome-relatedorganelles Biogenesis oflysosome-relatedorganelles. The IMMPACT REPORT • • • • 41 Currently, the team effectively translates new the team effectively Currently, of clinical made in the process Discoveries found of gene variants validation Biological with next generation sequencing using studies, cellular functional assays, and protein transgenic animal models; reverse of therapeutic targets to Identification disease phenotypes in cellular and transgenic animal models; and of imaging devices and imaging Re-engineering clinical utility of diagnostics. agents to improve scientists from across the Texas Medical Center Medical Texas the across scientists from partner apply with CMI members to effectively and clinical studies. diagnostics in preclinical literally NIRF molecular imaging technologies in “bench-to-bedside and back again,” from efforts embrace its division and clinical that Center and in the Medical Texas partners in the suburbs. Houston studies in “back to the bench” translation require the CMI include: • • • Ph.D. Sevick-Muraca, Eva & Professor Director Center and Rich Kinder Distinguished Chair in Nancy Disease Research Cardiovascular for in the NCI Network Center Director, Research Translational

he Mission of the Center for Molecular of the Center for Molecular he Mission and translate (CMI) is to develop Imaging new medical imaging technologies, The CMI houses a diverse, interdisciplinary The CMI houses a diverse, Biologics focuses Applied Division of The molecular imaging agents, and companion molecular imaging agents, and companion that advance diagnostics to accelerate discoveries molecular medicine. and develop team of scientists and engineers who and use multi-modality molecular diagnostics imaging, imaging techniques, including nuclear bioluminescence, X-ray computed tomography, fluorescence (NIR) near-infrared and fluorescence, disease to enable new understandings in several states. and engineering of antibody- upon development based diagnostics and therapeutics for high- and the affinity targeting of disease markers, Sequencing Generation of Next Division in bioinformatically associating specializes of disease-causing genotypes to enable discovery in translational studies. Biological gene variants lead of these disease-causing variants validation for newto the next steps of target discovery of unmet therapeutic and diagnostics in areas addition to having its own clinical need. In the projects, basic science and clinical research center and its divisions synergistically operate a and basic clinicians center where “collaboration”

MOLECULAR IMAGING FOR MOLECULAR CENTER T 42 • IMMPACT REPORT CENTER FORMOLECULARIMAGING human disease. in humandiseasesandanimalmodelsof of lymphangiogenesisandlymphaticstasis regulators associatedwithaberrantprocesses signalingpathwaysand include identifyingkey imaging studiesconductedbytheCMIteam the translationalNIRfluorescencelymphatic translational research. Discoveriesmadefrom and backagaininordertomakediscoveries technologies literallyfrom “bench-to-bedside” NIRimaging team effectivelytranslatesnew of Medicine, andtheMethodistHospital. Our engineers andscientistsatRice, BaylorCollege Pathology, andOtorhinolaryngology, aswell ogy, the UTHealth Vascular Anomalies Clinic, scientists inPediatrics, InterventionalRadiol- studies. We activelycollaboratewithclinical apply diagnosticsinpreclinicalandclinical withCMImemberstoeffectively Center partner basic scientistsfromacrossthe Texas Medical “collaboration” centerwherecliniciansand research projects, theteamalsooperatesa to havingitsownbasicscienceandclinical diseasestates.ings inseveral Inaddition understand - (NIR) fluorescencetoenablenew fluorescence, andourspecialty, near-infrared X-ray computedtomography, bioluminescence, molecular imagingincludingnuclearimaging, and engineerswhofocusuponmulti-modality sists ofaninterdisciplinaryteamscientists The CenterforMolecularImaging(CMI)con- Nancy andRichKinderDistinguishedChairinCardiovascularDiseaseResearch Professor andDirectoroftheCenterforMolecularImaging Eva MarieSevick-Muraca, Ph.D. 22034868 63: 217-31, 2012, Epub2011Oct27. PMID: emerging clinicalapplications,” AnnRevMed, infrared fluorescenceimagingtechnologies: E.M. Sevick-Muraca, “Translation ofnear- PMID: 24590275 Clinical Investigation, 124(3):905-14, 2014. technologies formouseandman,” Journal of Rasmussen, “Emerging lymphaticimaging Sevick-Muraca, E.M., Kwon, S.K., andJ.C. 2014, PMID:24972649 chylothorax patient,” Pediatric Cardiology, at thebedsideinapediatricpostoperative Muraca, “Investigational lymphaticimaging R., Bricker, J.T., Douglas, W.I., andE.M. Sevick- Tan, I.C., Balaguru, D., Rasmussen, J.C., Guilliod, 25383712. Plos One,10;9(11):e112548, 2014. PMID: contributing toalymphaticdysfunction,” containing 5’-inosotilphosphatase-2(SHIP2) Sevick-Muraca, “Evidence forSHdomain- Guilloid, R., Maus, E.A., King, P.D., andE.M. J.C., Tan, I-C., Aldrich, M.B., Darne, C., Fife, C.E., Agollah, G.D., Gonzalez-Garay, M.L., Rasmussen, Med. Imaging, 2015, PMID:25438307. a commercialmulti-modalscanner,” IEEE Trans. frequency-domain fluorescencetomographyin perimental comparisonofcontinuous-waveand Rasmussen, J.C., andE.M. Sevick-Muraca, “Ex- Lu, Y., Darne, C., Tian, I.C., Zhu, B., Rightmer, R., KEY PUBLICATIONS •Small animalimagingandtomography •Imaging lymphaticre-organizationinresponse •Imaging cancer-positivelymphnodeswith •Lymphatic deliveryoftherapeuticstargeting •Molecular imagingofMMP-targetedviralgene •Imaging chylo-andlymphothoraxinchildren RESEARCH PROJECTS Molecular imaginganddiagnostics to surgeryandradiation tive lymphnodedissection molecular imagingagenttoguideintraopera- a cancer-specificnear-infraredfluorescent the immunesystem disease delivery vectorsfortreatmentofheart defects with congenitalheart inheritable lymphaticcondition. biological roleofINPPL1mutationinthe evidencing complex denotedinred)further in lymphaticendothelialcells(INPPL1/VEGRF3 with VEGFR3 aknown growthfactorexpressed found fromproximityligationassaytoassociate (RIGHT) The genecandidateofINPPL1was mutations fromnextgenerationsequencing. lowing discoveryofcandidatecausativegenetic was usedtoaccuratelyphenotypethefamilyal- near-infrared fluorescencelymphaticimaging able non-syndromiclymphedemainwhich (LEFT) Pedigreeoffamilyharboringinherit- produced fromSevick-Muraca, etal.,JCI, 2014. to thelymphaticvesselsandlymphnodes. Re- Routes ofadministrationthatresultindelivery IMMPACT REPORT • • • • 43 26(8):1128-35. (a) lateral NIRF image of mouse lymphatics, (a) lateral NIRF image of mouse lymphatics, (b) pulsative movement of lymph bolus through lymphatic collector arms of subjects with breast cancer-related arms of subjects with breast near-infrared lymphedema as assessed by Biomedical Optics 2012. fluorescent imaging. Express 3:1256-65. Sevick-MuracaEM, Marshall MV, Aldrich MB, Uddin LV, Wang Culver J, Kotyk J, Lanza G, C, Contag Liao JC, Marnett LJ, Crews BC, J, Appelman H, Reisdorph B, K, Wang Crawford JM, Seeing it through: T. Wang Meyer C, KD, Turgeon translational validation of new medical imaging Biomedical Optics Express 2012. modalities. 3(4):764-776. XF, Huang, Lapteva N, Sanders D, Aldrich MB, SOCS1 downregulation in den- and Chen SY. T cell responses. dritic cells promotes memory 2008. Vaccine LAB MEMBERS Graduate students: co-advised Cynthia Davies- Pier-Anne Lachance Venn, Foot lymphatic vessel usage in a lymphedema patient before (left) and after (right) pneumatic compression therapy lymphatic function architecture and function in health and disease In addition to translational work, I am active I am In addition to translational work, KEY PUBLICATIONS Cytokines are Sevick-Muraca EM. Aldrich MB, systemic effectors of lymphatic function in Cytokine 64:362-9. 2013. inflammation. Rasmussen JC, Gonzales-Garay ML, Agollah GD, Guilliod R, Fife CE, Darne C, Aldrich MB, IC, Tan Evidence Sevick-Muraca EM. King PD, Maus EA, inositol phos- for SH2 domain-containing 5’ phatase-2 (SHIP2) contributing to a lymphatic PLoS One 9:e112548. 2014. dysfunction. Smith Maus EA, Fife CE, Guilliod RG, Aldrich MB, Lymphatic Sevick-Muraca EM. Rasmussen JC, L, abnormalities in the normal contralateral Validation in the context of translation •Validation •Inflammatory cytokine effects on systemic Imaging in immunologyImaging with research groups Working into the clinic. I served as the from several other institutions, Re- Translational leader of the NCI Network for and Clinical Studies Core that Validation search and a book chap- authored a consensus paper effortster describing some of the translation needed for validation of optical imaging devices This group was and molecular imaging agents. part of an effort sharing and by NCI to promote dissemination of translation practices amongst I published work In addition, researchers. describing processes for assessing parameters for which there was previously guidance no FDA available. in basic science investigations that employ the investi- I have technologies I work to translate. gated the effects of inflammation on lymphatic act and found that cytokines function in mice, as systemic mediators of lymphatic pumping Work through iNOS-associated mechanisms. by other groups has shown that inflammatory this but cytokines affect lymphatic function, study was the first to show that the effects are in and defines a role for inflammation systemic, are now extending We some lymphatic diseases. this work to study the role of lymphatic pumping dysfunction in rheumatoid arthritis and other inflammatory diseases. RESEARCH PROJECTS •Clinical studies of NIRF imaging of lymphatic Melissa B. Aldrich, M.B.A., Ph.D. M.B.A., Aldrich, Melissa B. Assistant Professor I bring a combination of expertise in Reimbursement from Medicare and medical I am formally and practically trained in trans- MOLECULAR IMAGING FOR MOLECULAR CENTER translational science and immunology to lead the program of imaging of the lymphat- is critical to the circulatory system which ics, Near- immune surveillance and response. infrared fluorescence (NIRF) imaging delivers low-cost images of lymphatic high-resolution, In disease vessel architecture and pumping. by manifested states such as lymphedema, NIRF imaging can provide severe swelling, limb information for diagnosis and evaluation of Our translational team’s treatment efficacy. study of NIRF images of breast cancer-related lymphedema arms revealed that the severity of the disease worsens over time not only in the and/or arms (that received surgical “affected” radiological treatment associated with breast but also in the contralateral cancer treatment), This work added evidence (“unaffected”) arms. to other studies suggesting that lymphedema is Our lab has disease. not just local, a systemic, also worked in NIRF imaging studies of primary, lymphedema and rare fat-associated or genetic, genetic disorders with lymphatic abnormali- recently identified severalWe previously ties. unknown causative mutations for lymphedema in families with members suffering from the disease. insurance companies for many clinical pro- as the health care cedures is under scrutiny, system tries to make the best use of health care recently surveyedWe different types of dollars. to using NIRF imaging, lymphedema patients, provide visual proof that one such procedure, is effective for pneumatic compression therapy, Similar NIRF studies by moving stagnant lymph. our group have now shown that this therapy is also efficacious for treating venous insufficiency and chronic wounds. Understanding concepts lation requirements. such as validation of imaging devices and batch release of imaging agents enables researchers to discern which types of laboratory tests are necessary for moving a medical device or drug 44 • IMMPACT REPORT CENTER FORMOLECULARIMAGING • • • • RESEARCH PROJECTS health. technologies intotheclinictoimprovehuman focus onrapidtranslationofdiscoveriesand tion techniques. Common toeachprojectisour boneforma- assessing theeffectivenessofnew of novelnanomaterialsforbiomedicaluse, and imaging of “good” fattissue, characterization cancersuch as collaborations inareasbeyond acterization hasallowedustoestablishdiverse inchemistry,expertise imaging, anddrugchar- ment protocols. Importantly, ourfundamental and visualizetheeffectstopersonalizetreat- platform tospecificallydelivertoxinstumors is notpossible, weaimtouseourchemistry In caseswherecancerhasspreadandsurgery would otherwisebemissedbythenakedeye. healthy tissues, andidentifysmalltumorswhich cancer fromnormaltissue, minimizeremovalof intraoperative imagesthatwilldistinguish potentially providesurgeonswithreal-time intraoperative imaging, respectively. This could specifically identifycancerbywhole-bodyand approachesto new has allowedustodevelop fluorescent labelsontotumor-seekingagents For example, theadditionofradioactiveand multiple labelsandthus, multipleapplications. we havetheabilitytoproducemoleculeswith of disease. Usingnovelchemistryplatforms, molecules forthevisualizationandtreatment andisfocusedondeveloping and biology cancer andotherdiseases Characterization ofimagingprobestargeting Tumor-specific chemotherapy erative imaging ofcontrastagentsforintraop- Development imaging andtherapy Application ofanovelchemistryplatformfor My laboratoryisattheinterfaceofchemistry Research Scientist:SukhenGhosh LAB MEMBERS PMID:25457653. conjugates. NucMedBiol. 42(2):177-83, 2015. Assistant Professor Ali Azhdarinia,Ph.D. NODAGA asChelatorsfor E.M., Azhdarinia, A. ComparisonofDOTA and B.R., Wilganowski, N., Gore, K., Sevick-Muraca, Ghosh, S.C., Pinkston, K.L., Robinson, H., Harvey, PMID:24628687. Nanomedicine (Lond). 9(16):2499-509, 2014. nanotube capsulesforinvivobimodalimaging. confinement ofPET&MRagentswithincarbon rinia, A., Sevick-Muraca, E.M., Wilson, L.J. Stable Cisneros, B.T., Matson, M.L., Law, J.L., Azhda- 4:2472, 2013. PMID:24045463 brown adiposetissueimaging. NatCommun. Muraca, E.M., Kolonin, M.G. Probesfortargeted S.C., Ghosh, P., Amaya-Manzanares F., Sevick- Azhdarinia, A.,Daquinag, A.C., C.,Ghosh, Tseng, 56(2):406-16, 2013. PMID:23214723. Fluorescence/Nuclear Imaging. JMedChem. Multimodal ChelationPlatformforNear-infrared E.M.,and Azhdarinia,B.,Sevick-Muraca, A. A Robinson, H., Hall, M.A., Dickinson, G., Harvey, Ghosh, S.C., Ghosh, P., Wilganowski, N., PMID:22872743. JNuclMed.53(9):1427-37,CT. 2012. contrast: NIRFimagingandquantitativeµPET/ cancer lymphnodemetastaseswithmultimodal mAbs targetingEpCAMfordetectionofprostate B.R., andSevick-Muraca, E.M. Comparisonof Arreguin, K., Kolonin, A.M., Chan, W., Harvey, Robinson,H.,Ghosh,P., Azhdarinia, A., Vasquez- Hall, M.A., Pinkston, K.L., Wilganowski, N., KEY PUBLICATIONS Molecular imagingprobedevelopment 64 Cu-labeled Immuno- Nanomedicine, 9(16):2499-509, 2014). nanotubes. (adapted fromCisnerosetal., after encapsulationofisotopewithinthecarbon images, whereas lunguptakeisonlyapparent Liver uptake(whitearrow)isobservedinboth ( Imagesofnormalmiceinjectedfreeisotope S.C. etal.,JMedChem, 56(2):406-16, 2013). Lu =lung, H=heart, M=muscle. (fromGhosh, ings. Arrow indicatesexcisedtumor. K=kidney, excised tissues(B)correlatedwithimagingfind- imaging (A)andfoundthatquantificationof excellent tumorvisualizationviawhole-body the tumor-specificimagingagent, weobserved infrared fluorescentlabels. Afterinjection of contrast agentpossessingradioactiveandnear- cells andusedtostudytheeffectivenessofa Mice wereimplantedwithprostatetumor 64 Cu) andradiolabeledcarbonnanotubes. IMMPACT REPORT • • • • 45 Pinkston KL*, Gao P*, Diaz-Garcia D, Sillanpää Sillanpää Diaz-Garcia D, Gao P*, Pinkston KL*, and Harvey BR. Murray BE, SR, Nallapareddy J, of Enterococ- “The Fsr quorum-sensing system display of cus faecalis modulates surface Ace through the collagen-binding MSCRAMM of Bacteriology, Journal regulation of gelE.” 21705589 PMID: Sep;193(17):4317-25 2011. LAB MEMBERS Peng Gao Research Instructor: Dr. Pinkston Research Coordinator II: Kenneth L. Graduate Student: Emily Stinemetz Modification of monoclonal antibody to decrease unwanted interactions with immune of EndoS treatment (deglycosylation) cells. mAb960 inhibits binding to macrophage as Preventing analyzed by flow cytometry analysis. unwanted antibody interactions decreases background labeling when antibodies are utilized as imaging agents (as in Figure 1). 2013, of Bacteriology 2013, Journal Molecularly targeted live animal imaging of heart infection for diagnostic use. PET/CT image of entero- PET/CT Molecularly targeted live animal imaging of heart infection for diagnostic use. coccal endocarditis (bacterial infection of the heart valve or inner lining) in live rats (A1-A3) imaged Signal increased labeled mAb targeting the bacterial surface. at 72 h post infection using Cu64-DOTA (yellow intensity) as infection grew units (CFUs)). in size (as measured by colony forming Therapeutic and diagnostic antibody development and diagnostic Therapeutic KEY PUBLICATIONS Robinson Wilganowski N, KL, Pinkston Gao P, Harvey Sevick EM, Zhu B, A, H ,Azhdarinia by EndoS for BR.“Deglycosylation of mAb Molecular improved molecular imaging. 2015 Mol Imaging Biol. Imaging and Biology.” PMID: 25135058 Apr;17(2):195-203. WilganowskiN, Gao P, Singh KV, KL, Pinkston, Sevick- A, Azhdarinia Ghosh SC, Robinson H, “Targeting BR. Harvey, Murray BE, Muraca EM, Infection Pili in Enterococcal Pathogenesis” PMID: Apr;82(4):1540-7. 2014 and Immunity, April 2014 (Featured article on 24452680. Cover). MR, Cruz A, Bourgogne Pinkston KL, Gao P, “Library Harvey BR. Murray BE, Garsin DA, Screen identifies Enterococcus faecalis CcpA, as an Effector A, the Catabolite Control Protein A of Ace, Collagen Adhesion Protein Linked to Virulence” PMID: 23974022. Oct;195(20):4761-8. Wilganowski, K.L.*, Pinkston, M.A.*, Hall, A., Azhdarinia, P., Ghosh, H., Robinson, N., Harvey, A.M., Kolonin, K., Vazquez-Arreguin, of “Comparison E.M.*, Sevick-Muraca, B.R.*, mAbs targeting EpCAM for detection of prostate cancer lymph node metastases with multimodal contrast: NIRF imaging and quantitative µPET/ Sep;53(9):1427-37 CT,” J Nuc Med, 2012. PMID: 22872743 Barrett Rowland Harvey, Ph.D. Harvey, Barrett Rowland Assistant Professor Technological achievements in antibody Technological static cancer models coccal infection bacterial infection engineering have made antibody drug development one of the fastest growing areas of the Successful design of pharmaceutical industry. antibody-based therapeutics or diagnostics requires both the ability to optimize the antibody and a clear understanding of the biology our laboratory this end, To of the target antigen. identify and build To has two main focuses: 1) a functional understanding of novel molecular as often utilizing custom antibodies targets, powerful tools to expedite the research and 2) to develophigh throughput strategies and engi- specific- neering methods to modify the affinity, recognition and Fc function epitope site ity, and diagnostic, of antibodies for therapeutic, imaging Utilizing molecular basic research use. antibody agent development can techniques, be monitored using in vivo models to predict and to validate targets prior specificity, efficacy, This line of research allows our to the clinic. laboratory to venture into a number of diverse currently with ongoing projects biological fields, focused in oncology and infectious disease. RESEARCH PROJECTS •Generation of surrogate antibodies for meta- •Molecular imaging for cancer staging •Virulence factor regulation governing entero- •Passive protection from hospital acquired MOLECULAR IMAGING FOR MOLECULAR CENTER 46 • IMMPACT REPORT •Multi-modal molecular imaging •Non-invasive imagingofgastrointestinal •Non-invasive characterizationoflymphatic RESEARCH PROJECTS disease. and inflammationindifferentanimalmodelsof molecular imagingofcancerandLNmetastasis fluorescent dye. We arecurrentlyconducting labeled withaPET/SPECTradiotracerandNIR and translatingimagingagents, which aredual- The CenterforMolecularImagingisdeveloping volve aroundmulti-modalitymolecularimaging. to therapy. provide diagnosticsandinformationinresponse and architectureindiseasepotentiallyto used toimagechangesoflymphaticfunction Therefore, non-invasiveNIRFimagingcanbe ing lymphaticfunctioninhealthanddisease. imaging methodsforandquantify- dynamic near-infraredfluorescence(NIRF) Recently, non-invasive, wehavedeveloped the lackofdiagnosticimagingapproaches. function hassignificantdifficulties, dueto recognized, non-invasiveimagingoflymphatic pathophysiological conditionshasbeenwell of thelymphaticsysteminphysiologicaland cancer metastasis. Although theimportance edema prevention, immunesurveillance, and rolein lymphatic systemplaysanimportant fluorescence opticalimagingtechniques. The inthelymphaticsystemusing particularly latory movementoffluidandmacromolecules, interest focusesoninvestigatingthemicrocircu- of gastrointestinaldisease. Mymainresearch of vasculardisease, withanemergingemphasis biological questionsinuniqueanimalmodels small animalimagingtechniquestoaddress CENTER FORMOLECULARIMAGING technique motility usingafluorescence opticalimaging response totherapeuticagents sion, cancer, andinflammationtracking lymphedema-like phenotypes,- hyperten function anddrainagepatternsinmicewith Other directionsofmyscientificinterestsre- andapplicationof I leadthedevelopment Assistant Professor Sun KukKwon, Ph.D. (IsLN) throughthesiteofPLNremovalwere visualizedatday20. flow resistance. However, once lymphaticcontinuitywasrestored, lymphaticvesselstotheischialLN At 2daysafterPLNremoval, alternatedrainagepathwaystotheILN weredetectedduetoincreased Cancer. 5;774-783, 2014. inflammatory breastcancermodel,” Journal of S. Kwon, “In vivolymphaticimagingofahuman D. A. Germaine, G. Wu, E. M. Sevick-Muraca, and 9; e106034, 2014. following lymphadenectomyinmice,” PLOSOne. lymphatic contractilityanddrainagepatterns Sevick-Muraca, “Spatio-temporal changesof S. Kwon, D. A. Germaine, G. Wu, andE. M. Investigation. 124;905-914, 2013. gies forMouseandMan,” Journal ofClinical Imaging Technolo- sen,Lymphatic “Emerging E. M. Sevick-Muraca, S. Kwon, andJ. Rasmus- Academy ofSciences, 110;8621-8626, 2013. mouse andman,” ProceedingsoftheNational are associatedwithRASA1genemutationsin M. Sevick-Muraca, “Lymphatic abnormalities Fife, S. Kwon, P. E. Lapinski, P. D. King, andE. sen, M. B. Aldrich R. Guilliod, E. A .Maus, C. E. P.E. Burrows, M. L. Gonzalez-Garay, J.C. Rasmus- KEY PUBLICATIONS lymphovascular disorders Functional lymphaticimaginginanimalmodelsof Research Coordinator: Grace Research Coordinator: Wu LAB MEMBERS publication, 2015. Worldfor Journal ofGastroenterology. Accepted impairs dermallymphaticfunctioninmice,” “Dextran sulfatesodium-inducedacutecolitis D. A. Germaine, G. Wu, H. Peng, andS. Kwon, image ofIBC(red). of thelymphaticsoveranon-invasiveiRFP reporter. (B) An overlayofaNIRFimage(green) tion ofIBCcellstransfectedwiththeiRFPgene NIRF imageofthelymphaticspriortoinocula- carcinoma (IBC)onlymphaticstructure. (A) A Images oftheeffectinflammatorybreast IMMPACT REPORT • • • • 47 Image of (A) the well-defined, linear lymphatics linear Image of (A) the well-defined, in the arm of a healthy subject and (B) the arm of a dermal lymphatics in the disorganized, subject with lymphedema. Rasmussen, J.C., Tan, I., Marshall, M.V., Adams, Adams, M.V., Marshall, I., Tan, J.C., Rasmussen, L., Smith, E.A., Maus, C.E., Fife, S., Kwon, K.A., “Hu- Sevick-Muraca, and E.M. K.R., Covington, and (dys)function man lymphatic architecture Translational imaged using NIR fluorescence,” 2010. 3(6):362-372, Oncology, Images illustrating the effect of a single session of sequential pneumatic compression (SPC) therapy on the ulcer draining lymphatics in sub- In some jects with chronic venous insufficiency. NIRF images acquired (A) before and subjects, (B) after treatment indicated that additional lymphatics had been recruited to aid lymphatic NIRF images in other subjects, drainage while, acquired (C) before and (D) after treatment illustrated the emptying of wound draining lymphatic vessels following SPC treatment. imaging systems analytical algorithms vascular diseases disease NIRF imaging KEY PUBLICATIONS Darne, I-C., Tan, M.B., Aldrich, J.C., Rasmussen, and C.E., Fife, T.F., O’Donnell, B., Zhu, C.D., “Lymphatic transport in E.M., Sevick-Muraca, patients with chronic venous insufficiency and venous stasis leg ulcers following sequential Journal of Vascular pneumatic compression,” Disorders, and Lymphatic Venous Surgery: http://dx.doi.org/10.1016/j. In Press, jvsv.2015.06.001; epub 7/16/15. Rasmussen, K.D., Cromwell, S.F., Shaitelman, and B.B., Lasinski, J.M., Armer, N.L., Stout, J.C., Progress in Cancer-Relat- “Recent J.N., Cormier, CA: Prevention,” and Treatment ed Lymphedema 65(1):55-81, for Clinicians, A Cancer Journal 2015. M.B., Aldrich, K.L., Herbst, J.C., Rasmussen, Fife, R., Guilliod, B., Zhu, I-C., Tan, C.D., Darne, “An E.M., Sevick-Muraca, E.A., Maus, C.E., abnormal lymphatic phenotype is associated with subcutaneous adipose tissue deposits 22(10): 2186- Obesity, disease,” in Dercum’s 2014. 2192, Gonzalez-Garay, P.E., Burrows, J.C., Rasmussen, Fife, E.A., Maus, R., Guilliod, M.B., Aldrich, M.L., and P.D., King, P.E., Lapinski, S., Kwon, C.E., “Lymphatic abnormalities Sevick-Muraca, E.M. are associated with RASA1 gene mutations in Proceedings of the National mouse and man,” 110(21):8621-8626, Academy of Sciences, 2013. •Application driven enhancement of NIRF •Development of automated NIRF image •Identification of genetic causes for lympho- •Etiology lymphedema of cancer-related vascular •Lymphatic involvement in peripheral Deviceimaging for lymphatic translation RESEARCH PROJECTS noninvasive •Nodal staging of cancer using John Rasmussen, Ph.D. John Rasmussen, Assistant Professor I am the faculty lead of the instrumenta- My work focuses upon the development of MOLECULAR IMAGING FOR MOLECULAR CENTER tion for translational fluorescence imaging. such as clinical imaging modalities, Traditional lack and ultrasound, MRI, X-ray, scintigraphy, the spatial and/or temporal resolutions needed to resolve fine lymphatic architecture and contractile function and/or require quantities of contrast agent not easily introduced into the research my fewthe past Over years, lymphatics. interest has focused upon the development and translation of near-infrared fluorescence (NIRF) optical imaging as a way to noninvasively image and characterize human lymphatics and quantify their contractile function in health and disease using microdose amounts of fluorescent contrast agent. NIRF imaging methodologies and its application to answer new biological and clinical questions - Specifi not addressed by other technologies. focuses upon using NIRF our program cally, imaging in translational clinical studies with partners across the Houston area to (i) study the growth and reorganization of the lymphatics, (ii) elucidate the termed lymphangiogenesis, role of the lymphatics in the development of such as lymphedema lymphovascular diseases, and cancer metastasis as well as in rare adipose disorders and chronic peripheral vascular diseases that may have a lymphovascular and (iii) identifying the lymphatic component, phenotype of genetic mutations that contribute My expertise involves to lymphatic disorders. the application of NIRF imaging instrumentation and development of software for clinical Specific projects focus on the applications. development of analytical tools to facilitate lymphatic image processing and analysis. 48 • IMMPACT REPORT molecular imagingdevices. the measurementsensitivityoffluorescence fluorescent irradianceforuseincharactering tocalibrateastable,ogy asolidphantomfor anddeploying a methodol- we aredeveloping Ad5-iRFP and Ad5-BMP-2 vector. Inaddition, with theinduciblecaspase9systemandan the osteoprogenitorcellsthataretransduced therapeutic forboneformationbyinjectionof draining lymphnodes, the and(ii)toevaluate implantation throughlymphaticvesselsandto siteofmammary tases fromanorthotopic track theinvivoprocessoflymphaticmetas- iRFP, (i)tolongitudinallyandnon-invasively are usingfar-redfluorescencegenereporter, optical imagingintotheclinic. Specifically, we with SIunitstoacceleratethetranslationof establishment oftraceableworkingstandards tion forpreclinicalandclinicalstudiesthe ofopticalimaginginstrumenta- development CENTER FORMOLECULARIMAGING My researchprogramfocusesuponthe Assistant Professor Banghe Zhu, Ph.D. Optical imaging, instrumentationandworkingstandards Co-advised: Grace Wu,JulieVoss Co-advised: LAB MEMBERS (2013). Molecular ImagingandBiology, 15:560-568, Validated iRFP”, byFar-RedGeneReporter tive, NIRFMolecular Imaging Targeting EpCAM E. M., “Tumor MarginDetectionusing Quantita- Azhdarinia, A., Harvey, B. R., andSevick-Muraca, N., Hall, M. A., Ghosh, S. C., Pinkston, K. L., Zhu, B., Wu, G., Robinson, H., Wilganowski, Express. 5(2):562-572(2014) ICCD andlaserdiode,” BiomedicalOptics ambient lightconditionsusingamodulated M., “Non-invasive fluorescenceimaging under Zhu, B., Rasmussen, J.C., andSevick-Muraca, E. to see?”, MedicalPhysics,41, 022105(2014). fluorescence molecularimaging:toseeornot intraoperative andnon-invasivenear-infrared E. M., “A matterofcollectionanddetectionfor Zhu, B., Rasmussen, J. C., andSevick-Muraca, 88:20140547(2015) applications,” BritishJournal ofRadiology, ing: ofdevices, performance,A review and clinical near-infraredfluorescenceimag- Zhu. B., andSevick-Muraca, E.M., “Update on 3351 (2015). discovery”, BiomedicalOpticsExpress,6:3346- preclinical models:atoolforacceleratingdrug imagingofcancermetastasisin gene-reporter Sevick-Muraca, E. M., “Longitudinal farred Zhu, B., Robinson, H., Zhang, S., Wu, G., and KEY PUBLICATIONS • • • • RESEARCH PROJECTS standards foropticalmedicalimaging working Collaborating withNISTtodevelop targeted boneformation Evaluating cell-basedgenetherapyfor reporter Tracking cancermetastasisusingiRFPgene for medicalapplications opticalimaginginstrumentation Developing gland byweeks6and7. mammary draining cellsfromtherightfourth the rightandleftlymphaticchannels(arrows) cer metastasisinpreclinicalmodels, indicating Longitudinal iRFPfluorescenceimagingofcan- (CID) ornot. treated withachemicalinducerofdimerization tensity, and(c)Radiographicalanalysisinmice tion overtime, (b) MeaniRFPfluorescencein- (a)iRFPfluorescenceimagingofboneforma- IMMPACT REPORT • • • • 49 small molecule drugs for in-depth proteomic or and metabolomics analysis of cells, tissues, proteins then servebiological fluids. These as targets for drug and nanomedicine development next- therapeutics and imaging agents, including generation X-aptamer reagents. Collaboration with the Center of Research Hubs include: Chemistry • Protein Monitoring Drug and Therapeutic • Proteomics Bioinformatics and UTHealth Biology • Systems LaboratoryCore Core Proteomics Translational • Clinical and Laboratory Diagnostics Molecular • CLIA ProteoPath Laboratory Cancer Computational in • NCI Programs and Nanomedicine Biology Facility Core Proteomics • UT System-wide Network Translational Clinical and / MDACC • UTHealth Technologies Center for Translational Ph.D. Gorenstein, David Director & Deputy Director, Center Professor, Distinguished Chair in Willerson T. James the Ewing from Tribute in Research Cardiovascular Foundation Halsell

iology connects campus-wide and state- efforts in systems biology, wide research clinical and translational sciences, While genomics can successfully catalog While genomics can successfully catalog mission of the Center for Proteomics One Facility is located Spectrometry Mass The nanomedicine, protein chemistry, genomics, chemistry, nanomedicine, protein bringing and bioinformatics by proteomics, intellectual exchange people together to promote in these key fields. nearly all drugs genetic variants, on the market products. today target their functional protein us starting sequences give points, but most Gene and processed extensively are cellular proteins regulation and To understand cellular modified. disease mechanisms, or to identify drug targets, that need detailed characterization of proteins we mass spectrometry through achievable are now technologies. and other proteomic the is to develop (CPSB) Biology and Systems experimental and analytical technologies that only The CPSB will not will make this a reality. a new technologies but also will provide develop for of centers and programs group coordinated and servicecollaborative for the UTHeath work protein community in cutting-edge proteomics, nanomedicine and systems biology chemistry, research. state-of-the-artin the IMM and houses seven the identification that allow mass spectrometers and and quantification of peptides, proteins,

PROTEOMICS AND SYSTEMS BIOLOGY SYSTEMS AND FOR PROTEOMICS CENTER B 50 • IMMPACT REPORT and E-Selectin. tissue andcellsexpressingCD44, Annexin A2 selection aredemonstratedfortargetingcancer the bead-basedthioaptamerandX-aptamer and diagnostics. Examples ofapplication crofluidics andmassspecchipsforproteomics cells andtumorvasculatureinvariousmi- peutics andbiomarkeridentificationtotumor as antibodysubstitutesinnanomedicinethera- X-aptamers andthioaptamersarebeingused these aptamerscanberapidlyselected. These Large bead-basedcombinatoriallibrariesof high affinityfordiversemolecularbiomarkers. be incorporatedtoselectX-aptamerswith cally expandsthechemicaldiversitythatcan conventional aptamers, thisapproachdramati- by oligonucleotidesequencing. Comparedto folding scaffoldthatcanbereadilyidentified sidechains, andsmallmoleculeleadsinaself- of aptamerbackbone, proteinaminoacid-like 5-X-dU positionandthusrepresentahybrid range ofchemical(X)modificationstothe targets. The X-aptamersalsoinclude alarge “X”-aptamers toanumberofdifferentprotein oligonucleotide “thioaptamers” andnext-gen methods toidentifyphosphorothioate-modified and split-synthesischemicalcombinatorial in vitroenzymaticcombinatorialselection and nanomedicine. both We havedeveloped target anumberofproteinsforproteomics selected fromlargecombinatoriallibrariesto modified DNAoligonucleotideaptamers CENTER FORPROTEOMICS AND SYSTEMSBIOLOGY We have developed novel,We havedeveloped next-generation Halsell Foundation James T. DistinguishedChairinCardiovascularResearch in Willerson Tribute fromtheEwing Professor andDirectoroftheCenterforProteomicsSystemsBiology Chair, ofNanoMedicineandBiomedicalEngineering Department Associate DeanforResearch David Gorenstein, Ph.D. 2015; doi:10.1038/mt.2015.45 Molecular Therapy,Cancer Metastasis, ofBreast metastatic NicheforthePrevention Hallgeir Rui, Takemi Tanaka, BlockingthePre- Suh, DavidG. Gorenstein, MassimoCristofanilli, Zheng, LichaoZhao, DanielZhao, K. Stephen Shin-Ae Kang, NafisHasan, Aman P. Mann, Wei PMCID: PMC4109393. PubMed PMID:24818768;Central jconrel.2014.04.057. Epub 2014May10. Release 187, 22-29(2014). doi:10.1016/j. tics formetastaticbreastcancer. JControl Bone marrowendothelium-targetedtherapeu- Gorenstein DG, LiuX, Ferrari, M., andShen, H. Xia X, Volk DE, LokeshGL, Thiviyanathan V, Mai J, Huang Y, MuC, ZhangG, XuR, Guo X, bi300471d. (Frontpagecover). binding” Biochemistry, 2012DOI:10.1021/ onto next-generationaptamersforenhanced for randomincorporationofdrug-likemoieties “X-Aptamers: A bead-basedselectionmethod Claudio N. Cavasotto, andDavidG. Gorenstein E. Volk, RossH. Durland, JohnnieEnglehardt, masunderam, Varatharasa Thiviyanathan,David Li, GaneshLakshmanaRaoLokesh, Anoma So- Weiguo He, Miguel-AngelElizondo-Riojas, Xin 10.1517/17460441.2011.537321 PMID: 21359096;PMCID:PMC3045091. doi: 6, Number1, January2011, pp. 75-87(13). OpinioninDrugDiscovery,Expert Volume gies forthediscoveryoftherapeuticaptamers, Xianbin Yang, LiNa, DavidG. Gorenstein, Strate- KEY PUBLICATIONS ofnovelX-aptamertargeting •Development •Nanomedicine targetingincancerandcardio- •Proteomics andmoleculardiagnostics for •Next-generation aptamerdevelopment RESEARCH PROJECTS cardiovasculature disease NanoMedicine andproteomicsincancer nanoparticles forimagingandtherapeutics nanoparticles vascular disease drug development 28 April Research Associate: XinLi, MS Research Scientists:LokeshRao, PhD, LiLi, PhD LAB MEMBERS 108, 1026-37(2015). tion ofDNAphosphate,” BiophysicalJournal, protein-DNA affinitybyoxygen-to-sulfursubstitu- and JunjiIwahara, “Entropic enhancementof Anderson, Mark A. White, DavidG. Gorenstein, Zandarashvili,Levani DanNguyen, M. Kurtis aptamer beadlibrary. randomly covalentlyboundtoacombinatorial X-aptamers inwhichsmallmoleculehitsare Schematic forselectionofNext-generation graft mousemodel. to E-selectiin. This isapancreatictumorxeno- Near-Infrared-Imaging-dye labeledthioaptamer Targeting witha ofgoldnanoshellnanoparticles IMMPACT REPORT • • • • 51 A) The end of the Raman fiber optic probe The end of the Raman fiber optic A) (right) that can be inserted through the biopsy channel of a clinical colonoscope (left) for non- A B) invasive detection of tissue composition. scheme of the Raman spectroscopy integrated colonoscopy. A) Bone spectrum from in vivo transcutaneous measurement (blue) is in great agreement the proving with that from excised bone (red), potential of in vivo Raman for noninvasive Raman imaging (C) bone quality evaluation. on sectioned bone specimen provides spatial information of biochemical components in bone and correlates with the (mineral and collagen), white light images (B). detection bone quality deterioration Raman measurement (SORS) cancer, etc) cancer, •Raman imaging for pathogenesis KEY PUBLICATIONS K. Lebrun, D. Ding, H. Yang, J. Sun, X. Zhang, Q. Li, Z. Ambrose, CG RG Postier, Houchen, C. Ding, ZIP4 silencing improves bone loss in Li, M. Bi, X. 2015 Jul Oncotarget, 2015, pancreatic cancer, In press (2015) 20, Reverse-time Bi., and X. Lu, G. Ding, H. Wang, Z. IEEE 2015, migration based optical imaging, Aug 18, on Medical Imaging, 2015 Transactions In press (2015). Perrien, D.S. Sterling, J.A. Nyman, J.S. Ding, H. Development of Bi, and X. Mahadevan-Jasen, A. Assess Metastatic Raman Spectral Markers to of Biomedical Journal Bone in Breast Cancer, 19(11): 111606 (2014) Optics, Use of a Bi, and X. Lu, G. Ding, H. Wang, Z. mechanical iris based fiber optic probe for the Optics spatially offset Raman spectroscopy, 39(13):3790-3 (2014) Letter, J.S. Perrien, D.S. Merkel, A.R. Sterling, J.A. Bi, X. Prostate cancer Mahadevan-Jansen, A. Nyman, metastases alter bone mineral and matrix composition independent of effects on bone A quantitative study architecture in mice – Bone, using microCT and Raman spectroscopy. 56(2):454-60 (2013) LAB MEMBERS Post-doctoral Fellow: Hao Ding Guijin Lu Technician: •Biomarkers and circulating tumor cells Assessment of metastasis and disease caused •Assessment of metastasis and •Development of noninvasive transcutaneous Optical spectroscopy and imaging for medicine and Optical spectroscopy RESEARCH PROJECTS in situ (IBD, •Noninvasive optical diagnosis Xiaohong Bi, Ph.D. Bi, Xiaohong Assistant Professor Optical spectroscopy and imaging techniques RS exploits subtle changes in the molecular have extensive experience in quantifying We Another area of research involves developing have demonstrated great potential in providing Our research noninvasive in situ diagnosis. especially focuses on developing optical tools, problems for clinical Raman spectroscopy (RS), therapy such as early disease diagnosis, and guidance of surgery. response evaluation, composition of tissue and is sensitive to disease and aging associated biochemical are currently We changes in tissue environment. using an RS fiber optic system to test patients with inflammatory bowel disease (IBD) in clin- In vitro RS studies on colon biopsies have ics. shown over 99.7% accuracy in differentiating the two distinct yet often indeterminate forms The colitis. of IBD: ulcerative colitis and Crohn’s incorporation of RS to colonoscopy is expected Further to improve diagnosis accuracy in situ. application of RS in cancer diagnosis and surgical margin assessment is also being explored in our laboratory. which bone mineralization and composition, are important determinants of bone strength. The effect of genetic variations and disease on bone compositional properties and mechanical In ad- function is constantly studied in the lab. we have developed RS spectral markers dition, that are related to breast and prostate cancers These markers can induced bone alterations. be used to assess bone quality and to evaluate the response of metastatic bone to treatment. A noninvasive method is in development to test on animal model and patients based on the above findings. targeted imaging and biosensing methods using surface enhanced Raman spectroscopy (SERS). such By combining RS and nanotechnology, SERS methods can detect biomarkers in body fluid in up to fm scale. PROTEOMICS AND SYSTEMS BIOLOGY SYSTEMS AND FOR PROTEOMICS CENTER 52 • IMMPACT REPORT basis of a rational strategy for selecting clinical basis ofarationalstrategy module, may formthe andinvestigatehowthey now wishtoidentifythe genesthatdriveeach shown thatthesemoduleslinktodisease. We by geneexpressionprofiles. We havepreviously sub-components calledmodules, represented strategies todissectRasactivitiesintodiscrete structure ofRassignaling, wearedeveloping downstream effectors. To betterunderstandthe tumorigenic processesthroughmultiple on theRasnetwork. Rascontrols numerous ing thestructureofsignalingcascades, focusing metastasis inpreclinicalmodels. pathways thathaveshowntheabilitytoinhibit pounds andFDA-approved drugstargetingnovel We haveidentifiedaselectionofnaturalcom- exhibit phenotypesthatpromotemetastasis. proaches torepositiondrugstargetcellsthat this problem, we haveusedcomputationalap- not respondtotraditionaltherapies. To address metastasis, becausemetastaticcellsdo inpart that upto90%ofcancerdeathsaredue three areasoffocus: metastasis. and driveuncontrolledcellproliferation signaling programsbecomealteredincancer Specifically, wewishtounderstandhowthe circumstances, indisease. andhowitisrewired to explainhowthenetworkoperatesinnormal manage (abadthing). Therefore, achallengeis thing) aswelldiseasesthataredifficultto ity toproduceorganismslikeourselves(agood the cellsignalingnetworkprovidesitcapac- exhibit anetworkstructure. The complexity of interconnections amongsignalingmolecules crosstalk amongthem. Indeed, thedense entities, therealityisthattheresignificant are oftenthoughtofasindependentandlinear the expressionofgenes. While cellularpathways of reactions(pathways)thatultimatelychange ceptors inthecellmembraneinitiatecascades cancer phenotypeusinggenomics. Briefly, re- CENTER FORPROTEOMICS AND SYSTEMSBIOLOGY 2. Growthsignaling networks. We aredissect- 1. Breastcancermetastasis. Itisestimated Our researchprogramcanbegroupedinto Our labdeciphersthecomplexityof Assistant Professor Chang,Jeffrey Ph.D. inhibit metastasisin mice. can pounds thatcanalter cholesterolincancercellsandhavefound inpreclinicalstudiesthatthey roleincancermetastasis.We havediscovered thatcholesterolplaysakey We haveidentifiedcom- JT*, andManiSA*. A novelembryonicplasticity Soundararajan R, Paranjape AN, Barsan V, Chang KEY PUBLICATIONS •Intelligent computationalpipelinesforbioin- •Transcriptional regulatoryprogramsofE2F1- ofRassignaling •Genetic perturbations •Cancer metastasis, cancerstemcells, andthe RESEARCH PROJECTS biology, andbiochemistry. techniques includingbioinformatics, molecular arange of cell cultureasamodelandleverage as theworkingsofacancercell). We usehuman constitute complexbiologicalphenotypes(such thesimplefundamental unitsthat to reveal oncogenic pathways. ing aplatformSIGNATURE fortheanalysisof analysis ofgenesets,- andarenowdevelop theGATHER developed websitefor previously work istomakeourmethodsavailable. We have component, aspectofour andanimportant of ourprojectscontainsacomputational distribute ourcomputationalalgorithms. Each Lastly, infrastructureto wearedeveloping treatments. Genomic approachesforcancertherapies formatic analysis driven apoptosis epithelial-to-mesenchymal transition Across ourinvestigations, weusegenomics 3. Computationaltoolsforgenomicanalysis. Graduate Student: Kevin Zhu Graduate Student:Kevin Prjic, Ph.D., Post-doctoral Fellows: Weina Zhao, Ph.D., Sarah LAB MEMBERS pies. Nature439(7074):353-357, 2005. in humancancersasaguidetotargetedthera- JR.and Nevins Oncogenicpathwaysignatures A, OlsonJA, MarksJR, DressmanHK, West M, D, JoshiMB, HarpoleD, LancasterJM, Berchuck Bild AH, Yao G, ChangJT, Wang Q, Potti A, Chasse Networks. Molecular Cell 34(1): 104-114, 2009. Modules ofOncogenicPathwaySignaling JR.Nevins toElucidate A GenomicStrategy Wang Q, LucasJ, Potti A, FebboP, West M, and Chang JT, CarvalhoC, MoriS, Bild AH, GatzaM, databases. Bioinformatics28(8), 2012. and functionalprocessesfromgeneexpression Chang JT. Derivingtranscriptional programs * Mesenchymal Transition. wolf: CancerStemCellsandtheEpithelial-to- Chang JT*andManiSA*. Sheep, Wolf, or Were - 2015. *Co-Corresponding Authors tence andclinicaloutcome., ScientificReports gene signaturethatpredictsmetastaticcompe- Co-Corresponding Authors Cancer Letters, 2013. IMMPACT REPORT • • • • 53 (mean b killed by chemotherapy (i.e., pathologic killed by chemotherapy (i.e., kill Gottumukkala, M.A. Rozner, H. Shen, J.E. Lee, Lee, J.E. Shen, H. Rozner, M.A. Gottumukkala, Abbruzzese, Plunkett, J.L. W. Chen, Y. Wang, H. - Varad G.R. Ferrari, M. Maitra, A. Wolff, R.A. Intra-tumoral 2014. Fleming. and J.B. hachary, delivery and mass heterogeneity of gemcitabine Physical transport pancreatic cancer. in human 11:065002. biology. LAB MEMBERS Yan Post-doctoral Fellow: Huaming Wang), Students: Joseph Butner (jointly with Dr. Terisse Wang), Prashant Dogra (jointly with Dr. Wang) Brocato (jointly with Dr. of the tumor vessel radii), BVF (blood volume of the tumor vessel radii), and L (diffusion perfusion), i.e., fraction, which depend on cell penetration distance), The x-axis represents a pathological biology. PNAS 110 (35): (Pascal et al., surrogate for L. 14266-71). A mechanistic model of cancer cell kill from chemotherapy (red line) describes heterogeneous response within human cancer (symbols, corresponding to 50 data points from 8 patients with colorectal cancer metastatic to The model predicts the fraction of tumor liver). f response) as a function of parameters r pharmacodynamics (PKPD) models functionally linking biological behaviors at different scales) and invasion and drug response in local and metastatic cancers - Coupled drug pharmacokinetic-pharmaco KEY PUBLICATIONS KEY PUBLICATIONS cancer Taking Wang. Z. Koay, E.J. Cristini, V. out of the equation. International Innovation. 191:38-40. Deisboeck. T.S. Cristini, V. Butner, J.D. Wang, Z. Integrated PK-PD and agent-based mod- 2015. of pharmacokinetics Journal eling in oncology. 42:179-189. and pharmacodynamics. Kotsuma, M. Wang, Z. Smith, B.R. Frieboes, H.B. Mu- S.M. Flinders, C. Cahill, B. Day, A. Ito, K. Al-Fhaid, A.S. Simbawa, E. Mallick, P. menthaler, 2015. Cristini. V. Gambhir, S.S. Mahmoud, S.R. Predictive Modeling of Drug Response in Non- 10:e0129433. PloS one. Lymphoma. Hodgkin’s T.S. Cristini, V. Kerketta, R. Butner, J.D. Z., Wang, Simulating cancer growth with 2015. Deisboeck. Seminars in multiscale agent-based modeling. 30:70-78. cancer biology. V. Truty, M.J. Ondari, A. Baio, F.E. Koay, E.J. Y. Chatterjee, D. Chen, R. Thomas, R.M. Cristini, Tamm, E.P. Bhosale, P.R. Court, L. Zhang, J. Kang, V.N. Katz, M.H. Javle, M. Crane, C.H. Qayyum, A. •Spatiotemporal drug pharmacokinetics and Upscaling and downscaling framework (i.e., •Upscaling and downscaling framework (i.e., Translational modeling of cancer treatment modeling Translational “dynamic based on cellular and tissue scales, density functional theory.” Many PKPD models dynamic (PKPD) modeling. equations (ODEs) based on ordinary differential have been developed to describe the temporal response of tumor and normal cells to chemo- drug However, therapy or other therapeutics. resistance sometimes occurs due to limited penetration of drugs deep into the tumor, “space” but also “time” implying that not only factors have an impact on drug efficacy in both are investigating We normal and tumor tissue. a combined PKPD and spatiotemporal tumor modeling approach to study tumor response to chemotherapy. RESEARCH PROJECTS •Biophysical theories to predict the growth Vittorio Cristini, Ph.D. Vittorio Cristini, Professor Our group primarily focuses on developing Physical physical oncology. Translational Biological processes Multiscale methods. PROTEOMICS AND SYSTEMS BIOLOGY SYSTEMS AND FOR PROTEOMICS CENTER mechanistic biophysical models for predicting tumor response to various treatment methods in individual patients using standard clinical di- such as histopathology, agnostic measurements, have taken a multidisciplinary We and MRI. CT, resulted in which has approach in our projects, novel mathematical modeling algorithms and insights into how and why cancer behaves the goal is Our ultimate way it does in each patient. to bring our models to the clinic so that patient we have Currently, outcomes can be improved. three major research areas. processes such as transport mechanisms for drug molecules within tissue and the forces exchanged by cancer cells with the surrounding tissue determine cancer growth and treatment and physical apply engineering We outcome. sciences approaches to the modeling of complex normal and pathologic biological tissue. clinical translation of the mathematical Toward the effects we have been investigating models, and other transport perfusion, of diffusion, mechanisms on the rate at which tumors grow and spread and on resistance to drug based on input and other systemic therapies, have We from experimental and patient data. produced a series of pioneering modeling work on describing and quantifying physical mechanisms that play fundamental roles in the growth Through of cancer and in response to therapies. our joint work with pathologists and oncologists, we have made important discoveries on the role of physical transport in patient drug resistance. can occur across physical time and space forming a complex system with multiple scales, Advanced feedback and feed-forward loops. multiscale methods are therefore needed to simulate and predict the behavior of complex are developingWe methods biological systems. to address a significant challenge in multiscale the gaps between dif- bridging i.e., modeling, ferent modeling methods and between models to the from the molecular, at different scales, 54 • IMMPACT REPORT CENTER FORPROTEOMICS AND SYSTEMSBIOLOGY temperatures). Duringthepreparation(O to face(spatialvoid, absenceofgravity, extreme had the mosthostileenvironmentthatmanever assembly oftheInternationalSpaceStationin application wasthesuccessinhand-made ments frombasicresearchtoinnovativehuman O that wasusedtoforthesuccessfultwo-hour of activelimbsduringthedynamicexercise concentrations inmicrocirculatorynetworks and deoxygenatedhemoglobin/myoglobin instantaneous variationsoftotal, oxygenated spectroscopy (NIRS)allowedobservationof those achievements. Non-invasivenearinfrared awardsfromNASA for the teamreceivedseveral EVA preparationdowntotwohours. Membersof to NASA, suchasthedecreasefrom24-72hrs products andproceduresthatweredelivered North and several American institutionsledto between NASA This extensivecollaborative effort growing insitubycavitationortribonucleation. in blood, brainorothertissuesformingand nano-, micronucleiofgasesormicrobubbles its thirdmayleadtothepresenceofbonafide pressure to the decompressionfromsea-level prebreathe) forextra-vehicularactivities(EVAs), the roleofgases(O drug release. challengeswillbetostudy New of encapsulatedmicrobubbleshasaneffect on target tissues(tumors, …); cavitation-induced microbubbles asdrug-loadedliposomesto a non-detrimentalway, usesencapsulatedgas leads tootherapplications. NanoMedicine, in mechanisms. Experience ofmicrobubblesalso clei reduction, NOpathways, orothermolecular -inflammatory effectofexercise, gasmicronu- the protectionpossiblymediatedviaanti enhanced induced effectmayhavefurther N nisms thatregulateO and characterizesome molecularmecha- individual susceptibility genevariantstoanxiety functional MRI(fMRI), fNIRS, the evaluate on cerebralcirculationandvigilance), using stress onneuronaloxygenconsumption(effects 2 2 - One ofourextraordinaryscientificachieve -prebreathe. Our conclusionwasthatbesides tissuewashoutanotherunknownexercise- 2 , CO 2 -induced neurogenesis 2 ) potentiallygenerating 2

at cerebralCO diseases (e.g. Alzheimer’s). We arealsolooking and theirapplicationsinneurodegenerative neural andcancerstemcells) (space-microgravity, cognition, nanomedicine, nucleicacids, ofoxygen Innovative approachofthebiology Assistant Professor Philip Foster, M.D., Ph.D. ing pathways. PlosOne.2015. neurons invivovia ASK1 and p38MAPKsignal- Rosenblatt, K.P. Klothoprotectsdopaminergic R.,Brobey …, FosterPP, MakotoKuro-o, and PlosOne. 2015. ing complexinresponsetooxidativestress. monomerizing andbindingto ASK1 signal- Rosenblatt, K.P. Klothoregulates14-3-3zeta R.,Brobey …, FosterPP, MakotoKuro-o, and ing laterinmidlife.Neurosci. FrontAging 2015. and reducedneuralactivityformemoryprocess- Foster PP. Mildtraumaticbraininjury early inlife Neurosci. 2015. in brainnetworkconfiguration. FrontAging Foster PP. Roleofphysicalandmentaltraining KEY PUBLICATIONS plasticity • Cognitive • • RESEARCH PROJECTS hypoxia ontumorigeniccells. regulate cancercellssuchastheeffectsof mechanisms andotherphysicalvariablesthat is alsotostudytheO treatments. goal An extensionoftheprevious lung cancer)withthegoaltodesignspecific cally targetingcancercells(ependymomaand is todeterminespecificX-aptamersspecifi- Doppler.artery-transcranial Another challenge ing systems, NIRscanning, andophthalmic response functionusingcustom-madebreath- by observingtheoncerebralhemodynamic relationship tocerebral-ophthalmiccirculation cancer Therapeutic targetsforependymomaandlung research) Spatial environment(basicandoperational 2 levels andtheirinfluenceonthe levels 2 -dependent molecular the neurofibrillarylesionsof AD. order purple arrows)andreplicatesinreversed (redspiraland toward theentorhinalcortex of long-rangewhitemattertractsisdirected myelinationmaturation The inwardcortical white mattertracts. crossing andblockingthepathoflong-distance purple spiralsandarrows). Those lesions are (blue- towardthecortex the entorhinalcortex gression ofneurofibrillarylesionsspreadsfrom In Alzheimer’s disease(AD), theoutwardpro- IMMPACT REPORT • • • • 55 A. Murine models of human breast cancer Murine models of human breast A. bone Growth of MDA-MB-231 bone metastasis. metastatic human breast cancers monitored by Aptamer- bioluminescence (A) and histology (B). siRNA delivery (D) can reduce targeted STAT3 the cancer burden and prolong life. 23(6), 1044- 23(6), sistance large data sets cancer targeting and directed drug deliver 1054, 2015. DOI: 10.1038/mt.2015.45. 2015. 1054, Bone marrow endothelium-targeted therapeutics for metastatic breast cancer, Guo, X. Xu, R. Zhang, G. Mu, C. Huang, Y. Mai, J. Thiviyanathan, V. Lokesh, L. G. Volk, D.E. Xia, X. Shen, and H. Ferraria, M. Liu, X. Gorenstein, D.G. 187:22-29. 2014, Controlled Release, J. Aptaligner: Automated Software for Align- ing Pseudorandom DNA X-Aptamers from Lu, E. Next-Generation Sequencing Data. Volk, E. and D. Chang T. J. Elizondo-Riojas, M.-A. 53(22):3523-3525. 2014, Biochemistry, A Bead-Based Selection Method X-Aptamers: for Random Incorporation of Druglike Moieties Aptamers for Enhanced onto Next-Generation G. Elizondo-Riojas, M.-A. Li, X. He, W. Binding. Thiviyanathan, D.E. V. A. Somasunderam, Lokesh, Cavasotto, C. Englehardt, J. Durland, R. Volk, Biochemistry 2012, Gorenstein, and D.G. 51(42):8321-8323. LAB MEMBERS Ph.D., Research Scientist: Lokesh Rao, Ph.D. Li Li, M.S., Associates: Xin Li, Research Ph.D. Ana Maria Zaske, Post-doctoral Fellow: Medical Student: Linda Nguyen KEY PUBLICATIONS Pre- Adhesion Cascade at the Blocking the metastatic Niche for Prevention of Breast Can- Hasan, S.-A., Kang, T., Tanaka, cer Metastasis, Suh, D., Zhao, L., Zhao, W., Zheng, A.P., Mann, N., M., Cristofanilli, D.G., Gorenstein, D., Volk, K.S., Molecular Therapy, H., and Rui, •Provide biostatics and programming as- •Develop of novel software for the analysis •Develop for next-generation X-aptamers (DNA) Targeting cancer with X-aptamers and nanoparticle with X-aptamers cancer conjugates Targeting RESEARCH PROJECTS ovarian and pancreatic tumor imaging •Breast, David Volk, Ph.D. David Volk, Assistant Professor The focus of my lab is to develop novel Another focus of the lab is to provide cancer targeting agents using combinatorial, pseudo-random X-aptamer reagents that combine drugs or protein side-chains with DNA targeted reagents can provide These aptamers. directed delivery of anti-cancer medications to tumors while avoiding damage to other tissues. they By conjugating them with nanoparticles, offer the ability to provide for the slow release thereby of anti-cancer drugs at the tumor, further reducing unwanted collateral damage have developedWe several to remote tissue. and CD44, X-aptamers targeting E-selectin, proteins that are over-expressed A2, annexin on the surface of tumors or tumor associated (Mai et al. In a recent publication vasculature. we showed that as part of a multistage 2014), our aptamer targeting E-selectin, vector ESTA1, directed anti-cancer siRNA to the bone marrow for the treatment of breast cancer metastasis, leading to significantly increased survival rates. bioinformatics support and to develop novel software for the analysis of next-generation NGS data files often sequencing (NGS) data. and the contain millions of DNA sequences, therefore We analysis of them is not trivial. a 2014), Aptaligner (Lu et al. developed completely automated program with easy-to-use noise-reduction filters, graphical user interfaces, and statistical analysis DNA length error filters, packages for the analysis of many X-aptamer projects contained in a single NGS data file. bioinformatics services through also provide We the UTHealth Bioinformatics Service Center for the analysis of biological data related to and genetics. metabonomics, proteomics, PROTEOMICS AND SYSTEMS BIOLOGY SYSTEMS AND FOR PROTEOMICS CENTER 56 • IMMPACT REPORT mathematical toolsbased onbiophysical ing practical(relatively simpleyetpowerful) algorithms.parameter perturbation adverse effects), basedonsingle-andmultiple- efficacy andsafetywhileminimizingunintended and multi-targettherapeutics(highinboth methods foridentifyingpotentialdrugtargets cross-scaledrugtargetevaluation developing erful thanfocusingonlyoncellsignaling. We are potentially morerealisticandhencepow- multicellular-, and tumor-scalebehaviorsis tion ofdrugtargetsthataccountformolecular-, ing networks). However, selectionandidentifica- (i.e., ongenes, proteins, andlarge-scalesignal- targets todatefocusonthemolecularlevel ematical modelsusedinidentifyingcancerdrug tumor scales. affect tumorgrowthbehaviorsatthetissueand percolateacrossand at themolecularlevel These modelsexaminehowchangesoccurring data analysiswiththemathematicalmodels. vitro andinvivoexperimentspairedwithpatient have successfullyintegratedacombinationofin cellular, multicellular, andtissuescales. We also the gapsbetween, andlinking, themolecular, bridging current systemsmodelingofcancer: of research projectsaddressachallengingpart and theirconstantlychangingenvironment. Our between individualcells, andbetweencells scales resultingfromdynamicinteractions spans multiplespatialandtemporalbiological is anemergent, integratedphenomenonthat research areas. medical hypotheses. We havethreespecific outcomes, andtestrefinetheirbiological/ optimize, andpredictclinicaltherapies scientists tosimulateexperimentalprocedures, use ourmodelstohelpbiologistsandmedical progression andinvasion. We areworkingto data analysistoquantitativelystudytumor with experimentalinvestigationsandpatient ematical, physical, andstatisticalmethods CENTER FORPROTEOMICS AND SYSTEMSBIOLOGY Translation cancermodeling. - We aredevelop Cross-scale drugtargetdiscovery. Mostmath- Multiscale cancermodeling. Cancergrowth Our groupfocusesonintegratingmath- Associate Professor Zhihui (Bill) Wang, Ph.D. cancer biology. 30:70-78. multiscale agent-basedmodeling. Seminarsin Deisboeck. 2015. Simulatingcancergrowthwith Z. Wang, J.D. Butner, R. Kerketta, V. Cristini, T.S. Hodgkin’s Lymphoma. PloSone.10:e0129433. Predictive ModelingofDrugResponseinNon- S.R. Mahmoud, S.S. Gambhir, V. Cristini. 2015. menthaler, P. Mallick, E. Simbawa, A.S. Al-Fhaid, K. Ito, A. Day, B. Cahill, C. Flinders, S.M. Mu- H.B. Frieboes, B.R. Smith, Z. Wang, M. Kotsuma, 42:179-189. pharmacokinetics andpharmacodynamics. agent-based modelinginoncology. Journal of Deisboeck. 2015. IntegratedPK-PDand Z. Wang, J.D. Butner, V. Cristini, and T.S. 191:38-40. out oftheequation. InternationalInnovation. V. Cristini, E.J. Koay, Z. Wang. Taking cancer KEY PUBLICATIONS •Predictive modelingofcancertreatment ofadynamicmolecular target •Development • RESEARCH PROJECTS modeling method. general applicabilityandbroaderimpactofhis on localdiffusionproperties, demonstratingthe cancer inanycasewheredrugdeliveryrelies thecontext of is alsolikelytobeusefulbeyond clinical sciences. The conceptofthisapproach transport, arebroadlyapplicabletothe they based onfundamentalprinciplesofmass actual treatment. Sincethesetoolsarederived outcome foreachindividualpatientpriorto and PDE-basedmodelstopredicttreatment experimental/clinical investigators, weuseODE- outcome. Together withDr. Cristiniandother withtumorprogressionandtreatment transport ofdrug theories tocorrelatephysicalproperties Multiscale modelinganddrugtargetdiscovery ing identification methodwithmultiscalemodel- and breastcancerinitiation study normalmammaryglanddevelopment A hybridmultiscalemodelingapproachto (Wang etal.,IETSystBiol8(5):191-7). MLRA: Multivariatelinearregressionanalysis. tumor growth. ANOVA: Analysis ofvariance. determined tobemorepromisingininhibiting multiscale cancermodel. Targeting ERKwas analysisofa global parameterperturbation Parameter rankingofmoleculartargetsfrom (Frieboes etal.,PNAS10(6):e0129433). drug-resistant (bluesymbols)tumorsinvivo. samples ofdrug-sensitive(greensymbols)and those measuredfromthehistopathological predicted bythemodelareinagreementwith as afunctionofbloodvolumefraction(BVF) fraction oftumorkilled(f response inlymphoma. The resultsofthe values foramathematicalmodelofdrug Model predictionschangewithparameter kill ) fromchemotherapy IMMPACT REPORT • • • • 57 Scanning electron microscopic image of panc-1 cell after electroporation demonstrating several nanopores in the cell membrane. Atomic force microscopy image of the surface of a panc-1 cell after electroporation demonstrating severalnanopores in the cell membrane.

in Rabbit VX2 rabbit animal model using VX2 rabbit animal model in Rabbit vascular normalization gemcitabine nanoparticle formulation on tumoral response in a pancreatic adenocarcinoma nude mouse model cellular membrane pore formation after electroporation in pancreatic adenocarcinoma the radiogenomic evaluation of childhood neuroblastoma chemotherapy in the treatment of pancreatic adenocarcinoma of coupling local tumor electroporation of coupling local tumor electroporation on patients with hepatocellular TACE with carcinoma carcinoma genetic and metabolomic tumor carcinoma genetic and metabolomic therapies response to minimally invasive Yue Zhang, Sarah B. White, Jodi R. Nicolai, Nicolai, Jodi R. White, Sarah B. Zhang, Yue Dong-hyun Kim, West, Derek L. Zhuoli Zhang, Omary, A. Reed Miller, Frank H. Lee Goodwin, A. Imaging to Multimodality Larson. Andrew C. Assess Immediate Response to Irreversible Model. Tumor Electroporation in a Rat Liver 271: 721-729. 2014, Radiology, Jose West, Derek L. Bui, T. James Shah, Rajesh P. Martel- Joan N. Hatipoglu, A. Betul Oberholzer, A Case of Pancreatic Owens. A. Charles lotto, in a Patient with Transplantation Islet Cell Situs Ambiguous: Anatomical and Radiological Semin Intervent Radiol. 2007 Considerations. March; 24(1): 43–46. KEY PUBLICATIONS Omary AC, Larson Zhang Z, White SB, DL, West Therapy Optimization of Catheter Directed RA. Int J Nano- Animal Model. VX2 Rabbit in Rabbit medicine. 2014; 9: 4169–4176. •Optimization of catheter directed therapy •Evaluation of effects of electroporation and •Correlation of diffusion weighted MRI to •Use of magnetic resonance spectroscopy in •Evaluation of safety and efficacy of electro- An investigation on the therapeutic efficacy •An investigation on the therapeutic Interventional oncologyInterventional research RESEARCH PROJECTS •Prospective evaluation of hepatocellular Derek Lamont West, M.D., M.S. M.D., West, Derek Lamont Assistant Professor One treatment that my research centers on is Another treatment that my research is PROTEOMICS AND SYSTEMS BIOLOGY SYSTEMS AND FOR PROTEOMICS CENTER applica- Electroporation is the electroporation. tion of electrical fields across cells to produce is a procedure Electrochemotherapy nanopores. that combines electroporation and systemic chemptherapy for the treatment of malignant administration of In this treatment, neoplasia. a chemotherapeutic drug is followed by local Elec- application of electroporation pulses. troporation (at low field strength) transiently thus permeabilizes tumor cell membranes, enabling diffusion of a chemotherapeutic drug into the cells and increasing its cytotoxicity. My laboratory research involves optimizing electroporation and electrochemotherapy for the treatment of pancreatic adenocarcinoma. focused is on radiogenomics in hepatocellular correlation Radiogenomics is the carcinoma. of imaging phenotypes to genomic genotypes. Hepatocellular carcinoma (HCC) is the sixth most prevalent cancer and the third most The frequent cause of cancer-related death. majority of HCC patients are diagnosed at the advanced tumor stages and are treated such as with minimally invasive treatments, ablation transarterial chemoembolization, cryoablation or microwave, (radiofrequency, or yttrium 90 brachytherapy. electroporation), many While these treatments are effective, It has been patients have tumoral recurrence. suggested that one reason for failure is that post-treatment recurrent tumors are genetically different than the primary tumors and are more the labora- examine this, To resistant treatment. tory examines tumor genetic and metabolic As beginning and end-points in expression. this examination can help cellular processes, determine if specific tumoral genetic expres- in sions lead to specific metabolic expressions, hopes of better understanding the genetic and and TACE metabolite changes associated with microwave ablation (MWA) protocols and their prognostic implications. 58 • IMMPACT REPORT A CENTER FORSTEMCELL AND REGENERATIVE MEDICINE within UTHealth, to the Centeralsoserves effectively withotherprograms and institutions emanating from theCenter. By interfacing stem cellbasedtranslational research initiatives as well asthequalityandappropriateness of and innovation ofresearch withintheCenter, tissue engineeringtopromote theexcellence rigor inthedisciplinesofstemcellbiologyand appropriate breadth andscientific ofexpertise facultywiththe and retained amultidisciplinary translation. TheCenterhassuccessfully recruited forclinical tools andmethodologiesnecessary coupled withaclearfocusondevelopment of excellence infundamentalstemcellresearch, that suchanendeavor have atitsfoundationan for currently intractabledisorders. It isessential novel cellulartherapies fortissueregeneration into translate theiruniquebiologicalproperties ofstemcellsandto fundamental properties and Regenerative Medicine istostudythe cells: stemcells. ofspecializedon theuniquebiologicalproperties of adamagedtissueand/ororganisthereliance regenerative medicineapproaches totherepair and applicationof implementation, the successfuldesign, function. Implicit in or establishnormal or organstorestore human tissuesand/ or regeneration of goal thereplacement Medicine hasasits joints). Regenerative degeneration ofthe oraging(e.g. injury), (e.g. spinalcord deficiencies), trauma anemia, orimmune such assicklecell diseases oftheblood, (e.g. inheritedgenetic organs losttodisease human tissuesand for therestoration of effective therapies the development of Our focus withintheCenterforStem Cell medicine is contemporary major focusof University Chair The G. Harold andLorineG. Wallace Distinguished ProfessorAssociate andCenter Director Brian R.Davis, Ph.D. Research. Senator Lloyd andB.A.Bentsen Center forStroke the academicandadministrative homeforthe our research as activities.Our Centeralsoserves significantly increase thebreadth anddepthof and translationalCenterfacultyinorder to recruiting additionaloutstanding basicresearch Diseases; andLung Diseases. We are currently Hernia; Blood Diseases; Cancer;Musculo-Skeletal Stroke; Traumatic Brain Diaphragmatic Injury; the following diseaseareas: Spinal Cord Injury; research fortherapeuticapplicationtargeting Neurosurgery, andPediatric Surgery are pursuing appointments intheIMM, faculty withprimary of diseasesanddisorders. At present, Center of novel cellulartherapiesforawiderange stimulate thedevelopment andimplementation IMMPACT REPORT • • • • 59 S, Mountford J, Guenechea G, Gouble A, Bueren A, Gouble Guenechea G, J, Mountford S, Generation of Healthy Segovia JC. BR, Davis JA, Knock-in mediated TALEN Erythroid cells by Deficiency patient- Gene edited Pyruvate Kinase Stem Cells (Stem specific Induced Pluripotent in press) 2015, Cell Reports, Zhao J, Matthias N, Q, Yan Oda H, Umeda K, Expandable Long-Term Nakayama N. Davis BR, SOX9+ Chondrogenic Ectomesenchymal Cells Stem Cell from Human Pluripotent Stem Cells. 2015. Reports 4:712-726, Quintana-Bustamante O. Davis, B.R. Garate, Z. New Frontier in Regenerative Segovia. and J.C. Medicine: Site-Specific Gene Correction in Patient-Specific Induced Pluripotent Stem Cells. 2013 Human 24:571-583, Gene Therapy HIV/AIDS: modified Davis BR. Soto C, Amarijo E, - Cellular and Molecu stem cells in the spotlight. 2014 14: 2641-2649, lar Life Sciences. LAB MEMBERS Nadine Dr. Crane, Ana M. Research Staff: Dr. Matthias Leila Rouhigharabaei Post-doctoral Fellow: Dr. Applications of Cystic Fibrosis iPS Cells Genetically corrected stem cells for treatment of inherited stem cells for treatment of inherited Genetically corrected blood and lung diseases KEY PUBLICATIONS Chung, W.J. Bui, J.H. Kramer, P. Crane, A.M. Liao, W. Hawkins, F. M Gonzales-Garay, Li, X.S. D.Y. Paschon, D.E. Sun, H.C. Wang, J. Mora, D. Holmes, M. Kotton, D.H. Gregory, P.D. Guschin, Correction Targeted Davis. B.R. Sorscher, E.J. and Restored Function of the CFTR Gene in Cystic Fibrosis Induced Pluripotent Stem Cells. 2015. Stem Cell Reports 4:569-577, AM, Crane Quintana-Bustamante O, Garate Z, Kung C, Kosinski P, Galetto R, Poirot L, Oliver E, Alberqui- Cerrato L, Orman I, X, Aguirre Bazinet J, Garcia-San- Fusaki N, Rodriguez-Fornes F, lla O, Jin Prosper F, Sevilla J, Ribero M, T, Maia chez F, Brian Davis, Ph.D. Brian Davis, Associate Professor Regenerative Medicine Center for Stem Cell and Director of the Chair Distinguished University Wallace Lorine G. Harold and C. NKX2.1 GFP Reporter Line. We are utilizing an NKX2.1 GFP reporter to specifically identify and isolate We NKX2.1 GFP Reporter Line. we As shown in the right hand panels, lung cells derived from Cystic Fibrosis and corrected iPS cells. can use such GFP-expressing cells to generate lung-specific organoids and bronchospheres in the Hawkins). Kotton and F. laboratory (in collaboration with D. My laboratory has as its primary objective from inherited lung diseases (cystic fibrosis, surfactant protein-B deficiency) cells and blood stem cells from inherited blood disorders (Wiskott-Aldrich Syndrome, Pyruvate Kinase Deficiency) mutation resulting in correction of inherited Wiskott-Aldrich Syndrome defects the sequence-specific genetic correction of mutations in the chromosomal DNA of induced pluripotent stem (iPS) cells derived from patients with inherited disorders affecting the goal with the ultimate lung or blood system, of developing stem/progenitor cell-based have utilized Zinc We therapeutic approaches. Finger Homology Nuclease-mediated Directed Repair to correct the most common genetic mutations in iPS cell lines derived from patients with Cystic Fibrosis or Surfactant Protein B Deficiency – with the objective of demonstrating genotypic/phenotypic correction in lung epithelial cells derived from these corrected iPS cells. The second project in the laboratory focuses on the site-specific correction of gene mutations responsible for inherited blood disorders (e.g. Wiskott-Aldrich Syndrome) in patient-specific iPS cells – with subsequent differentiation to The third blood stem cells for transplantation. gene “natural laboratory project focuses on when spontaneous mutations that is correction,” arising in blood cells bearing inherited genetic mutations result in functional restoration of the followed by in vivo selection for defective gene, This gives rise to the revertant corrected cells. the phenomenon of revertant somatic mosa- are presently examining this natural We icism. gene correction particularly as it occurs in vivo Wiskott-Aldrich Syndrome. in patients with the RESEARCH PROJECTS •Correction and lung differentiation of iPS cells •Correction and blood differentiation of iPS •Characterization of spontaneous gene STEM CELL AND REGENERATIVE MEDICINE REGENERATIVE AND CELL FOR STEM CENTER 60 • IMMPACT REPORT CENTER FORSTEMCELL AND REGENERATIVE MEDICINE a potentiallyautologous cellsourcesforthe induced pluripotentstem cells(hiPSC)offer cell sources, also, need tobeoptimized. Human pansion anddifferentiationofclinicallyrelevant widespread clinicaluse, protocolsfortheex- pendent concentrations. multiple bioactivesignalingpeptidesatinde - novel syntheticpolymerscapableofdisplaying of stem cellstoneuronsandthedevelopment laminin fragmentsforthedifferentiationof on optimizingtheconcentrationofbioactive scaffolds. Preliminarystudies havefocused farbetterthantraditional tissue development native tissuemicroenvironmentandsupport these hybridscaffoldscanbegintoemulatethe bioactive signalingandcontrolledarchitecture, respectively. When combinedwithadditional andcellularresponse,in scaffoldproperties signaling andbatchtoinconsistency their disadvantages, namely lackofbioactive bioactive signaling)polymers, whilemitigating and cellularresponse)natural(natural of bothsynthetic(consistencyinfabrication scaffolding systemsmaximizetheadvantages into advancedhybridscaffoldingsystems. These combinatorial methods, willbeintegrated they concentrations havebeenidentifiedusing cal properties, andbioactivesignalingpeptide ity, featuresize, fiberalignment, etc.), mechani- Once optimalcomposition, architecture(poros- integration withthesurroundinghosttissue. extracellular matrixfacilitatetissuerepairand seek tounderstandwhichaspectsofthenative sources foruseinstemcelltherapy. cell and theexpansionofclinicallyrelevant laboratory seekstooptimizescaffolddesign science. Utilizingengineeringapproaches, the stem cellbiology, chemistry, andmaterial involving techniquesfromcell, molecular, and laboratory usesaninterdisciplinaryapproach traumatic braininjury, defects. andcartilage The toward clinicaltreatmentsforspinalcordinjury, tissueengineeringapproaches developing In ordertoadvancetissueengineering By examiningcell-materialinteractions, we The researchinmylaboratoryfocuseson Primary Amine-derivatized Surfaces.Primary Amine-derivatized tiation andNeuriteExtension ofmouseESCon ML. ConcentrationDependentNeural Differen- Smith CallahanLA, Ma Y, StaffordCM, Becker 34(36): 9089-9095, 2013. Functionalized with YIGSR Peptide. Biomaterials. Stem Cellon Aligned Poly(lactide)Nanofibers and NeuriteExtensionofmouseEmbryonic Dove AP, BeckerML. DirectedDifferentiation Smith CallahanLA, XieS, BarkerIA, ZhengJ, gradients. ActaBiomaterialia . 21:55-62, 2015. continuous Ile-Lys-Val-Ala-Val concentration stem cellsusinghydrogelsfunctionalizedwith neural differentiationofmurineembryonic LA. Optimization ofadhesiveconditionsfor Yang Y-H, KahnZ, MaC, LimHJ, SmithCallahan KEY PUBLICATIONS •Synthesis ofnovelbiomaterialsforspinal matrix •Identification ofoptimalartificial •Optimization ofculturesurfacesforthediffer- ofmulti-component scaffoldsto •Development RESEARCH PROJECTS onhiPSClineagechoice.properties gradient tostudytheeffectofmechanical of hydrogelswithcontaininga Young’s Modulus the covalenttetheringofproteinstosurface from hiPSC. Preliminarystudieshavefocusedon stem cellsandoligodendrocyteprogenitor cell culturesurfacesfordifferentiationofneural culture systemstooptimizetwodimensional to extendourknowledgeofthreedimensional to hiPSCdifferentiation. The laboratoryseeks have notbeenstudiedsignificantlyinrespect significantly affectcellulardifferentiation, but the cellculturesurfacehavebeenshownto of biochemical andmechanicalproperties differentiation protocolsisextremelylow. Both cells fortransplantproducedfromcurrent nervous system. However, thenumberofviable treatment oftraumaticinjuriestothecentral the centralnervoussystem ofhybridtissueengineeringscaffoldsforusein Development Assistant Professor Laura A. SmithCallahan, Ph.D. cord, brain, discrepair andvertebral combinatorial approaches using concentration ormechanicalproperties suchasbioactivesignalingmoiety properties progenitor cells cells toneuralstemandoligodendrocyte entiation ofhumaninducedpluripotentstem replication ofthenativeextracellularmatrix facilitate tissueregenerationthroughbetter Biomateri- thew Mosley andZaraKhan Mosley thew Undergraduate Students(RiceUniversity):Mat- Student: AlexanderAria Medical and ThomasWilems Perera Hiran Post-doctoral Fellows:HyunJuLim, Thuduwage LAB MEMBERS Biomaterials 31(21):5526-5539, 2010. Differentiation withNano-fibrousScaffolding. of HumanEmbryonicStemCellOsteogenic Smith LA, LiuX, HuJ, MaPX. The Enhancement 6095–6104, 2013. Gradient inModulus. ActaBiomaterialia. 9(4): PEGDM HydrogelsPossessingaContinuous Phenotype MaintenanceusingRGDderivatized Chondrocyte ExtracellularMatrixFormationand EP, Weiner, SD, BeckerML. PrimaryHuman Smith CallahanLA, Ganios AM, Childers als Science.1(5):537-544, 2013. stemcelltherapytobesuccessful.supported for appropriatecelltypesinordermatrix extracellular matricesandmustbeoptimized significant roleincellularresponsetoartificial gradient of IKVAV. Peptide concentration plays a ylene glycolhydrogelscontainingacontinuous and 3D(encapsulated)cultureusingpolyeth- stem cellneuriteextensioninboth2D(surface) derived peptideIKVAVformouseembryonic Optimization oftheconcentrationlaminin IMMPACT REPORT • • • • 61 Survival and neuronal differentiation of grafted neural stem cells derived from human induced pluripotent stem cells after SCI. tome in acute/subacute phases: a resource tome in acute/subacute phases: for understanding the pathology at the systems * co-corresponding PLoS One (in Press). level. authors. Luo Xiong K, Cheng XX, Chen D, H, Wang Fan CL, XG and Cao QL (2014) Effect of type-2 astrocytes on the viability of dorsal root ganglion neurons and length of neuronal processes. Res. 9: 119-128. Regen. Neural LAB MEMBERS Associate: Post-doctoral Fellow Research Yiyan Michelle Wang, Zheng Assistant: Jun Li Senior Research Student: Chrystine Gallegos Specific expression of green fluorescence pro- AAV8-Flex-GFP injection tein in astrocytes using in GFAP-cre mice. protection agents for spinal cord injury functional neurons astrogliosis and the functions of astrogliosis traumatic brain injury, after spinal cord injury, or stroke using conditioned knockout mice models regulators for oligodendrocyte differentiation and remyelination after spinal cord injury of hiPSC-derived neural stem or precursor cells for spinal cord injury and stroke KEY PUBLICATIONS KEY PUBLICATIONS Howard RM, Cheng XX, YP, Wang He Q, Cao QL, Magnuson Shields CB, WH, DeVries YP, Zhang Whittemore SR (2010) Kim DH and Xu XM, DSK, of CNTF-expressing adult Transplantation oligodendrocyte precursor cells promotes remyelination and functional recovery after spinal J Neurosci 30: 2989-3001. cord injury. Whittemore Kim DH, He Q, Cheng XX, YP, Wang Astrocytes from the SR and Cao QL (2011) contused spinal cord inhibit oligodendrocyte differentiation of adult OPCs by increasing the J expression of bone morphogenetic proteins. Neurosci 31(16):6053– 6058. Luo XG, Bu P, Qi XB, XX, Cheng YY, Zheng Fan CL, Transplantation Kim DH and Cao QL (2013) of D15A-expressing glial-restricted-precursor- derived astrocytes improves anatomical and Int J locomotor recovery after spinal cord injury. 2013;9(1):78-93. Biol Sci. GD, Yang YY, Zheng Lu HZ, Deng SY, Chen KN, RNA-Seq Wu JQ* (2013). Cao QL* and Kim DH, characterization of spinal cord injury transcrip- •Screening and identification of novel neuro- •In situ reprogramming of astrocytes into •The molecular mechanisms to regulate •Identification and characterization of key Stem cells for neurological diseases for neurological Stem cells cells will astrocytes into neuronal precursor to promote axonal decrease astrocyte inhibition The newly reprogrammed neuronal regeneration. the lost neurons precursor cells could replace may These two mechanisms after SCI or stroke. great functional work synergistically to promote goal Our long-term recovery after SCI or stroke. is to develop novel stem cell-based therapies to treat human SCI or stroke. RESEARCH PROJECTS •The long-term therapeutic efficacy and safety Qi Lin Cao, M.D. Qi Lin Cao, Associate Professor Transplantation of neural stem cells (NSCs) Transplantation is a promised therapeutic approach to promote functional recovery after neurological diseases, including spinal cord injury (SCI) and stroke. is no consensus as to which there However, central Human NSC resource is optimal for SCI. nervous system stem cell isolated from fetal cadaver brain tissue and neural progenitor cells derived from human embryonic stem cells (hESCs)-derived have been approved for these However, clinical trials for SCI patients. cells are associated with ethical controversy hESCs Cells derived from and graft rejection. have additional risk of teratoma forma- stem cells Human induced pluripotent tion. (hiPSCs) are recently developed remarkable from ESC-like cells reprogrammed pluripotent, adult somatic cells by over-expression of four developmental/pluripotency transcription hiPSCs offer Compared with ESCs, factors. significant additional advantages in terms of availability of source material without ethical the and especially concerns of embryo use, ability to generate isografts without the need have developedWe of immunosuppression. neuro- protocol to differentiate and purify NSC, nal precursor cells or glial precursor cells from Our results show that hiPSC-derived hiPSCs. NSCs can proliferate over long time in vitro and be induced to differentiate into functional Im- astrocytes and oligodendrocytes. neurons, hiPSC-derived NSCs can survive and portantly, differentiate into both neurons and glias after transplantation into the contused spinal cord These studies and promote functional recovery. suggest that transplantation of hiPSC-derived NSC is an effective therapy to preserve and we are Currently, restore neurological functions. testing the therapeutic efficacy and long-term or glial precursor cells neuronal, safety of NSCs, to identify the optimal cell graft for SCI and we are testing whether we can Recently, stroke. directly reprogram the astroglial cells in the injured spinal cord or stroke brain into neurons. Astroglial scar are the major inhibitor for axonal In situ reprogramming active regeneration. STEM CELL AND REGENERATIVE MEDICINE REGENERATIVE AND CELL FOR STEM CENTER 62 • IMMPACT REPORT cell productionforuseinourclinicalprotocols. treatments. These facilitiesallow clinical grade ties thatenableustotranslatediscoveryinto which arecGMPandcGTPcellprocessingfacili- tory andtheHoffbergerStemCellLaboratory, beds. use invivomodelsofinjuryandvitrotest inquestion.specific tothepathophysiology We delivery aswelltiming), whichmaybevery regimens (numberofcells, typeandrouteof these typesofdatatohelpusdeterminedosing macrophage phenotypeinthebrain. We use upregulation andmodulationofthemicroglia/ reticuloendothelial systemhaveresultedin Treg term. Cell-cellinteractionsintheperipheral without significantengraftmentinthebrainlong how cellulartherapieshavebeensuccessful of theinnateimmuneresponseto TBI, and TBI. We havebeeninterestedinthemodulation injuries, principallytraumaticbraininjury, or the useofcellulartherapiesforneurological CENTER FORSTEMCELL AND REGENERATIVE MEDICINE Our teamdirectstheGriffinStemCellLabora- Our currentresearchprogramfocuseson George andCynthiaMitchellDistinguishedUniversityChair Professor Charles Cox, Jr., M.D. 23020860 J Neuroinflammation 9:228-240, 2012. PMID: injury: Modulationofmicroglia/macrophages . progenitor celltherapyfortraumaticbrain Pati S, CoxCS. Intravenousmultipotentadult H, HamiltonJA, SmithP. Thomas CP, MaysRW, Walker PA, BediSS, ShahSK, Jimenez F, Xue 4: 161ra150, 2012. PMID:23175708 the solublefactor TIMP-3. Science/Transl Med in traumaticbraininjurythroughproductionsof stem cellsregulateBloodBrainBarrierintegrity Holcomb JH, DashPK, PatiS. Mesenchymal J, PengZ, XueH, KozarR, CoxCS, Khakoo A, Zhang J, LeToumeau P, RedellJ, ShenL, Wang Menge T, Zhao Y, ZhaoJ, Wataha X, GerberM, Translational Medicine. 2:953-960, 2013. improves spatiallearningafter TBI. StemCells/ T, MaysRW, CoxCS. IntravenousMAPCtherapy Smith P, XueH, Aroom K, UrayK, Hamilton Bedi SS, HetzR, Thomas C, Olsen A, S,Williams 225:341-352, 2010. via interactionwithsplenocytes. ExpNeural brain injury:Preservingtheblood-brainbarrier tent adultprogenitorcelltherapyfortraumatic Pati S, DashPK, CoxCS. Intravenousmultipo- H, CutroneR, HamiltonJA, MaysRW, DeansRA, Walker PA, ShahSK, JimenezF, GerberMH, Xue children. Neurosurgery 68:588-600, 2011. traumaticbraininjury in nuclear cellsforsevere clinical trialofautologousbonemarrowmono Day MC, LeeD, JimenezF, Gee A. 2010. Phase1 Walker PA, ShahSK, Ewing-CobbsL, HasenK, Cox CS, JE, Baumgartner MT, Harting Worth L, KEY PUBLICATIONS •Imaging ofmicroglialactivationinvivo •Novel deliverysystemsforstemcellsin •Amniotic fluidderivedMSCsforthetreatment •IND-enabling studiesusing APCs fortrau ofPhase1and2Clinical •Development Trials RESEARCH PROJECTS Cellular therapiesforneurologicalinjury neurological injury bypass/hypothermic circulatoryarrest diseaseandcardiopulmonary genital heart of neurologicalinjuryassociatedwithcon- brain injury traumatic braininjury using non-ESCstem/progenitorcellsfor matic

patients. (Liao, etal.PedsCritCareMed, 2015) matory mediatededemaaftercelltherapy in observations ofareducedcerebralinflam - rodent model. These datacoincidewithclinical rier permeabilityaftertraumaticbraininjury ina demonstrate improvementinblood-brainbar- The useofarangestemandprogenitorcells injury sites. implantation intofocalcavitaryneurological with MAPCsandNSCsasacompositegraftfor Electrospun PLGAnanofiberscaffoldseeded SufiraKiran GMP-QA Director: Fabio GMP CenterDirector: Triolo, Ph.D. Research Scientist:HasanXue, M.D. Medical Student:HenryCaplan, B.S. Flow Cytometry Technician: PhillipaSmith, M.S. Post-doctoral Fellow:SuchitSahal, Ph.D. NIH T32 Fellow:MargaretJackson, M.D. Glassell FoundationFellow:Ben Aertker, M.D. KarenUray,Assistant Professor: Ph.D. SupinderBedi,lnstructor: Ph.D. LAB MEMBERS IMMPACT REPORT • • • • 63 Human PAX7 reporter iPS cells show the Human PAX7 expression of the GFP reporter following PAX7 VP160 mediated activation using a dead Cas9 gene activator. PAX7 Skoglund G, Lainé J, Darabi R, Fournier E, Fournier E, Darabi R, Lainé J, Skoglund G, Physiological and N. Tabti Perlingeiro R, induced ultrastructural features of human cell-derived pluripotent and embryonic stem Acad Sci Proc Natl skeletal myocytes in vitro. Jun 3; 111(22):8275-80. 2014 USA. Derivation of Skeletal Perlingeiro RC. Darabi R, Myogenic Precursors from Human Pluripotent Stem Cells Using Conditional Expression of Methods Mol Biol. 2014 Nov 18. PAX7. Skeletal Darabi R. Wu J, Hunt SD, Matthias N, muscle perfusion and stem cell delivery in muscle disorders using intra-femoral artery Exp Cell Res. 2015 Sep 1. canulation in mice. LAB MEMBERS Nadine Wu, Post-doctoral Fellows: Jianbo Matthias Hunt Samuel D. Technician: Research A B A. NIRF image demonstrates high efficiency A. hindlimb perfusion following intra-arterial (intra- femoral artery) canulation/perfusion in mouse. Human iPS-derived myogenic progenitors en- B. graft and restore the missing protein/dystrophin (red) in a mouse model of Duchenne muscular dystrophy (DMD) following intra-arterial perfu- Asterisks and arrows mark the presence of sion. human nuclei (green) in perfused region. models for skeletal mass loss injuries cell therapy in muscular dystrophies progenitors from human ES/ iPS cells progenitors from human ES/ survival and engraftment of human ES/ iPS survival and engraftment of in skeletal derived myogenic progenitors muscle Disease modeling and skeletal muscle regeneration using muscle regeneration and skeletal Disease modeling human ES/iPS cells Radbod Darabi M.D., Ph.D. M.D., Radbod Darabi Assistant Professor KEY PUBLICATIONS Kamath S, Bachoo MR, Gehlbach K, Darabi R, Perlingeiro RCR. Kyba M, Kam KE, Osawa M, Functional skeletal muscle regeneration from Nature differentiating embryonic stem cells. 2008; 14 (2): 134-143. Medicine, Darabi R (equal contribution), AL, Ramos Dineen SP, Catanese J, Borges L, Akbarloo N, Analysis Clonal Perlingeiro RC. Brekken RA, Reveals Progenitor for Endothelial, a Common in Umbilical and Lymphoid Precursors Myeloid, 2010 Dec Research, Circulation Cord Blood. 10; 107(12):1460-9. Koene W, Pan A, Filareto Santos FN, Darabi R, Assess- Perlingeiro RC. Kyba M, Rudnicki MA, R, ment of the myogenic stem cell compartment following transplantation of pax3/pax7-induced Stem embryonic stem cell-derived progenitors. 2011 May;29(5):777-90. Cells, Grskovic Dimos JT, Irion S, Arpke RW, Darabi R, Human ES- and Perlingeiro RC. Kyba M, M, iPS-Derived Myogenic Progenitors Restore Dystrophin and Improve Contractility upon Cell Stem in Dystrophic Mice. Transplantation 610-619. 2012 May; 10 (5), Cell, Iacovino Borges L, Darabi R, Parker S, A, Filareto Ervasti Chamberlain J, T, Mayerhofer T, Schaaf M, An ex Perlingeiro RCR. Kyba M, Scott McIvor R, J, vivo Gene Therapy Approach to Treat Muscular Nature Communica- Dystrophy Using iPS cells. tions, 2013; 4:1549. Toyama Y, Zhang TL, Mader Darabi R, Arpke RW, Perlingeiro RC, Lowe DA, Nash N, Lontree CL, A, A New Dystrophin- Deficient Immuno- Kyba M. Provides The NSG-mdx4cv Mouse, Model, Evidence for Functional Improvement Following Stem Transplantation. Allogenic Satellite Cell 2013 Aug; Cells, 31(8):1611-20. •Using bio-scaffolds for cell delivery in mice •Systemic/arterial cell delivery approaches for Generation of integration-free/safe myogenic •Generation of integration-free/safe My lab’s main interest is using pluripotent My lab’s the lab focuses on the Here at IMM, Using high throughput screening (HTS) to Our lab research also includes derivation My lab is currently funded by a research grant MYF5) lines for early myogenic genes (PAX7, CRISPR/Cas9 system stem cells for skeletal muscle regeneration. we have developed During the last few years, novel methods for using mouse/human embryonic stem cells (ES cells) and induced pluripotent cells (iPS cells) for cell therapy in mice models for different types of muscular dystrophies (MDs). approaches to use stem cell therapies for our Currently skeletal muscle regeneration. lab uses cutting-edge gene editing technologies (CRISPR/Cas9) for generation of knock-in reporter lines in human ES/iPS cells for early and MYFF5. myogenic genes such as PAX7 This will allow studying the emergence of early myogenic progenitors from human ES/iPS cells; a crucial step to identify and isolate these cells for future iPS cell based therapies. identify important inducers of myogenesis in iPS cells and evaluation of in vivo regeneration potential of the cells in mice models for MDs and muscle mass injuries are other major aims of the lab. of iPS cells from MD patients; in vitro gene correction of iPS cells; optimizing cell delivery and engraftment; study mechanisms involved in cell homing into the muscle after systemic/ arterial cell delivery; as well as exploring the effect of local tissue perfusion in cell survival and engraftment. Association award from Muscular Dystrophy period of three years to develop over a (MDA) methods using stem cells to regenerate skeletal muscle tissue in a mouse model of Duchenne muscular dystrophy (DMD). RESEARCH PROJECTS •Generation of knock-in human ES/iPS cell •Gene correction of MD iPS cells using •Study the role of local tissue perfusion on STEM CELL AND REGENERATIVE MEDICINE REGENERATIVE AND CELL FOR STEM CENTER 64 • IMMPACT REPORT that can be partially corrected by rapamycin.that canbepartially migration andlearning andmemorydysfunction TSC2 inradialgliacauses abnormalneuronal School ofMedicine, wehaveshownthatlossof with Dr. MichaelGambelloofEmoryUniversity spectrum disorder(ASD). Incollaboration learning andmemoryimpairmentsautism characterized bytheformationofbraintumors, activation andtuberoussclerosis, adisease Mutations inthesegenescausemTOR over- are encodedbytheTSC1andTSC2 complex, theprotein componentsofwhich is negativelyregulatedbythetuberoussclerosis act, inpart, throughthispathway. mTOR activity and thatmemoryenhancerssuchasglucose plays anobligatoryroleinmemoryformation, that mammaliantargetofrapamycin(mTOR) neurodegeneration after TBI. We haveshown to learningandmemoryimpairmentsand/or if dysregulationofthesecascadescontributes critical formemoryformationanddetermine identify thesignalingcascade(s)thatare attenuate neurodegeneration. pathway canreduce TAU phosphorylation, and exploring thepossibilitythattargetingthis improves learningandmemory. We arecurrently ment reducesexperimental and TBI pathology lithium, wehavefoundthatpost-injurytreat- Using thepharmacologicalinhibitorofGSK3 to TAU hyperphosphorylationandaggregation. kinase 3(GSK3), anditsdysregulationcanlead that phosphorylates TAU isglycogensynthase largely unknown. Oneoftheproteinkinases trigger TAU and Aß aggregationafterrmTBIare the cellularandmolecularmechanismsthat protein TAU andamyloid-beta(Aß). However, the neuronalproteinsmicrotubule-associated CTE arecharacterizedbythedepositionof traumatic encephalopathy(CTE). Both AD and such as Alzheimer’s disease(AD)andchronic ofneurodegenerative diseases development repeated concussion), isariskfactorforthe repetitive mildtraumaticbraininjury(rmTBIor that traumaticbraininjury(TBI), especially CENTER FORSTEMCELL AND REGENERATIVE MEDICINE Another focusofourlaboratoryisto indicates Clinical andexperimentalevidence genes. 2015. memory. LearningandMemory22:239-246, mTORC1 signalingandimpairslong-term hippocampal glutamineadministrationinhibits J, Moore AN, GambelloMJandDashPK. Intra- Rozas NS, Redell JB, Pita-AlmenaraJ, McKenna epub) 2015. traumatic braininjury. JNeurotrauma, (April4 and improveslearningmemoryfollowing of eIF2αphosphatasereducestissuedamage Redell JB, Tsvetkov AS andMoore AN. Inhibition Dash PK, HylinMJ, HoodKN, Orsi A, ZhaoJ, Neurotrauma 30:716-726, 2013. visuospatial memoryandcomplexlearning. J mild closedheadinjuryimpairsshort-term Redell JB, Moore AN andDashPK. Repeated Hylin MJ, OrsiSA, RozasNS, HillJK, ZhaoJ, Outcome. PLoSOne. 6(9):e24648, 2011. Pathway in andNeurocognitive TBI Pathology of theGlycogenSynthaseKinase-3Signaling Zhao J, GrillRJ, Moore AN, PatiS. Involvement Dash PK, JohnsonD, ClarkJ, OrsiSA, ZhangM, nism. Neuroscience, 159:483-491, 2009 foratime-dependent mecha- injury: evidence memory dysfunctionfollowingtraumaticbrain Kobori, N. andDash, P.K. Persistentworking Hoskison, M.M., Moore, A.N., Hu, B., Orsi, S.A., KEY PUBLICATIONS ofstrategiestoreduce protein •Development utilizationintheinjuredbrainand •Energy •Role ofsystemicinflammationin TBIpathol- RESEARCH PROJECTS TBI-elicited learningandmemoryimpairments. been examiningifthiscascadecontributesto on mTOR’s roleinmemoryformation, wehave ofautistic-likebehaviors.development Based kinje cellscausesneurodegenerationandthe Interestingly, lossofTSC2incerebellarPur- Nina andMichaelZilkhaDistinguishedChair, NeurodegenerativeDiseaseResearch Professor Pramod Dash, Ph.D. dysfunction Cellular andmolecularmechanismsofmemoryits neurodegeneration. aggregation inthebrainandattenuate crisis strategies tomitigateenergy andoutcome ogy disruptions. (cing) canbeobserved, suggestingaxonal of thefibersinipsilateral (ipsi)cingulum a shamandmTBIrat. An apparentshortening passing throughtheareaofcingulum from imaging (DTI)demonstratingtheaxonalfibers Tract tracingsobtainedfrom diffusiontensor Jacalyn S. MacGowan Research Assistants: KimberlyHood, M.A., Research Coordinator: Anthony N. Moore, B.S. Ph.D. Post-doctoral Fellow:Karthikeyan Tangavelou, Ph.D. Research Scientist:NobuhideKobori, M.D., Sr. Research Scientist:JingZhao, M.D., Ph.D. Ph.D. Research JohnB.Assistant Professor: Redell, LAB MEMBERS merged image. axons. Areas ofoverlapappearyellowinthe (MBP; green)indicatingdamagetomyelinated (red) colocalizedwithmyelinbasicprotein compared tosham-operatedcontrols. APP fibers precursor protein(APP)immunoreactivity losum (cc)asindicatedbyincreasedamyloid mTBI causesaxonalinjuryinthecorpuscal- IMMPACT REPORT • • • • 65 Identification of the THSD1 R450X Mutation in Large Family with IA and the Spectrum of THSD1 Rare Variants. IA001. A shows the pedigree of family Panel circles females, Squares represent males, symbols white black symbols affected persons, gray symbols unknown unaffected persons, carri- plus symbols R450X mutation phenotype, Slashes and negative symbols non-carriers. ers, B shows the Panel denote deceased individuals. in unrelated Variants spectrum of THSD1 Rare (blue) individuals control in (red)and patients IA domain and in relation to exons (numbered), THSD1 control variants presented organization. here are from the NHLBI GO Exome Sequenc- Panel C shows a protein sequence ing Project. alignment displaying evolutionary conservation of substituted amino acids in THSD1 orthologs. Panel D shows images of aneurysms in patients with a THSD1 mutation. Advancing the field of neuroscience Advancing the field Dong Kim, M.D. Dong Kim, Chairman Professor and Department Smith of Neurosurgery Vivian L. Institute Mischer Neuroscience Director, Memorial Hermann Hospital–TMC RESEARCH PROJECTS cord injury •Stem cell therapy for spinal •Genetic aneurysm research •Clinical trials KEY PUBLICATIONS FM, Waldman A, Bollen Mohapatra G, Kim DH, Feuerstein BG: Chromosomal abnormalities in glioblastoma multiforme and glioma cell lines detected by comparative genomic hybridization. 1995. 60:812-815, Int J Cancer. Feuerstein BG: Detection Kim DH, Mohapatra G, of multiple gain and losses in ten glioma cell lines by comparative genomic hybridization. 1995. 13:86-93, Cancer. Chrom, Genes, The Milewicz DM: Ginhoven G, Van Kim DH, incidence of familial intracranial aneurysms in 200 patients: comparison between Caucasian, and Hispanic populations. African-American, 2003. 53(2):303-8, Neurosurgery. Milewicz DM: Familial Ginhoven G, Van Kim DH, aggregation of both aortic- and cerebral aneu rysms: evidence for a common genetic basis in 56:655-61, J Neurosurg. a subset of families. 2005. McDonough Ju KL, Depalma SR, T, Santiago-Sim Kim DH: Genome- Seidman JG, Seidman CE, B, wide linkage in a large Caucasian family maps a newchromo- to aneurysms intracranial for locus 2009. Stroke. 40[suppl 1] S57-S60, some 13q. How- Xiaoxin, Cheng, Yaping, Wang, He Q, Cao Q, William DeVries, Yiping, Zhang, Russell M., ard, Kim D, Xu X, Magnuson DSK, Shields CB, H., of Ciliary Neuro- Transplantation Whittemore SR: Adult Oligodendrocyte trophic Factor-Expressing Precursor Cells Promotes Remyelination and J Functional Recovery after Spinal Cord Injury. 2010. Neuroscience. 30 (8):2989-3001, Nalbach S, Hennessy M, Fang X, T, Santiago-Sim McDonough B, Greenway S, Lee MS, DePalma S, Qualmann Colosimo S, Patek K, Hergenroeder G, Seidman Dymecki S, MacRae C, Milewicz D, K, Kim DH: Mutations in THSD1 as Seidman JG. C, (submitted) Aneurysm. a Cause of Intracranial : 2015. Professor and chair of the Department of devel- My research has focused on the origin, Report’s World Named to the US News and A graduate of Stanford and the University I specialize in the following diseases: STEM CELL AND REGENERATIVE MEDICINE REGENERATIVE AND CELL FOR STEM CENTER I Neurosurgery at McGovern Medical School, also lead the clinical neuroscience efforts for the Memorial Hermann Healthcare System (MNI) as director of the Mischer Neuroscience the group includes over 100 Currently, Institute. faculty and residents/fellows. Our and treatment of brain aneurysms. opment, group recently identified the first gene defect proven to cause intracranial aneurysms in I also work to develop neural familial patients. stem cells for implantation into the brain and spinal cord. Surgeons, Top America’s and 1% Doctors, Top I have received grants from the National Stroke American Institutes of Health and the Association. School of San Francisco (UCSF) of California, training I completed general surgery Medicine, Prior to then neurosurgery at UCSF. at Harvard, I held positions at Harvard Texas, coming to Hospital, Women’s Brigham and Medical School, Cornell Uni- the Dana-Farber Cancer Institute, Hospital York The New versity Medical College, and Memorial Sloan Kettering Cancer Center. •Intracranial aneurysms benign and malignant •Brain tumors, •Skull base tumors and meningiomas neuralgia •Trigeminal •Chiari malformations 66 • IMMPACT REPORT models toregenerativemedicineapplications. regenerative to transferourlearningfromnewt limbregrowth.cesses ofnewt Ourexpectationis accelerate regenerationbyduplicatingthepro- using murinedigitastheamputationmodelto to humantissueregeneration. Currently, weare in thenewts, therelationship(s) andascertain mechanism(s) behindtheregenerativeprocess healing process. Ouraimistodeterminethe with fibroticscartissueattheendof including thatofhumans, isusuallyreplaced after injury. However, injuredmammaliantissue, (suchaslimbs,body parts liver, lensandheart) modelthatcanrebuildmost missing a newt a classictissue/organregenerationmodel, e.g. diseases andtraumaticinjuries. We havesetup and engineeredtissuefortreatingcongenital study andclinicalapplicationofstemcells The laboratoryisalsointerestedintranslational repair ofchildren’s diaphragmhernia(CDH). defects withscarlesshealing, whichinclude use ofbioengineeringtissuestorepairwound and 3) candidate genesforagingprevention; the musculoskeletalsystemanddetecting the mechanismbehindagingprocessesin junction (NMJ)reconstruction;2)studying healing, includingvascularandneuromuscular (iMuSCs) andtheirnaturalroleduringtissue of theinjuryinducedpluripotentstemcells areas ofstudy:1)exploringtheproperties major animal-based studiestofocusonfew novel techniquesformolecular, cellular, and CENTER FORSTEMCELL AND REGENERATIVE MEDICINE This research team has developed several several This researchteamhasdeveloped Associate Professor Yong Li, M.D., Ph.D. Pluripotent stemcellandregenerativemedicine 2015;331(1):200-210. expansion. ExperimentalCellResearch muscle regenerationthroughsatellitecell and muscle Arnt-like 1promotesskeletal Chatterjee S, HS,Yin NamD, Li Y, Ma K. Brain (2015, accepted) Derived StemCell-LikeCells. ScientificReports of anInjuryInducedPopulationMuscle- Vojnits K, PanHY, MuXD, Li Y. Characterization 292. mice. engraftment followingimplantingintomdx/SCID expression enhancesmyoblastmigrationand YG, HuardJ, CoxC, LallyKP, Li Y. MMP1gene Pan HY, Vojnits K, MengFW, Liu T, Yang L, Wang KEY PUBLICATIONS •Neuromuscular junction: We havebuilta •Aging study: ourspecificmurineaging With Studies:Investigate andPrevention •Fibrosis •Injured DerivedStemCells: Tissue The project •Children’s RegenerativeMedicine: The project RESEARCH PROJECTS muscle atrophy, drugscreening studies, suchasneuromusculardiseases, (NMJ) indish, whichcanbeusedasseveral model ofhumanneuromuscularjunction protein profilingwithinthemodelsystem screening genome-widetranscriptomeand nisms andbiomarkersforagingrepressionby and determinethespecificmolecularmecha- model, wewillidentifytheanti-aginggenes tissue formation andtreatmentoffibrous scar for prevention after injuriesanddiseases, andseekmethods the mechanismbehindfibrosisprocess tissue injuryandrepair stem cells, toclarifytheiressentialroleduring the pluripotencyofinjuredtissuesderived aims toidentifythecharacteristicsincluding sue compoundforwoundtissuerepair proteins andcellstocreateafunctionaltis- are alsobuilding3Dprinterbyusingnatural as children’s diaphragmatichernia(CDH). We tissues forrepairofpediatricdefects, such bioengineering scaffoldstobuildfunctional will usevariouscellsourcescombinedwith Cell Adhesion &Migration 2015;9(4):283- Justin Pham Medical students:ChenFu, ParshaForouzan, Fanwei Meng. Postdoc researchfellows:Dr. Kinga Vojnits; Dr. HaiyingPan Lab seniortechnician/manager: StephanieBaca Administrator: LAB MEMBERS Stem Cell2014;9(1):1-16. stem cell, typeofstemcell. anew Journal of Vojnits K, ZhanM, CoxCS, Li Y. Smallembryonic Regenerative Medicine2014;9(2):231-242 method inmodulatingfingerregeneration. Choi YH, CoxCS, LallyKP, Li Y. and The strategy stem cell-likecells(iMuSCs). The characteristicsofinjuredmusclederived ization ofratdiaphragms. an extracellularmatrix(ECM)fromdecellular - Scanning ElectronMicroscope(SEM)visualizes IMMPACT REPORT • • • • 67 Directed neural differentiation of human Neural induced pluripotent stem cells (hiPSCs). rosettes (A) express neural stem cell markers Sox1 (B) and Pax6 (C) Jul 18;5:4430. doi: 10.1038/ncomms5430 Jul 18;5:4430. (*Corresponding authors) and T., Truong, L., Sun, B., Long, H., Xue, S., Li, (2014) Efficient generation of hiPSC neu- Y. Liu, reportersral lineage specific knockin using the CRISPR/Cas9 and Cas9 double nickase system. J Vis doi:10.3791/52539. Exp. e52539, W.* (2011) Olig and Deng, P., Jiang, Y.*, Liu, gene targeting in human pluripotent stem cells for motor neuron and oligodendrocyte differen- (*Corresponding 640-655. Nat Prot. 6, tiation. authors) LAB MEMBERS Post-doctoral Fellow: Shenglan Li Associate: Haipeng Xue Research Li Bo Long, Yang, Visiting Seung H Scientists: Lihua Luo Sun, Down syndrome using patient derived iPSCs and neural populations progenitors for cell-based therapy in spinal cord injury and stroke CRISPR/Cas9 mediated gene targeting CRISPR/Cas9 mediated gene iPSCs A Neurogenin 2 knockin hiPSC reporter line made using the CRISPR/Cas9 system. NEUROG2-mCherry NEUROG2-mCherry A Neurogenin 2 knockin hiPSC reporter line made using the CRISPR/Cas9 system. hiPSC clones are induced as embryoid bodies (EBs) which glow red under the fluorescence microscope signal) expression faithfully native NEUROG2 antibody staining (green) confirms that mCherry (red, (A). C). reflects the endogenous NEUROG2 expression along the differentiation pathway (B, KEY PUBLICATIONS (2015) Reverse Engineering W. and Deng Y. Liu, Human Neurodegenerative Disease Using Plu- Research, Brain Technology. ripotent Stem Cell in press. (2014) Genetic Y. Liu Rao MS, Li S, J, Wu Xue H, modification in human pluripotent stem cells by homologous recombination and CRISPR/Cas9 [Epub 2014 Mar 11. Methods Mol Biol. System. ahead of print] PMID:24615461 Tran, S., Peterson, H., Xue, P., Jiang, C., Chen, Pleasure, S., Li, M., Parast, A., McCann, H.T., and Y.*, Liu, F., J. Loring, L.C., Laurent, D.E., (2014) Role of astroglia in Down W*. Deng, Syndrome revealed by patient-derived human Nat Commun. induced pluripotent stem cells. •Characterization of the role of OLIG genes in Identification of optimal neural lineage •Identification of optimal neural Creation of neural lineage hiPSC reporters•Creation of neural lineage hiPSC by RESEARCH PROJECTS integration-free •Generation of patient-specific, Human pluripotent stem cells in cell-based therapy in cell-based stem cells Human pluripotent for CNS injury Ying Liu, Ph.D. Ying Liu, Assistant Professor We have been pursuing basic and translation- We By transient overexpression of four transcrip- al research in the following two areas: (i) stem and cell biology and regenerative medicine, (ii) pathogenesis of neurodegenerative disease Our research entails the use of and CNS injury. combined genetic and molecular and cellular biological approaches applied to in vitro and in the neural focus on dissecting We vivo models. developmental pathways and the corresponding pathogenesis in spinal cord injury and stroke. Our long-term goal is to identify therapeutic targets for the treatment of CNS injury and neurodegenerative diseases. KLF4 and C-MYC, SOX2, OCT4, tion factors, somatic cells such as dermal fibroblasts, can be and blood cells, keratinocytes, reprogrammed to human induced pluripotent iPSCs provide Most critically, stem cells (iPSCs). theo- which autologous materials for patients, retically omit the need for immune suppression. have optimized the more clinically relevant, We integration-free hiPSC generation protocol and performed directed differentiation of patient- neuronal specific iPSCs into neural stem cells, as well as mature cell and glial progenitors, transplantation types for disease modeling, and neural regeneration and repair, studies, Recently we have drug screening and testing. adapted the highly efficient genome editing tool CRISPR/Cas9 system in creation of neural lineage reporters and gene corrections of patient These neural lineage specific cells are iPSCs. applied to in-depth study of signal transduction in disease and development. STEM CELL AND REGENERATIVE MEDICINE REGENERATIVE AND CELL FOR STEM CENTER 68 • IMMPACT REPORT •MM cellsandmicroenvironmentniche: We RESEARCH PROJECTS with microenvironmentisunderway. acterization ofPKH+cellsandtheirinteraction the quiescentPKH+populationsandchar- mice. We conductedgeneprofilinganalysesfor tumor formationinthesecondaryxenograft drug resistanceandcontributedtoincreased marrow compartment. These cellswerealso disease duetotheirmanifestationinthebone with microenvironment. MM isheterogeneous in multiplemyeloma(MM)andtheirinteraction lineofresearchstudying anew have developed generate reactiveoxygenspecies(ROS). We also shuttling influencesresistancetodrugsthat discovered thatBACH2nucleo/cytoplasmic that isinvolvedindrugresponsesMCL. We homology), whichisanothertranscriptionfactor of PAX5 signalingisBACH2(BTBandCNC direct translationalapplications. Downstream target drugresistancepopulationsinMCLhave drug resistant, discoveryofcompoundsthat cells. GiventhatPAX5 silencedcellsarehighly target thesurvivalpathwaysofPAX5 silenced libraries. We discoveredthatselectcompounds of NCIoncology, customclinical, andprestwick using librariescomprisedof3991compounds conducted highthroughputdrugscreening PAX5 isapotentialtumorsuppressor. We also in vitroandxenograftmice, indicatingthat in MCLleadstoincreasedtumorformation development. We discoveredthatPAX5 silencing 5 (PAX5), adeterminantofnormalBcelllineage progression. Oneofthosefactorsispairedbox are involvedinmalignantBcellinitiationand determine normalBcelllineagedifferentiation understanding howtranscriptionfactorsthat to malignantBcells. We areinterestedin selective growthandsurvivaladvantages to deciphermolecularpathwaysthatconfer CENTER FORSTEMCELL AND REGENERATIVE MEDICINE We willcontinueto characterizefunctionsof osteoblastic, vascularandspleenniches. genes expressedinquiescent MMcellsfrom conducted microarrayanalysestoidentify The majorgoalsofmyresearchprogramare multiple myeloma pathogenesis anddrugresistanceinhumanlymphoma Deciphering molecularandcellularmechanismsof Jerold B. KatzDistinguishedProfessorshipinStemCellResearch Associate Professor Nami McCarty, Ph.D. Blood 123:2204-2208, 2014. growth ofquiescentmultiple myelomacells. McCarty, N. the Osteoblasticnichesupports Chen, Z., Orlowski, R.Z., Wang, M., Kwak, L., and doi:10.1371/journal.pone.0069126, 2013. mantle celllymphoma. PLOSone8(8):e69126. modulates ROSmediatedcytotoxicresponses in of B-cell-specifictranscriptionfactor, BACH2, Kwak, L., andMcCarty, N. Nucleartranslocation L., Wang, M., Neelapu, S.S., McLaughlin, P., Chen, Z., Pittman, E.F., Romaguera, J., Fayad, 159:243-246,Hematology 2012. oxygen speciesproduction. BritishJournal of mantle celllymphomaviainductionofreactive in tically enhancescytotoxicityofbortezomib Kwak, L.W., andMcCarty, N. Verapamil synergis- Chen, Z., Romaguera, J., Wang, M., Fayad, L., 87:1057-1064, 2012. (MCL). AmericanJournal ofHematology (ATO) inmantlecelllymphoma withbortezomib antiproliferative effectsofarsenictrioxide Jung, H-J., Chen, Z., andMcCarty, N. Synergistic 119:2568-2578, 2012. of tissuetransglutaminaseactivities. Blood mantle celllymphoma(MCL)viamanipulation resistancein blockers decreasethebortezomib guera, J., Kwak, L.W., andMcCarty, N. Calcium Jung, H-J., Zheng, C., Wang, M., Fayad, L., Roma- KEY PUBLICATIONS •Delineating transcriptionfactornetworkson ofsmallmoleculeinhibitors •Development patient survival determine drugresistanceoutcomeand BACH2 sub-cellularlocalizationinthecell collaborators atMDACC todeterminewhether pathogenesis. We alsowillcloselyworkwith to addressrolesforPAX5 signalinginMCL drug resistantlymphomas: We willcontinue test itsefficaciesinthepatients models forpre-clinicalstudiesandplanto andtestthesecompoundsinanimal develop bone marrowcompartment. We willfurther selectively targetMCLcellsthathometothe screening toidentifythecompoundsthat have conductedhighthroughputchemical for targetingadvancedlymphomas: We marrow microenvironment these genesintheMMinteractionwithbone Post-doctoral Fellows:JenniferHan, JimmyLin Senior Research Associate: JudyChen LAB MEMBERS leu.2015.140. [Epubaheadofprint], 2015 dent ofCCND1. Leukemia15. doi:10.1038/ a poorprognosisinMCLpatientsindepen- progression anddrugresistance, andpredict promotemalignantB-cellinfiltration,levels Medeiros, L.J., andMcCarty, N. DifferentialPAX5 Teo, A.E., Chen, Z., Miranda, R.N., McDonnell, T., tioned medium)methylcellulosemedium. (phytohemagglutinin humanleukocytecondi- (MCL) cells(5x103)wereculturedinPHA-LCM Jeko, SP53andZ138mantlecelllymphoma into NOD/SCID mice to evaluate engraftment.into NOD/SCIDmicetoevaluate the fluorescentdyePKH26andwereinjected shRNA cells(control, 5x105)werelabeledusing CXCR4 shRNAMCLcells(CXCR4KD)orcontrol in vivouponCXCR4silencing. GFP-positive Decreased quiescentPKH-positivecellrecovery IMMPACT REPORT • • • • 69 KEY PUBLICATIONS KEY PUBLICATIONS (2015) et al. N., Matthias, H., Oda, K., Umeda, chondrogenic “Long-term expandable SOX9+ human pluripotent ectomesenchymal cells from Stem Cell Reports 4:712-726. stem cells” et al. K., Umeda, M., Ikeya, K., Yokoyama, of iPS cells “Enhanced chondrogenesis (2015) from neonatal-onset multisystem inflammatory disease occurs via the caspase-1-independent Arthritis Rheumatol. cAMP/PKA/CREB pathway” 67: 302-314. et al. M., Tanaka, S., Trilok, S., Li, J., Zhao, specification “Small molecule-directed (2014) of sclerotome-like chondroprogenitors and induction of a somitic chondrogenesis program 141: Development, from embryonic stem cells” 3848-3858. (2013) al. et F., Shioda, A., Shono, S., Mae, “Monitoring and robust induction of nephro- genic intermediate mesoderm from human 4:1367 , Nat Commun pluripotent stem cells” (2012) et al. P, Simmons, J., Zhao, K., Umeda, “Human chondrogenic paraxial mesoderm, directed specification and prospective isolation 2:455. , Sci Rep from pluripotent stem cells” LAB MEMBERS BA. Assistant: John Lee, Research Animal Special- Associate and Senior Research DVM ist: Nadine Matthias, Eight weeks after transplantation of cartilage particles generated in vitro from hiPSC-derived ectomesenchymal cells under two different culture conditions: Stable cartilage-forming condition (upper panels) and growth plate-like unstable cartilage-forming condition (lower panels). plantation model for cell-based articular cartilage repair the hPSC-derived chondroprogenitors and joint progenitors to elucidate the molecular basis of joint chondrogenesis sion of joint progenitors from hPSCs using fluorescence reporter PSC lines term expansion without loss of chondrogenic activity of the hPSC-derived chondroprogenitors expansion of three embryonic chondropro- limb mesenchyme and genitors (sclerotome, ectomesenchyme) from hPSCs We have established culture conditions that We •Establishment of an orthotopic xenotrans- •Defining the process of chondrogenesis from Generation, detection, isolation and expan- detection, •Generation, •Elucidation of the molecular basis of long- extent, expand joint progenitor-like cells that extent, precursor) express syndetomal (ligament progeny. markers from the paraxial mesoderm we have recently discovered a way Furthermore, to generate cartilage particles from the hPSC- that show no to derived chondroprogenitors, very limited mineralization/bone formation after currently focusing both are We transplantation. on the characterization of joint progenitor-like cells and on the elucidation of critical signaling mechanism for stable cartilage formation, aiming to establish cellular and humoral environment suitable for reproducibly generating joint-type stable cartilage. maintain and expand the sclerotomal and ectomesenchymal chondroprogenitors for an of their without loss extended period of time, stable expansion of Such chondrogenicity. chondrogenic activity is currently very hard to are also focus- We achieve adult MSCs. with ing on genome-wide molecular search (e.g. epigenome analyses) proteome, transcriptome, aiming in these expandable chondroprogenitors, to understand the mechanistic basis that may be applied to improve the expansion culture method for adult MSCs and the hPSC-derived join progenitor-like cells in future. RESEARCH PROJECTS prospective isolation and •Specification, Pluripotent stem cell differentiation and lineage specification Pluripotent stem Naoki Nakayama, Ph.D. Naoki Nakayama, Associate Professor in Stem Cell BiologyGraham Distinguished Chair Annie & Bob The cartilage of joints is not spontaneously Permanent cartilage formation – proper There has been repaired after injury in humans. considerable interest in the clinical application of stem cells to the repair of damaged current cell therapies using cartilage; however, chondrocytes and mesenchymal stromal cells (MSCs) face the problems of low yield of cells and their tendency to yield unsuitable and/ Joint or unstable cartilage after expansion. Therefore, is formed during embryogenesis. we hypothesize that the embryonic cell-type responsible for limb and vertebral- joint forma the common precursor joint progenitor, tion: i.e. of synovial joint components including articular and meniscal chondrocytes and ligaments, would be the best for the regeneration of adult stem cells (PSCs), Pluripotent joint cartilage. induced or whether derived from an embryo, are expected to differentiate from adult cells, into any somatic cell-type in culture through processes that mimic embryogenesis in vivo, making human (h)PSCs a promising source of embryonic cells for regenerative medicine. The major challenges have been to direct their differentiation toward the cell type of interest to obtain progeny of the right quality), (i.e. and to isolate them in large quantities without introducing transgenes and mutations. have previouslyWe signaling and right cell type: developed and purified from hPSCs paraxial two of mesoderm and neural crest progeny, the three embryonic origins of chondrocytes, with the capacity to expand and differentiate into sclerotomal and ectomesenchymal have We respectively. chondroprogenitors, recently established a condition to generate the third chondrogenic lateral plate mesoderm, from hPSCs, embryonic origin of chondrocytes, All these progeny give rise to hyaline-like too. cartilage particles in culture under our standard most of them are unstable However, condition. in vivo and are mineralized and turned into bone when ectopically transplanted into im- have discovered We munocompromised mice. a way to selectively generate and to a limited STEM CELL AND REGENERATIVE MEDICINE REGENERATIVE AND CELL FOR STEM CENTER 70 • IMMPACT REPORT CENTER FORSTEMCELL AND REGENERATIVE MEDICINE from theprimarytumor. Byapplicationofbio - contribute tothesystemic spreadofcancer and activationofoncogenic pathwaysthat the impactoffluidforceoninvasivepotential stress experiencedbycancercellsandevaluate lymphatic vasculature, wemodulatetheshear biomimetic microchipsdesignedtomodel the metastatic potentialofcancercells. Using vascular shearstressesmayplayinregulating it remainsunclearwhatrolelymphaticor have beenimplicatedintumorbiology, but stimuli. readouts ofstemcellresponsetomechanical and animalmodelsoftraumaticbraininjuryas permeability. We utilizeculture-basedassays ability tomodulateinflammationandvascular ofmesenchymalstemcells,ogy includingtheir determine howmechanicalforcealtersthebiol- Consequently, oursecondareaofinterestisto jury andchronicstatesofimmunedysfunction. response whichcanaccompanytraumaticin- inflammatory signalingandinnateimmune gests thatmesenchymalstromalcellsreduce umbilical cord, andfat. Currentresearchsug - out thebody, includingthebonemarrow, can befoundinavastarrayoftissuesthrough- for regenerativemedicine. These stem-like cells great dealofattentionaspotenttherapeutics mouse genetics, andtransplantationassays. opment, includingmicrofluidics, pharmacology, - theirroleinblooddevel proaches toevaluate signaling cascade, andweemployvariousap- playersmediatingthis investigation askey and biochemicalpathwaysarecurrentlyunder stem cellsforclinicaluse. A numberofgenetic to expandimprovedsourcesofhematopoietic we mightusethisinformationinthelaboratory duringembryogenesis andhow development address howfrictionalforcepromotesblood cell potentialandbehavior. ture andlymphflowinthelymphaticsimpacts force generatedbybloodflowinthevascula- Finally, fluidflowandhydrostaticpressure Mesenchymal stromalcellshaveattracteda One armofourresearchisdesignedto Members ofourlabstudyhowbiomechanical migration underfluid flow. of filopodialextensionsindicativeactive cell development ning electronmicroscopyreveals Utrastructural analysisofcancercellsby scan- HSCs. DevCell,29:621-628. graftment definesthemostnascentembryonic Yoder, M.C., Daley, G.Q. (2014)Neonatalen- Ross, S.J., Kim, P.G., Chou, S., Yoshimoto, M., Arora, N., Wenzel, P.L., McKinney-Freeman, S.L., embryo development. Blood,125:1418-1426. endothelial cellsduringmouseandzebrafish Foxc2 toinducespecificationofhemogenic C.E., andDaley, G.Q. (2015). Notch1actsvia P.G., Schlaeger, T.M., Zon, L.I., Bigas, A., Burns, T., Datta, S.M., Arora, N., Guiu, J., Lagha, M., Kim, ang, I.-H., Lu, Y.-F., Zhao, L., Wenzel, P.L., Kume, J Stem Cells. MethodsMolBiol,1212:183-193. of HemogenicEndotheliumandHematopoietic of FluidMechanicalForcetoEmbryonicSources Li, N., Diaz, M.F., Wenzel, P.L. (2015). Application 212: 665-680. andcoverart. Featuredarticle E2 andthecAMP-PKASignaling Axis. JExpMed , through Prostaglandin mote BloodDevelopment Wenzel, P.L. (2015). BiomechanicalForcesPro- Ingber, D.E., Daley, G.Q., García-Cardeña, G., and D.A., Morris, S.A., Naveiras, O., Murthy, S.K., S.M.,H.E., Willey, Arora,N., Torisawa, Y., Vickers, Diaz, M.F., Li, N., Lee, H.J., Adamo, L., Evans, KEY PUBLICATIONS •Fluid flowininitiationofmetastasis •Modulation ofanti-inflammatoryprogramsin ofblooddevelopment •Mechanobiology RESEARCH PROJECTS ment optionsforpatientsaffectedbycancer. cancer biology, treat- wehopetoidentifynew engineering approachestomicroenvironmental Assistant Professor Pamela Wenzel,Ph.D. Biomechanical forceregulatescellpotentialandbehavior mesenchymal stemcells lymphatic vasculature. may experienceduringmetastasisthrough the activity underconditionsthatacancercell cancer cellmigrationandmonitoringofgene microchip permitsreal-timevisualization of lymphatic flow. Thedesignofthisbiomimetic is pushedthroughthemicrochanneltomimic thelial cellsandcancercells. Culturemedium followed byintroductionofmicrovascularendo- microchannel istreatedwithcollagenmatrix, ded withinasmalltransparentdevice. The create acylindricalculturesurfaceembed- Elastic polymeriscastbysoftlithographyto (Pediatric Surgery) Administrative Assistant: StephanieBaca Student: Alexander Student: KatherinePrice Research Assistant:AbishekVaidya,M.S. Postdoctoral Fellow:HyunJungLee, Ph.D. Research Associate: MiguelDiaz LAB MEMBERS 92-103. pro-hematopoietic signaling. Differentiation89: extrinsicphysicalcuesdrive development: P.L. (2013)Biomechanicalforceinblood Lee, H.J., Li, N., Evans, S.E., Diaz, M.F., Wenzel, IMMPACT REPORT • • • • 71 TCF7, together with RUNX1, regulates a tran- together with RUNX1, TCF7, scriptional regulatory network. “Isoform specialization”--Splicing diversity is the highest in hESCs and decreases when cells commit to neural differentiation. Zhang, Y., Chen, K., Sloan, S., Bennett, M., M., Bennett, S., Sloan, K., Chen, Y., Zhang, Guarnieri, H., Phatnani, S., O’Keeffe, A., Scholze, Liddelow, S., Deng, N., Ruderisch, C., Caneda, P., Barres, T., Maniatis, R., Daneman, C., Zhang, S., and splic- An RNA-Seq transcriptome J.Q. Wu, B., cells glia and vascular ing database of neurons, 34(36): Neurosci., J. of the cerebral cortex. PMID:25186741 11929-11947. Application The J.Q. Wu, and X., Dong, K., Chen, of single-cell sequencing in dynamic transcrip- (ed.), Wang X. Single Cell Sequencing. tomes. Single Cell Sequencing and Systems Immunol- 5. Springer Bioinformatics Translational ogy, 41-63 Publisher Dordrecht 2015. LAB MEMBERS Ph.D., Post-doctoral Fellows: Xiaomin Dong, Ph.D. Raquel Duran, Ph.D., You, Yanan Ph.D. Research Associate: Han Yan, Undergraduate Students (Rice University): Fanny Ashkan Huang, Rohani and global network integration of genomic and proteomic data precursor cell self-renewal and differentiation regulatory RNAs, and modulate key regulators regulatory RNAs, and to improve differentiation efficiency transplantation safety Immunofluorescence labeling of neurons derived from H1 human embryonic stem cells red) labels both beta-tubulin (TujIII (hESCs). Nuclei (blue) immature and mature neurons. are stained by DAPI. KEY PUBLICATIONS Qiu, A., Szekely, P., Noisa, L., Habegger, Q., J. Wu, M., Egholm, H., Lin, D., Raha, S., Hutchison, C., and Snyder, M., Gerstein, W., Cui, S., Weissman, during Transcriptomes Dynamic (2010). M. Neural Differentiation of Human Embryonic Stem Cells Revealed by Integrating Short, 107: and Paired-end Sequencing. PNAS. Long, 5254-5259. M., Hariharan, V., Schulz, M., Seay, Q., J. Wu, Ger- J., Z. Ye, M., Shi, J., Du, J., Lian, D., Tuck, (2012). S. Weissman, and M., Snyder, M., stein, Tcf7 is a key of the self-renewal regulator and differentiation switch in a multipotential hema- PLoS Genet 8(3): e1002565. topoietic cell line. G., Yang, Y., Zheng, H., Lu, S., Deng, K., Chen, RNA-Seq (2013). J.Q. Wu, and Q., Cao, D., Kim, characterization of spinal cord injury transcriptome in acute/subacute phases: a resource for understanding the pathology at the systems PMC3739761 8(8):e72567. Plos One. level. •Network analysis of stem cell differentiation Identify the molecular switch of hematopoietic •Identify the molecular switch •Identify key transcription factors and Gene transcription and regulation of stem cell differentiation of stem and regulation Gene transcription Jiaqian Wu, Ph.D. Jiaqian Wu, Assistant Professor My laboratory combines stem cell biology The other area of our research interest lies cord injury and neurological diseases neural differentiation STEM CELL AND REGENERATIVE MEDICINE REGENERATIVE AND CELL FOR STEM CENTER and systems-based approaches involving and bioinformatics, proteomics, genomics, functional assays to unravel gene transcription and regulatory mechanisms governing stem cell our group One major focus of differentiation. is investigating stem cell neural differentiation and developing effective and safe treatment for We spinal cord injury and neurological diseases. are studying gene expression and the regulation of transcription factors and regulatory RNAs using next-generation sequencing technologies, These studies including RNA-Seq and ChIP-Seq. are crucial in understanding the molecular mechanism of stem cell neural differentiation is to iden- Our goal and its clinical implications. tify and modulate key regulators as therapeutic targets to direct the differentiation of stem cell into neural cells more efficiently and to increase transplantation safety. in the studies of the regulatory networks of hematopoietic precursor cell self-renewal and differentiation using multipotent EML (erythroid, and lymphocytic) cell as a model myeloid, are using integrated genomic and We system. proteomic approaches to identify key compo- have identified We nents that control the switch. together with RUNX1 are important regu- TCF7, Future study will generate lators in this process. a global interaction network and a novel and comprehensive view of the regulation of early stages of hematopoietic precursor self-renewal This study can serve as a and differentiation. model for the analysis of cell self-renewal and differentiation in general and provide insight for efficient expanding and manipulating hema- including topoietic precursor and stem cells, reprogramming partially differentiated cells to return them to a self-renewing state. RESEARCH PROJECTS •Characterize molecular signatures of spinal •Investigate gene expression during stem cell 72 • IMMPACT REPORT from highlymalignant humancarcinomassuch efficiently clonethe elusive “cancerstemcell” isthatwecan the moststrikingdevelopment possibility ofpreemptivetherapies. Butperhaps appear decadesbeforefrankcancersoffers the intestinal metaplasia, andPanINsthatoften lesions suchasBarrett’s esophagus, gastric Our abilitytoclonestemcellsofprecursor of theovary, esophagus, pancreas, andlung. andlethalityofepithelialcancers evolution totheproblemof cell cloningtechnology pancreas, andkidney. invariably lethaldiseasesofthelung, liver, editing, approachestochronicand offersnew and colon, coupled withadvancesingenome in lungand3-Dvillistructuresoftheintestine for complexepithelialstructuressuchasalveoli these stemcellsalsopossesstheinformation signaling thatlikelydrivesthesediseases. That alteredinflammatory stem cellsarerevealing diseases). Ouranalysesofpatient-derived the gastrointestinaltract(inflammatorybowel diseases oftheairways(asthma, COPD)andof epithelial stemcellstochronicinflammatory interestedinthecontributionof is particularly regenerate complexepithelia. Mylaboratory diseasemechanismsandfaithfully reveal these clonedstemcellsaretheirabilityto cells. derived, allofwhichdefinethemastruestem were committed tothetissuesfromwhichthey 2) aremultipotent, and3)areabsolutely marked by1)unlimitedproliferativepotential, thatthesecells are Our analysesarerevealing lung, gastrointestinaltract, pancreas, andliver. and diseasedhumanepithelialtissuessuchas technologies forcloningstemcellsfromnormal states. robust Mylaboratoryisdeveloping while clonogenic, arefarremovedfromdisease onic andinducedpluripotentstemcellsthat, clone thesecells. This isincontrasttoembry - medicine hasbeenlimitedbyourinabilityto modeling criticaldiseasesandforregenerative CENTER FORSTEMCELL AND REGENERATIVE MEDICINE Finally, wehaverecentlyadaptedourstem Perhaps themostremarkablefeaturesof The potentialoftissue-specificstemcellsfor CPRIT Scholar Assistant Professor Wa Xian, Ph.D. malignancies Personalizing stemcellsfromchronicdiseasesand 109:10516-10521. of cervicalcancer. ProcNatl Acad SciUSA. junction cellsimplicatedinthepathogenesis A discretepopulationofsquamocolumnar S, McKeonFD, Xian W*, CrumCP*. (2012). MR, McLaughlin-DrubinME, MüngerK, Feldman M,Herfs Yamamoto Y,Laury A,Nucci X, Wang 616-620. are essentialforlungregeneration. Nature517, (2014). distalairwaystemcells p63+/Krt5+ Lessard, M., Crum, CP, Xian, W*, McKeon, F*. Yamamoto, Y., Wang, X., Lim, SW, Vincent, M., Zuo, W., Zhang, T., Wu, D.Z-A, Guan, SP, Liew, AA, 173-178. ground stateintestinalstemcells. Nature522, and Xian, W*. (2015)Cloningandvariationof R, Lacy, D.B., Ho, KY, Crum, CP, McKeon, F*, N, Sylvester, FA, Hyams, JS, Devers, T, Bronson, Y, Khor, CC, Chevalier, B, Bertrand, D, Nagarjan, Howitt, B, Farrow, MA, Kern, F., Gang, N, Hong, X,Wang, Yamamoto, Y,LH,Zhang, Wilson, T., KEY PUBLICATIONS •Models forlungstemcelltransplantationin •Stem cellalterationsinpatientswithCOPD •Gastrointestinal stemcellsfrompatientswith •Evolution ofcancersviametaplasticprecur- •Cancer stemcellsfromuppergastrointestinal •Libraries ofcancerstemcellsfromhigh-grade RESEARCH PROJECTS outcome forpatientswiththesediseases. anticipate thatsuchapproacheswillalterthe population ofresistantcancerstemcells. We disease mounted, inlargepart, bythisminority patient-specific therapiestomitigaterecurrent is nowbeingemployedtoidentifyoptimal standard-of-care chemotherapy. This knowledge stem cellsthatpossessintrinsicresistanceto cell heterogeneityincludingsubsetsofcancer remarkablefeaturesofcancerstem revealing and genomicanalysesoftheselibrariesis “library” ofthousandssuchcells. Functional that eachpatient’s tumorisrepresentedasa acute andchroniclungdamage and asthma inflammatory boweldisease sors, dysplasia, adenocarcinoma tract cancers ovarian cancer cal section. culture enface. Bottom, H&Estainedhistologi - to air-liquidinferfaceconditions. Top, of view colonic stemcellpedigreefollowingexposure Right, Colonicepitheliaderivedfromasingle stem cellsonirradiated3T3feeder(left). Colonies ofapproximately500-1000colonic Senior ResearchScientist:Xia Wang, Ph.D. LAB MEMBERS cating author metaplasia. Cell145, 1023-1035. *Communi - embryonic cellsasprecursorsofaBarrett’s-like Crum CP, Xian W*, McKeonF*. (2011). Residual TS, DagherR, M,Vincent LuX, Bellizzi AM, HoKY, Wang X, OuyangH, Yamamoto Y, KumarPA, Wei influenza infection. Cell147, 525-538. and duringlungregenerationfollowingH1N1 Distal airwaystemcellsrenderalveoliinvitro Crum, C.P., Xian, W*, andMcKeon, F*. (2011). Zielonka, E.M., Wang, D.Y., Lim, B., Chow, V.T., Wei, T.S., Mu, D., Sun, Y., Lim, S.J., Dagher, R., Kumar, P.A., Hu, Y., Yamamoto, Y. Neo, B.N., expansion andCNVanalysis(right). case (left). Fourof96CSCclonessampledfor library ofclonesfromhigh-gradeovariancancer Derivation ofpatient-specificcancerstemcell IMMPACT REPORT • • • • 73 Our research programs are focused on methods are programs research Our will research The successful completion of this degenerative diseases in the elderly has led to the diseases degenerative hypothesis that aging is caused, in part, the by loss of functional stem cells necessary for tissue rejuvenation. these and techniques to ameliorate and reverse changes. for approaches of novel in the development result cells or rejuvenating the use of stem/progenitor functional stem/progenitor from factors, derived We cells, to extend human health and lifespsn. in investigators also establishing numerous are our can synergize so we of aging research the area research. efforts on tissue engineering and aging Ph.D. Huard, Johnny Professor Wallace & Distinguished Director Center Department of Orthopaedic Chair for Research, Vice Surgery Center for Director of Chief Scientific Officer & Medicine Sports Regenerative Vail, Institute, Research Philippon Steadman Colorado

he demographics of the American an increasing shifting toward population are elderly population, placing extraordinary A universal characteristic of aging is the loss A universal demands on our health care system. Aging demands on our health care attrition of homeostasis in the progressive results and functional reserve of all organ systems. As a consequence, the incidence of numerous debilitating diseases including neurodegeneration, sensorineural defects, sarcopenia, osteoporosis, disease, diabetes, and cancer cardiovascular the molecular with age. Understanding increases strategies for basis of aging and developing or delaying age-associated diseases preventing two of the most fundamental and therefore are challenges that the medical research pressing of nature The precise community faces today. for aging-related the damage that is responsible ill-defined but may changes remains degenerative attrition, include mitochondrial damage, telomere accumulation of genetic nuclear dysmorphology, or DNA, protein mutations, and cumulative membrane damage. potential, which leads to of tissue regenerative and to stress, ability to respond an impaired the as a consequence, dramatically increases This and the risk of morbidity and mortality. of numerous incidence exponentially increased

TISSUE FOR TISSUE CENTER AGING AND ENGINEERING RESEARCH T 74 • IMMPACT REPORT TISSUE ENGINEERING AND AGING RESEARCH ENGINEERING ANDAGING CENTER FORTISSUE and internationalconferences. abstracts acceptedforpresentationatnational papers, 82book chapters, andhavehad757 team haspublishedover300peerreviewed with naturalagingandprogeria. Myresearch thephenotypicchangesassociated alleviate MDSCs asasourceforparacrinefactorsto nerve regenerationusingMDSCs;theuseof regeneration, peripheral andfibrosisprevention; tion; cardiacandskeletalmuscleinjuryrepair, regeneration throughstemcelltransplanta- cartilage transplantation; boneandarticular muscular dystrophy(DMD)throughMDSC degeneration associatedwithDuchenne’s ofthemuscular characterization; alleviation skeletal musclestemcellisolationandtheir Our currentmajorresearchinterestsinclude: have volunteeredforthisstemcelltherapy. more than400patientsinCanadaandtheU.S and myocardialinfarction. As ofthisdate, for thetreatmentofurinarystressincontinence my teamarecurrentlyundergoingclinicaltrials industry. The MDSCsthathavebeenisolatedby that wehavedeveloped, havebeenlicensedto international recognition, andthetechnologies of tissues. Myteamhasreceivednationaland aid inthehealingandregenerationofavariety andtheirtranslationtoclinic cell biology My primaryareasofinterestareinbasicstem the useofmuscle-derivedstemcells(MDSCs). regenerative medicineapplicationsbasedon areas ofgenetherapy, tissueengineering, and The focusofmyresearchprogramisinthe muscle. red) infiltrationwere detectedindystrophic ated musclefibers)andmacrophages(F4/80, Embryonic myosin(green, regener- showingnewly Cell StemCell, 2008Sep11;3(3):301-13. chymal stemcellsinmultiplehumantissues. Huard J, PéaultB. Perivascularoriginofmesen- Andriolo G, Bühring HJ, LazzariL, GiacobinoJP, Yap S, NorotteC, CorselliM, Traas J, DeasyB, Crisan M, Park, TS Casteilla, L, SunB, ZhengB, PMID: 17767154 25(9):1025-34. J. Huardcorrespondingauthor. human skeletalmuscle. NatBiotech2007Sep; identification ofmyogenicendothelialcellsin ibeh B, DeasyB, HuardJ, PéaultB. Prospective Yap S, Pollett, JB, L, Drowley Cassino T, Ghara- Zheng B, CaoB, CrisanM, SunB, LiG, Logar A, potential. NatCellBiol2003Jul;5(7):640-6. hematopoietic lineagesbutretainmyogenic Z, HuardJ. Musclestemcellsdifferentiate into M, DeasyB, CumminsJ, EpperlyM, Qu-Petersen Cao B, ZhengB, JankowskiRJ, KimuraS, Ikezawa KEY PUBLICATIONS ofbiologicalapproaches to •Development •Cell autonomousandnon-autonomous •Biomimetic coacervatedeliveryofmuscle asanew •The useofcoacervatetechnology •Bone abnormalitiesandhealingdefectin RESEARCH PROJECTS Director, IMMCenterfor Engineeringand Tissue Aging Research Surgery ofOrthopedic Department Distinguished Wallace Professor& Research ChairforOrthopedic Vice Johnny Huard, Ph.D. resistance mechanismstotherapeuticantibodytreatment Heterogeneity oftumormicroenvironmentandcancer ment syndromeinjury - improve functionalrecoveryaftercompart mechanisms ofstemdefectswithaging stem cellstoimprovecardiacrepair tissue repair drug deliverysystemformusculoskeletal muscular dystrophy (Red) invitro, NucleistainedwithDAPI (Blue). ated intoMyosinheavychainpositivemyotubes Muscle derivedstemcellsareabletodifferenti - DE: Defect:DE, BR:Brain. Duramater: planted inacriticalsizecranialdefectmodel. endochondral boneformationwhentrans- with GFP(donorcells)duringMDSCmediated and 14dayspost-implantation)colicalizes expressed highlyinthechondrocytestage(7 cell (MDSC)mediatedboneregeneration. Itis formusclederivedstem Cox-2 isimportant 1745-56. PMID:24642464 generation.” JClinInvest. 2014 April 1. 124(4). promote functionalmurineperipheralnervere- “Human muscle-derivedstem/progenitorcells A, StolzDB, ClarkKA, SunB, PéaultB, HuardJ. Lavasani M, Thompson SD, PollettJB, Usas A, Lu Commun 2012Jan;3:608. PMID:22215083 and lifespaninamurineprogeriamodel. Nat progenitor celldysfunctionlimitshealthspan Niedernhofer L, HuardJ. Muscle-derivedstem/ J, Ahani B, J,Tilstra FeldmanC, RobbinsP, Lavasani M, Robinson A, Lu A, SongM, Feduska IMMPACT REPORT • • • • 75 Human muscle-derived stem cells transduced with lenti-BMP2 regenerated functional bone that constitute bone and bone marrow with three lineages of hematopoietic stem cells in the critical size bone defect model and are as efficient as bone marrow MSCs (Biomaterials. ). 2014 Aug;35(25):6859-70. Mechanisms of murine BMP4GFP-transduced MDSCs mediated bone regeneration: differentiation and interaction with host cells via paracrine 2014 Aug;28(8):3792-809.). J. effect (FASEB ties in muscular dystrophy disease mode and osteoarthritis repair using ex vivo gene therapy and biomaterial scaffold. gene therapy for the bone tissue repair Muscle derived stem cells for bone and cartilage stem cells Muscle derived regeneration and repair Xueqin Gao, M.D., Ph.D. M.D., Xueqin Gao, Assistant Professor KEY PUBLICATIONS Tan J, Schneppendahl Lu A, Y, Gao Tang X, Usas A, Tuan Cummins JH, B, Wang AM, Kozemchak J, A comparison of bone regenera- Huard J. RS, tion with human mesenchymal stem cells and muscle-derived stem cells and the critical role 2014 Aug;35(25):6859- Biomaterials. of BMP. PMID:24856105 70. Cummins JH, A, Lu Proto JD, A, Usas Gao X, Role of donor and Huard J. CW, Chen A, Proctor host cells in muscle-derived stem cell-mediated paracrine ef- bone repair: differentiation vs. 2014 Aug;28(8):3792-809. J. fects. FASEB PMID:24843069 Chen CW, B, Wang Y, Tang A, Lu A, Usas Gao X, BMP2 Huard J. Cummins JH, JC, Tebbets Li H, is superior to BMP4 for promoting human muscle-derived stem cell-mediated bone regeneration in a critical-sized calvarial defect 2013;22(12):2393-408. Cell Transplant. model. PMID:23244588 Sekiya N, Gao X, Proto JD, Okada M, Chen CW, A, Saparov Crisan M, Corselli M, Beckman SA, Human pericytes Huard J. Péault B, K, Tobita 2013 Stem Cells. for ischemic heart repair. PMID: 23165704 Feb;31(2):305-16. •Exploring the mechanism of bone abnormali- •Human muscle derived stem cells for cartilage investigating how the muscular dystrophy affects investigating how the muscular bone interact the bone and how muscle and are hoping to unveil We in this mouse model. as a strategymechanisms that can be used to benefit DMD patients. RESEARCH PROJECTS •Utilizing human muscle derived stem cells and I am a member of Dr. Johnny Huard’s research Johnny Huard’s I am a member of Dr. Human muscle derived stem cells for bone Large segmental bone defect and non-union Human muscle-derived stem cells for age- Exploring the muscle and bone interaction in Muscular dystrophy is a deadly muscle laboratory Huard’s My research in Dr. team. focuses on using muscle-derived stem cells and I gene therapy for bone and cartilage repair. am doing translational studies to use muscle- derived stem cells for the treatment of bone de- and age-related bone fracture non-union, fect, and cartilage conditions such as osteoporosis also working on the I am and osteoarthritis. bone biology of a disease model – muscular dystrophy. regeneration cancer fracture caused by traumatic injury, resection represent major issues in orthopaedic and growth factors, Using stem cells, clinic. scaffold to regenerate bone tissue to replace traditional autografts and allografts to treat these disease is a new trend and has achieved new are exploring We vectors, a lot of progress. growth factors to mediate human muscle also We derived stem cells ex vivo gene therapy. are investigating the age of human muscle- derived stem cells and host on the human muscle-derived stem cells mediated bone repair. related cartilage injury or osteoarthritis Much progress has been made in using stem including murine muscle derived stem cells, cells for osteochondral defect or osteoarthritis we will explore ex vivo In this project, repair. gene therapy including using viral vector and biomaterial to deliver growth factors for cartilage repair especially osteoarthritis using both in vitro and in vivo model. a muscular dystrophy model Patients disease that inflicts 1 in 3,000 boys. often become wheelchair bound in their second have found bone abnor- We decade of life. malites in a dystrophin /utrophin double knock out (dko) model that closely mimic the clinical are We manifestation of human DMD patients. TISSUE FOR TISSUE CENTER AGING AND ENGINEERING RESEARCH 76 • IMMPACT REPORT TISSUE ENGINEERING AND AGING RESEARCH ENGINEERING ANDAGING CENTER FORTISSUE organs. inhibits chondrogenesis. into chondrocytesorwhetherthehighpressure sure canenhancethehMDSCstodifferentiate microenvironment andwhetherthehighpres - whether hMDSCscantolerateahighpressure ronment ofthejoint. The projectwillelucidate to mimictheinvivohighpressuremicroenvi - acontinuouspressureculturesystem developed To utilizestemcellsforOAtherapy, wehave the highpressuremicroenvironmentisunknown. can survive, proliferate, anddifferentiateunder stem cellregeneration. Whether thestemcells inpatients,ticular cartilage whichmayprevent effects ofcontinuouscompressionlivingar- of ajointinforcedpositioncausedfromthe degeneration associatedwithimmobilization rently nocure. Inclinical physical therapy, joint musculoskeletal diseaseforwhichthereiscur- repair. cartilage MDSCs toenhancearticular cytokines orgrowthfactors. enhance angiogenesisthroughthereleaseof potential oftheMDSCs, butalsosynergistically retention, survival, andthecardiacregenerative of researchthatcouldnotonlypromotecell vehicle fortheMDSCs, area representsanew or growthfactor-coacervate, asanoveldelivery potential oftheMDSCs. The useofcytokines genic potentiallimitsthecardiacregenerative and survival;moreover, MDSCslowcardiomyo- in PBS), whichleadstolimitedcellretention ering thecells(directintramyocardialinjection limitations, suchasapoorapproachesfordeliv- improve cardiacfunction. However, several approach torepairinjuredmyocardiumand for cardiacrepairandregeneration. of research. diseases. Currently, wehavethreemajorareas related cell therapyfortreatinghumansport ofgenemodificationandstem development 3) MDSCsandregeneration indigestivetract (OA)isadebilitating Osteoarthritis 2) The effectsofcontinuouspressureon Cellular cardiomyoplasty(CCM)isapromising 1) BiomimeticcoacervatedeliveryofMDSCs Our labisfocusingonthediscoveryand of development,2014(134):67-79. pancreas.lium inthedeveloping Mechanisms Gittes G. Barrierfunctionofthecoelomicepithe- ersch J, GaffarI, Tulachan S, El-Gohary Y, SongZ, Guo P, PreuettB, KrishnaP, XiaoX, ShiotaC, - Wi Protocols, 2014, (12):2719-24. viral vectorsand/orcell-taggingdyes. Nature pancreatic ductalinfusionofadeno-associated manipulations inmultiplecelltypesusing tracing, lineagetaggingandtargetedgenetic Gohary Y, HusainSZ, GittesGK., Pancreaticcell L, S, Fischbach SongZ, GaffarI, J,Wiersch El- Xiao X, GuoP, PrasadanK, ShiotaC, Peirish KEY PUBLICATIONS in •hMDSCs potentialtherapeuticdevelopment •In vitrocontinuouspressurecultureof • RESEARCH PROJECTS tis ordiabetes. into pancreaticcellsinordertotreatpancreati- or couldrecruitotherhostcellstodifferentiate capacity todifferentiateintopancreaticcells, we proposethatMDSCsmayhavethepotential nerve, bone, cartilage, muscle, andheart. Here that MDSCsarecapableofdifferentiatinginto have notbeenreported. Ourlabhasshown (MDSCs) fortreatingpancreatitisanddiabetes however, theuseofmusclederivedstemcells pancreatic diseasesinanimalsandpatients; are beingstudiedfortheirefficacyatimproving the nativepancreas. Manysourcesofstemcells that couldbeplacedinaBiohub, whichmimics and hencecouldbeasourceofpancreaticcells anykindofcell potential tobecomevirtually havethe tremendous potentialbecausethey controller ofbloodsugarlevels. Stemcellshold ofthedigestivesystemandacritical vital part forathletes.and energy The pancreasisa and, ofcourse, forprovidingessentialnutrients Assistant Professor Ping Guo, Ph.D. diseases related Gene therapyandtissueengineeringfortreatingsports type Idiabetesmouse enveroment forchondrogenicdifferentiation hMDSCs tomimicinvivohighpressuremicro- and regenerationininfarctionmice livery ofmusclestemcellsforcardiacrepair hMDSCs andmMDSCspluscoacervatede- The digestivesystemiscriticalforhumanlife virus-infected pancreaticducts. GFP expressionspecificallyin AAV6-Sox9-GFP Technicians: Andrea Liebowiz, ElizabethMorris andsmallanimalsurgery. ogy biol- with experienceincellbiology/molecular and iscurrentlyseekingpostdoctoralfellows SurgeryinJune2015 ofOrthopedic Department Post-doctoral Fellow:Dr. PingGuojoinedthe LAB MEMBERS cal methods.2012, 183(2):139-46. nant adeno-associatedvirus. Journalofvirologi- Rapid andsimplifiedpurificationofrecombi- X, J,Wiersch ParedesJ, Tulachan S, GittesGK., Guo P, El-Gohary Y, PrasadanK, ShiotaC, Xiao 93(11):1241-53. creatic ducts. Laboratory investigation, 2013, recombinant viralinfusionintomousepan- and labelingofpancreaticductsbytargeted Gaffar I, EsniF, GittesGK., Specifictransduction Prasadan K, RymerC, ShiotaC, J,Wiersch Guo P, XiaoX, El-Gohary Y, Criscimanna A, (PNAS), 2014, 111(13):E1211-20. of SciencestheUnitedStates America SMAD7. ProceedingsoftheNational Academy beta-cell proliferationbyup-regulationof Shiota C, GittesGK., M2macrophagespromote S, PeirishL, SongZ, El-Gohary Y, PrasadanK, Xiao X, GaffarI, GuoP, J,Wiersch Fischbach IMMPACT REPORT • • • • 77 Embryonic myosin (green, showing new regener- Embryonic myosin (green, ated muscle fibers) and macrophage (F4/80, red) infiltration were detected in dystrophic muscle. Muscle derived stem cells are able to differentiate into myosin heavy chain positive myotubes (red) in vitro. LAB MEMBERS I started with the University my employment Houston in Health Science Center at Texas of and I am currently seeking to hire May 2015, one postdoc with one research technician and experience of cell biology. mice, for evaluate muscle and heart mice, investigate the cell- cell interaction be- To tween myogenic cells and non-myogenic cells from different transgenic mice for PO1 aging projects models, including mdx/mTR and dystrophic including mdx/mTR and models, dog for stem cell therapy for DMD Identification of therapeutic muscle derived stem cells for muscle derived of therapeutic Identification disease and accelerate aging treatment of muscular Aiping Lu, M.D. Aiping Lu, Assistant Professor • KEY PUBLICATIONS Aiping YingXueqin Arvydas Tang, Gao, Usas, Adam Schneppendahl, Johannes Tan, Jian Lu, James H Cummins, Wang, Bing Kozemchak, A comparison of Johnny Huard. Tuan, Rocky S bone regeneration with human mesenchymal stem cells and muscle-derived stem cells and 2014 Aug; Biomaterials, the critical role of BMP. 35(25):6859-70. Proto, Jonathan D. Arvydas Usas, Xueqin Gao, Alexander Proctor, James H Cummins, Aiping Lu, Role of and Johnny Huard. Chen Chien-Wen donor and host cells in muscle derived stem cell mediated bone repair: differentiation vs. 2014 Aug; Journal. The FASEB paracrine effects. 28(8):3792-809. Mu Sohn J, Proto JD, Y, Tang Poddar M, A, Lu Rapid Depletion Huard J. B, Wang Oyster N, X, of Muscle Progenitor Cells in Dystrophic mdx/ Hum Mol Genet. 2014 Sep utrophin-/- Mice. 15; 23(18): 4786-800. A, Usas Pollett JB, Thompson SD, Lavasani M, Huard Péault B, B, Sun Clark KA, Stolz DB, A, Lu Human muscle-derived stem/progenitor J. cells promote functional murine peripheral Apr 1; nerve regeneration. J Clin Invest. 2014 124(4):1745-56 Paraboisis between dystrophic mice and WT •Paraboisis between dystrophic mice and •Isolation and characterization of MDSPCs that had a tissue-specific deletion of the ERCC that had a tissue-specific deletion DNA damage gene and investigated if increased was sufficient to in the skeletal muscle niche also will use We induce MDSPC dysfunction. aging affects parabiosis to determine whether impacting stem cell func- the stem cell niche, tion via a nonautonomous mechanism. RESEARCH PROJECTS •Muscle stem cell depletion in different animal Our group focuses on identification of thera- The Role of Muscle Progenitor Cell Exhaustion Duchenne muscular dystrophy (DMD) patients analyze the cell non-autonomous effects To demonstrated that muscle have already We peutic muscle-derived stem cells for treatment of muscular disease and accelerate aging, including Duchenne Muscular Dystrophy (DMD) we have two major Currently, and progeria. research areas. in the Rapid Disease Progression in Dystrophic Mice. muscle lack dystrophin from birth; however, weakness only becomes apparent at 3-5 years with the which happens to coincide of age, depletion of the muscle progenitor cell (MPC) whether the progres- are investigating We pools. sion of muscular dystrophy is a consequence of are working We the decline in functional MPCs. believeWe that on different animal models. alleviating depletion could represent an MPC approach to delay the onset of the histopa- thologies associated with DMD patients. induced by aging and stress on MDSPCs using heterochronic parabiosis and tissue-specific PO1) (NIH, inactivation of ERCC1. derived stem/progenitor cells (MDSPCs) isolated from the skeletal muscle of naturally aged and progeroid ERCC1- deficient mice have a reduced ability to proliferate and differenti- and an impaired regenerative capacity ate, compared to MDSPCs isolated from young wild Although this dysfunction is type (WT) mice. consistent with the concept that the accumulation of damage directly to the stem/progenitor cells (cell autonomous effect) contributes to the loss of tissue regeneration and homeostasis a potential defect in the associated with aging, stem cell niche or circulating factors could also affect MDSPC function in a non-autonomous It remains unclear if aging-related loss manner. of adult stem cell function is primarily driven by increased DNA damage) cell autonomous (e.g. and/or non-autonomous mechanisms (aged In microenvironment or circulating factors). we utilized a mouse model the current study, TISSUE FOR TISSUE CENTER AGING AND ENGINEERING RESEARCH 78 • IMMPACT REPORT TISSUE ENGINEERING AND AGING RESEARCH ENGINEERING ANDAGING CENTER FORTISSUE by reducingfibrosis or applicationofstemcells. other factors(i.e., BMPs, Notch, and NF-κB). cord injuries, aswellsitscorrelationwith the hipsandlegs, andtraumaticbrainorspinal mediator ofHOcausedbysurgeryortrauma to as towhetherRhoAactivationcouldbea mediator.was foundasakey Questionremains affected dystrophicmice, andRhoAsignaling skeletal muscleandcardiacofseverely toboneosteoporosis. and itsrelevance etal muscle, cardiacmuscle, andbloodvessels, heterotopic ossification(calcification)inskel- aging andcellsenescence. regulatorsoftissue all beenshownasimportant of Rapamycin(mTOR) signalingpathwayshave tors: Notch, Wnt, RhoA, andmammalian Target dystrophin deficiency). stemcellregula- Thekey degenerative muscledisorderinboyscausedby and DuchenneMuscularDystrophy(DMD, a such asthatofprogeria(acceleratedaging) been observedinthediseasedskeletalmuscle, of regenerationpotentialstemcellshave regenerating muscles. toreducefibrosisandimprovefunctionof effort cell defectindiseasedandagedmuscles, inan lecular regulatorymechanismsofmusclestem following areas: topic ossification, osteosarcoma, andsoon). cancer cachexia-relatedmuscleatrophy, hetero- disorders (i.e., DuchenneMuscularDystrophy, and themechanismofvariousmusculoskeletal of stemcellsenescenceandprematureaging, especially interestedinstudyingthemechanism (osteosarcoma).cancer biology Currentlyweare muscular dystrophy, prematureaging, and ogy, wound regeneration, fibrosisprevention, and ourstudieshaveinvolvedstemcellbiol- Surgery, ofOrthopedic center intheDepartment ofDr.group isapart JohnnyHuard’s research 3). Improvementofsofttissuewound healing extensiveHOinthe reported I havepreviously 2). The cellularandmolecularmechanismof Accelerated exhaustion, senescence, and loss 1). Understandingofthecellularandmo- Currently Iamfocusingmyresearchinthe Our “Stem Cellin Aging andCancer” research heterotopic ossificationinthecardiacmuscles ofmusculardystrophicmice. Inactivation ofRhoA/ROCKsignalingreduced theintramyocardiallipidaccumulation, fibrosis, and •Role ofNotchand Wnt signalinginregulating •Improvement ofsofttissuewoundhealingby •The cellularandmolecularmechanismofhet- •Understanding ofthecellularandmolecular RESEARCH PROJECTS stem cells. application ofrelaxin, MMPs, andmultipotent regeneration ofdiseasedsofttissueswiththe fibrosis, Iwouldnextfindouthowtohelpthe muscle) isusuallyaccompaniedwithexcessive of diseasedskeletalmuscle(i.e., dystrophic muscle oramputateddigits. Sincethehealing during thehealingprocessofinjuredskeletal offibrosis relaxin andMMPsintheprevention theeffectsofhormone reported previously been observedinscarlesswoundhealing. Ihave (MMPs) totheirtissueinhibitors(TIMPs)has Assistant Professor Xiaodong Mu, Ph.D. tissues andprematureageddiseasemodels Stem cellsenescenceanddysfunctionindiseasedmuscle well asthecancerstemcells animal incontrasttonormalaginganimal, as stem cellsenescenceofacceleratedaging reducing fibrosisorapplicationofstemcells toboneosteoporosis and itsrelevance muscle, cardiacmuscle, andbloodvessels, erotopic ossification(calcification)inskeletal defect indiseasedandagedmuscles regulatory mechanismsofmusclestemcell A higherratioofmatrixmetalloproteinases Pathology, 2010;177:2399-2410. both youngandagedmice. AmericanJournal of lite cellmobilizationduringmusclehealingin regulates MMPexpressionandpromotessatel- Mu X, UrsoM, MurrayK, FuF, andLi Y. Relaxin dics andRelatedResearch. 2013, May. genes inosteosarcomacells.- ClinicalOrthopae Embryo Extractdemethylatestumorsuppressor Schott T, GrecoN, HuardJand Weiss K. Chick Mu X, SultankulovB, Agarwal R, Mahjoub A, Oncology. 2013, May. rine OsteosarcomaCells. FrontiersinPediatric And An Aggressive MetastaticPhenotype InMu- ALDHActivity Is AssociatedWith Signaling Notch Mu X, IsaacC, GrecoN, HuardJ, and Weiss K. phic muscleofmice. FASEB J.2013, May. function andheterotopicossificationindystro- Huard J. RhoAmediatesdefectivestemcell X,Usas A,Mu Tang Y,Lu A,B,K,and Wang Weiss Hum MolGenet. 2015Feb12. inmusculardystrophy.onset ofhistopathology muscle progenitorcelldepletionandtherapid Weiss K, HuardJ. The roleofNotchsignalingin X,Mu Tang Y,Lu A,K,Usas A, TakayamaB, Wang KEY PUBLICATIONS IMMPACT REPORT • • • • 79 Osterix serves as molecular switch for gene activation in osteoblasts (JBMR, Epub 2013 Sep 23) Sinha K, Yasuda H, Zhou X and deCrombrug- H, Yasuda Sinha K, Osterix and NO66 histone ghe B (2013). architecture demethylase control the chromatin osteoblast of Osterix target genes during Epub Research. J Bone Mineral differentiation. 2013 Sep 23. Krahe R, H, Deng J,Yasuda Sinha K, Chen Q, Behringer R and de Crombrugghe B (2015). Mesenchymal Deletion of Histone Demethylase J NO66 in Mice Promotes Bone Formation. Epub 2015 Feb 26. Research, Bone Mineral Jerman E, O’Connell D, Gambero G, Brien G, Wynne Jurgens M, Dunne E, Egan C, S, Turner Sinha K, Shi X, Ferguson N, A, Lohan Piao L, K, A (2012).The and Bracken, Cagney G, Loftus J, Polycomb PHF19 binds H3K36me3 and recruits PRC2 and the NO66 demethylase to hitherto active genes during embryonic stem cell dif- Nature Structure and Molecular ferentiation. 2012 Dec;19(12):1273-81. Biology, deCrombrugghe Zhang D, Zhou X, Wu M, Y, Tao Structural Insights Zang J (2013). Sinha K, B, into the Interactions of Histone Demethylase - Transcrip NO66 with Osterix to Repress Gene 288, of Biological Chemistry 2013, Journal tion. 16430-16437. sion and bone metastasis remodeling process using mouse model Osterix by lysine methylation in regulation of Osx activity during osteoblast differentiation and bone formation identify epigenetic signature marks of histone methylations and DNA methylation in the key osteoblast genes and to better study the multiple pathways/mechanisms involved in normal and malignant bone formation Histone demethylase NO66 has an oncogenic The fact that bone is the most susceptible Epigenetic control of bone formation and bone metastases formation and bone control of bone Epigenetic Krishna Sinha, Ph.D. Krishna Sinha, Assistant Professor The long-term goal of my research is to understand the role of epigenetic regulators and develop therapies for better bone health in age-related bone disorders. KEY PUBLICATIONS Dent SR and H,Coombes MM, Yasuda Sinha KM, Regulation of the osteoblast- de Crombrugghe B. a specific transcription factor Osterix by NO66, EMBO J, Jumonji family histone demethylase. e-Pub 11/2009. •Oncogenic function of NO66 in PCa progres- To study the in vivo role of NO66 in bone •To To study the post-translational modification of •To differentiating osteoblasts, these genes become differentiating osteoblasts, and are bound with Runx2, hypomethylated, other active histone modifiers during and Osx, then demonstrated with We their activation. in mesenchy- conditional inactivation of NO66 of NO66 in mice mal cells that loss of function leads to increased bone mass phenotype. role in PCa and bone metastasis. organ for metastases by nearly all types of skeletal cancer primarily by prostate and breast, metastases further lead to severe defects NO66 is in bone architectures during aging. also found that We up-regulated in lung cancer. NO66 levels are elevated in prostate cancer patient samples and xenografted samples. Our data indicates that NO66 overexpression promotes proliferation and invasion of prostate femurs In xenograft studies, cancer cells. of male SCID mice implanted with NO66- overexpressing PC3 cells have significant bone loss compared with mice with control PC3 cells, suggesting that NO66 plays an oncogenic role in PCa progression and bone metastasis. RESEARCH PROJECTS •Use of Next-Gen sequencing approach to My research interests are focused on epi- Epigenetic control of bone formation by Differentiation of osteoblasts commences Osterix acts as the molecular switch from during early embryonic showed that We genetic control of cell-fate determination with specifics on critical factors responsible for bone well as as formation and bone homeostasis, prostate cancer-induced skeletal metastases. Johnny Huard’s The current research plan in Dr. lab is to identify and study gene function in age- pivotal which provide related bone disorders, information in considering preventive measures and treating bone disorders for better bone health and life. Osterix and histone demethylase NO66. from mesenchymes with stage-specific gene which supports maturation and activation, ap- Using a proteomic function of osteoblasts. regulator NO66 I identified a chromatin proach, a JmjC- NO66 is as Osterix-interacting protein. domain histone demethylase specific for lysine is an Osterix (Osx) 4 and 36 of histone H3. essential transcription factor required for bone and tooth formation and controls activation of a repertoire of genes in osteoblast and osteo- NO66 inhibits Osterix activity through cytes. interactions with Osx and through demethyl- ation of histone at specific euchromatin regions we reported the crystal Later on, in osteoblasts. structure of NO66 and its interface required which provides useful for interactions with Osx, structural insights while designing or developing NO66 interacts with inhibitors against NO66. EZH2 (enhancer of zeste homolog 2) containing which PRC2 (polycomb-repressive complex 2), has histone K27 methyl transferase activity and inhibits gene activation during ES cell differentiation. gene repression to gene activation. the promoters stages of mouse development, of Osx, Col1a1 and Bsp are hypermethylated in mesenchyme cells and are also bound with methylation-induced repressors complex which prevents gene activation including NO66, In in those cells as well as in Osx-null cell. TISSUE FOR TISSUE CENTER AGING AND ENGINEERING RESEARCH 80 • IMMPACT REPORT T TEXAS THERAPEUTICS INSTITUTE TEXAS THERAPEUTICS include: 1)signaling mechanismsofreceptors and from CPRITin2015. four majorgrantstotalingmore than $7million that noteworthy investigators wereTTI awarded and Research Institute of Texas It (CPRIT). is Institutes ofHealth, and theCancerPrevention & Johnson, Merck andPanaMab, theNational the biotechnologyindustry, includingJohnson significant fundingfrom thepharmaceuticaland development. investigators haveTTI brought in in theareas ofcancerbiologyandbiologicsdrug center andmadesignificantscientificdiscoveries into acomprehensive discovery academicdrug therapeutics. and establishmentofproof-of-principle for targets, identification and validation ofdrug Research conductedatthecenter focusesonthe therapeutic agentsanddiagnostictools. development, andcommercialization of Science CenteratHouston forthediscovery, System, andThe University of Texas Health Technology Fund, The University of Texas in 2010withfundingfrom the Texas Emerging Current research activities at TTI-IMM During thepastfive years,TTI hasgrown Medicine wasestablished (TTI-IMM) Foundation Institute ofMolecular exas Therapeutics Institute atThe Brown Chemistry A. Robert Welch Distinguished University Chairin Professor &Center Director Zhiqiang An, Ph.D. discoveriesTTI intolifesavingmedicines. significant venture capitalfundingtotranslate two UTHealth spinoffbiotechcompanieswith discovery center,drug facultyhave founded TTI other Texas-based institutions.Asanacademic from theIMM, Texas Medical Center, and collaborative projectssupport withscientists support internalprojects, butthey also TTI discoveryPlatform. platformsnotonly Thedrug Products andSmall Molecular Drug Discovery Development Platform and2)theNatural Monoclonal AntibodyLeadOptimization and discovery platforms:1)theTherapeutic drug research programs, isbuildingtwomajor TTI to experimentalvaccines. antibodies from animalsandhumansinresponse discovery; and3)characterizationof for drug activity ofthesetargetsaspotentialleadmolecules synthetic smallmoleculesthatmodulatethe 2) discovery ofbiologics,naturalproducts, and enzymes thathave criticalroles inhumandiseases; In additiontothebasicandtranslational IMMPACT REPORT • • • • 81 Trastuzumab-mediated phagocytosis of cancer Trastuzumab-mediated (Journal of Immunology, cells by macrophages. 194:4379-4386). 2015, Differential intensity of tumor associated macrophage (CD68+) infiltration in breast tumor 2015; doi: tissues (Clinical Cancer Research, 10.1158/1078-0432.CCR-15-1057). Yun Shi, Xuejun Fan, Hui Deng, Randall Brezski, Randall Brezski, Hui Deng, Xuejun Fan, Shi, Yun William Strohl, Robert Jordan, Michael Rycyzyn, An. Ningyan Zhang and Zhiqiang Quanming Zou, killing triggers phagocytic Trastuzumab 2015. interaction with of high HER2 cancer cells by of Immunology Journal FcγRIV on immune cells. 194:4379-4386. Pooja Hui Deng, Brezski, Randall J. Xuejun Fan, Songlin Anneliese Gonzalez, Shi, Yun Dhupkar, Dr. Strohl, William R. Michael Rycyzyn, Zhang, and Zhiqiang Ningyan Zhang, Jordan, Robert E. A novel therapeutic strategy to rescue 2015. An. the immune effector function of proteolytically- inactivated cancer therapeutic antibodies. Molecular 14(3): 681-691. Cancer Therapeutics LAB MEMBERS Weixu Salameh, Ahmad S. Post-doctoral Fellows: Xun (Mark) Gui, Leike (Simon) Li, (Ella) Meng, Yue Qun Schultz, Robbie D. Hao Ching Hsiao, Li (jointly with Dr. Yan Bills), (jointly with Dr. Bills) Li Chen (jointly with Dr. Bills), Student: Jingnan An ACS Chemical ACS ment natural products drug discovery responses in preclinical animal models and humans development of HER3 targeting monoclonal antibodies for cancer therapy Therapeutic monoclonal antibody drug Therapeutic monoclonal antibody Apr 23. [Epub ahead of print]. Biology Apr 23. KEY PUBLICATIONS Anneliese Xuejun Fan, Hui Deng, Ningyan Zhang, Randall J Brezski, Songlin Zhang, Gonzalez, William R Michael Rycyzyn, Byung-Kwon Choi, 2015. An, and Zhiqiang Jordan, Robert E. Strohl, Dysfunctional antibodies in the tumor microenvironment associate with impaired anticancer (Published Clinical Cancer Research immunity. 2015; doi: 10.1158/1078- OnlineFirst July 29, 0432.CCR-15-1057). Xuejun Fan, Xiong, Wei Leike Li, Meng, Weixu Aimin Zhifeng Chen, Andrew J Bett, Hui Deng, Daniel C. Joyce, Joseph G. Cox, Kara S. Tang, Danilo R. Fu, Tong-Ming Thoryk, Elizabeth Freed, Zhiqiang Vora*, Kalpit Ningyan Zhang, Casimiro, Efficient generation of monoclonal 2015. An*. antibodies from single rhesus macaque anti- mAbs 7(4): 707—718. body secreting cells. Xing-Zhong Liu, Yue, Qun Li; Li Chen, Yan Genetic 2015. and Gerald Bills. An, Zhiqiang Manipulation of the Pneumocandin Biosynthetic Analogues and Evalu- Pathway for Generation of ation Activity. of Their Antifungal •Therapeutic antibody discovery and develop- •Biocombinatorial chemistry approach for •Evaluation of vaccine-induced antibody discovery platform. Supported by a grant from discovery platform. Fund and as part Technology Emerging Texas the our group Therapeutics Institute, Texas of the has been building a comprehensive antibody drug discovery platform with a focus on such antibody lead optimization technologies, deep sequencing as antibody phage display, of antibody encoding genes from individual maturation affinity antibody expressing B cells, and humanization. RESEARCH PROJECTS •HER3 mediated cell signaling and the with improved oral availability or broader with improved oral availability spectrum of antifungal activities. Discovery and development and of therapeutic antibodies antibiotics Zhiqiang An, An, Ph.D. Zhiqiang Therapeutics Institute Texas Co-Director of the Professor and Chair in Chemistry Distinguished University Welch A. Robert , a lipohexapeptide produced , 0 Our group focuses on the discovery and HER3 mediated cell signaling and HER3 Antibodies response to experimental HIV, Pnemocandin biosynthesis and biocom- by a fungus. In collaboration with Dr. Gerald In collaboration with Dr. by a fungus. we are studying the pneumocandin group, Bills’ biosynthesis pathway using a combination of and chemical approaches. genetic, genomic, Elucidation of the pneumocandin biosynthetic pathway will pave the way for designing experimental procedures to engineering analogues development of therapeutic antibodies and including antibiotics against human diseases, we Currently, cancer and infectious diseases. have four major research areas. Ablated targeting antibodies for cancer therapy. regulation in the HER/ErbB family receptor signaling has been implicated in various Agents targeting EGFR and HER2 cancer types. exhibited clinical benefits for the treatment is but drug resistance of some cancer types, of the drug Current understanding widespread. resistance mechanisms is limited and HER3 has been implicated in the resistance to current is working Our group EGFR and HER2 therapies. on: 1) HER3 mediated cell signaling; 2) the role HER3 plays in resistance to current anti-HER2 and EGFR antibody therapies; and 3) generation of HER3 targeting antibodies and their mode of actions. Design of highly im- and CMV vaccines. dengue, munogenic peptide and protein based vaccines that induce neutralizing antibodies against a broad range of clinical isolates is one of the approaches in developing effective HIV and have an ongoing project to We dengue vaccines. aid the design of HIV and dengue vaccines by profiling antibody response to the experimental also have a project in We vaccines in rhesus. the discovery and development of neutralizing antibodies against the human cytomegalovirus (HCMV). binatorial chemistry approach for natural The antifungal therapy products drug discovery. caspofungin is a semi-synthetic derivative of pneumocandin B TEXAS THERAPEUTICS THERAPEUTICS TEXAS INSTITUTE 82 • IMMPACT REPORT candin B echinocandin antifungals, includingpneumo- biosynthetic pathwaysresponsibleforthefamily human diseases. Forexample, wecharacterized forpharmaceuticalintervention in lies relevant model organisms, especiallybiosyntheticfami- of microorganismsusingfilamentousfungias predict geneticallyencodedchemicaldiversity drug discovery. changing theapproachabilityofnaturalproduct availability ofnoveldiseasetargetsarerapidly cal massspectrometry(MS)andNMR, andthe products viabioengineering, improvedanalyti- strategies, engineeringunnaturalnatural metagenomic approaches, improvedculturing such asmeasuringmicrobialdiversityfrom andotherconvergingtechnologies,This view throughgenomic mining.have beenrevealed naturalproduct that natural productsandnew of pathwaysmanyhistoricallyimportant ity hasledtothefunctionalcharacterization imagined.previously Experimentaltractabil- diverse andexperimentallytractablethan product biosyntheticpathwaysarevastlymore thatnatural view genomics haveledtoanew TEXAS THERAPEUTICS INSTITUTE TEXAS THERAPEUTICS pathways, andinotherhumantherapies. Texas modulation ofhuman molecularsignalling appropriate forintervention incancerbiology, chemical collectionfocusedonmetabolites are intheearlystagesofbuildingamicrobial using filamentousfungiasmodelorganisms. We regulating unexpressedbiosyntheticpathways physiological methodsforexpressingand un- products withimprovedtherapeuticproperties. from thesepathwaystoproducehybridnatural pathways willenableustorecombinegenes costs. Access tothegenesofechinocandin kinetics, whilereducingfermentation production broaden thespectrumandimprovepharmaco- purity, increasedpotency. We ultimatelywishto analoguesthathaveimprovedproduct new pneumocandin biosynthesistoproduce fungal drugCANCIDAS. We haveprogrammed Our labemploysgenomicstointerpretand Technological breakthroughsinmicrobial We are developing new geneticand new We aredeveloping 0 , moleculefortheanti- thestarting Kay and Ben Fortson DistinguishedChairinNeurodegenerativeDiseaseResearch Kay andBenFortson Professor Gerald Bills, Ph.D. metabolites forinfectiousdiseasesandcancertherapies Genome mining, biosynthesisanddiscoveryofmicrobial (Fungi, Ascomycota,Leotiomycetes). metabolic geneclustersinGlarealozoyensis Liu. 2015. Functionaloperonsinsecondary Xiang, S. Lia, Y. Che, C. Wang, X. Niu, Z. An &X. Yue, Q., L. Chen, Y. Li, G.F. Bills, X. Zhang, M. Spotlight Article). (Editor’s fungal metabolites.EukaryoticCell14:698-718. diversity intheechinocandinlipopeptideanti- Z. An &G.F. Bills2015. Evolutionofchemical Yue, Q., L. Chen, X. Zhang, K. Li, J. Sun, X. Liu, KEY PUBLICATIONS ofanaturalproducts •Development ‘chemical ofmethodsforreprogramming •Development •Understanding howechinocandinfungiresist •Biosynthesis andpathwayengineeringof RESEARCH PROJECTS diseases. andforinterventioninhuman in cellbiology chemicalsasprobes centers resultinginnew will bepromotedamong Texas-based screening microbial resourcesavailablefrom Texas and Therefore, ourcollectionwillemphasizethevast is theUSA’s secondmostbiodiversestate. (Interviewed for (Interviewed Podcast) ACS ChemicalBiology activity. ACS 10:1702–1710. ChemicalBiology oftheirantifungal analogues andevaluation candin biosyntheticpathwayforgeneration of 2015. Geneticmanipulationofthepneumo- Li, Y., L. Chen, Q. Yue, X. Liu, Z. An &G. F. Bills. species. PLoSOne 10:e0125221. Pseudomonas HIF pathwayproducedbyanew peptide MDN-0066, anovel inhibitorof VHL/ Vicente. 2015. Identificationofthelipodepsi- I. González, G. Bills, F. Reyes, O. Genilloud&F. da Sousa, J. Martín, I. Pérez Victoria, F. Asensio, Cautain, B., N. dePedro, C. Schulz, J. Pascual, T. 6:e00703-15. elsewhere investigators withintheUTSystem, Texas, and resource platform’ fordrugdiscoveryother human diseases naturalproductsuseful totreat discover new transcription ofbiosyntheticgenesfungito their ownantifungalmetabolites antifungals the echinocandinlipopeptidesforimproved mBio pneumocandins A Candida albicans, thantheparent compounds, more potentagainstthehumanpathogen, pneumocandinsFandGwere eight-fold New candins byinactivatingdifferentpathwaygenes. derivativesofthepneumo- We havemadenew biology, Chinese Academy ofSciences). Li Chen(allvisitingfromtheInstituteofMicro- Research Associates: Dr. Qun Yue, Dr. Yan Li, Dr. LAB MEMBERS 16:549–565. (featuredcoverarticle). mutualism. CurrentOpinioninMicrobiology underexplored arenaofmicrobialdefensive fungi: Antibiotic discoveryandfunctionsinan Bills, G.F., J.B. Gloer&Z. An. 2013. Coprophilous 0 andB 0 . IMMPACT REPORT • • • • 83 Anti-LGR5-ADC treatment significantly decreases colon cancer cell survival (left) and has no effect in cells manipulated to no longer express antibody Unconjugated anti-LGR5 LGR5 (right). has no effect on cell survival. The anti-LGR5-ADC is highly specific and forms a complex with the LGR5 receptor at the cell The LGR5-ADC surface of colon cancer cells. complex (red) is then internalized and transported to the lysosome compartment (LAMP1, green) where the drug can be released to yellow). cells (co-localization, the cancer destroy Proc Natl Acad Sci U S Proc Natl cancer and drug resistance conjugates KEY PUBLICATIONS KEY PUBLICATIONS Liu Q. A., Thomas Yi J., Gong X., Carmon K.S., (2014) RSPO-LGR4 functions via IQGAP1 to potentiate Wnt signaling. 111(13): E1221-9. A. Liu, A. Thomas, X., Gong, Q., Lin, K.S.*, Carmon, (2012) LGR5 Interacts and Co-Internalizes Q*. Wnt/beta- Wnt Receptors to Modulate with Mol Cell Biol. 32(11): 2054- catenin Signaling. 2064. (*Corresponding Authors) Liu, A. Thomas, Q., Lin, X., Gong, K.S., Carmon, (2011) R-spondins function as ligands of the Q. orphan receptors LGR4 and LGR5 to regulate Acad Sci Proc Natl Wnt/beta-catenin signaling. 108 (28): 11452-7. USA. (2010) Develop- D.S. and Loose, K.S., Carmon, Wnt Binding ment of a Bioassay for Detection of Anal Affinities for Individual Frizzled Receptors. 401(2): 288-94. Biochem. (2008) SFRP4 D.S. and Loose, K.S., Carmon, and Wnt7a Signaling Pathways Two Regulates Inhibits Proliferation in Endometrial Cancer Mol Cancer Res. 6 (6): 1017-1028. Cells. •Investigation of LGR5 signaling and its role in Development conjugates for targeting cancer of antibody-drug stem cells A series ADCs into CSCs. and specific delivery of ADCs are being generated using of anti-LGR5 and distinct functionally different antibodies Using the toxins. chemical linkers to incorporate we are testing the cutting-edge techniques, ADCs to precisely bind and destroy ability of the LGR5-positive cancer cells and evaluating their anti-tumor effects in xenograft tumor mouse we are further investigating Additionally, models. the signaling mechanism(s) of LGR5 and its role This research in cancer and chemoresistance. will provide preclinical proof-of-concept for the feasibility of the future development of A CSC-targeted ADC anti-LGR5 ADCs. could be a breakthrough treatment to eradicate residual and more importantly, tumors and metastasis, prolong overall quality of life and survival for a large number of cancer patients. RESEARCH PROJECTS •Development of anti-LGR5 antibody-drug Kendra S. Carmon, Ph.D. Carmon, Kendra S. Assistant Professor Emerging evidence has shown that within G The LGR5 (Leucine-rich repeat-containing, My research is focused on the develop- several malignant tumors types there different exists a subpopulation of cancer cells that These cancer behave like normal stem cells. cells, or tumor-initiating stem-like cells (CSCs), can renew and sustain the cancer, themselves much like normal stem cells repopulate and can drive CSCs maintain our organs and tissues. and resistance to metastasis, tumor growth, often not Since CSCs are anti-cancer therapies. entirely eliminated by conventional treatments, they the tumor and potentially can regenerate patient leading to a decline in metastasize, Thus it is essential quality of life and survival. to developa new generation of novel therapies that can ultimately target and destroy CSCs. protein-coupled receptor 5) receptor is a bona fide marker of normal adult stem cells in the intestine and multiple other epithelial tissues. (Jim) Qingyun’s As a postdoctoral trainee in Dr. I discovered that LGR5 func- laboratory, Liu’s tions as a receptor of the secreted growth fac- a Wnt signaling, tors R-spondins to potentiate key regulatory pathway in stem cell survival and numerous reports Since then, tumorigenesis. have shown that LGR5 expression is significantly including elevated in several tumor types, major and ovarian carcinomas. gastric, liver, colon, Recent evidence has further demonstrated that human cancer cells with high levels of fueling tumor growth, LGR5 behave like CSCs, These findings and drug resistance. metastasis, suggest the potential of LGR5 as a promising new target for the development of CSC-based therapies. ment of innovative anti-LGR5 antibody-drug “seek and destroy” conjugates (ADCs) that will similar to LGR5-expressing tumors and CSCs, ADCs will incorporate These guided missiles. a chemical toxin that is only released once it enters target cells with high levels of LGR5. My previous work has shown that LGR5 is continuously and rapidly internalized into the making it an exceptional transit for fast cell, TEXAS THERAPEUTICS THERAPEUTICS TEXAS INSTITUTE 84 • IMMPACT REPORT TEXAS THERAPEUTICS INSTITUTE TEXAS THERAPEUTICS pain andinfections. diseases includingcancer, diabetes, chronic exploring theirpotentialusesinvarioushuman specific EPAC inhibitorsandagonistsin secondgeneration isoform in developing rickettsioses. Currently, weareactivelyengaged andtreatmentoffatal tors intheprevention have identifiedapotentialuseofEPAC inhibi- signalingmolecules.of important Recently, we ofthisfamily functions anddiseasesrelevance mouse modelstostudythephysiological EPAC selectiveinhibitorsandEPAC knockout ment ofhumandiseases. and controlledpharmaceuticallyforthetreat- ecules sothattheirfunctionscanbeexploited signalingmol- modulators fortheseimportant EPAC proteinsandtodesignpathwayspecific goals aretounravelthesignalingintricaciesof proteins directlyactivatedbycAMP(EPAC). Our function ofafamilycAMPsensors:exchange cal biology, tounderstand thestructureand andchemi- withpharmacology and cellbiology approaches, couplingbiochemistry, biophysics of humandiseases. We applymultidisciplinary forthedevelopment jor stresssignalimportant associated withsecondmessengercAMP, ama- Our laboratory has developed first-in-class Our laboratoryhasdeveloped Our laboratorystudiesintracellularsignaling Walter andMaryMischerDistinguishedProfessorinMolecularMedicine Professor Xiaodong Cheng, Ph.D. Wei, J., Patrikeev, I., Hao, D., Stutz, S. J., Dineley, Yan, J., Mei, F. C., Cheng, H. Q., Lao, D. H., Hu, Y., .Molecular Pharmacology 83:122-128, 2013. pancreatic cancercellmigrationandinvasion. X. A novelEPAC-specific inhibitorsuppresses Zhou, J., Sastry, S. K., Schwede, F., andCheng, Almahariq, M., Tsalkova, T., Mei, F. C., Chen, H., 110:19615-19620, 2013. fatal rickettsioses. Proc. Acad. Natl. Sci. USA. directly activatedbycAMPplayscriticalrolein Walker, D. H., andCheng, X.*Exchangeprotein G., Boor, P, J., Bouyer, R., Popov, V., Chen, J., Chang, Q., Hu, Y., Wakamiya, M., Ksiazek, T. Gong, B.*, Shelite, T., Mei, F., Ha, T., Xu, G., 25:60-71, 2014. Trends andMetabolism. inEndocrinology Sensor EPAC Homeostasis. ProteinsandEnergy Almahariq, M., Mei, F., andCheng, X. cAMP and Therapeutics. Mammalian rapgef3Gene:Structure, Function Directly Activated bycAMPEncodedthe Banerjee U. andCheng, X. ExchangeProtein 87:142-149, 2015. Metastasis invivo. MolecularPharmacology. Knockdown ofEPAC1 ReducePancreaticCancer X. PharmacologicalInhibitionandGenetic M. R., Patrikeev, I., Motamedi, M., andCheng, Almahariq, M., Chao, C., Mei, F. C., Hellmich, KEY PUBLICATIONS •Examine therolesofEPAC proteinsinmajor ofnextgeneration isoform •Development ofnoveldrug •Preclinical development ofinvivochemicalprobes •Development • RESEARCH PROJECTS cAMP -mediatedcellsignalinganddrugdiscovery out mousemodels pain, diabetesandobesity, usingEPAC knock- human diseases, suchascancer, chronic with NIHChemicalGenomicsCenter(NCGC) specific EPAC modulators, incollaboration bacteria Rickettsia, fundedbyNIH of microbialinfectionscausedbytick-borne candidates targetingEPAC forthetreatment cancer metastasis, fundedbyNIH targeting EPAC forsuppressingpancreatic (EPAC), fundedbyNIH exchange proteinsdirectlyactivatedbycAMP Structural andfunctionalanalysesofthe Gene. 570:157-167, 2015. protects micefromhigh-fatdietinducedobesity Loss ofEPAC1 increasesleptinsensitivityand spectrometry byamide hydrogenexchangemass revealed Molecular mechanism ofEPAC activation Student: YaohuaHu Robichaux Wang, William Post-doctoral Fellows:UpasanaBanerjee, Hui Research Scientist: Zhu Yingmin Research FangMei Assistant Professor: LAB MEMBERS .Molecular CellularBiology 33:918-926, 2013. protein directlyactivatedbycAMPisoform1. cose homeostasisinmicelackingofexchange sensitivity, reducedadiposityandimprovedglu- K. A., Chen, J.*, andCheng, X*. Enhancedleptin K. T., Motamedi, M., Hommel, J. D., Cunningham, IMMPACT REPORT • • • • 85 Baldwin A, Li W, Grace M, Harlow E, Münger E, Harlow Grace M, W, Li A, Baldwin Genetic Interac- II. K and Grueneberg DA. Alterations in Kinase tion Screens Identify E7 Expression Requirements Following HPV16 Proceedings of the National in Cancer Cells. 2008 Oct Academy of Sciences USA (PNAS). 28;105(43):16478-83. W*, Li Pearlberg J*, Degot S*, Grueneberg DA*, Doench J, W, Endege A*, Baldwin Davies JE*, Harlow Munger K, Xian J, Boyce F, Y, Hu Sawyer J, Kinase requirements across Comparing I. E. Proceedings of the National Cell types. Various 2008 Oct Academy of Sciences USA (PNAS). contrib- *these authors 28;105(43):16472-7. uted equally (co-first author) LAB MEMBERS Dayong Zhang, Yan Post-doctoral Fellows: Zhi Li, Zheng Student: Mohit Hulsurkar Ph.D. We discovered that human kinase GRK3 pro- discovered We motes prostate primary tumor growth and lung metastasis in mouse xenografts. 2015 Jun; GRK3 mechanism in aggressive therapy- GRK3 mechanism in aggressive cancers induced neuroendocrine prostate New regulators for EMT and their mechanisms in cancer progression Development on of precision medicine based genetic profiles and drug sensitivities of patient samples and patient-derived-xenografts (PDXs) growing in mice 150:33-46. *corresponding author 150:33-46. CDKL2 W*. Li Du G, Chang JT, Liu C, Li L, promotes epithelial-mesenchymal transition Oncotarget and breast cancer progression. *corresponding 2014 Nov 15; 5(21):10840-53. author Ai Wang Vrbanac V, S, Bhattacharya Li W*, N, N, Gravdal Boyce FM, Y, Hu Signoretti S, Collins M, Harlow Akslen LA, Nalwoga H, Halvorsen OJ, K, GRK3 is essential for metastatic RS. Watnick E*, cells and promotes prostate tumor progression. of Sci- Academy Proceedings of the National ences USA (PNAS) 2014 Jan 28;111(4):1521-6. *corresponding author E. Davies JE and Harlow, W*, Li Grueneberg DA*, kinase requirements shRNA screens identify IV. in human cells: differential kinase requirements in cervical and renal human tumor cell lines. Academy of Scienc- Proceedings of the National 2008 Oct 28;105(43):16490-5. es USA (PNAS). *these authors contributed equally (co-first author) Molecular mechanisms of cancer metastasis mechanisms of Molecular RESEARCH PROJECTS • • • KEY PUBLICATIONS Epithelial-mesenchymal transition in W*. Li Li L, human cancer:reprogramming comprehensive - epigenetics and differentia of metabolism, Pharmacology tion. & Therapeutics Wenliang Li, Ph.D. Li, Wenliang Assistant Professor My research is to study novel molecular to the spread of tumor Cancer metastasis, a Epithelial-mesenchymal transition (EMT), mechanisms of cancer metastasis with the goal of identifying new biomarkers and drug targets for the development of better therapeutics for human cancers. responsible for is body, other parts of patient’s it is still However, over 90% of cancer death. poorly understood and the current approaches to prevent treat human metastatic diseases or Through genomics, are mostly unsuccessful. lab my RNAi and cDNA functional screens, has identified several critical but previously unknown regulators for cancer metastasis. Signaling pathways and molecular mechanisms of these genes are under investigation with genomic, biochemical, cellular, molecular, and mouse models. proteomic approaches, These studies will yield new insights for cancer metastasis and may facilitate the development of new and biomarkers. therapeutics is believeddevelopmental to play a key process, organ drug resistance, role in cancer metastasis, Another and stem cell phenotypes. fibrosis, exciting research program in the lab is involved in identifying and studying human kinases as Kinases play central novel regulators for EMT. roles in many aspects of signaling transduction, They are also and diseases. cell physiology, one of the most important gene families for Our literature search cancer drug development. indicated that the majority of >700 kinases Our in human genome are still poorly studied. lab is employing unbiased functional screens against hundreds of human kinases to identify novel regulators for EMT and linking them to stem cell phenotypes and cancer metastasis. Investigation of the molecular mechanisms of these kinases will have a significant impact in expanding our knowledge in the crossroad of ex- such as development, citing and critical areas, and metastasis. drug resistance, stem cell, These kinases may also become new biomarkers and cancer drug targets for the development of novel therapeutics for human cancer. TEXAS THERAPEUTICS THERAPEUTICS TEXAS INSTITUTE 86 • IMMPACT REPORT TEXAS THERAPEUTICS INSTITUTE TEXAS THERAPEUTICS for colonandlungcancers. ing theRSPO-LGRsystemaspotentialtreatment identifying andcharacterizingdrugleads target- carcinomas. arefocusedon Ourcurrentefforts major roleintheaggressivenessoflungadeno- recently, weuncoveredthatRSPO3-LGR4hasa to regulatecellgrowthandmigration. Most elucidated howRSPOsandLGRsworktogether survival andgrowthofstemcells. We havenow R-spondins (RSPOs)thatareessentialforthe receptors ofagroupstemcellfactorscalled ously, wediscoveredthatLGR4-6functionas of normalstemcellsandtumorcells.- Previ (LGR4-6) thatplaycriticalrolesinthesurvival surface receptorscalledLGR4, LGR5, andLGR6 function andmechanismsofagroupcell cancer treatment. therapeutics forregenerativemedicineand ofnovel to thediscoveryanddevelopment cancer stemcellswillprovidecrucialknowledge anddifferentiationof normaland self-renewal the mechanismsthatgoverncontrolof profile andregulatorysystems. Understandingof similarity tonormalstemcellsinmolecular cells ortumor-initiatingcells, oftenbeargreat cancercells,renewing designatedcancerstem that makeupthebulkoftumor. These self- andprovidedaughter cells cells canself-renew heterogeneous, in which only a subpopulation of indefinitely. Furthermore, tumortissuesarealso areprogrammedtodivide of cancerasthey tobethecells-of-origin for manytypes believed repair afterinjury. However, thesecellsarealso rate, suchasthegutandskin, andfortissue the maintenanceoftissueswithhighturnover the stemcellsreside. areessentialfor They types ofdifferentiatedcellsinthetissuewhere andgiverisetoalltheother can self-renew Our researchisfocusedondelineatingthe Adult stemcellsarespecializedthat roles intheregulationofcellmigration. LGR4 isco-localizedwithIQGAP1attheleadingedgeofcellmigration. LGR4andIQGAP1playcritical Janice DavisGordonChairforBowelCancerResearch Professor andCo-Directorofthe Texas Therapeutics Institute (Jim)Liu,Qingyun Ph.D. of cancertherapeutics Investigation ofnormalandcancerstemcellsforthediscovery mor Suppressor. PLoSOne7:e37137-e37146. R-Spondins andPotentiallyFunctionsasa Tu- Liu Q(2012). LGR6IsaHigh Affinity Receptorof Gong X, CarmonKC, LinQ, Thomas A, J,Yi and Signaling. MolCellBiol32:2054-2064. Modulate Wnt/beta-Catenin Receptors To Wnt (2012). LGR5InteractsandCointernalizeswith Carmon KC, LinQ, GongX, Thomas A, andLiuQ KEY PUBLICATIONS •Identification ofleadmoleculestargeting •Investigation oftherolesaberrantexpres- •Determination ofthefunctionandmechanism •Delineation ofsignalingmechanismsstem RESEARCH PROJECTS therapeutics the RSPO-LGRsystemasnovelanticancer metastasis oflungandcoloncancer sion oftheRSPOsincontroltumor cancer cellgrowth of thereceptorsincontrolnormaland cell receptors Technician: AnthonyThomas Post-doctoral Fellow:Jing Yi XingGong Instructor: LAB MEMBERS E1221-E1229 (2014). Wnt signaling. ProcNatl Acad SciUS A. 111: RSPO-LGR4 functionsviaIQGAP1topotentiate Carmon, K. S., Gong, X., Yi, J., Thomas, A., Liu, Q. 8:e78144-e7850 (2013). tion inhumanandmousetissues. PloSOne, and Liu, Q. Analysis ofLGR4ReceptorDistribu- Yi, J., Wei, X., Gong, X., Bellister, S., Ellis, L.M., U S A, 108:11452-11457(2011). Wnt/beta-catenin signaling. ProcNatl Acad Sci orphan receptorsLGR4andLGR5toregulate Liu, Q. R-spondinsfunctionasligandsofthe Carmon, K.S., Gong, X, Lin, Q., Thomas, A., and IMMPACT REPORT • • • • 87 Scheme of selective killing of pathogenic bacteria. Shibata, T.; Tsuchikama, K. Recent advances in K. Tsuchikama, T.; Shibata, Org.enantioselective [2+2+2] cycloaddition. 1317-1323. 6, Chem. 2008, Biomol. Highly T. Shibata, Y.; K.; Kuwata, Tsuchikama, of a chiral Spiro- Enantioselective Construction cyclic Structure by the [2 + 2 + 2] Cycloaddition of Diynes and exo-Methylene Cyclic Compounds. 13686-13687. 128, Soc. 2006, Chem. Am. J. almost Ir-catalyzed K. Tsuchikama, T.; Shibata, perfect enantioselective synthesis of helical polyaryls based on an axially-chiral sequence. 6017-6019. Commun. 2005, Chem. LAB MEMBERS Ph.D., Post-doctoral Fellows: Upasana Banejee, Ph.D. Shimamoto, Yasuhiro TTI in July 2014 and Tsuchikama joined Dr. is currently seeking postdoctoral fellows with experience in organic chemistry/medicinal chemistry. Scheme of target identification using antibacterial metal probes. 2015, 2015, J. Org. Org. Lett. Lett. Org. based on antimicrobial metal complexes using untraditional chemical agents KEY PUBLICATIONS A., C. Lowery, K., Tsuchikama, C., K. Collins, AI-2-mediated Dissecting D. K. Janda, J., Zhu, quorum sensing through C5-analogue synthesis and biochemical analysis. Tetrahedron •Proteomic profiling using chemical probes in press. Modulat- D. K. K.; Janda, Tsuchikama, X.; Cai, Through Hapten Potency Vaccine ing Cocaine 135, Soc. 2013, Chem. Am. J. Fluorination. 2971-2974. A.; Kaufmann, C. J.; Lowery, K.; Zhu, Tsuchikama, A Potent Class C4-Alkoxy-HPD: D. K. Janda, F.; G. of Synthetic Modulators Surpassing Nature in 2012, Soc. Chem. Am. J. AI-2 Quorum Sensing. 13562-13564. 134, D. K. A.; Janda, C. K.; Lowery, Tsuchikama, Autoinducer-2 Based Quorum Sensing: Probing The Biological Consequences of Molecules Traverse Equilibrium Unable To States. Selected as a 6981-7294. 76, 2011, Chem. Article and featured on the front cover Featured Issue 17. 76, Volume of K.; Endo, Y.; K.; Hashimoto, Tsuchikama, Selective Synthesis Iridium-Catalyzed T. Shibata, of 4-Substituted Benzofurans and Indoles via Catal. Synth. Adv. Directed Cyclodehydration. 2850-2854. 351, 2010, K.; Shi- M.; Endo, K.; Kasagawa, Tsuchikama, “Cationic Ir(I)-Catalyzed sp3 C-H Bond T. bata, Alkenylation of Amides with Alkynes”, 1821-1823. 11, 2009, Chemical probes based on such antimicrobial Chemical probes based on such us to identify their metal complexes will enable novel insights protein targets and thus provide and drug into pharmacological mechanisms design for developing innovative antimicrobial we Throughout these projects, therapeutics. hope to drive our efforts toward innovative medical cures to save people suffering from serious diseases. RESEARCH PROJECTS •Selective killing of drug-resistant bacteria Development of chemical agents, tools, and strategies for com- and strategies tools, Development chemical agents, of diseases bating infectious Kyoji Tsuchikama, Ph.D. Kyoji Tsuchikama, Assistant Professor Antibiotics are powerful agents for the my lab is With this background in mind, TEXAS THERAPEUTICS THERAPEUTICS TEXAS INSTITUTE However, treatment of infectious diseases. their strong pharmacological effect poses evolutionary pressure on pathogenic microbes, leading to the development of drug resistance. The emergence of drug-resistant bacteria such as methicillin-resistant Staphylococcus aureus (MRSA) and multidrug-resistant Pseudomonas aeruginosa is an increasingly serious problem According to the report from for human health. the Centers for Disease Control and Prevention antibiotic resistance in the 2011), April (CDC, United States costs us approximately twenty billion dollars per year in excess health care costs and more than eight million additional days alternative antimi- Therefore, spent in hospital. crobial strategies based on a novel mechanism of action has been pursued to overcome this clinical challenge. working on two research projects by taking ad- vantage of the power of chemical synthesis and synthesize, we design, Firstly, chemical biology. and evaluate untraditional antimicrobial agents that could potentially circumvent the drug Our molecular design stems resistance problem. “delivering catastrophic from the concept of We agents only to target pathogenic cells.” are developing and evaluating various types and “targeting” of molecules consisting of we have Based on this concept, motifs. “killing” succeeded in synthesizing such a molecular which “RoseBengal/C11-AHL”, conjugate named gratifyingly exerted >2-fold greater bacteri- are We cidal activity than Rose Bengal alone. currently optimizing this structure to achieve greater potency and broader antibacterial we are conducting proteomic Secondly, spectra. profiling using synthetic chemical probes based In the past on antimicrobial metal complexes. metal complexes have attracted decades, increasing attention as potential drugs for the autoimmune diseases, treatment of cancer, infectious diseases caused more recently, and, in However, by drug-resistant pathogens. their molecular targets are still unclear. general, 88 • IMMPACT REPORT TEXAS THERAPEUTICS INSTITUTE TEXAS THERAPEUTICS cancer immune evasion andresistanceto cancer immuneevasion and mousetumormodels toinvestigate cancer cells/immune cellsco-culturesystem therapeutic antibodies. We haveestablished roleincancerresistanceto plays animportant bodies bymatrixmetalloproteinases(MMPs) hypothesize thatproteolyticcleavageofanti - breast cancerpatients. The findingsleadusto compromised immunityintumortissuesfrom existenceofimpairedantibodieswith prevalent effector cells. a Ourrecent work revealed Fc gammareceptorsexpressedonimmune the antibodyFcinteractionwithactivating dependent cellularcytotoxicity(ADCC)through immune effectorfunctionsuchasantibody extracellular domainshedding, andtriggering inhibition ofHER2signaling, ofHER2 prevention trastuzumab havebeenproposed, including antibodies. Multiple mechanismsofaction and theirroleinresistancetotherapeutic mechanismsofcancer explore immuneevasion antibody trastuzumabasamodelsystemto vironment. We areusingtheHER2targeting the contextofheterogeneoustumormicroen- antibodies targetingEGFRfamilymembersin resistance mechanismstocancertherapeutic present asignificantchallengeintheclinic. and tic antibodieshavebeenwidelyreported innate andacquiredresistancetothetherapeu- ment ofdifferenttypescancer. However, both HER2, arecurrentlyusedintheclinicfortreat- against EGFR andtrastuzumabpertuzumab such as, cetuximab, panitumumabtargeting multiple therapeuticmonoclonalantibodies well documentedinmanytypesofcancerand and overexpressionofEGFRHER2are and signaling. Abnormal geneamplification rolesincellgrowth HER4) andplaysimportant kinase receptors(EGFR/HER1, HER2, HER3and closely relatedtype1transmembranetyrosine factor receptor(EGFR)familyconsistsoffour types ofcancer. Humanepidermalgrowth shown clinicalsuccessfortreatmentofvarious drug modalityforcancertreatmentandhave My researchinterestistounderstandthe Monoclonal antibodiesarebecomingamajor Oncogeneing. doi:10.1038/onc.2014.56. andSignal- tively RegulatesHER3/ErbB3Level (2014) The E3ubiquitinLigaseNEDD4Nega- Mukherjee S, OwitiN, Zhang*NY, and An Z Huang Z, ChoiBK, MujooK, FanX, FaM, 194:4379-4386. FcγRIV onimmunecells. Journal ofImmunology, of highHER2cancercellsbyinteractionwith (2015) Trastuzumab triggersphagocytickilling Jordan R, Strohl WR, ZouQ, Zhang*NYand An Z Shi Y, FanX, DengH, BrezskiR, RycyzynM, Cancer Therapeutics cancer therapeuticantibodies. Molecular effector functionofproteolytically-inactivated torescuetheimmune novel therapeuticstrategy Jordan RE, Zhang* NY, andZhiqiang An (2015) A Gonzalez A, ZhangS, RycyzynM, Strohl WR, Fan X, BrezskiRJ, DengH, DhupkarP, Shi Y, 10.1158/1078-0432. Cancer Research, ClinicalCancerResearch, doi: with impairedanticancerimmunity. Clinical bodies inthetumormicroenvironmentassociate Jordan RE, and An Z(2015)Dysfunctional anti- S, BrezskiRJ, ChoiBK, RycyzynM, Strohl WR, Zhang* NY, DengH, FanX, Gonzalez A, Zhang KEY PUBLICATIONS - •Discovery novelcancertargetsfordevelop •Role ofproteolytichingecleavageantibody RESEARCH PROJECTS for activationofimmunityagainstcancer. design effectiveantibodytherapeuticstrategies immune cellsintumormicroenvironmentandto dynamic interactionbetweencancercellsand moleculartargetsthatgovern the identify key (FACS). The long-termgoalofourresearchisto analysis andfluorescenceactivatedcellsorting imaging, massspectrometry, multi-color flow in ourstudiessuchashighcontentfluorescence patients. technologiesareused State-of-the-art and studieswithclinicalsamplesfromcancer 2D and3Dcellculture, mousetumormodels, of experimentalapproachesincludinginvitro antibody therapeutics. We employawidearray Associate Professor Zhang,Ningyan Ph.D. resistance mechanismstotherapeuticantibodytreatment Heterogeneity oftumormicroenvironmentandcancer ment ofantibodytherapeutics. antibody therapeutics andresistance to in cancerimmuneevasion 14(3): 681-691. doi: 10.1158/1078-0432.CCR-15-1057) subclassess. (ClinicalCancer Research, 2015; class iscalculatedbasedonthesumofallIgG breast cancerpatients. Percentage ofeachsub- subclass thanthatinthepairedplasma(B) samples. Tumor tissues(A)showedlowerIgG1 Profiles ofIgGsubclassesintumorandplasma co-culture condition Immune cells/cancercellsinteractionina page. see completelistofmembersinDr. An’s lab Joined teamwithDr. Zhiqiang An’s laboratory, Wei Xiong, Ph.D. Xuejun Fan, M.D., Ph.D. Hui Deng, M.S. LAB MEMBERS Breast CancerResearch 16(2):R33. tion incancercellsthroughSTAT1 activation. cells bytrastuzumabinducesHER2downregula- An Z(2014)Engagementofimmuneeffector Shi Y, FanX, Meng W, DengH, Zhang*NY, and IMMPACT REPORT • • • • 89

enter tional a C ransl e ervic T

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Current trends in biomedical research are are in biomedical research trends Current designed are The basic services provided Navigation Laser Oriented Image The linical eomics rot P C focused on translational studies not increasingly processes only for the understanding of disease and and therapies but also for disease diagnosis These of therapeutic efficacy. the evaluation analyses of research extensive studies often require and biological specimens for the differential in and modification of proteins expression ServiceOur Center sample populations. different state-of-the-art the servicesprovides to UTHealth, other UT community and Center, Medical Texas external organizations. and their to identify and quantitate proteins research range of modifications in a broad samples to simple purified protein specimens from in complex and verification discovery biomarker such as cell and tissue extracts, plasma, mixtures and/or other biofluids. The service center contains instrumentationthe latest and most advanced and sample preparation trained personnel to provide and analysis servicesspecimens. This of research type of instrumentation is highly sophisticated both in terms of the mechanics of operation as the extraction and and maintenance as well of the data. interpretation (ION LMD) offers a Device Microdissection to of UV laser microdissection unique feature from groups of cells efficiently isolate single or tissue sections without changing morphology or LMD integrity of the biological content. Using of quality material for a wide variety technology, analyses can be obtained DNA, RNA, and protein and accurate molecular assays, such as for sensitive sequencing, next-generation sequencing, Sanger the optical PCR. Beyond and quantitative external funding to continue development of continue development external funding to with the therapeutic antibodies the optimized to novel basic research ultimate goal of translating therapies.

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Antibody therapeutics represent a major Antibody therapeutics represent To accomplish IMM’s strategic goal of strategic goal accomplish IMM’s To ntibody xpression A diseases, in combating human breakthrough the pharmaceutical though Even including cancer. in the center and biotechnology industries are stage of drug and development, discovery engaged increasingly are academic researchers new antibody drug candidates. in discovering of some of the promising advancement However, from antibodies in the early stage of discovery laboratories is often hindered academic research the lack of access to the expertiseby and for antibody engineering required infrastructure antibody Our key technologies. and other related serviceengineering and expression center will fill the gap of the much needed expertise for early of monoclonal antibodies and lead to discovery and drug communities. discovery the research of the serviceThe objective center is to provide technical support and services to molecular and advance cloning and antibody expression development. antibodies to the stage of preclinical the service generated from center will Results to attract ability the collaborators’ strengthen IMM Service Centers Service IMM protein technologies and animal models. Our animal models. Our technologies and protein the latest service goal is to provide center highest quality servicestechnology and the to our while operating in a colleagues and customers Service Centers are IMM’s manner. cost-effective expertsresearch technologies in the by top staffed offered. support high quality and effective providing initiated have we services capacity, for our research our to furthera systematic process improve to our faculty and and to provide infrastructure The customers access to cutting–edge technology. establishment of key service centers at UTHealth- commitment. IMM is a critical component of this

E T 90 • IMMPACT REPORT gather information on very rare populations gather information onvery timeframeand number ofsamples inashort allowinstruments scientisttoevaluate alarge UTHealth orexternalorganizations.These chargetoallresearch investigatorsservices from and aLuminex 200. FACS Calibur, BDFACS AriaII,BCIFC500, Building BD andmaintainsfourinstruments: on thesixthfloorof Fayez S. Sarofim Research F in molecularimaging. to maximize benefitfrom thelatestdevelopments Generalized protocols are available toinvestigators unique imagingagents/genereporter systems. fluorescence thatispaired instrumentation with and anarrayofcustombioluminescence with customfluorescence tomographycapabilities Siemens hybridPET/CT smallanimalscanner vascular disease, andothers.Facilities includea nanotechnology, chronic woundcare, peripheral autoimmune disorders, gastrointestinal disorders, translational projects incancer, discovery, drug researchers acrossand industry thenationon clinicians, clinician-scientists,aswell asacademic tointeractwith utilizes this existingexpertise formed Molecular Imaging “collaboration” center technologies intoclinicaltrials.Thenewly imaging and pioneeringtranslationofnew of targetedprobes, innovative algorithms, design,andchemistry instrumentation, new different aspectsofmolecularimaging,including faculty memberswhoseresearch focuseson and ledby 7engineeringandbasicscience with. TheCenterisdirected by Dr. Eva Sevick translational imagingstudiesshouldbeacquainted are orwishtobeinvolved insmallanimal/ a facilitythatallresearchers atUTHealth who S C cells ortissue. enables ustomicrodissect anyfluorescence labeled microscope function,ourfluorescence lightsource ervice low oll These instruments areThese instruments available onafeeper The Flow Centerislocated Service Cytometry The IMMCenterfor Molecular Imaging is abor C C ytometry entr tion I ma S ervice ging C entr analysis and cell sorting. analysis andcellsorting. forboth andtechnicalexpertise instrumentation, centerprovidesof cells.Theservice training, • • • • centrifuge tubforDNA,RNAstudies • stain) • provided: both paraffinand fresh frozen-tissue. equipment forthepreparation ofthinsections; houses thelaboratory histopathology services, the growth ofresearch activitiesthatrequire from UTHealth orexternalorganizations. With of research projects toallresearch investigators insupport services and immunohistochemistry provides in-houseroutine histology, special stain, S T technical support, training, and consultation technical support, basisby providingfee-for-service microscopy from UTHealth orexternalorganizationsona the research needsofallresearch investigators post-acquisition analysisofimagingdata. Amirasoftwarecomputer workstation running for a Zeiss AxioScopebrightfieldmicroscope, anda Eclipse wide-field microscope,TE2000E inverted with conventional andresonant scanner, aNikon a Leica TSC SP5uprightconfocalmicroscope and imageanalysis.Thefacilityisequippedwith fluorescence microscopy, brightfieldmicroscopy, assistance inconfocalmicroscopy, wide-field M ervice issue icroscopy Immunohistochemistry andspecialstain Immunohistochemistry Blood Smear stain Frozen tissueembeddingandsectioning Multi-tissue embeddingandsectioning Section cutrolled andplacedinmicro Routine histology(process, embed,cutand is A fullrangeofhistopathologyservices The Tissue Center Histopathology Service The Microscopy Centerwillsupport Service The IMM Microscopy Centerprovides Service H C istop entr S a ervice thology C entr IMMPACT REPORT • • • • 91

es ervic S ell C tem S gnostics ia and

D enter ar C Our Immunology and Autoimmune Diseases Diseases and Autoimmune Immunology Our been derived The stem cell lines that have Our Molecular Diagnostic Laboratory, Laboratory, Diagnostic Molecular Our olecul ransgenic e ervic Cells and Stem Transgenic Center operates a in 1998. which was established service center, 750 new transgenic and has generated over It all research knock-out mouse animal models for and external UTHealth from investigators organizations on a fee-for-service basis. in the laboratory for the highly effective are and for generation of knock-out/ knock-in mice In addition to the studies. cell differentiation cryopreservation,production, and re-derivation and rats, the mice of genetically-engineered services of the facility also include gene targeting, of new cell lines and intellectual/ derivation aspects of technical support in different and stem cells research. cell culture, microsurgery, T M S testing in a CLIA diagnostic provides ProteoPath, certified laboratory investigators research to all on a or external organizations UTHealth from fee-for-service mass testing includes Major basis. Monitoring Drugspectrometry based Therapeutic Monitoring D Vitamin (TDM) and program and anomalies trends for monitoring program as metabolites as well during clinical treatment, also provide We testing. analysis for research ELISA assays and customized manufactured serve as a diagnostic We of targets. for a variety Brown site for The technology development Medicine, of Molecular Institute Foundation other Clinical Laboratories, physicians, and external organizations. 92 • IMMPACT REPORT FY 11 IMM By theNumbers 46 Number ofFaculty FY 12 50 FY 13 53 17% 48% FY 14 52 Total Expenses Supporting Research 56 FY 15 15% $10,000,000 $12,500,000 $15,000,000 $17,500,000 $20,000,000 Sponsored Projects Service Center Endowment/Gifts $2,500,000 $5,000,000 $7,500,000 Total FundsSupporting Research 14% 6% 0 2011 Service CentersandEndowments/Giftsbasedonexpenses Sponsored Projectsbasedonawardreceived Excludes all ARRA funds Note - 2012 Service Centers Industry Foundations State Government Federal Government 2013 2014 2015 IMMPACT REPORT • • • • 93

Gary Mercer Gary Irma Gigli, M.D. Gigli, Irma Hobby Family Foundation Family Hobby Estate of Marjorie Poyner of Marjorie Estate Gift Report Gift Estate of Herbert F. Poyner F. of Herbert Estate Frois Family Foundation Inc. Foundation Family Frois Patricia and John McDonald and John Patricia

Anne and Don Fizer Foundation Fizer Anne and Don Susan Atkins and Steven Gordon and Steven Atkins Susan We are deeply grateful to UTHealth benefactors deeply grateful to UTHealth are We who generously made a gift of $1,000 or greater to the or greater made a gift of $1,000 who generously during fiscal year 2015, September 1, 2014 – August 31, 2015. August 31, 2015. – September 1, 2014 year 2015, during fiscal George and Mary Josephine Hamman Foundation Hamman Josephine and Mary George Thank you to all of our supporters! New Gifts and Bequests Fiscal Year 2015 and Bequests Fiscal New Gifts Brown Foundation Institute of Molecular Medicine for the Prevention of Human Diseases of Human for the Prevention Medicine of Molecular Institute Foundation Brown 94 • IMMPACT REPORT The University of Texas Health Science Center at Houston (UTHealth) Leadership Giuseppe N. Colasurdo, M.D. President Kevin Dillon, M.B.A., C.P.A. Senior Executive Vice President, Chief Operating & Financial Officer George M. Stancel, Ph.D. Executive Vice President for Academic and Research Affairs Barbara J. Stoll, M.D. Dean, McGovern Medical School

IMM Senior Administrators John F. Hancock, M.A., M.B., B.Chir., Ph.D., Sc.D. Executive Director David G. Gorenstein, Ph.D. Deputy Director

IMM Advisory Council Members Dudley Oldham, Chair Alan Baden David J. Beck John Beckworth Louana H. Frois Ted Frois Irma Gigli, M.D. Steve Gordon Rodney Margolis John E. McDonald Brad McWilliams Beth Robertson Clive Runnells Ralph Thomas

The University of Texas System Board of Regents Paul L. Foster, Chairman Jeffery D. Hildebrand, Vice Chairman R. Steven Hicks, Vice Chairman Ernest Aliseda David J. Beck Alex M. Cranberg Justin A. Drake Wallace L. Hall, Jr. Brenda Pejovich Sarah Martinez Tucker Office of Communications 6431 Fannin Street MSB B.340 Houston, Texas 77030