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

FISCAL YEAR 2017 About the cover

It’s a beautiful day at McGovern Medical School’s Brown Foundation Institute of Molecular Medicine for the Prevention of Human Diseases.

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 Jonathan Garris IMM Faculty

Design: Roy Prichard

Photography: Dwight Andrews Maricruz Kwon

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

OOC-ThomasPrintworks-1175-3/18 IMMPACT REPORT 1 • • • •

8 14 12 practice Cheering in the fight on the IMM against disease genome territory therapeutics into Moving promising Moving Entering uncharted Entering

6 11 it starts research in the lab Gene editing – Gene into lung disease Breathing new life Breathing 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 Biomedicine Center for Precision 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 31 40 48 57 74 81 90 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 ChairinPhysiology andMedicine Executive Director, Institute ofMolecular Medicine John Hancock, M.A.,M.B.,B.Chir., Ph.D., Sc.D. calendar becauseitisagreat tovisittheIMM. opportunity answered. Full detailsare inthisreport; pleasemark thedateinyour the future ofmedicineandhealthcare, andtohave all your questions research storiesdirectly from ourfaculty, todiscussitsimplicationsfor physicians. Thesymposiumisanexcellent tohearexciting opportunity panelcomprisingthe speakersplusUTHealthquestion timewithanexpert be slightlydifferent from previous years inthat we willhave anextended gene editingtechnologiestocorrect inheriteddiseases.Theformatwill will beheldonApril 18,2018,andwillfeature talksonhow tousenew the James T. Willerson, M.D.Discovery Hall. This year thesymposium talks intheBeth Robertson Auditorium andattendedareception in our annualIMMpactsymposium.Lastyear 165guestslistenedtothree talk withyou personally. Alternatively, Iwouldbedelightedtoseeyou at like toinvestigate how you canalsobeinvolved, Iwouldbedelightedto in thecontinuedgrowth anddevelopment oftheIMM.If you would council,whichplaysakeyrole work anddedicationoftheIMMadvisory human disease.In thisregard, we are deeplyappreciative ofthestrong of ourmissiontotranslatemoleculardiscoveries therapies for intonew our relationships withallinourcommunitywhovalue theaspiration of ourfriendsanddonors. collaborations,and,mostimportantly,industry thecontinuinggenerosity from alternativeattracting support sources, includingresearch foundations, said, fullimplementationofourmissionremains heavilydependenton research fundsfrom whatisanever-diminishing nationalpool.That creativity ofourscientiststhattheIMMremains sosuccessfulinattracting increase over theprioryear. It isatestament totheremarkable qualityand percent over thepreceding year, whichinturnhadseenaconsiderable financial grantsandcontracts year justended,our new were upsome 30 from theNIH.Despite this,IMMfaculty have excelled again.Over the scientific research fundingcontinuestobeextremely challenging,especially for therecruitment ofhighlysought-afterscientists.Theenvironment for STARs awards from The University of Texas System, whichare reserved recruits, whosestoriesareof ournew featured, alsosecured prestigious bring withthemexciting research ideasandinnovative technologies. Two describing theirresearch programs. our facultyanddonorsplusanaccountfrom eachIMMfacultymember colleagues. Inside thereport onsomeof you willfindin-deptharticles basic andtranslationalscientistswhocollaboratecloselywithourclinical medicine thatbenefitspatients. To thisend, we have teamsofoutstanding mission istodeliver translationaloutcomesfrom research inmolecular institute thatisembeddedwithinMcGovern Medical School.TheIMM In additiontoadvancing scienceandmedicine,we wishtodevelop This year we have faculty, additionaloutstandingnew recruited who of Human Diseases (IMM).TheIMMisastand-alone research Brown Foundation Institute ofMolecular Medicine forthePrevention ’m pleasedtointroduce thelatestannualIMMpactreport forThe T potential forinnovative medical cell biologyhave enormous human diseases. strategies fortheprevention of and, wherever possible, methods ofrationaltherapy institute endeavors todesign area ofclinicalresearch. The in theincreasingly important interest forfuture planning and progress are ofparticular mechanisms. Thisdevelopment technologies toelucidatedisease levels, using DNAandprotein the cellularandmolecular causes ofhumandiseasesat that seekstoinvestigate the (IMM) isaresearch institute Prevention ofHuman Diseases

Mission Advances inmolecularand Medicine forthe Institute ofMolecular he Brown Foundation of molecularand cellbiology of theIMM. promises tofulfillthemission investigators isinspiringand The research oftheinstitute’s and immunologicdiseases. illnesses, andinflammatory disorders, majormental vascular disease, hypertension, medicine, suchasheart unsolved problems ofmodern clarifying thecausesofmany will playamajorrole in molecular andcellbiology diseases. infectious disorders andgenetic used todeterminethecausesof have been mostsuccessfully therapies. Theseapproaches of medicinewithmore novel research andthefuture practice Because theapplications However, itisclearthat and internationally. collaboration locally, nationally, for research excellence and strives tosetthe example Prevention ofHuman Diseases Molecular Medicine forthe Foundation Institute of Medical School,theBrown opportunities. topharmaceutical properties discoveries andintellectual various industriestoapplyits own research activitiesand collaborations between its linksand form important the potentialanddesire to industry, the IMMhas and thepharmaceutical development inbiotechnology toproduct major importance to medicalpracticeare of As aninstituteofMcGovern

• PACT REPORT PACT 3 IMM • 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, S AV E

Distinguished U C. Ha R Dir Brian Davis,P egener THE ector oftheCenterforS rold andLorineG. ative M edicine niversity Chair D hD W tem Celland AT allace E 1825 Pr Fa I Pr M I e Bro W 4:00-6:30 p.m. A nstitute of olecular M ev yez S.SarofimResear pril 18,2018 ention ofH ednesday and R Assistant D Radbod Darabi,MD, P essler St wn F egener oundation edicine forthe Pr ofessor r ative M uman D eet , CenterforS edicine ch B iseases uilding tem Cell

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IMMPACT REPORT Profiles in research A fibrosis, ageneticlung disease are from patientswith cystic blood stemcells. Thelungcells to correct damagedlungand replacing itwithahealthy gene. offending geneticmutation and scientists toprecisely trim the as “molecular scissors,” allowing four typesoftechnologyknown effector-based nucleases,are the transcription activator-like ases, zincfingernucleases,and as CRISPR-Cas,meganucle- disease. New technology, such mia orearlyonsetAlzheimer’s child, suchassicklecellane- passed down from parent to to DNAmutationsthatare genetic disease. could potentiallybecured ofthe healthy gene-corrected cells patients. Apatientinfusedwith els sothatitcansafelymove to animalmod- fully inlaboratory to completethisprocess success- to thesamepatient. tion. Return thecorrected cells the identified,inheritedmuta- from apatient. Precisely correct ageneticdisease cells carrying something likethis:Remove patients healthyagain. the errors in hopesofmaking to modifygenes–weeding out and editinginthelaboratory uses theseconceptsofpruning Cell &Regenerative Medicine, of theIMM’s CenterforStem ing keepscopysharp. all goodwritersknow thatedit- Dr. Davis’ labisworking Dozens ofdiseasesare linked Dr. Davis’ goalistobeable In simplifiedtermsitgoes Brian Davis, Ph.D., director healthy garden. Just as iskeytoa that pruning ll green thumbsknow it starts inthelab Gene editing– we needandreturning organ- and create the typeofstemcell from anadulthairorblood cell – whereby we cantakeacell induced pluripotentstemscells to create stemcellsthrough imagine sucharobust method sible. has madetheseadvances pos- progress inthefieldofstemcells Hunter syndrome. patient withtherare disease California inNovember ina live geneeditingtookplacein eyesight. Andthenation’s first with thegoalofsavingpatients’ such asmaculardegeneration, retinal degenerative diseases, cells are inclinicaltrialtotreat ment. Induced pluripotentstem be damagedby cancertreat- replenish theirblood,whichcan of stemcellsallows patientsto apy orradiation.The renewal ceive high dosesofchemother- in somecancerpatientswhore- stem celltherapyisbeingused patients. For example,blood therapies usingstemcellsin approach will bethebestbet.” in vivo. We don’t know which sidering targetinggeneediting stem cell,sowe are alsocon- the easiestplacetoreintroduce a vis says.“Thelungmightnotbe healthy tothepatient,” Dr. Da- anddeliver themback airway stem cellsastheyexistinthe isolate, expand,andeditthe from recurrent infections. deficiency patientswhosuffer blood cellsare from immune people intheUnited States. The that affectsmore than30,000 “Ten years agowe didnot Dr. Davis saystherapid There are already approved “Our goalforthelungisto Davis explains. funding’ ismostvaluable,” Dr. this iswhere philanthropic ‘seed principle’ demonstrations –and typically likestosee‘proof of providing theirgrantfunding, al Institutes ofHealth before off in5to10 years. The Nation- ment, somethingthatwillpay as awisedevelopment invest- philanthropic donorswhoseeit offorward-thinking the support work. thelaboratory to support the patients,fundingisrequired DNA.” versely affect thechromosomal to dosomethingthatwouldad- to beasprecise aspossiblenot ence andmedicinehave learned technology ismoving fast,sci- working on this. While the of scientistsaround theworld ideas, andthere are manyteams idly,” hesays.“It’s amarket of diseases. “It’s moving rap- very years and even soonerforblood genetic lungdiseasesin8to10 applications forpatientswith see geneeditingmove toclinical says. genetic correction,” Dr. Davis cut andpastetomakeaspecific “We canhighlightanarea to coming more usefulandprecise. cally correct genedefectsare be- cells, thetoolsusedtospecifi- rejecting thecell.” individual, minimizingthebody donor stemcellsfrom another “This eliminatestheneedtouse to thatpatient,” heexplains. specific cells(e.g.lungcells) “Cutting-edge research takes Before theresearch moves to Dr. Davis sayshehopesto In additiontocreating stem

Dr. Brian Davis usesgeneeditingtechniques tocorrect damagedlungandblood stemcells.

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IMMPACT REPORT Profiles in research “ N discovery. of drug sourcesinsupport funding fromfederal, state,andindustry The TTIhasbeensuccessful inattractingmorethan$30 millionin development. forfurther biotech industry tothe and spinalcord injury candidatestargetingcancer drug and sofarhaslicensedoutfive two biotechspinoffcompanies discovery.”of drug sources insupport and industry in fundingfrom federal,state, tracting more than$30million hasbeensuccessfulinat- TTI An, theinstitutedirector. “The research community,” saysDr. totheacademic gy andexpertise development technolo- cal drug death at by providingTTI criti- Molecular Medicine. centerattheInstitute for ery discov anacademicdrug - (TTI), the Texas Therapeutics Institute was recruited in2009tolead Research Laboratories,Dr. An ogy industry, mostlyatMerck maceutical andbiotechnol- worked onbothsides. Zhiqiang An,Ph.D., has turned academicresearcher, falltothewayside. drugs new is where mostgoodideasfor development discovery anddrug The institute’s areas of The institutehasestablished “We’re bridgingthevalley of After 15years inthephar- developerA commercial drug gap between academic death” by insiders,the icknamed the“valley of therapeutics into practice ” — Dr. ZhiqiangAn Moving promising gies available to Texas-based antibody platformtechnolo- technologies. specialized protein engineering not have accesstothehighly is thatmanyresearchers do tion, according toDr. An, basic discovery stage.” ries donotadvance beyond the generated inacademiclaborato- of thetherapeuticantibodies treatments,” Dr. Ansays.“Most their basicdiscoveries intonew From Bench toClinic.” peutic Monoclonal Antibodies resource onthetopic—“Thera- research inthisarea. is recognized worldwideforhis cine, according toDr. An,who ofmedi- advances in thehistory represent oneofthesignificant University ChairinChemistry. Robert A. Welch Distinguished and efficacy,” says Dr. An,the safety profile, targetspecificity largely duetotheirdesirable antibody-based therapiesis and cancer. Therapidriseof they fightinfectiousdiseases natural immunesystem,and discovery. includeantibodydrug expertise The TTI ismakingthese One reason forthislimita- “We helpscientiststranslate In fact,heeditedascientific Therapeutic antibodies “Antibodies are ofour apart State of Texas,” Dr. Ansays. inHouston and the industry and inadvancing thebiotech ley ofdeath’ discovery indrug critical role inbridgingthe‘val- will continuetogrow andplaya ization expertise. development andcommercial- nies ontheotherhandhave the discovery andinnovation. An seesacademiaexcelling in ing promising treatments, Dr. disease biomarkers.” sensitive and specifictothe says. “Antibodies are extremely in thehumanbody,” Dr. An biomarkers incells,tissues,and conditions by detectingdisease also usedtodiagnosemedical ment ofdisease,antibodiesare payload. seeking missilewithadrug kill cancercells. toxins andradioisotopesto carry dition, theycanbedesignedto and circulating proteins. In ad- ing receptors oncellsurfaces to binddiseasetargetsinclud- researchers andbeyond. “We are confidentthat TTI Pharma and biotechcompa- When itcomestoadvanc - “Not limitedtothetreat- Picture aminiaturized heat- Antibodies canbeengineered So how dotheywork?

Dr. ZhiqiangAn leadsthe IMM’s Texas Therapeutics Institute, whose chargeistotranslate scienceintotreatments.

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10 • IMMPACT REPORT Dr. Sarah Huang’s inhumanpluripotent stemcellscouldbekeyin thefightagainstlungdisease. expertise I can beintegratedintotheor - in thebodyandmakesure they says. “How doyou putthecells tion ofthecells,” Dr. Huang have tothinkabouttheintegra- oftheresearch.was thecrux platform forstudyingdiseases cal environment andactasa maturity tobeusedinaclini- Generating cellswiththeright different typesofepithelialcells. stem cells(hPSCs)tobecome directing humanpluripotent step-wise differentiation strategy platform wasthecreation ofa The firststepindeveloping this cell typesforstemtherapy. generate clinicallyapplicable disease andcelltherapy tissue modelsforstudiesoflung ing waystodevelop invitro lung siderable numberofcelltypes. architecture andinvolves acon- organ hasthemostcomplex for lungsspecificallyisthatthe stem celltherapy. Thechallenge models forlungdiseasesand the difficultiesindeveloping erative Medicine, understands Center forStem Cell&Regen- Ph.D., assistant professor inthe the fightagainstdiseases. cells asanemergingplatformin turning theirattentiontostem researchers attheIMMare board research. forfurther Now, platforms touseasaspring- are always looking for new are alwayslookingfornew disease andcancer, scientists n thebattleagainstlung “For stemcelltherapy, we Dr. Huang’s labisresearch - Sarah Huang, M.B.B.S., into lung diseaseresearch Breathing new life in Beijing. Initially, shehad sity Health ScienceCenter program atPeking Univer- joining themasterofscience focusing onlungdiseasesafter in China2000andstarted University College ofMedicine M.B.B.S. from Xi’an Jiaotong the cancercell,” Dr. Huang says. of thegenesandorigin see thesource ofthemutation genome editing,andwe could PR technology, we coulddo groups. fully adoptedby otherresearch and herwork hasbeensuccess- epithelial cellsfrom hPSCs, of anumberlungandairway versity showed thegeneration atColumbiaUniLaboratory - her timeatDr. Hans Snoeck’s cer. Dr. Huang’s research during origin forspecifictypesofcan- whether ornotacelltypeisthe cells, researchers couldmodel stem sources like embryonic rates. high survival applications astheydon’t have to beinadequateforcelltherapy have been shown inmanycases therapy. Adult progenitor cells such aspost-natalcells,for that are completelymature, that doctorsmaynotneedcells up tousfigure out.” suitable forcelltherapy?Thisis maturity ofthecellsthatismost always thinkaboutiswhatthe gan? Theotherchallengepeople Dr. Huang received her “In combinationwithCRIS- With cellsgeneratedfrom Research currently shows Huang says. do itinahumanculture,” Dr. but itwouldbeinteresting to how itproliferates. labeling themandobserving cells inthevitro cultures, ent viralstrainsandfollowing also involves comparingdiffer- This the humaninfluenzavirus. have different susceptibilityto different typesofcellscould form tohelpcharacterize how of a plat- epithelial cellsaspart culture ofthelungandairway complex, three-dimensional has interest indeveloping a laborations,” Dr. Huang says. laborations insidetheIMM. Medicine, andvarious othercol- Cancer Center’s Pulmonary Rice University, MDAnderson bio-engineering group from They includeworkinga with variety of collaborative efforts. in Houston, hasestablisheda after spendingaboutayear Medical Schoolinstrideand, ing herpositionatMcGovern after gettingcancer.” wereoff afterbirth turnedon ment pathwaysthatwere turned me,” shesays.“A lotofdevelop- and stemcellswasintriguingto therapy. found herselfdrawntostemcell been studyingcancer, butlater “This has been done in mice, “This hasbeendoneinmice, In thelong-term,Dr. Huang “I cannotstopmakingcol- Dr. Huang hasbeentak- “The linkbetween cancer Profiles in research

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IMMPACT REPORT Profiles in research D small fraction of RNAtran- sequence. Many thinkonlya encoded andyou getthe protein up whataminoacidisbeing can reference thetableand look the codinggene,researchers once you know theframeof the helpofcodontable- reading thecodingregions with genome sequence,” hesays. know how toread mostofthe lot ofpeople,butwe stilldon’t already beendiscovered. tions lurking beneathwhathas there are stillunanswered ques- the humangenomesequenced, technology. Althoughwe have next generationofsequencing of genomesequencingandthe the lastdecadewithadvent become more complicatedover might function. coding genesandhow they and by exploring potentialnew examine RNAbindingproteins development toolsto ofnew to changethatby exploringthe assistant professor atIMM,aims genetic variation. common diseasesarisefrom way tofundamentallygrasphow tion ingeneexpression canbea mystery. Understanding regula- tion leftcloudedinrelative the post-transcriptionalregula- from complete,withmuchof current understandingisfar Researchers are adeptat “I’m sure thisisclichétoa Dr. Wang saysthefieldhas Sidney H. Wang, Ph.D., Entering uncharted genome demanding task.Our of generegulation isa iving intotheworld fight against disease territory inthe almost 5,000 new coding genes. codinggenes. almost 5,000new and researchers have found already assembled, Dr. Wang peptides.” beingmadeinto some portion lations, andIwasabletoverify regions withsignatures of trans- actually findalotofthesetiny But whatIfoundisthatwe can frame isnotconsidered real. 200 basepair, theopenreading explains. “If it’s smallerthan computation issue,” Dr. Wang filtered outbecauseofthis protein. how smallistootobea cutoff point,whichdetermines some ofthesefalloutsidethe number ofpotentialproteins, scripts, and,duetothesheer researchers have examinedtran- been overlooked isduetohow Part ofthereason theyhave strange, “non-typical processes.” have been overlooked dueto be makingproteins butmay RNA’s. Some maystillactually closely atso-callednon-coding actually coding,orby looking re-evaluate how manyRNAsare coding genes,Dr. Wang can Dr. Wangsays. be abitbiggerthan2percent,” postdoc career isthatitmight but onethingIfoundover my only 2percent ofthegenome, different belief. research hasledhimtohave a into protein, butDr. Wang’s scribed by thegenomeismade Just from thetranscripts “A lotoftheseare getting By examiningpatternsof “Most peoplethinkthat’s for collaborators.” says. “People are alwayslooking environment ingeneral,” he eager tohelponeanother. with researchers andprofessors by adiverse environment, filled the IMMhasbeenbolstered Dr. Wang sayshispositionwith distinguished Dr. Sarah Elgin. of the Louis inthelaboratory Washington University inSt. obtained aPh.D in2012from double majored inphysics.He of scienceinlifeand sity in2003withabachelor’s National Dong HwaUniver- protein translation. studying direct regulation of be effective forhis research in developing toolsthatwill new transcription, Dr. Wang willbe ing ontranslationratherthan man health.However, infocus- could have implicationsforhu- processes potentially important in out thosewhichparticipate tial codingregions andsingling by mostpeople,” hesays. protein coding genesaccepted currently there are only20,000 bers, it’s pretty strikingbecause research labattheIMM. of themainareas ofDr. Wang’s and whattheyare doingisone genes are, infact,functional and verifying thatthesecoding genes are madeintopeptides, 25 percent ofthosecoding Current estimatesshow about “It’s collaborative justavery Dr. Wang graduatedfrom poten- The discovery ofnew “If you lookatthenum-

Dr. Sidney Wang’s research couldleadtonewcodingregions inthehumangenomeby re-evaluating so-callednon-coding RNAs.

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14 • IMMPACT REPORT sharing theIMM visionwiththebroader Houston community. John McDonald, Council leadershipand left,and DudleyOldham theIMMthrough Advisory support “ T a breakthrough discovery. without bureaucratic oversight,withthegoaltomake the bestscientistsare recruitedtoworkontheirdrivingpassion, What Ihavealways admiredabouttheIMMisthatita placewhere 2016. – whichhedidinSeptember of person tosucceedhimaschair realizing hewouldbetheideal years council memberafew ago, then recruited McDonald asa chairing itforseven years. He counciland first formaladvisory decade –buildingtheinstitute’s with theIMMforabouta Oldham says. make abreakthrough discovery,” cratic oversight, withthegoalto driving passion,withoutbureau- are recruited towork ontheir place where thebestscientists about theIMMisthatita IMM. their scientificsightsonthe recent years, theyhave focused of sciencecourses.In more Dudley recalls hetook66hours The University of Texas –where other sincetheirstudentdaysat McDonald have known each of Human Diseases (IMM). lar Medicine forthePrevention Foundation Institute ofMolecu- theBrownmission tosupport technology intoaphilanthropic “I knew that his enthusiasm thathisenthusiasm “I knew Oldham hasbeeninvolved Ihave alwaysadmired “What Dudley Oldham andJohn passion for science and passion forscienceand friends have vaulted their wo formercollege Cheering ontheIMM still anactive memberofthe stepped down aschair, heis that growth.” toensureresources necessary ing theIMMandattracting hisvisionin growis tosupport - Dr. John Hancock, andourgoal close working relationship with energetic supporters. We have a Councilwithadditional visory can. We are enlargingtheAd- mission toasmanypeoplewe introducing theIMMandits need to‘get theword out’ by medical research institute,sowe for IMM. done andactsasacheerleader the innovative research being throughout thecommunityfor encourages financialsupport IMM,” McDonald says,“It group forthe as thesupport the community. increase theIMM’s exposure in that aschairhewantstohelp large Texas accountingfirm,says committee of Weaver, LLP, a and memberoftheexecutive oftheHoustonpartner office bright &Jaworski. retired atFul seniorpartner - be infectious,” saysOldham, a for the IMM would and support ” Although Oldham has “Awareness ishugeforany Council theAdvisory “I view McDonald, retired managing — Dudley Oldham, Advisory Councilmember — DudleyOldham, Advisory director, IMM. Dr. John Hancock, executive advocate onourbehalf,” says ourmissionandtirelessly port as Dudley andJohn whosup- are gratefultohave suchleaders andwe community supporters, and care,” Oldham explains. vancement ofmedicalresearch our desire thead- tofurther professionals atUTHealth, and of thescientistsandhealthcare the encouragementandsupport dent Dr. Giuseppe Colasurdo, leadership ofUTHealth Presi - our esteemfortheoutstanding Research. tion Endowment forDepression Medical School’s Fizer Founda- in establishingtheMcGovern Don Fizer Foundation Board as amemberoftheAnneand Oldham alsowasinstrumental (McGovern Medical School). White Oldham Scholarship and theD.Dudley and Judy UTHealth Graduate School), Faculty Award (MDAnderson and Judy White Oldham search Fund (IMM),D.Dudley and Judy White Oldham Re - faculty, and students:D.Dudley research,lished fundstosupport and hiswife,Judy, have estab- Council.HeIMM Advisory “The IMM depends upon its “The IMMdependsuponits “These giftsare a of reflection Donor spotlight

15 • PACT REPORT PACT IMM IMMPACT REPORT • • • • 17 cal trial to test efficacy of ATPase modulator ATPase cal trial to test efficacy of tolerance on improve symptoms and exercise hypertrophicin patients with obstructive cardiomyopathy EXPLORER study: An industry –sponsored clini- •EXPLORER study: KEY PUBLICATIONS Suppression of Activated FOXO Transcription Factors in the Heart Prolongs Survival in a Auguste G, Mouse Model of Laminopathies. Willerson JT, Coarfa C, Lombardi R, Gurha P, doi: 10.1161/ 2018 Jan 9. Circ Res. AJ. Marian PMID: 29317431 117.312052. CIRCRESAHA. A Distinct Cellular Basis for Early Cardiac The Cardinal Manifestation of Arrhythmias, and the Skin Arrhythmogenic Cardiomyopathy, Phenotype of Cardiocutaneous Syndromes. Lombardi R, Miyake CY, Zhou QQ, Karmouch J, Clevers H, Bannier-Hélaouët M, Kretzschmar K, Res. Circ AJ. Marian Willerson JT, XH, Wehrens PMID: 29018034 2017 Oct 10. A Potential Oligogenic Etiology of Hypertrophic A Classic Single Gene Disorder. Cardiomyopathy, Marian Willerson JT, Y, Tan Bainbridge MN, Li L, 2017 Mar 31;120(7): 1084-1090. Res. Circ AJ. PMID: 28223422 LAB MEMBERS Ph.D.; Auguste, Post-doctoral Fellows: Gaelle Sirisha C Marreddy Ph.D.; Jennifer Karmouch, M.S. Associate: Grace Czernuszewicz, Research RN Tan, Yanli Research and Clinical Nurse: and sudden cardiac death including etic and transcriptomic changes, non-coding RNAs and histone modifications in hereditary cardiomyopathies molecular pathways that link the genetic mu- tations to the clinical phenotype in patients with cardiomyopathies including delinea- tion of the mechanical signaling pathways regulated at the intercalated discs N-Acetylcysteine in Hypertrophic Cardio- myopathy) clinical trial (ClinicalTrial.Org NCT01537926) Approach to Research Programs on Cardiomyopathies AJ Marian, M.D. AJ Marian, Cardiovascular Genetics Director of the Center for Professor and WillersonResearch Chair in Cardiovascular Distinguished T. James of and treatment pathogenesis, genomics, Molecular genetics, hereditary cardiomyopathies genomic remodel- link the causal mutations to re- The ing and to the pathogenic pathways. are identified sponsible molecular mechanisms studies in ge- through molecular mechanistic models and cultured netically modified animal then The mechanistic discoveries are cells. utiliz- utilized to intervene in model organisms, ing genetic and pharmacological approaches order in that target the pathogenic pathways, to prevent evolving the phenotype and reverse These or attenuate the established phenotype. findings in the model organisms are extended to human studies through pilot randomized placebo-controlled double-blind clinical trials. are pursued through if favorable, The findings, collaborative large-scale clinical trials. RESEARCH PROJECTS •Identification of causal genes for heart failure •Identification and characterization of epigen- •Identification and characterization of the (Hypertrophic Regression with •HALT-HCM Our long-standing research objectives have ACM Arrhythmogenic Cardiomyopathy (ACM): Hypertrophic is Cardiomyopathy (HCM): HCM Dilated Cardiomyopathy (DCM): DCM is The overall approach entails an integrated CARDIOVASCULAR GENETICS FOR CARDIOVASCULAR CENTER been to delineate the molecular genetics, and pathogenesis of hereditary genomics, cardiomyopathies in humans and apply the discoveries to prevent the evolving and reverse the established phenotypes of heart failure and active research have We sudden cardiac death. programs in three common forms of hereditary cardiomyopathies: is an enigmatic form of hereditary cardiomy- opathies that clinically presents with cardiac heart and sudden cardiac failure arrhythmias, A unique particularly the young. in death, feature of this disease is a gradual replacement There of cardiac myocytes with fibro-adipocytes. ACM. is no 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 HCM is the most common cause of 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 pharmacologi- cal and non-pharmacological therapies for DCM but currently there is no cure for DCM. approach that includes human molecular genet- ic studies through high throughput whole exome and genome sequencing to identify the causal followed by genomic stud- genes and mutations, ies including transcriptomics and epigenetics to define molecular remodeling of chromatin in The aim is to the presence of causal mutations. Genomics studies of human heart failure studies of human heartGenomics failure Therapeutic targeting of dysregulated The center participates Clinical Studies: II. III. IV. probands and family members. These studies have and family members. These studies have probands identification of new disease-causing genes and the genetic causes of heart advanced failure. have additional probands recruiting actively are We and family members. The and mouse models of cardiomyopathies: to DCM and ACM relate studies predominantly by and included whole transcriptome analysis RRBS, DNA methylation analysis by RNA-Seq, of ChIP-Seq by remodeling and chromatin The integrated specific histones and proteins. and therapeutic used for preventive findings are targeting. Dysregulated pathways in cardiomyopathies: integrated genomics pathways identified through pharmacological genetic and targeted through are interventions in model organisms and their function, and clinical effects on survival, cardiac programs active Several analyzed. outcomes are underway. current are single center pilot clinical in investigator–initiated as industry-sponsoredtrials as well multi-center clinical trials in hereditary cardiomyopathy. randomized double blind An NIH-sponsored in patients with pilot study (HALT-HCM) completed. The center also HCM was recently participates in industry clinical trials in sponsored cardiomyopathies. M.D. AJ Marian, & Professor Director Center Willerson Distinguished T. James Research Chair in Cardiovascular

Human molecular genetic studies of Human he IMM Center for Cardiovascular Center for Cardiovascular he IMM in 2006, established Genetics, elucidation of molecular focuses on

Mission: To prevent and treat and treat prevent To Mission: Ph.D., M.D., Raffaella Lombardi, Faculty: programs: theme of the research General are programs research The Programs: Research I.

CARDIOVASCULAR GENETICS FOR CARDIOVASCULAR CENTER

genetics, genomics, and pathogenesis of genomics, and pathogenesis genetics, diseases with the objective cardiovascular and to prevent of utilizing the discoveries humans. The diseases in cardiovascular treat services clinical specialized center provides cardiovascular to patients with genetic Genetic Cardiovascular at the disorders Research also has a Clinic. The center research for clinical Clinic, which is utilized NIH- and industry-activities, including clinical trials. sponsored diseases in humans through cardiovascular identification and targeting of the pathogenic genes and pathways. Ph.D., Gurha, Priyatansh assistant professor; M.D., professor AJ Marian, assistant professor; at the Center starts programs research The aimed with human molecular genetic studies at identifying the causal genes for human diseases. The focus is primarily on cardiovascular important which are hereditary cardiomyopathies, death and heart failure. causes of sudden cardiac analysis is performed exome whole Genetic by discoveries and genome sequencing. Genetic then coupled with the genomic studies to are coding and non- expressed identify differentially pathways, coding transcripts and dysregulated methylation and DNA remodeling, chromatin is The integrated approach in cardiomyopathies. pathogenic used to identify the key dysregulated and therapeutic genetic pathways for preventive and pharmacological interventions. The findings to human extended in the model systems are placebo- pilot randomized patients through double-blind studies clinical trials. The control the platform for large-scale findings provide multi-center efficacy clinical trials. as follows: a repository have of We cardiomyopathies: cases and their family members hundred several including hypertrophic with cardiomyopathies, (HCM), dilated cardiomyopathy cardiomyopathy (DCM), and arrhythmogenic cardiomyopathy are and causal variants Pathogenic (ACM). sequencing in the by whole exome identified T

IMM PACT REPORT PACT • • • • 16 18 • IMMPACT REPORT CENTER FORCARDIOVASCULARGENETICS mutation, anLMNA-deficient(Lmna with anLMNA By studyingthehuman heart specific diseasephenotypes inlaminopathies. and mechanisticdetailsthatleadstotissue identify andcharacterizemolecularcomponent Laminopathies. Objectiveofthisstudyisto transcription ensuingcardiacphenotypein ming theepigeneticcodethatgovernsgene regulatory roleofLamin(LMNA)inreprogram - rescuetheadipogenicphenotype. partially adipogenesis andoverexpressionofthismiRNA suppression ofmiR-184leadstoenhanced miR-184 expressioninheart. We showedthat DNA methylationtranscriptionallysuppress network encompassingE2F1pathwayandCpG cells. We alsoshowedthatin ACM anepigenetic expressed incardiacmesenchymalprogenitor andwaspredominantly regulated inmiceheart down show thatmiR-184isdevelopmentally down-regulated miRNA(~10-fold)in ACM. We sion dataweidentifiedmiR-184asthemost By integratingRNA-seqdatawithmiRNAexpres- replacement ofmyocytesbyfibro-adipocytes. pathogenicfeaturesof key ACM isgradual failure,heart andsuddendeath. Oneofthe clinically manifestswithcardiacarrhythmias, a primarydiseaseofthemyocardiumthat in Arrhythmogenic Cardiomyopathy(ACM), miRNAsthataredifferentially expressed several fibrosis. Furthermore, ourresearchidentified and role instress-inducedcardiachypertrophy Purine RichBindingProteinB(PURB)thatplaya miR-22regulatedcandidate geneslike several and fibrosis. Mechanistically, weidentified regulatorofcardiachypertrophy miR-22 isakey miRNA intheheart. We demonstratedthat cal roleofmiR-22, oneofthemostabundant failure. Previously, weidentifiedapathologi- in pathogenesisofcardiomyopathiesandheart functionbutarealsoinvolved in properheart clear thatnon-codingRNAsnotonlyplayarole failure.cardiomyopathies andheart Itisnow mechanisms involvedinthepathogenesisof ter understandtheroleofgeneregulatory Recently, wehavebeguntoinvestigatethe The broadgoalofmyresearchistobet- -/- ) mouse Assistant Professor Priyatansh Gurha, Ph.D. expressed Lmna causal relationofKDM5inlaminopathieswere- not beendocumentedsofar. To determinethe andinlaminopathieshas role ofKDM5inheart leading tosuppressionofgeneexpression. The of lysine4histoneH3(H3K4me3), often methylase thatremovestri-anddi-methylations of genedysregulation. KDM5isahistonede- KDM5) asmostinducedupstreamregulator and specificallyidentifiedKDM5AB(or differentially expressedgeneinlaminopathies tomic analysis, weidentifiedadiversesetof model, andisolatedcardiacmyocytetranscrip- failure heart Molecular mechanismsandfunctionsofnon-codingRNAsin gene involved in energy metabolism/Mitochondrial function. gene involvedinenergy Pathway analysisofgenesuppressedinhuman DCMthataretargetsofKDM5showingenrichment heart failure. heart determine ifinductionofKDM5ispathogenicin and theelusiveroleofKDM5inheart ascertain contribution ofKDM5inlaminopathiesto investigating thetissueandcelltypespecific wide arrayofgenomicapproaches, weare and increasedoverallsurvival. Currentlyusing of theKDM5networkanddecreasedapoptosis and foundthatthiswasassociatedwithrescue WT inLmna -/- mouse (by AAV9) mouse Gurha P* 31(3): 249-54 .Current OpinioninCardiology 2016May; Gurha P. MicroRNAsincardiovasculardisease. KEY PUBLICATIONS • •Role oflncRNAsinthepathogenesis AC RESEARCH PROJECTS authors). (*Authors contributedequally, 1Corresponding In Vitro.Circulation Research 119(6), 731-750. Pathway andLeadstoEnhanced Adipogenesis Hypermethylation andSuppressionoftheE2F1 2 DownregulatesmiR-184 Through CpG JT andMarian AJ failure. pathways incardiomyopathiesandheart non-coding RNAsandepigeneticregulatory molecular mechanismsandfunctionsoflong Identification andcharacterizationof 1 , ChenX*, LombardiR, Willerson 1 . KnockdownofPlakophilin originates fromtheepicardium. tion thatfibroadiposis inthepatientstypically for thisstudyhasderivedfromtheobserva - factor] inthepathogenesisof AC. The impetus expression of Wt1 (Wilms Tumor 1)transcription cardial progenitorcells[EPCs, identifiedbythe Association. Beginning Grantin Aid from the American Heart in theheart. ForthisprojectIhavereceiveda cardiac myocytes, toinclude non-myocytecells of AC, commonlyrecognizedasadiseaseof in AC. The findingsexpandthecellularspectrum proteins andareamajorsourceofadipocytes pocyte progenitors(FAPs)expressdesmosome from theItalianSocietyofCardiology. Award Harvey the William and Association Heart Prize for Young Investigatorsfromthe American Arnold M. KatzBasicScienceResearch Award I wasnominatedforandwontheLouisN adipocytes in AC. Becauseofthesediscoveries, of mutantdesmosomeproteins, differentiateto set ofcardiacprogenitorcellsinthepresence tracer miceandsuccessfullyshownthatasub- ated andcharacterizedanumberoflineage ofpatientswith in theheart AC. Ihavegener- fication ofthecellularoriginfibro-adipocytes myocardium. by theprogressivefibrofattyreplacementof and suddencardiacdeath, andpathologically clinically bycardiacarrhythmias, failure heart in desmosomalgenes. AC ischaracterized familiar cardiomyopathycausedbymutation esis of Arrhythmogenic Cardiomyopathy(AC), a causal mutationsinfamilieswithHCM. Cardiomyopathy (HCM)andidentifiednovel ofpatientswithHypertrophic physiopathology death intheyoung. causesofsuddencardiac which areimportant pathogenesis ofhereditarycardiomyopathies, CENTER FORCARDIOVASCULARGENETICS I havegeneratedamouse modelof AC Recently, Iaminvestigatingtheroleofepi- Recently Ihaveshownthatcardiacfibroadi- My researchprogramisbasedontheidenti- Lately Iamstudyingthemolecularpathogen- In myearlierwork, Istudiedthecardiac My researchfocusisthedelineationof Phenotype oftheWt1-CreETR2:Dsp Dsp diac fibroblastsandadipocytesinthe Wt1Cre: disease. However, onlyasmallfractionofcar- and prematuredeath, resemblingthehuman increased adipogenesis, cardiacdysfunction transgenic mouseshowsepicardialfibrosis, causal gene, hasbeendeletedintheEPCs. The in whichDesmoplakin(Dsp), aknown AC hereditary cardiomyopathies Molecular geneticsandpathogenesisof Assistant Professor Raffaella Lombardi, M.D., Ph.D. adipogenesis. epicardialfibrosis(inred).shows severe Oil Red O(ORO)staininginthelowerpanels shows increased •Identification andmolecularcharacteriza- •Delineation ofthesignalingpathwaysinvolved RESEARCH PROJECTS therapeutictargets.and possiblynew it mayleadtothediscoveryofnovelbiomarkers This studyhasahightranslationalpotential, as which mediatefibrosisandadipogenesisin AC. tion oftheparacrinefactorssecretedbyEPCs Hence, Iamcurrentlyfocusingontheidentifica- tion ofcardiacresidentcelltypestoadipocytes. secreted byEPCsmaymediatethedifferentia- mutant EPCs, suggestingthatparacrinefactors opathies in thepathogenesisofhereditarycardiomy- w/f :Eyfp w/f micedirectlyoriginatefromthe w/f : Eyfp w/f mice. PicrosiriusRed(SR)stainingin theupperpanels •Paracrine roleoftheepicardiumincon- MS Research Assistant: GrazynaZCzernuszewicz, LAB MEMBERS contributed equallytothiswork) opathy. Circ Res . 2014;114:454-68. (*Authors Cardiomy- in Arrhythmogenic For Adipogenesis Pathway Is Activated And IsaCausalMechanism J T,Ruggiero, Marian AJ.Hippo Willerson The Chen SN*, GurhaP*, LombardiR*, Alessandra Circulation Research. 2016;119(1):41-54. pocytes upondeletionofthedesmoplakingene. proteins andpreferentiallydifferentiatetoadi- fibro-Adipocyte progenitorsexpressdesmosome GZ,nuszewicz JT,Willerson Marian AJ. Cardiac Lombardi R, ChenSN, Ruggiero A, GurhaP, Czer- KEY PUBLICATIONS Arrhythmogenic Cardiomyopathy tion ofcellularsourcesfibro-adipocytesin text of Arrhythmogenic Cardiomyopathy

19 • PACT REPORT PACT IMM IMMPACT REPORT • • • • 21 Figure 1: When we replace just 0.7% of the Figure 1: genome of injury susceptible SHR-A3 (red) with equivalent sequences from SHR-B2 (green) two important measures of renal injury (Glom TI Injury) are reduced to the same level as and The parts of the in resistant SHR-B2 (mauve). genome replaced affect a) blood pressure and b) the formation of antibodies. Dmitrieva, R.I. S. M. Cranford, and P. A. Doris. Ge- Doris. A. and P. Cranford, M. S. R.I. Dmitrieva, levelsnetic control of serum marinobufagenin in the spontaneously hypertensive rat and the Heart Amer. J. relationship to blood pressure. Aug 2017 Association. 6(10): In Press, The genetics of hypertension: an P.A. Doris, assessment of progress in the spontane- Physiological Genomics, ously hypertensive rat. 2017. 49(11):601-617, LAB MEMBERS Ph.D. Post-doctoral Fellows: Isha S Dhande, Research Assistants: Yaming Zhu, Sterling Kneedler We have discovered a genetic variation We Our genome contains templates on which Figure 2: When strongly stimulated, T lymphocytes limit inflammation by spontaneously dying (SHR-B2, T lymphocytes limit inflammation by spontaneously dying (SHR-B2, When strongly stimulated, Figure 2: T loss of gene function causes loss of In SHR-A3, right peak is dying cells). left blue peak is live cells, sustaining inflammation(only right blue peak). lymphocyte death responses, for immune cell that damages a protein vital It plays a key role in helping immune responses. believeWe cells respond to novel antigens. contributes to end that this genetic defect also this protein seems In particular, organ disease. T and B-lymphocytes communicate to help to prevent development of antibodies of low T and B cell inter- The protein alters specificity. can become allowing antibodies that actions, The reactive to normal tissues and injure them. genetic variant has several other effects on lymphocyte function. Complex Genomic Regions Structurally effective antibodies can be developed to permit recognition of an infinite variety of antigens. This arsenal must undergo adaptation in B cells. arsenals that each of us the individual However, divergence Similar possess are highly divergent. in the rat models we study alters disease sus- these highly complex to define Tools ceptibility. template regions of the genome are now emerg- have begun to apply them to describe We ing. how these genes vary and how they become We hope to determine adapted in inflammation. which specific template variations create the predisposition toward end organ injury. KEY PUBLICATIONS S.E. Hicks, M.J. M.C Braun, Zhu, Y. I, Dhande, Mycophenolate Doris. A. P. and Wenderfer mofetil prevents cerebrovascular injury in stroke-prone spontaneously hypertensive rats. 2017. Physiological Genomics 49:132-140, Peter A. Doris, Ph.D. Doris, A. Peter Director Professor/Center HoldsworthMary Elizabeth and Inflammatory University Chair in Metabolic Distinguished Disease Research end organ injury Genetics of cardiovascular Elevated pressure (BP) is the main blood One small region of chromosome 17 is HUMAN GENETICS FOR HUMAN CENTER driver of death from heart and kidney disease to But BP alone is not enough and stroke. also Susceptibility create these diseases. requires genetic factors that permit injury are We of the organs to advance to disease. uncovering which biological processes contain this genetic variation and how this leads to of hypertension use rat models that We illness. We differ in susceptibility to BP-induced injury. have learned that genetic susceptibility to organ genes Immune injury lies within immune genes. provide a defensive arsenal against attacks These pathogens evolve rapidly from pathogens. have to Our defenses to overcome our defenses. have enough pre-wired variety to quickly recog- nize pathogens and adapt immune responses can Imperfect adaptation specifically to them. allow our tissues to become damage targets. Increased BP triggers this damage by initiating variation de- but immune genetic inflammation, termines whether the result is an attack on our a dangerous Evolution has created own organs. in which our disease an arms race, equilibrium, resistance to pathogens requires some risk of harming ourselves. RESEARCH PROJECTS Mapping of Hypertensive End Organ Disease Loci responsible for elevation of BP in the injury SHR-A3, prone spontaneously hypertensive rat, compared to the injury resistant SHR-B2. Genome sequencing shows which genes may and several interesting genes be involved, One is involved in salt balance. have emerged. Another is part of the signal mechanism that permits B-lymphocytes to steer antibodies to We are subdividing this small specific targets. region in our animals in order to determine which part of this region of chr 17 contains the actual disease-causing genetic variation. Identified in Variants of Genetic Testing Disease Pathways All of us have had, or will have, one of our had, or will have, All of us have We have shown that kidney injury shown associated have We the from results blood pressure with increased emergence of auto-antibodies that damage understanding of developed have We tissues. and the interplay susceptibility to atherosclerosis new between drug targets, such as PSCK9, and population cells. Our by uptake lipoprotein global leaders in their field, geneticists are study in the making especially notable progress decline and age-related of susceptibility to stroke A significant fraction function. in cognitive rhythm from death results of sudden cardiac disruptions in that arise in genetic variation the electrical activity processing the proteins recruited a we have within the heart.year This new faculty member who is an emerging leader with a major proceeding We are in this field. to identify additional genetic new initiative disease and contributing to Alzheimer’s variation extending our neuro-degeneration, age-related cardiovascular studies of the interactions between function and brain disease in this new and critical direction. in life disrupted common by close relationships for Human the Center disease. In cardiovascular the opportunity for have to work we Genetics from change, pushing forward the knowledge which and from medicine draws which current medicine will be advanced. future Ph.D. A Doris, Peter & Professor Director Center Distinguished Holdsworth Mary Elizabeth and Inflammatory Metabolic Chair in University Disease Research

he Center for Human Genetics Genetics for Human he Center on generating squarely is focused new the about understanding

An emerging concept developing in our An emerging concept developing in the laboratories of our investigators Progress

HUMAN GENETICS FOR HUMAN CENTER

genetic risk for common cardiovascular for common cardiovascular genetic risk that information diseases and using therapies for to identify effective blood pressure these diseases. High that amplifies is a unifying element stroke, disease risk from cardiovascular heart diseases and kidney disease. These later life and emerge in middle and intricately linked to the normal so are variation genetic of aging. The processes and that has been that makes us unique impacts our parents passed to us from work our risk of these diseases. Our targets the identification of genes that diseases common contribute to cardiovascular in middle and later life and the mechanisms the in these genes re-shape variation which by which disease biological pathways through emerges. laboratories is that an important element of is system disease of the cardiovascular chronic a persistent state of that these diseases involve For example, in atherosclerosis inflammation. immune cells by wall is invaded the blood vessel plaques and the danger posed in atherosclerotic inflammation in of the ongoing level may reflect need a better understanding of these We them. in which our inflammation” of “sterile processes to in response immune systems become activated need We the emergence of damage to our tissues. that understanding of the genetic variants greater determine whether these inflammatoryresponses The advance. or even active subside or remain is challenge of identifying these genetic variants is a the fact that there complex by made more affecting in our immune lot of genetic variation to be able to adapt to the order In responses. continuous and rapid mutation of pathogens, genetic our immune systems harbor extensive us a head- can provide variation Such variation. pathogens. to newstart or evolving in responding risk of disease later in life. also can create it But and our increased have As our living standards provided lengthened, the advantages have lives to our health earlier in life can turn into threats cardiovascular our risk of chronic increasing by disease. and important insights. continues to yield exciting T

IMM PACT REPORT PACT • • • • 20 22 • IMMPACT REPORT cal age. We haveshownthatindividualswhose aging andcompared ittohis/herchronologi- methylation asamarker ofaperson’s biological ences andaging. Recently, wehaveusedDNA lifetime, andinresponsetoenvironmentalinflu- Some ofthemcanchangethroughoutthe DNA methylationmarksarenotfixedat birth. gene expression. UnlikeDNAsequencevariants, epigenetic mechanismusedbycellstocontrol tion andthesediseases. DNA methylationisan tion. glial proliferativepathwaysandneuroinflamma- matter lesionsonMRIanduncoveredaroleof genesinfluencingwhite ies haveidentifiednew risk forthesedisorders. Our recentgeneticstud- variants, eithercommon orrare, whichinfluence nome sequencingtechnologiestoidentifyDNA in theUnitedStatesandEurope, weapplyge- abnormalities. In collaborationwithresearchers for stroke, Alzheimer’s disease, andbrainMRI to discovernovelgenesinfluencingtherisk subjects. We usepowerfulgenometechnologies population samplesfromyoungadultstoelderly in itsclinicalandpre-clinicalformslarge neurodegenerative diseaseofthebrainboth genetics andgenomicsofvascular functional impairment, dementia, anddeath. with anincreasedriskofstroke, cognitiveand beginning inmiddleage, whichareassociated have consistentlydetectedbrainabnormalities such asmagneticresonanceimaging(MRI) diagnosis. Indeed, neuroimagingtechniques before manifestationofsymptomsorclinical these disordersbeginyears, ifnotoverdecades reach 16million. that There isgrowing evidence disease. By2050, thisnumberis projected to than 5million Americans arelivingwiththe now thesixth-leadingcauseofdeathandmore term disabilityforadults. Alzheimer’s diseaseis States andisamajorcauseofseriouslong- the fifth-leadingcauseofdeathinUnited public healthproblemsofourtime. Strokeis and dementia, areamongthemostsignificant CENTER FORHUMANGENETICS We alsostudythelinksbetweenDNAmethyla- My researchprograminvestigatesthe Diseases oftheagingbrain, suchasstroke on MRI.tionandthese brainabnormalities. cal ageissignificantly olderthantheirchronologicalageare at increasedriskofbrainabnormalities DNA methylationprofiling canbeusedtoestimateaperson’s biologicalage. Individualswhosebiologi- E., Mosley, T.H., &Fornage, M. Cerebral White tesman, R.F., Guan, W., Pankow, J.S., Boerwinkle, Raina, A., Zhao, X, Grove, M.L., Bressler, J., Got- KEY PUBLICATIONS •Discovering novelgeneticvariantsinfluencing •Discovering novelgeneticlociforcardiovascu- •Discovering novelepigenetic(DNAmethyla- •Discovering commonandraregeneticvariants RESEARCH PROJECTS disease progression. orslow therapeuticstoprevent ment ofnew - disease mechanismsandleadtothedevelop function impairment. brainabnormalitiesandcognitive developing their chronologicalageareatincreasedriskof methylation profileissignificantlyolderthan biological ageestimatedfromtheirDNA ofbrainvasculardiseaseandaging Molecular epidemiology The LaurenceandJohannaFavrotDistinguishedProfessorshipinCardiology Professor Myriam Fornage, Ph.D. ancestry in adultsofEuropean, African, andHispanic cognitive functionand Alzheimer’s disease genetic variationtoinfluencebloodpressure. how depressionandanxietyinteractwith pathway analysis. Inparticular, discovering lar traitsusinggene-lifestyleinteractionsand cognitive outcomes small vesseldiseaseanditsrelatedneuro- tion) variantsthatinfluenceriskforbrain genome sequencing ties usinglarge-scalegenotypingandwhole influencing MRI-definedbrainabnormali- insightsinto These discoveriesmayyieldnew cian; Ping Wang, Ph.D., Research Associate Research Assistants: RuiXia, Ph.D., Biostatisti- Graduate Students:DaokunSun, B.S. Melissa Richard, Ph.D. Post-doctoral Fellows:XueqiuJian, Ph.D.; LAB MEMBERS e44-e71, 2017 with silentcerebrovasculardisease. Stroke48, Hypertension. ofstrokeinpatients Prevention ics and Translational andCouncilon Biology; and Intervention;CouncilonFunctionalGenom- Council; CouncilonCardiovascularRadiology behalf ofthe American Heart Association Stroke Higashida, R.T., Kasner, S.E., Seshadri, S.;on F.N., Fornage, M., Gorelick, P.B., Greenberg, S.M., Smith, E.E., Saposnik, G., Biessels, G.J., Doubal, vessel stroke. AnnNeurol81:383-394, 2017 Variation at16q24.2isassociatedwithsmall tional StrokeGenetics, Consortium. Genetic Group oftheCHARGEConsortium, &Interna- Stroke GeneticsNetwork, Neurology Working STROKE, UK Young LacunarDNAStudy, NINDS Traylor, M., Malik, R., etal. onbehalfofMETA - 2017 Communities Study. Clin. Epigenet., 9:21-29, in Risk Epigenetic Aging:TheAtherosclerosis of Matter HyperintensitiesonMRIand Acceleration in endothelialcells. Thus, PCSK9contributes atherosclerosis anddown regulationof TGF-β ester andphospholipids, resultinginincreased of cholesteryl levels LDL containingelevated lipogenesis, whichproducedmoreatherogenic 2).(Figure PCSK9 modulatedSREBP-1c of both AMPK andULK1kinasecomplexes including activationofp-AKTandinduction signalingpathway PCSK9 regulatedautophagy 1).LDL cholesterolconcentrations(Figure that islipolyticprocessedtoincreasedplasma VLDL (verylowdensitylipoprotein)secretion pathway. This resultedinincreasedhepatic degradation viatheautophagosome/lysosome its cytoplasmic apoBinawaythatprevents for PCSK9-i.e. PCSK9 interactsdirectlywith Figure). We discovered anovelintrahepaticrole secretion, andendothelial cellfunction(Cover rosis development, apolipoproteinB(apoB) we showedthatPCSK9modulatesatheroscle- /-Pcsk9-/-). Usingthesetwomousemodels, knockout mousemodel(LTp=Ldlr-/-Apobec1- PCSK9 genefromLDbmicetogenerateatriple and prematurecoronarydisease. We deleted haveincreasedplasma cholesterol PCSK9 levels gene forhyperlipidemia. Patientswithelevated identifiedcausative kexin type9)isanewly females. than atherosclerosis fasterandmoresever development. Moreover, malemicedevelop high-fat dietacceleratestheiratherosclerosis age.erosclerosis asthey Feedingona Western Ldlr-/-Apobec1-/-). ath- These micedevelop LDL receptorandanRNAeditingenzyme(LDb; with hyperlipidemiabydeletingbothgenesof made amousemodelthatmimicshumans between atherosclerosisandimmunity. We have sclerosis andtoelucidatethecross-regulation ofathero - understanding thedevelopment vasculature.arterial Ourresearchfocuseson maladaptive immuneresponsesthataffectthe metabolism thatleadstohyperlipidemiaand age. The diseaseincludesimbalance lipid aswe thatincreasesitsseverity in theaorta CENTER FORHUMANGENETICS PCSK9 (proprotein convertase subtilisin/ PCSK9 (proproteinconvertase Atherosclerosis isaninflammatorydisease tion. sinusquantifica - compared withLDbmice asdeterminedby(A)enfacequantification and(B)aortic 2.Figure PCSK9modulatesatherosclerosis. LTp significantlylessatheroscleroticlesions micedevelop •Using genetictoolsandproteomicstoidentify •Using CRISPR/Cas9techniquetogenerateIL- •The roleofPCSK9inautophagy, inflammation, RESEARCH PROJECTS of diseases therapeutic approachestocombatprogression process. efficient Itwillprovidebasistodevelop of physiologicalandpathologicaldisease will provideinsightintotheunderstanding modulate diseasedevelopment. Ourdiscovery mechanisms bywhichproatherogenicfactors Cas9 todefinethecellularandmolecular including RNA-Seq, ATAC-Seq, RPPA andCRISPR/ lipid metabolismandcellularimmunefunction. to atherosclerosisthroughitseffectonhepatic atherosclerotic vasculardiseases ofgeneticandcelltherapiesforthetreatment development Pathogenesis ofatherosclerosisandimmunitythe DiseaseResearch Distinguished ProfessorshipinHeart The JerryandMauryRubensteinFoundation Professor Ba-Bie Teng, Ph.D., FAHA genes associatedwithatherogenesisandto on atherosclerosis. 17 RCtripleknockoutmicetostudyitseffect and atherosclerosis. Our laboratoryisusingcurrenttechnologies Christine Huang;NithyaNarayana Summer InternshipStudents:SaxonHancock; Research Assistant: ZhengmingXu Scientist: YunlongPh.D. Wang, Visiting Research Scientist:HuaSun, Ph.D. LAB MEMBERS Dec, 2017). 648timeontheJLRwebsite inNovand viewed Research asshownbelow). was This article wasselectedastheCoverforJ.article Lipid Ph.D., 2017, J. LipidResearch , inpress. (This M.D.; Jeffrey T. Chang, Ph.D. andBa-Bie Teng, Dysfunction: HuaSun, Ph.D.; RonaldM. Krauss, Apolipoprotein BSecretion And Endothelia PCSK9 DeficiencyReduces Atherosclerosis, KEY PUBLICATIONS suppresses theautophagy. pathway; itincreasesp-AKTandp-AMPK (4). (2). signaling PCSK9regulatesautophagy on endothelialcells, affecting atherogenesis differences inLDLinducesdifferentreaction LDL compositions(3). The consequenceof ing theproductionof VLDL andalteringthe degradationpathwaybyincreas- autophagy (1). PCSK9interactswith ApoB, modulatesthe tophagy, altersLDLandaffectsatherogenesis. PCSK9 interactswith ApoB, modulatesau - atherosclerotic vasculardiseases. genetictherapyforthetreatmentof develop

23 • PACT REPORT PACT IMM IMMPACT REPORT • • • • 25 The center recently established a robust recently established a The center Asthma and Sinusitis Retinopathy Diabetic & Autoimmunity Immunology Mucosal Disease Infectious Microbial Injury and COPD Lung Acute Deficiencies Surfactant Lung Degeneration Macular Medicine Regenerative Pulmonary erythematosus. focused on the development program research of of stem cell therapeutics for the treatment lung diseases and for genetic acute and chronic function as deficiencies that affect normal lung diseases, including macular as for major eye well degeneration and diabetic retinopathy. include: interests Research • • • • • • • • Ph.D. Rick Wetsel, & Professor Director Center and Irma M.D., Ph.D. J. Müller-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 CENTER FOR CENTER AUTOIMMUNE AND IMMUNOLOGY DISEASES 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 their ligands in modulating immune and 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 T KEY PUBLICATIONS YI, Li A, Harpak Shim H, SH, Wang A, Raj Pritchard Y, Gilad Stephens M, Engelmann B, of novel translated open Thousands 2016. JK. by ribosome reading frames in humans inferred eLife 5: e13328. footprint profiling. multiple tissues using ribosome profiling and CRISPR-CAS9 knockout screens. ing to investigate translational regulation of ing to investigate translational protein synthesis stress response Genotype of a genetic variant is associated with uORF regulation of protein translation at HMSD locus Negative correlation in the levels of protein translation between the two Open Reading in HapMap LCL. Frames at HMSD locus is clearly shown through stratifying ribosome profiling data by genotype. •Identification of functional smORFs across Using RNA binding protein footprint sequenc- •Using RNA binding protein footprint Deciphering the regulatory code: a functional genomics Deciphering the regulatory translation approach to protein RESEARCH PROJECTS by uORF in •Regulation of protein translation Sidney Wang, Sidney Wang, Ph.D. Assistant Professor Regulation of gene expression is fundamental

From to a wide range of biological processes. cell fate determination during development to malignant transformation during tumori- precise control of gene expression genesis, current Our forms the basis of these processes. however, understanding of gene regulation is, that Most published studies far from complete. profile gene expression are transcript-centric they focus on measuring mRNA levels and (i.e. While levels factor binding). of transcription these efforts revealed intricate networks of cooperativity amongst transcription factors in much of shaping complex biological processes, the post-transcriptional regulation are left unex- the process It remains unclear whether plored. of protein translation is regulated by a network of factors to an extent of complexity similar to ask questions such We transcription regulation. proteins “Do sequence specific RNA binding as (RBP) cooperate in controlling translation?” networks there translational regulatory “Are that orchestrate critical biological processes?” Our research program focus on addressing these questions in biological contexts that are Our immediate goals relevant to human health. are to develop novel tools to systemically study RBP binding; to investigate regulatory functions of upstream Open Reading Frames (uORFs); and to integrate these functional genomics in annotations with results from genetic studies, order to fine map the regulatory variants and to provide mechanistic understanding for disease associated variants. HUMAN GENETICS FOR HUMAN CENTER

IMM PACT REPORT PACT • • • • 24 26 • IMMPACT REPORT IMMUNOLOGY AND AUTOIMMUNE DISEASES IMMUNOLOGY ANDAUTOIMMUNE CENTER FOR pathogenesis ofvarious diseasepathologies. numerous biological roles ofcomplementinthe of thesemicehasfacilitated thediscoveryof mouse embryonicstemcells. The generation targeting andhomologousrecombination using receptors havebeenselectivelyablatedby gene complement proteins, regulators, andcell mice inwhichthegenesencodingspecific ment, wehavegeneratednumerous “knock-out” andbiologicalfunctionsofcomple - importance asthma andCOPD. To determinetheoverall a majorcontributortolungdiseasessuchas been discoveredasamajorcauseof AMD and dysregulation ofthecomplementsystemhas these novelcomplementbiologicalfunctions, of thecentralnervoussystem. Inadditionto cells including T-cells, andnormaldevelopment tissue regeneration, polarization ofimmune and viruses. These otherfunctionsinclude biological functionsotherthankillingbacteria inother complement systemisveryimportant inthepastdecade thatthe become evident plasma proteinsandcellularreceptors. Ithas and viralpathogens. Itiscomprisedofover30 being thefirstlineofdefenseagainstbacterial innate immunesystemandiswellknownfor The complementsystemisamajorarmofthe has involvedstudiesofthecomplementsystem. pathological conditions. Muchofthisresearch duringbothnormaland the lungandeye the inflammatoryandimmuneresponsesin moleculesthatmediate delineating thekey yearsfocused on tory hasforthepastseveral of thelungandeye. Accordingly, ourlabora- for theeffectivetreatmentofchronicdiseases ment ofnoveltherapeuticsthatcouldbeused - and repairprocesseshasslowedthedevelop immunity,regarding lungandeye inflammation, paucity ofcellularandmolecularknowledge normal tissuerepairmechanisms. However, the pathologies duetofibrosis, andimpairmentof of healthytissue, establishmentofdebilitating toxic substances, resultinginthedestruction and inflammatoryresponsetopathogenicor often theresultofdysregulationimmune Chronic diseases of the lung and eye are Chronic diseasesofthelungandeye Cells; Red-C3aRorC5aR1. endothelial cellsfollowingstimulationwith eitherC3aorC5a. Blue-cellnucleus; Green-Endothelial Expression ofthecomplementanaphylatoxin receptors(A. C3aRandB. C5aR1)byhumanvascular Develop hES-retinalpigmentepithelial cells •Develop •Evaluate thetherapeuticpotentialofgene •Generate “universal donor” embryonicstem •Determine howthefunctionofvascularand RESEARCH PROJECTS deficiency ofsurfactantproteinB. bornwithgenetic gene therapyfornewborns thelium inacutelunginjury, andforcellbased AMD, forregenerationofthedamagedlungepi- derived cellsforrepairofdamagedretinain (hES) andinducedpluripotent(iPS)stemcell investigating thetherapeuticuseofembryonic ofthisoverallresearchprogram,As part weare therapeutic targetforthetreatmentofasthma. and thereforemayprovetobeanexcellent asthma, includingairwayhyperresponsiveness, hallmarksof mediatorofkey is animportant the complementanaphylatoxinpeptideC3a C3a receptorwasdeleted, wediscoveredthat For example, instudiesusingmicewhichthe stem celltherapeuticsfordiseasesofthelungandeye 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. therapeutics fortreatmentof AMD surfactant proteindeficiencies corrected iPScell-derivedlungcellsfor transplantation plantable cellsthatwillnotberejectedafter cell linesthatcanbedifferentiatedintotrans- complement duringtheimmuneresponse lymphatic endothelialcellsareimpactedby Senior Research Assistant: Yi-Dong Li Research Assistant:Yi-Dong Senior PoojaShivshankar,Research Instructor: Ph.D. Post-doctoral Fellow:JohnL. Mazzilli, M.D. Research Scientist:KenSimmons, Ph.D. Ph.D. Mueller-Ortiz,Senior ResearchScientist:Stacey LAB MEMBERS 221: 1407-1417(PMID:27476791). monocytogenes 2016: infection.Immunobiology complement inthehostresponsetoListeria nate andadaptiveimmunologicfunctionsof Calame DG, SL, Mueller-Ortiz Wetsel RA. In- 2017: 18:37-40(PMID:28395800). retinal pigmentepithelialcells. StemCellRes. stem celllineCR-4:differentiationtohuman and characterizationofthehumanembryonic Garcia CA, Wetsel RA, ZsigmondEM. Derivation Mazzilli JL, Domoshirov AY, SL, Mueller-Ortiz 3237-3244 (PMID:28275134). surveillance pathway. JImmunol.2017: 198: of thecyclicdinucleotide-activatedcytosolic sponse toListeriamonocytogenes byinhibition C5a andC3a, suppressIFN-βproductioninre- Wetsel RA. The complementanaphylatoxins, SL,Mueller-Ortiz CalameDG, ShenoiN, Li Y-D, KEY PUBLICATIONS Investigation of adenosine transport in acute •Investigation ofadenosine transport •Examining theroleof A2B adenosinereceptor RESEARCH PROJECTS treating chroniclungdisease. approaches helpusidentifynovelstrategies for the Texas MedicalCenter. These translational obtained followinglungtransplantationherein work extensivelywithhumanexplantedlungs celltypesand experiments indiseaserelevant and signaling. We alsoconductmechanistic mice ofcomponentsadenosinemetabolism in chroniclungdisease. This includesknockout mice toexaminetheroleofadenosinesignaling context oflunginflammationandremodeling. ing influencestheactivitiesofthesecellsin ing themechanismsbywhichadenosinesignal- projects inmylaboratoryfocusonunderstand- cells andsmoothmusclecells. Mostofthe inflammatory cells, fibroblasts, airway epithelial adenosine receptorsontargetcellssuchas ing. Adenosine signalsbyengagingspecific promote airwayinflammationandremodel- accesspathwaysthatserve to in thelungthey increase is ourbeliefthatasadenosinelevels is generatedinresponsetocelldamageandit of lunginflammationanddamage. Adenosine the signalingmoleculeadenosineisanamplifier strategies. noveltherapeutic with theintentofdeveloping that regulatethechronicityofthesedisorders goal ofmylaboratoryistoidentifypathways chronic natureofthesediseases. The major signaling pathwaysthatservetoregulatethe have beendescribed, littleisknownaboutthe associated withthegenesisofthesediseases nary hypertension. Although signalingpathways disease (COPD), pulmonaryfibrosisandpulmo- such asasthma, chronicobstructivepulmonary prominent featuresofchroniclungdiseases DISEASES IMMUNOLOGY ANDAUTOIMMUNE CENTER FOR and chroniclunginjury the progressionofpulmonary fibrosis expression onpulmonarymacrophagesduring We makeextensiveuseofgeneticallymodified A centralhypothesisofmylaboratoryisthat Inflammation andremodelingresponsesare Adenosine signalingandtheregulationofchroniclungdisease 874-883. PMID:26527068 exacerbation ofpulmonaryfibrosis. FASEB J. 30, areassociatedwiththeprogression and levels Blackburn, M. R. (2016)Extracellularadenosine Volcik, K. A., Liu, H., Xia, Y., Eltzschig, H. K. and Quintana, H., Molina, J. G., Davies, J., Philip, K., Luo, F., Le, N. B., Mills, T., Chen, N. Y., Karmouty- PMID: 12871304 pulmonary fibrosis. FASEB J. 31, 4745-4758. activated macrophagesandcontributesto regulates the ADORA2B receptoronalternatively H. K. andBlackburn, M. R. (2017)HIF1Aup- Amione-Guerra, J., Sinha, N., Guha, A., Eltzschig, Karmouty-Quintana, H., Luo, F., Molina, J. G., Philip, K., Mills, T. W., Davies, J., Chen, N. Y., KEY PUBLICATIONS approachestounderstand •Systems Biology •Understanding novelmechanisticrolesfor •Examination ofthehypoxiaasanamplifier •Novel regulationofmRNApolyAtailsinthe Executive PresidentandChief Vice Academic Officer, UTHealth S.William KilroySr., ChairinPulmonaryDisease Professor and ofBiochemistryandMolecularBiology ChairmanDepartment Vice The Universityof Texas GraduateSchoolofBiomedical Sciences atHouston Dean andJohnP. McGovernDistinguishedProfessorofBiomedicalSciences Michael R. Blackburn, Ph.D. fibrosis (BLEO). Increased expression(browncolor)ofproteinases inpulmonarymacrophagesmicewith the progressionofchroniclungdisease IL-6 signalinginpulmonaryfibrosis chronic lungdisease Obstructive PulmonaryDisease regulation ofpulmonaryfibrosisandChronic from geneticallymodifiedmice. Primary typeIIalveolarepithelialcellsisolated Graduate Student:JoshKo, Ph.D. Research Scientist:JoseMolina, Sr. Research Associate: Ning-Yuan Chen Senior ResearchScientist:Kelly Volcik, Ph.D. Ph.D. Weng, Tingting Professor: Assistant LAB MEMBERS sion. FASEB J. 29, 50-60. PMID:25318478 - dampens lungfibrosisandpulmonaryhyperten (2015) Deletionof ADORA2B frommyeloidcells R. A., Xia, Y., Eltzschig, H. K. andBlackburn, M. R. L., Le, Bao, Bunge, I., Volcik, K., Le, T., Johnston, Weng, T., Molina, J. G., Luo, F., Davies, J., Acero, Karmouty-Quintana, H., Philip, K., Chen, N. Y.,

27 • PACT REPORT PACT IMM 28 • IMMPACT REPORT IMMUNOLOGY AND AUTOIMMUNE DISEASES IMMUNOLOGY ANDAUTOIMMUNE CENTER FOR separating thesinuses fromtheintracranialand of sinuscavitiesand ultimately erosionofbone ties. This trapped mucuscancauseexpansion between thenasalpolypsandwithinsinus cavi- laden withfungalhyphaeandeosinophils with anaccumulationofthickentrapped mucus clinical subtypeofCRSwNPthatisassociated treatment. phenotypes, allowing forpossiblepersonalized has identifiedendotypeswithintheseclinical using clusteranalysisofgeneticinformation 5, andIL-13. However, recentstudybyourlabs expression oftype2cytokinessuchasIL-4, IL- of eosinophils, mast cells, andbasophils with nasalpolyps(CRSwNP)hashighpresence helper celltype1(Th1)cytokines, whileCRS predominance ofneutrophilsandelevated T CRS withoutnasalpolypsischaracterizedby logic profilesoftheinflamedsinustissue. generallybyimmuno- has beensupported polyps (seeimage1). This clinicalclassification defined bytheabsenceorpresenceofnasal tion ofthepatientanddiseasedmucosa. - regularly inclinicwhichallowsperiodicevalua harvest criticaldiseasedtissueandareseen to undergo surgeryprovidinganopportunity matory respiratorydiseases. CRSpatientsoften research modelforstudiesinchronicinflam- not curative. CRSrepresentsanidealhuman surgeries andanti-inflammatoryagents, are ment options, whichtypicallyinvolverecurrent about itspathophysiology, andcurrenttreat- this healthcareburden, muchremainsunknown chronic illnessesintheUnitedStates. Despite diseases representsomeofthemostprevalent Together, CRSandasthmaaschronicrespiratory from CRSoftenarediagnosedwithasthma. 3 billiondollars. Inaddition, patientssuffering annual directhealthcaretreatmentcostofover 18-22 millionphysicianvisitsyearlywithan These symptomsultimatelydriveconservatively and pressure, nasalcongestionandobstruction. rhinosinusitis (CRS), whichcausesfacialpain Allergic fungalrhinosinusitis(AFRS)isa CRS isclinicallyclassifiedintotwogroups Over 40million Americans sufferfromchronic •Clinical characterization andidentificationof •Molecular signalingthroughrespiratory •Immunologic characterizationofimportant RESEARCH PROJECTS intersect thesetwoentitiesasitrelatestoCRS. elucidating thespecificsofpathwaysthat and inflammation. Studiesarefocusingonthe a diseaseatthecrossroadsofcoagulation us toourrecentinterestinCRSrepresenting response insomeCRSsubtypes. This hasled responsible forsignalingintheinflammatory 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 for sinonasalmucosal. Recentstudieshave first lineofdefenseagainsttheenvironment complications andblindness, respectively. orbital cavitieswhichcanresultinintracranial chronic airwayinflammation Environmental triggersregulatinginnateimmuneresponsesin –HeadandNeckSurgery Otorhinolaryngology Associate Professor, and CenterforImmunology of Autoimmune DiseasesandDepartment Amber Luong, M.D., Ph.D. immune response ing and/ormaintaining thecharacteristic Th2 environmental triggersresponsibleforinitiat - epithelial cellsoffungialoneandwithother response asameansofendotypingCRS cell typesinvolvedinthe Th2 immune In addition, mylab isinterestedinthe Given theappreciationofinnateimmunity Our labhasfocusedontheroleofIL-33in Respiratory epithelialcellsrepresentthe Epithelial cells Samantha McMichael, B.A. Caroline J. Padro Dietz, Ph.D. LAB MEMBERS Immunol, 2017Mar;139(3):1061-1064. in chronicrhinosinusitissubtypes.Clin JAllergy distincttype2inflammatorypatterns reveals Luong A. Largescalegeneexpressionprofiling etz CJ, HuX, CitardiMJ, FakhriS, Assassi S, and Tyler MA, RussellCB, SmithDE, RottmanJB, Di- Aug 134(2):325-331. Airway Disease. J Allergy ClinImmunol.2014 Surface MycosisinChronic Th2-Associated 1 Kheradmand F, CorryDB, andLuong A. Airway CL, CitardiMJ, FakhriS, ShawJL, Fothergil A, P,Porter LimDJ, MaskatiaZK, MakG, Tsai 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. Science.2013 Aug 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 KEY PUBLICATIONS nasal cavityofCRSwNP patient. Nasal polypsseenby nasalendoscopywithin biomarkers forCRSsubtypes polyps Nasal My research efforts arefocused on1)toisolate My researchefforts a goodmodeltostudy lungdiseaseprocesses. injuries andgeneticdisorders, butalsoprovide can betherapeuticallyusedtotreatdistal lung are notonlyapromisingsourceofcellsthat cell deriveddistallungstem/progenitorcells disease-specific inducedpluripotentstem (iPS) diseases. The embryonicstem(ES)cell/lung use ofstemcellsinthetreatmentpulmonary inthetherapeutic able interesthasdeveloped diseases. years, During the pastfew consider- mechanisms underlyingpathogenesisoflung targetsforexploringthe cells representthekey doubt,Without thedistallungstem/progenitor to facilitatethetreatmentoflungdiseases. ofnoveltherapies tive needfordeveloping notpositive.is unfortunately There isanimpera- limited timerange, andthe long-termoutcome symptoms andimprovelifequalitywithina for thosepulmonarydiseasesatbestrelease In addition, currently availabletreatments disease processesbothinvivoandvitro. model availableforstudyingthebiologicaland since thereisnoreliablebiomedicalresearch well asthecorrespondingrepairmechanisms, pathogenesis ofthesepulmonarydiseasesas cancer. littleisknownregardingthe However respiratory distresssyndrome(RDS)and fibrosis, asthma, CODP, cysticfibrosis, neonatal pulmonary diseases, includingpulmonary ofchronic orsevere role inthedevelopment deficiencies isthoughttoplayanimportant progenitor celltypesduetoinjuriesorgenetic Loss ofnormalfunctionsanythesestem/ branch point-associatedneuroepithelialbodies. at thebronchioalveolarductjunctionand and 3)asubsetofvariantClaracellslocated transient amplifyingbronchiolarClaracells; tions: 1)alveolarepithelialtypeIIcells;2)the maintaining distallungepithelialcellpopula- stem/progenitor celltypesresponsiblefor from theenvironment. There areatleastthree that areconstantlyexposedtoinjuriousstimuli airway andalveolarepithelialcellpopulations critical forthemaintenanceofhomeostasis DISEASES IMMUNOLOGY ANDAUTOIMMUNE CENTER FOR Lung epithelialstem/progenitorcellsare •Isolation andcharacterizationofES/iPScell RESEARCH PROJECTS cell-derived exosomemiRNApathways. and4)toexplorelungcancerstem sue repair; endogenous stem/progenitorcellsforlungtis- fortargetedactivationof a novelstrategy during thediseaseprocessesfordeveloping control distallungstem/progenitorcellfate characterize factorsorregulatorypathwaysthat for lungtissueregeneration;3)toidentifyand cell-basedgenetherapeuticstrategy developing studying pulmonarydiseaseprocessesandfor cal grade” lungdisease-specificiPScellsfor both invitroandvivo;2)togenerate “clini- derived distallungstem/progenitorcelltypes and characterizehumanmouseEScell trate thePI3K/Aktsignaling. StemCellRes Ther. 2017Jun 8;8(1):138-51 miR-371b-5p promotes proliferationoflungalveolarprogenitor typeIIcellsbyusingPTENtoorches- Quan Y., Wang Z., GongL., PengX., RichardM., ZhangJ., Fornage M., Alcorn JL., and Wang D. Exosome Lung stem/progenitorcellsanddiseases Assistant Professor M.D.Dachun Wang, derived distallungstem/progenitorcells •Therapeutic potentialofES/lungdisease-spe- •Characterization oflungcancerstemcell- Research Associate: Dr.Research Associate: Quan Yuan LAB MEMBERS Stem CellRes Ther. 2017Jun8;8(1):138-51 PTEN toorchestratethePI3K/Aktsignaling. of lungalveolarprogenitortypeIIcellsbyusing Exosome miR-371b-5ppromotesproliferation Zhang J., FornageM., Alcorn JL., and Wang D. Quan Y., Wang Z., GongL., PengX., RichardM., KEY PUBLICATIONS cells forthetreatmentoflungdiseases cific iPS-deriveddistallungstem/progenitor cancer cellgrowthandmetastasis derived exosomemiRNApathwayscontrolling

29 • PACT REPORT PACT IMM IMMPACT REPORT • • • • 31 How does the brain and circadian clock and circadian does the brain How metabolism lead to circadian does altered How glucose control does the brain How the functions of the genes mutated in What are of proteins does abnormal processing How disease impact Alzheimer’s does stress How laboratories Collaboration among the center’s to treat muscle diseases? to treat • energy balance? the body’s control • cancer? • homeostasis in type 1 and type 2 diabetes? • diseases? neurodegenerative • degeneration? cause neuronal • pathogenesis? increasing thereby synergy, research promotes The center’s and innovation. productivity members collaborate with pathologists, epidemiologists, and clinicians to translate for the benefit of patients with their discoveries diseases. metabolic and degenerative Ph.D. Kolonin, Mikhail & Associate Professor Director Center Chair University Distinguished Jr. E. Bovay, Harry Disease Research in Metabolic he Center for Metabolic and Degenerative and Degenerative he Center for Metabolic integrates eight laboratories Diseases aging-associated diseases, investigating How do progenitor cells in adipose tissue do progenitor How does dysfunction of adipose cells promote How and inflammation in adipose does fibrosis How is angiogenesis implicated in adipose and How Can cells in adipose tissue be targeted for What transcriptional pathways can be targeted METABOLIC AND DEGENERATIVE DISEASES DEGENERATIVE AND FOR METABOLIC CENTER including muscle wasting, neurodegeneration, including muscle wasting, neurodegeneration, the and type-2 diabetes. Obesity, cancer, focus underlying medical condition, is a research Aging and obesity-associated groups. of several changes in energy metabolism, cell signaling, homeostasis, and cell fate determination protein being that lead to physiological abnormalities are in animal models and studies on interrogated clinical specimens. The specific questions being faculty include the the center’s by addressed following: • adipogenesis? commit to white and brown • of diseases and aging? progression • tissue affect insulin sensitivity? • muscle tissue remodeling? • therapeutic purposes? • T degeneration models Retinal pigment epithelial cells were derived from the NIH-approved CR-4 human ES cell The cells were characterized for molecular line. tested for correct function and safety markers, in an animal model of age-related macular degeneration. RESEARCH PROJECTS macular •Stem cell therapy for age-related implantation •Patient-derived tumor xenograft KEY PUBLICATIONS Mueller-Ortiz, A.Y., Domozhirov, J.L., Mazzilli, E. and Zsigmond, R.A. Wetsel, C.A., Garcia, S.L., M.: Derivation and characterization of the hu- man embryonic stem cell line CR-4: Differentia- tion to human retinal pigment epithelial cells. 2017. 18:37-40, . Stem Cell Research LAB MEMBERS Senior Research Associate: Aleksey Domozhirov M.D. Post-doctoral Fellow: John Mazzilli, lines, including human ES cells approved for including human ES cells approved lines, NIH-funded research in the production of knock-out mice when using ES cells derived at the facility 23%) cells and MEF feeder layer cells cell lines preservation of mouse ES cells eggs eggs of knock-out and knock-in mice the production of transgenic animal models the production of transgenic Pronuclear microinjection of mouse zygotes for the production of genetically-engineered animal models. •100% success rate in re-derivation of mice ••Derivation of over 20 mouse and human cell •100% success rate of germline transmission Accomplishments: •Consistently high transgenic rates (average •Availability of germline-competent mouse ES •Derivation of novel mouse ES cells and other •CRISPR/Cas9-mediated genome editing cryo- expansion, selection, •Gene targeting, •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. Associate Professor Facility and Stem Cells Core Transgenic Director, The Transgenic and Stem Cells Core Facility and Stem Cells Transgenic The - cryopreserva In addition to the production, The embryonic stem (ES) cell lines derived in functional human RPE cells have derived We

provides a unique service to the UTHealth scientific community by generating animal models of specific human diseases in order to The laboratory was develop treatments. novel it established in 1998 and since that time, has generated more than 800 new transgenic, knock-out and knock-in animal models for as for as well investigators from UTHealth, scientists from numerous other academic institutions. 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 cell culture and stem of animal microsurgery, cells research. the Core Laboratory are highly effective for stud- a current In ies involving cellular differentiation. human our laboratory is using research project, ES cell-derived retinal pigment epithelial (RPE) cells as a therapeutic strategy for the treatment of age-related macular degeneration (ARMD). ARMD is a leading cause In the United States, The aim of our study is to use RPE of blindness. cells derived from human ES cells in a clinical trial of sub-retinal transplantation into patients ARMD for the reversal of the visual loss with associated with the disease. in our laboratory and are currently testing the efficacy and safety of these cells in animal we will In preparation of clinical trials, models. examine the long-term viability of the trans- ARMD planted cells in murine animal models of and we will generate transplantable human RPE cells in a GMP-certified facility. CENTER FOR CENTER AUTOIMMUNE AND IMMUNOLOGY DISEASES

IMM PACT REPORT PACT • • • • 30 32 • IMMPACT REPORT CENTER FORMETABOLIC AND DEGENERATIVE DISEASES recently showedthatD-WAT targetshuman In collaborationwithbariatricsurgeons, we obesity andsuppressestumorgrowthinmice. publications demonstratethatD-WAT prevents experimental drug(D-WAT) targeting ASCs. Our are available. thefirst We havedeveloped in fattissueforobesityorcancerindications progression. Currently, nodrugstargetingcells that tumorsrecruitthese ASCs thatfuelcancer are generated. Ourlaboratoryhasdiscovered In obesity, increasednumbersofwhitefat ASCs are maintainedbyadiposestemcells(ASCs). replaced asweage, andtheirpoolsinfattissue white andbrownadipocytesarecontinuously theonsetofmetabolicdisease.prevent Both and cancer. Incontrast, activebrownfatcan causing diabetes, cardiovasculardiseases, becomes inefficientinholdinglipids, hence the bodywarm. Inobesity, overgrownwhitefat while brownadipocytesburnlipidsofftokeep scarcity,and releasethemintimesofenergy lipids differently. White adipocytesstorelipids are twotypesoffatcells(adipocytes)thatuse acting likeaspongebyabsorbinglipids. There can causemetabolicdiseaseiffattissuestops source,molecules servingthebodyasanenergy the functionofotherorgans. Lipids, the Blue: nuclei. genes.lineage tracingofprogenitor cellsbyusinggreenandredreporter Distinct lineagesofadipocytes identifiedinmousefattissue through Health reliesonfattissuethatcoordinates report, 15(12), 3144-3150, 2017. metastasis tobone. CancerResearch . Priority and proteinase3mediatesprostatecancer Interaction betweentumorcellsurfacereceptor Kolonin M.G., etal., PasqualiniR, Arap W. KEY PUBLICATIONS • Cancer-associatedadipocytesasthesource • Aging-associated exhaustionofadipocyte • Adipocyte progenitors:lineages, andfunction RESEARCH PROJECTS the consequencesofD-WAT treatmentinaging. forming studiesinmousemodelstoestablish models. To ensuresafetyofD-WAT, weareper- This hypothesisisnowbeingtestedinanimal it involvesexcessivefatcellnumberexpansion. leads toprematureexhaustionof ASCs because and losereplicativepotentialthatobesity out because ASCs become ‘exhausted’ withage the elderly. We hypothesize thatadipocytesrun ders accountingforcancerandorganfailurein can causeinflammationandmetabolicdisor- lipids, spillingintootherorgans. whichstart This the deficientfattissuefailstoeffectivelyabsorb aging. As weage, fatcellnumbersdecreaseand is oninvestigatingtheroleof ASCs inhealthy metabolic benefit. Thefuturefocusofourwork brown adipocytesandenablesashort-term spares brownfat ASCs, leadstogenerationof ASCs. indicatethatD-WAT Ourreports treatment Adipocyte progenitorcells:dysfunctionindiseaseandaging Harry E. Bovay, Jr. DistinguishedUniversityChairinMetabolicDiseaseResearch Associate Professor&Director, CenterforMetabolicandDegenerativeDiseases Mikhail Kolonin, Ph.D. of lipidsdrivingcancerprogression progenitors anditsroleinmetabolicdisease in health, andtargetingdisease nuclei. cells. Green:intratumoraladipocytes and adiposestromalcells. Blue: Immunofluorescence ofmousebreastadenocarcinoma. Red:malignant Research Assistant: Wanqiao Cao Research Assistant:Wanqiao Graduate Student:ShraddhaSubramanian Research Scientist:ZhanguoGao, FeiSu Senior ResearchScientist: Alexis Daquinag LAB MEMBERS Research, 77(18):5158-5168, 2017. prostate cancerinobesemice. Cancer CXCR4/CXCR7 signalingaxisdrivesMyc-induced and DiGiovanniJ. ProinflammatoryCXCL12- Saha A., Ahn S., BlandoJ., SuF., KoloninM.G. 6:1-9, 2017. Stromal Cells, Oncolytics. MolecularTherapy ed DecorinisaDrug Target onHuman Adipose Daquinag A.C. etal., KoloninM.G. Non-glycanat- animal metabolism. in individualtissuesandtheimpactonwhole us tostudytheintricaciesofcAMPsignaling drug.otherwise inert These toolsareallowing Instead, wecancontrolcAMPsignalingusingan hormones ordrugsthatregulatealltissues. different tissuesinlivingmicewithoutuseof study andvisualizecAMPpathwayfunctionin mousetoolsto new have recentlydeveloped conditions. Inconjunction withthiswork, we has onSIK1functioninstarvationandobesity muscle andwhatinfluencethecAMPpathway itseffectsinskeletal the SIK1enzymeexerts resistance. Ourongoingwork addresseshow inpeoplewithinsulin even blood glucoselevels thatSIK1inhibitorscouldreduce evidence This isexcitingbecauseitprovidespre-clinical byaparallelpathway.blood glucoselevels muscle insulintargets. Instead, SIK1regulates that SIK1doesthisnotthroughtheusual blood glucoseconcentration. We discovered this enzymeactsinskeletalmuscletoregulate After creatingmicelackingSIK1, wefoundthat is theenzymesaltinduciblekinase1(SIK1). links cAMPwithinsulinsignalingwithincells that bothregulatetobloodglucoselevels. pathways inmajortissues(liverandmuscle) studying themolecularintersectionsofthese and thecAMPpathwayisincreased. We are type 2diabetes, theinsulinpathwayisreduced balanced yin-yangrelationship. Inpeoplewith regulated hormonalpathwaysarenormallyina the fightorflightresponse. InsulinandcAMP- more glucosewhenneededduringstarvationor pathways regulatedbycAMPsignalingprovide other hand, instarvation, opposinghormonal normally reducesbloodglucose. Onthe and liverbecomeresistanttoinsulin, which in theseindividualsisthattissueslikemuscle pre-diabetes andtype2diabetes. Oneproblem ofinsulinresistance,contribute todevelopment obese, manychangesinhormonalpathways CENTER FORMETABOLIC AND DEGENERATIVE DISEASES One enzymeactivatedbycAMPsignalingthat When peoplechronicallyovereatandbecome Mol CellBiol. 201737(9):e00584-16. temporal stimulationofcyclic AMP signaling. Gs-DREADD knock-inmicefortissue-specific, Rajendran K, RossiM, Wess J, BerdeauxR. Akhmedov D, Mendoza-RodriguezMG, activity. PloSONE.201611(6):e0158274. miceforinvivomonitoringofCREB reporter MG, andBerdeauxR. Knock-inluciferase Akhmedov D, RajendranK, Mendoza-Rodriguez KEY PUBLICATIONS •Study ofhowcAMPsignalingregulates ofgeneticallymodifiedmiceto •Development •Identification oftherolesSIK1inmuscle RESEARCH PROJECTS we proposethatshapechangesinSIK1mutantscontributetoincreasedglucoseuse bymuscles. their structurechanges. Images showmusclemitochondriafromwild-type(WT)andSIK1mutantmice; Mitochondria useglucoseandfattogenerate ATP. Indiabetes, mitochondriabecomedamagedand pathways Regulation ofmusclemetabolismbycellularsignaling Associate Professor Rebecca Berdeaux, Ph.D. skeletal muscleregenerationinaging animals regulate andmonitorcAMPpathwaysinliving and livermetabolisminobesity Research Assistants: MariaMendoza-Rodriguez Graduate Student:RandiFitzgibbon Research Scientist:Dmitry Akhmedov LAB MEMBERS 2(14): 93367. by increasingO2supply. JCIInsight2017 expenditure contribute todiet-inducedenergy Tong Q. Redblood cellb-adrenergicreceptors Chung Y, Xia Y, Xu Y, LiF, SunK, BerdeauxR, O’Brien W, Xu Y, Mangieri, LR, Zhu Y, LeeCC, Kim ER, FanS, Akhmedov D, SynK, LimH,

33 • PACT REPORT PACT IMM 34 • IMMPACT REPORT can be reversed byrestoringwholebodyinsulin can bereversed diet-induced circadian changesinthesetissues bolic diseasessuchas typeIIdiabetes. Highfat humans contributestoweightgainandmeta - suggest thatthiscircadianmisalignment in zone, perse, thanthebrain. Studiestodate these clocks, puttingthemonadifferenttime tissues. Forexample, ahighfatdietcandisrupt rhythmicity intheliver, adipose, andmuscle quality isastrongdeterminantofcircadian nucleus. the centralpacemaker, thesuprachiasmatic back tothebrainandaffectfunctionof how disruptedperipheralclockscommunicate zeitgeber properties. We arealsointerestedin function andthemechanismsunderlyingtheir fortissue-specificclock are mostimportant whichzeitgebersthose designedtoreveal increases. Ourcurrent experimentsinclude body aremisaligned, riskformetabolicdisease such asfood. When circadianclocks acrossthe influenced byotherzeitgebers (“time-givers”) oscillations inperipheralorgansareheavily is predominantlyentrainedbylight, circadian produces theseeffects. trying tounderstandwhycircadiandisruption cancer, andmetabolicimbalance. We are outcomes result, includingacceleratedaging, cally orenvironmentally, deleterious several When thecircadianclockisdisruptedgeneti- clock genemutationsleadtosleepdisorders. white andbluelightsources. Inaddition, some or rotatingshifts, andlightcontaminationby across timezones(jet-lag), workinganightshift disrupt thisalignment. Examplesincludetravel hasprovidedamplewaysto modern technology with the24-hr. onitsaxis, rotationoftheearth hormone section, etc. adapttoandarealigned cycle, foodintake, internalbodytemperature, in allcellsofthebody. While oursleep/wake ing oscillationsof24-hrperiodicity, arepresent rhythms, whichareendogenous, self-perpetuat - circadian clockinhealthanddisease. Circadian CENTER FORMETABOLIC AND DEGENERATIVE DISEASES To date, wehavediscoveredthatnutrient While thecentralpacemakerofbrain The goalsofmylabcenterontherole staining=DAPI nuclearstain.) HNF4α protein(red), whichdisruptsthecircadian clockinthosecells. (Greenstaining=tubulin, Blue heterogeneity inexpressionofthe 3D organoidsgrown from humanandmouselivercancercells reveal • • • RESEARCH PROJECTS altogether. tumorgrowth be usedtodelayorprevent for thiscancer-specificHNF4αisoformmight or alternatively, timedapplicationofligands endogenous circadianrhythmsincancercells, are studyingwhetheragentswhichimprove on itsuniqueexpressionincancercells. We which expressthiscircadianproteinbased totreattumors therapies couldbedeveloped model. Thus, weaimtodeterminewhether death andimpairedtumorgrowthinarodent expression oftheBMAL1proteinresultsincell growth oflivertumors. Experimentallyrestoring homeostasis ofcancercellsandpromotesthe nuclear factor4(HNF4α)disruptsthecellular of thiscancer-specificisoformhepatocyte rodent livercancercells, wefindthatactivity drivers, BMAL1, inthecell. Usinghumanand circadian and downregulatesoneofthekey not normaltissue, hasuniquecircadianactivity which isexpressedonlyinlivercancercellsbut nuclearreceptorisoform,ered thataparticular and livercancer. Specifically, wehavediscov- linksbetweencircadiandisruption new reveal in thesetissues. diet andthecircadianclockisinsulinsignaling sensitivity, linkbetween indicatingthatakey Circadian clocksinhealthanddisease Assistant Professor Kristin Eckel-Mahan, Ph.D. cancer formation Mechanisms linkingcircadiandisruptionto balance forenergy dian oscillatorsanditsimportance Coordination oflight-vs. food- drivencirca- dian clockinmetabolictissues Mechanisms linkingnutrientsandthecirca- In additiontometabolicdisease, ourdata control thesleep/wakecycle. those ofthecentralnervoussystem, which the circadianclockofperipheraltissuesvs. the animals’ rest period. HFdifferentiallyaffects constant darkconditions). Graybarsrepresent (CD) vs. highfatdiet(HF)in “free running” (i.e. Circadian wheelrunningofmiceonalowfat Research Assistant: CorrineBaumgartner Ribas-Latre Post-doctoral Fellows:BaharanFekryand Aleix LAB MEMBERS Pages 133-152. 2016 Mahan KL, MolecularMetabolism, Volume 5, Metabolic Health” Ribas-Latre, A andEckel- for the CircadianClockSystem:Importance “Interdependence ofNutrientMetabolismand KL, EBioMedicine, Volume 20, Pages5-6. 2017 vascular Disease?” Fekry, BandEckel-Mahan “Clocks andCholesterol:Co-AgonistsinCardio- roscience, Volume 11, Pages369-378. 2017 Dyar, KAand Eckel-MahanKL, FrontiersinNeu- “Circadian Metabolomicsin andSpace”Time KEY PUBLICATIONS • humans Links betweenweightlossandchronotypein 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 causes Alzheimer’s diseasetoprogressfaster. have shownthatstressandexcessCortisol including Alzheimer’s disease. We andothers also negativelyimpactotherongoingdiseases, human patients. of thesepathwaystomodulatesymptomsin anxiety-related diseaseswillallowthetargeting 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 Stress signalinginpsychiatricanddegenerativedisease Assistant Professor Nicholas Justice, Ph.D. co-corresponding) doi: 10.1038/nn.4491. Epub2017Jan30. (*- stress. NatNeurosci. 2017Mar;20(3):385-388. sential forHPA axisregulationfollowingchronic Justice N*, Chen A*. HypothalamicCRFR1ises- Ramot A, JiangZ, JB,Tian Nahum T, Kuperman Y, KEY PUBLICATIONS •The roleofstressintheprogression •Neural circuitsthattransitthebasalganglia populationof •Investigations ofanew RESEARCH PROJECTS surrounding CRFR1neurons(red). CRF neuronsarehighlyinterconnectedwith with thatneuron(green). In thehypothalamus, infection ofalltheneuronsthatsynapse neurons(yellow,tion ofstarter arrow), allows Rabies viraltracingofneuralcircuits. Genera- Alzheimer’s disease to impactmovementinresponsestress and behavioralstressresponses signaling toimpactendocrine, autonomic, hypothalamic neuronsthatmediatelocalCRF related memorylossand Alzheimer’s disease. release. Chronic stress hasbeenlinkedtoage- formation, responds tostressbyactivatingCRF pocampus, formemory abrain regionimportant CRF neuronsinthehippocampus. The hip- ickam, Ph.D.; ZhiyingJiang, Ph.D. Post-doctoral Fellows:ShivakumarRajaman- LAB MEMBERS e20142. doi:10.7554/eLife.20142. GABAergic inhibition. Elife.2017Jan5;6. pii: KCC2 expressionandfunctioninhippocampal Wang Y, ZhengH, LongC, Yang L. APP modulates Wang K, RanX, Li Y, HuoQ, ZhangJ, LiH, LuN, Chen M, Wang J, JiangJ, ZhengX, JusticeNJ, 88. doi:10.1523/JNEUROSCI.1720-15.2016. rons. JNeurosci.2016May18;36(20):5472- Pallidal Neurons Target StriatalProjectionNeu- Huang TY, LuoM, JusticeNJ, ChanCS. Npas1+ HS,EC,Hernández VM,N, AugustineVerma Glajch KE, KelverDA, HegemanDJ, CuiQ, Xenias

35 • PACT REPORT PACT IMM 36 • IMMPACT REPORT CENTER FORMETABOLIC AND DEGENERATIVE DISEASES •Activation ofmusclestemcellsbynuclear •Therapeutic roleofERR’s inischemicmuscle •Regulation ofapoptosis, and autophagy •Regulation ofmusclemetabolism, vasculariza- RESEARCH PROJECTS and structureiscommonlycompromised. as musculardystrophies, wheremusclefunction medicine, diabetes, andorphandiseases such Our workhastherapeuticimplicationsinsports capacity throughregulationofgeneexpression. etal muscleendurance, size, andregenerative are interestedinhowthesefactorscontrolskel- muscle functionanddiseases. Specifically, we tors andtheirco-regulatorproteinsinskeletal receptors and DuchenneMuscularDystrophy muscle massbyPGC1-beta tion andendurancebyERR-alpha/gamma We areinvestigatingtheroleofnuclear recep- 2017 and musclewasting. SciRep. 7(1):10237, overexpression leadstoapoptosis, autophagy MS, GG, Rodney Narkar VA. Long-termPGC1β Sheth M, LorcaS, IK,Vila KimER, Tong Q, Song Sopariwala DH, Yadav V, BadinPM, LikhiteN, 2085. Muscle. Endocrinology. 2017;158(7):2084- Exercise andMetabolicRegulationinSkeletal and NucleosomeRepositioning:InsightsInto Narkar VA.PGC1αPromoterMethylation KEY PUBLICATIONS Nuclear receptor&co-activatorsignalinginskeletal muscle Associate Professor Narkar,Vihang Ph.D. muscle. and electronmicroscopyimagesofPGC1-beta mediatedautophagosomeformation(arrows)(D)in transcriptionalprogram(A),activation ofautophagy biomarkergenes(B)and proteins(C), autophagy PGC1-beta inducesmusclewastingbyactivating aprocesscalledautophagy. Panels (A-D)represent Research Assistant: MeghaSheth Neah Likhite Post-doctoral Fellows:DaneshSopariwala, LAB MEMBERS 12723, 2016 of mitochondrialfunction. NatCommun.7: SIRT1-dependent transcriptionalregulation integrates lipiddropletlipolysiswithPGC-1α/ sky W, Narkar VA, BickelPE. NuclearPerilipin5 CM, Yang C, McAfeeJL, HahnerL, HochK, Dubin- Gallardo-Montejano VI, SaxenaG, Kusminski collagen 6α3andproducesendotrophin, pre-existing unhealthy adiposetissue, itdigests hypoxia; Ontheother hand, inthecontextof thepathologicalconditionscausedby relieving esis incombinationwith VEGF-A andleptin, thus collagenous proteinsandstimulatesangiogen - at earlystagesofobesity, MT1-MMPcleaves metabolic contextdependent:Ontheone hand, regulation ofECMflexibilitybyMT1-MMP isalso note, ourrecent research suggeststhatthe ofextracellularmatrix(ECM).of thelevels Of MMP (MMP14)playsacriticalroleinregulation cally dysfunctionaladiposetissueandMT1- found fibrosisisthehallmarkinmetaboli- obese anddiabeticanimalmodels. Indeed, we in of adiposetissueremodelingatotherlevels of obesity-relatedmetabolicdisorders. andtreattheprogression approach toprevent foranoveltherapeutic the greatopportunity HIF1α and VEGF-A inadiposetissuemayoffer adipocytes. Ourfindingssuggestthattargeting ments inmetabolismbyablatingdysfunctional tissue, antiangiogenicactionleadstoimprove- contrast, inpathologicallyexpandedadipose “browning” phenotypeinwhiteadipocytes;In improving vascularizationandinducinga angiogenesis ismetabolicallybeneficialby at theearlystageofobesitydevelopment, dichotomous andmetaboliccontextdependent: angiogenesis inwhiteadiposetissuecouldbe demonstratedthat we further VEGF-A induced flammation inadiposetissue. Moreimportantly, which ultimatelyleadstolocalfibrosisandin- hypoxic andHIF1αinductionistheinitialstep discovered thatobesefatpadsarefrequently and obesitydevelopment. Specifically, we connections betweenadiposetissueremodeling paradigm-shifting findingsaboutthetight in humanobesityandinsulinresistance. them ascriticalfactorswithclinicalsignificance andpinpoints ing duringobesitydevelopment tors tothedynamicsofadiposetissueremodel- essential contributionsofadipocyte-derivedfac- CENTER FORMETABOLIC AND DEGENERATIVE DISEASES We exploredthefine-tunedregulation further Inthepastyears, wehavepublishedmany Research inourlaboratoryexaminesthe Reversibility ofadiposetissuefibrosis bynovel •Reversibility •VEGF-A stimulatedmetabolicbenefitsduring •Hypoxia inducedpathologicalchangesin 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 vironment tosustainmetabolicflexibility. ultimately leadingahighlyunfavorablemicroen- which acceleratesfibrosisandinflammation, Targeting adiposetissuefortreatmentofobesityanddiabetes Assistant Professor Kai Sun, M.D., Ph.D. by anti-UCP- Adipose Tissue-derived VEGF-A InducesBrowningof White Adipocytes: Immunofluorescent(IF)staining arrows indicate “crown-like” structuresformedbymacrophage accumulation. of endotrophintransgenic mice(right)andtheirlittermatecontrols (left)(Scalebars, 100µm). The cal (IHC)stainingby anti-Mac2 antibody(themarkerofmacrophages) insubcutaneousadiposetissue Adipose EndotrophinStimulatesLocalMacrophage Tissue-derived Accumulation: Immunohistochemi- transgenic mice(right)andtheirlittermatecontrols(left)(Scalebars, 50µm). anti-fibrotic therapies adipose tissuehealthyexpansion adipose tissue More recently, weusemoleculartools 1 antibody(themarkerofbrownadipocytes)insubcutaneousadiposetissue VEGF-A Visiting Scholar: ShuhongPeng Scholar: Visiting Sr. Research Assistant: XueGu Post-doctoral Fellows: Yueshui Zhao, XinLi LAB MEMBERS 20;2(14). by increasingO2supply. JCIInsight.2017Jul expenditure contribute todiet-inducedenergy Tong Q. Redbloodcellβ-adrenergicreceptors Chung Y, Xia Y, Xu Y, LiF, SunK, BerdeauxR, O’Brien W, Xu Y, MangieriLR, Zhu Y, LeeCC, Kim ER, FanS, Akhmedov D, SunK, LimH, Diabetes. 2017Jun;66(6):1479-1490. Metabolic Improvements(*contributedequally). Transplantation andConfersIL-4-Independent Rapid BeigingandEnhancedSurvival After Shows PE. VEGF-A-ExpressingAdiposeTissue Park J*, KimM*, SunK*, An YA, GuX, Scherer E963. AJP-Endo &Metab.2016Dec1;311(6):E952- Different CellPopulationsin Adipose Tissue. Sun K. DivergentFunctionsofEndotrophinon Zhao YS, GuX, ZhangNY, KoloninMK, An ZQ, KEY PUBLICATIONS

37 • PACT REPORT PACT IMM 38 • IMMPACT REPORT (bottom panels)compared tocontrols(toppanels). Arc: arcuateregion ofthehypothalamus. 3V:thethirdventricle. LepR:leptinreceptor. Dramatic increaseof p-AMPkinasesignal(green)inthehypothalamic neuronsofthebrainthatexpress leptinreceptor(red)inmicewithtype1diabetes CENTER FORMETABOLIC AND DEGENERATIVE DISEASES used inthelab. chemogenetics andinvivoliveimagingare techniquesincludingoptogenetics,of theart animal modelsincombinationwiththestate of obesityanditsassociateddiabetes. Various andtreatment effective targetsforprevention regulation, thereforeprovidingconceptualand causes fordefectivefeedingandbodyweight driving fundamental mechanisticinsightsonkey Our researchisdirectedtounderstandthe but withnoapparenteffectivetherapeutics. edented challengestosocietyandmedicine, ated metabolicsyndromehaveimposedunprec- The currentobesityepidemicanditsassoci- 26;4(3). pii:ENEURO.0083-17.2017. Diet FeedingRegulation. eNeuro, 2017May Midbrain LeptinReceptorNeuronsinHigh-fat Giros Band Tong Q. DopamineReleasefrom Xu Y, Lu Y, XuP, MangieriLR, IsingriniE, Xu Y, KEY PUBLICATIONS •Erythrocytes asanoveltherapeutictargetfor •Brain mechanismsmediatingbloodhormone •Brain efferentpathwayscontrollingperipheral •Novel neuronsandneuralpathwaysforfeed- RESEARCH PROJECTS Brain controloffeeding, bodyweightandglucosemetabolism Cullen ChairinMolecularMedicine Associate Professor Qingchun Tong,Ph.D. metabolic disorders genesis involvement inobesityanddiabetespatho- andglucose,action onenergy andtheir metabolism states ing regulationanditsrelationwithemotional Instructor: Instructor: Yuanzhong Xu, M.D., Ph.D. Cassidy, CanjunZhu(visiting) Graduate Students:LeandraMangieri, Ryan Huang, Ph.D. (visiting) Post-doctoral Fellows: Yungang Lu, Ph.D.; Xugen LAB MEMBERS Nature Communications, 2017, accepted. Aergic andGlutamatergicLH→PVHProjections. Feeding andSelf-groomingBehaviorsbyGAB- Arenkiel BRand Tong Q. Antagonistic Controlof Mangieri LR, Lu Y, Xu Y, Cassidy RM, Xu Y, insight.93367 Jul 20;2(14). pii:93367. doi:10.1172/jci. by IncreasingO2Supply. JCIInsight,2017 Expenditure Contribute toDiet-inducedEnergy Tong Q. RedBlood Cellβ-adrenergicReceptors Chung Y, Xia Y, Xu Y, LiF, SunK, BerdeauxRand O’Brien W, Xu Y,. MangieriLR, Zhu Y, LeeCC, Kim ER, FanS, Akhmedov D, SunK, LimH, CENTER FORMETABOLIC AND DEGENERATIVE DISEASES malian systems, westudyhowchaperonesand tools frommodelorganism Drosophilatomam- nonfunctional inagingneurons. mechanisms, whichoftenbecomeinefficientor have beenlinkedtotheseself-protective diseases andformationofpathogenicdeposits toxic cellularmaterials. Bothneurodegenerative Greek) thatcleansupandrecyclesworn-out or shape, (meaning andautophagy “self-eating” in chaperones thatfacilitateproteinstostayin robust self-maintenancemachines, including affected brains. Cellsnormally operateseveral often knownastanglesandplaques, inthe is thepresenceofabnormalproteindeposits, of almostalltheneurodegenerativediseases and externalinsultsfordecadestocome. cell tostayhealthandwardoffinternalcrisis robust self-clearancemachineriesinsidethe Accordingly, theselong-livedneuronsrelyon cannot bereplacedfortherestoftheirlife. units, losetheirabilitytoregenerateand they born andmatureintointerconnectedfunctional are challenge:oncethey face oneparticular ing andbeingreplenishedeachday, neurons the body, suchasskin, thatareconstantlydivid- ing, andresponses. Unlikemostothercellsin connections thatunderlieoursenses, reason- significant lossofneuronsandtheirfunctional brain diseases. utilize tostayhealthybutaredisruptedinthese molecular machineriesthatneuronsnormally tive treatmentstrategies, ourlabisstudyingthe well-being ofoursociety. To helpdiscovereffec- their familiesandposeapressingthreattothe emotional andfinancialtolltopatients ing maladies, whichinflictunbearablyhigh againstanyofthese debilitat- or preventions disease. Currently, thereisnoeffectivecure Parkinson’s disease(PD)andHuntington’s disorders, including Alzheimer’s disease(AD), affected byaging-relatedbraindegenerative longer lifeexpectancy, morepeoplearebeing Using biochemical, cellbiology, andgenetic Indeed, onecommon pathologicalhallmark Neurodegenerative disordersaredueto As oursocietyisenjoyinganunprecedented leads toaspectrumofdisordersincluding AD, pects ofcellularfunctions, andtheirdisruption related organelles(LROs), controldiverseas- such assynapticvesicles(SVs)andlysosome- brain diseases organelles andtheirdysfunctionin Biogenesis ofspecializedcellular approach. abnormality mightofferaneffectivetherapeutic protective mechanism, andcorrectionofsuch causing mutationscaninterferewiththis raises anintriguingpossibilitythatthedisease- . calledselectiveautophagy autophagy This roleinasubtypeof itself playsanimportant protein. Recently, wefoundthatHuntingtin tations (polyglutamineexpansion)inHuntingtin brain disorder, causedbyauniquetypeofmu- cellular healthyandlongevity. machinery worksinsideneuronstopreserve aggregates. We arestudyinghowthischaperone of dismantlinglargeandtightlypackedprotein machine calleddisaggregase thatiscapable a majorcomponentofthepotentmolecular abundant proteinsinthebrainandisalso ance onneuronalsurvival Protein folding, aggregation andclear- related diseases. protection machineriestofightagainstaging- toemploytheseinternal astrategy to develop and protectingnormalconstituents. Ourgoalis efficiently cleartoxicmaterials, whilesparing operateinthecelltorecognizeand autophagy Molecular mechanismsofneurodegenerativediseases Becker FamilyFoundationProfessorinDiabetesResearch Assistant Professor Sheng Zhang, Ph.D. neurons withnormal autophagy. butarecompletely absentin man diseasebrainsare prominentinneuronswithdisruptedautophagy In neurons(green)of Drosophilabrains, abnormalaggregates(reddots)similar asthatfoundinhu- In neurons, specialized cellularorganelles, Huntington’s fatal diseaseisadevastating Chaperone Hsp110isoneofthemost •Mechanisms ofproteinfoldingandclearance RESEARCH PROJECTS links tobraindiseases. these specializedcellularorganellesandtheir are studyingtheformationandregulationof SVs fortheirproperstorageandfunction. We mally packagedinsidethemembrane-enclosed in PD, canbe highlyunstablechemicalsnor- dopamine thatisproducedinneuronsaffected PD, andschizophrenia. Signalcarriers, suchas •Endogenous functionsofHuntingtinandits Lili Ye Research Assistants: JuanChen, Ph.D., Mrs. Ph.D.,Ph.D. AntonioTito, Post-doctoral Fellows:BoliHu, Ph.D., GangLi, Shiyu Xu,Instructor: Ph.D. LAB MEMBERS alized Experiments(118). doi:10.3791/55128. Drosophila brains”. (2016)TheJournal of Visu- protocol forwhole-mountpreparationofadult Sheng Zhang*.“A simpleone-stepdissection Antonio J Tito, ShebnaCheema, MianJiang, (2016); DevCell.36(2):139-51. for the TTT andPontin/ReptinComplexes.” Regulates mTOR Activity by Acting asan Adaptor Duraine, ShengZhang*, HugoJ. Bellen*. “WAC moto, BoXiong, KeZhang, HectorSandoval, Lita Wu-Lin Charng, ManishJaiswa, Shinya Yama- Gabriela David-Morrison, ZhenXu, Yan-Ning Rui, KEY PUBLICATIONS •Biogenesis oflysosome-relatedorganelles pathways inbraindegenerativedisorders perturbation inHuntington’sperturbation disease

39 • PACT REPORT PACT IMM IMMPACT REPORT • • • • 41 Sevick-Muraca, E.M., Kwon, S.K., and J.C., and J.C., S.K., Kwon, E.M., Sevick-Muraca, “Emerging lymphatic imaging Rasmussen, of Journal for mouse and man,” Technologies 2014. 905-14, 124(3): , Clinical Investigation PMID: 24590275 Lymphatic outflow imaged in head and neck J.C., Reproduced from Rasmussen, cancer. E.A., Maus, M.B., Aldrich, S., Naqvi, I.C., Tan, Sevick-Muraca, and E.M. R.J., Karni, A.I., Blanco, “Longitudinal monitoring of head and neck lymphatics in response to surgery and radia- 2017 39(6): 1177-1188, Head and Neck, tion,” PMID: 28263428. the immune system fluorescent a cancer-specific near-infrared guide intraopera- molecular imaging agent to tive lymph node dissection rheumatoid arthritis and its treatment with biologics surgery in head and neck cancer under microgravity conditions Diagnostic Imaging and Delivery of Therapeutics directed by Therapeutics and Delivery of Diagnostic Imaging NIRF imaging The lymphatics in health and disease. Reproduced in part from O’Donnell, T.F., Rasmussen, J.C., and J.C., Rasmussen, T.F., Reproduced in part from O’Donnell, The lymphatics in health and disease. Journal of Vascular “New diagnostic modalities in the evaluation of lymphedema,” Sevick-Muraca, E.M. 2017 PMID: 28214496. Mar; 5(2): 261-273, Disorders, and Lymphatic Venous Surgery: Eva Marie Sevick-Muraca, Ph.D. Sevick-Muraca, Eva Marie Molecular Imaging Director of the Center for Professor and Research Chair in Cardiovascular Disease Kinder Distinguished Nancy and Rich Imaging cancer-positive lymph nodes with •Imaging cancer-positive lymph to progressive •Imaging lymphatic responses •Imaging lymphatic responses to radiation and •Assessing CSF outflow into the lymphatics •Small animal imaging and tomography KEY PUBLICATIONS and E.M. J.C., Rasmussen, T.F., O’Donnell, “New in diagnostic modalities Sevick-Muraca, - Journal of Vas the evaluation of lymphedema,” Disorders, and Lymphatic Venous cular Surgery: 2017 PMID: 28214496. Mar; 5(2): 261-273, S., Kwon, F.C., Velasquez, M.A., Aldrich, Chan, B.R., Harvey, K., Pinkston, A., Azhdarinia, Sevick- and E.M. R.F., Ross, J.C., Rasmussen, W., delivery of etanercept via “Lymphatic Muraca, nanotopography improves response to collagen- - Arthritis And Treat Research induced arthritis,” 2017 PMID: 28566090. ment, 19(1): 116, The Center for Molecular Imaging (CMI) with congenital heart defects delivery vectors for treatment of heart disease MOLECULAR IMAGING FOR MOLECULAR CENTER consists of an interdisciplinary team of scientists and engineers who focus upon multi-modality molecular imaging including X-ray computed tomogra- nuclear imaging, and our fluorescence, bioluminescence, phy, fluorescence (NIRF) to near-infrared specialty, enable new understandings in several disease my team was the Between 2005-2007, states. first to dual label imaging agents with nuclear and NIR fluorescence agents to demonstrate the potential for pre-surgical PET imaging and intraoperative NIRF imaging for reduced tumor my In addition, burden in surgical oncology. team adapted NIRF tomography within the CT gantry of a PET/CT device to demonstrate hybrid small animal PET/CT/NIRF tomography. we translated into the clinical setting Finally, unique NIRF instrumentation and trace dose of NIRF Imaging agent to demonstrate for the first of the “pumping” time to visualize the active immune which mediates lymphatic vasculature, response and maintains fluid homeostasis actively collaborate with clinical We (Figures 1). Interventional Radiol- scientists in Pediatrics, Anomalies Clinic, Vascular the UTHealth ogy, as well as and Otorhinolaryngology, Pathology, Baylor College engineers and scientists at Rice, Our and the Methodist Hospital. of Medicine, team effectively translates new NIR imaging “bench-to-bedside” technologies literally from and back again in order to make discoveries Fluorescence lymphatic in translational NIR. imaging studies conducted by the CMI team include identifying key signaling pathways and regulators associated with aberrant processes of lymphangiogenesis and lymphatic stasis in human diseases and in animal models of hu- man disease. RESEARCH PROJECTS •Imaging chylo-and lymphothorax in children •Molecular Imaging of MMP-targeted viral gene •Lymphatic delivery of therapeutics targeting In addition to having an assembly of faculty- addition In enable visualization of the lymphatic system using enable visualization of the lymphatic and photonics technologies for better diagnosis Conditions such as vascular treatments. directed anomalies, congenital heart peripheral disease, and head and cancer, disease, breast vascular our using under investigation neck cancer are Translational imaging technologies. investigational visualization of brain activities further explore models of function in neonates,in preclinical into the lymphatics, human disease, CSF outflow detection of lymph node and intraoperative team metastases and tumor margins. Our focuses upon translating new NIRF Molecular that standards imaging agents using validated photonics device different can be applied across platforms. independent basic science and clinical driven the center synergistically projects, research clinicians center where operates a “collaboration” partner apply to effectively and researchers and translate imaging diagnostics to investigate therapeutics. novel 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, A highlight of the CMI is the basic science/

MOLECULAR IMAGING FOR MOLECULAR CENTER

molecular imaging agents, and companion molecular imaging agents, and companion to medicine. diagnostics to accelerate discoveries 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 and near-infrared fluorescence, (NIRF) to enable new understandings of conditions. Sponsored disease and chronic and federal research philanthropic, industry, funding focuses upon autoimmune disorders, cancer metastases, hemo- and neuroinflammation, diseases, and lymphedema. The lymph-vascular team has experts in instrumentation, imaging antibody engineering, animal agent development, models of human disease, and translational and inventions moves science that effectively and when “bench to bedside” discoveries, “bedside made in the clinic, from are discoveries back to bench.” clinical translational team that engages clinicians and at partnering institutions in at UTHealth Center and in the Houston Medical Texas the clinical studies suburbs. These FDA approved T

IMM PACT REPORT PACT • • • • 40 42 • IMMPACT REPORT CENTER FORMOLECULARIMAGING suggest appropriate oral therapeuticsforLE. and identificationof serumbiomarkerscould LEdevelopment, reverse often deterandeven have shownthatearlytreatmentofLEcan early LEdevelopment. studiesbyothers Several to identifyimagingandserumbiomarkers of LE,are ahighriskfordeveloping andIhope Center atMD Anderson. 40%ofthesepatients patients attheNellieB. ConnallyBreastCancer and bloodimmunecellsinbreastcancer changesinlymphaticfunction now surveying approval forresearch)tothebedside, andIam This imagingmodalityhasbeentranslated(FDA provide real-timemoviesoflymphaticpumping. the onlyimagingsysteminworldthatcan red fluorescencelymphaticimaging(NIRFLI), is available. compression bandagingandmassage)—nocure only palliativetreatment(daily, time-consuming currently areburdenedwithLE, andthereis (LE) results. Over15millionpeopleintheU.S. and apermanentdiseasecalledlymphedema pain, infection, andothermorbidities arise, after cancertreatment, forexample, swelling, When lymphatictransitisdisrupted, ashappens lymph passthroughthethoracicductdaily. 12 litersofinterstitialfluid, and4litersof veins andthoracicduct. blood circulatorysystemthroughthesubclavian numerous lymphnodesbeforereunitingwiththe collector lymphvessels, andpassesthrough capillaries. Lymphthenproceedstolarger enters theunidirectional, blind-endedlymphatic enter venouscapillaries, andimmunecells, cellular waste, moleculesthataretoolargeto the other10%ofinterstitialfluid, containing via venouscapillaryuptake. Lymphformswhen spaces returnstothebloodvascularsystem tissues, 90%oftheresultingfluidininterstitial cell transit. pumpsbloodto After theheart absorption, immunesurveillance, andimmune system toensurefluidhomeostasis, dietarylipid tory systemandworkswiththebloodcirculatory My colleagues recently developed near-infra- My colleaguesrecentlydeveloped An average-sizedhumanadultbodycontains The lymphaticsystemisthe “other” circula- breast cancerpatient. Leaky lymphaticsindicateearlylymphedemadevelopment. Near-infrared fluorescencelymphaticimage(NIRFLI)ofaxillarylymphnodebasin/upper armina J.C., RossR.F., C.E., Fife Sevick-Muraca, E.M. A., PinkstonK., B., Harvey Chan W., Rasmussen Aldrich M.B., Velasquez F.C., KwonS., Azhdarinia KEY PUBLICATIONS •Clinical studyofpneumaticcompression •Clinical studyoflymphaticdysfunctionin •Phase 1clinicalstudyofrheumatoidarthritis •5-year longitudinal, prospectivestudyof RESEARCH PROJECTS Lymphatic functionalandimmuneaberrationsinlymphedema Assistant Professor Melissa B. Aldrich, M.B.A., Ph.D. and cancer-associatedLEpatients cancer survivors, primary/geneticLEpatients, treatment efficacyinLEheadandneck peripheral venousdisease medication vialymphaticdelivery patients receivingananti-inflammatory funded) breast cancerpatientsathighriskforLE(NIH (PMID: 28356336), 2017. lymphedema. Pediatrics 139(4):e20154456 lymphatic imagingofatoddlerwithcongenital and RasmussenJ.C. Near-infraredfluorescence Greives M.R., Aldrich M.B., E.M., Sevick-Muraca March 2017 1650049 doi:10.1142/S1793545816500498, Innov OpticalHealthSciences, Vol. 10, No. 02, near-infrared fluorescencelymphaticimaging. J edema-affected extremities, asassessedby sion therapyonlymphmovementin- Rasmussen, J.C. Effectofpneumaticcompres- Aldrich M.B., GrossD., MorrowJ.R., C.E., Fife Res Ther19:116,arthritis. 2017. Arthritis ography improvesresponsetocollagen-induced Lymphatic deliveryofetanerceptviananotop- CENTER FORMOLECULARIMAGING is notpossible, weaimtouseourchemistry In caseswherecancerhasspreadandsurgery would otherwisebemissedbythenakedeye. healthy tissues, andidentifysmalltumors, which 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 My laboratoryisattheinterfaceofchemistry sues (B)andnear-infrared fluorescenceimagingofresectedtissues (C). Nuclear imagingofa tumor-bearingmouseinjectedwithamultimodality, tumor-seekingcontrast agent(A), tis- corresponding uptakeinkey Molecular imagingprobedevelopment John S. DunnResearchScholarIII Assistant Professor Ali Azhdarinia,Ph.D. hyaluronic acid with orthogonal chemistrygraft- hyaluronic acidwithorthogonal Callahan, L.A. Responsetodi-functionalized Zheng, Y., Azhdarinia, A.,Caoab,Q.,Smith Lim, H.J., Perera, H., Wilems, T.S., Ghosh, S., KEY PUBLICATIONS •Receptor-targeted deliveryofchemotherapy ofcontrastagentsfor real-time •Development RESEARCH PROJECTS improve humanhealth. discoveries andtechnologiesintotheclinicto to eachprojectisourfocusontranslationof antibody-based cancertreatments. Common and assessingtheeffectivenessofemerging of novelnanomaterialsforbiomedicaluse, imaging of “good” fattissue, characterization cancer,collaborations inareasbeyond suchas acterization hasallowedustoestablishdiverse inchemistry,expertise imaging, anddrugchar- ment protocols. Importantly, ourfundamental and visualizetheeffectstopersonalizetreat- platform tospecificallydelivertoxinstumors agents fortreatmentofcancer surgical guidance Research Scientist:SukhenGhosh Julie Voss Research Assistant: Graduate Students:ServandoHernandez Vargas LAB MEMBERS PMID:28740605 ACS MedChemLett.6;8(7):720-725, 2017. Analog forSomatostatinReceptor Targeting. of aFluorescentlyLabeled68Ga-DOTA-TOC T.,Schonbrunn, A., Azhdarinia,Synthesis A.* M., Kossatz, S., Voss, J., Carmon, K.S., Reiner, Ghosh, S.C., Hernandez Vargas, S., Rodriguez, ahead ofprint]. PMID:28572490. Receptor Targeting. Retains forSomatostatin Agonistic Properties A.* A ModularDualLabelingScaffold That J.,N.L.,Schonbrunn, Wilganowski, A., Azhdarinia, Ghosh, S.C., Rodriguez, M., Carmon, K.S., Voss, injury. J. Mater. Chem. B,4, 6865, 2016. models ofneuraldifferentiationandspinalcord ing atindependentmodificationsitesinrodent J. NuclMed.2017[Epub

43 • PACT REPORT PACT IMM 44 • IMMPACT REPORT CENTER FORMOLECULARIMAGING Research Instructor: PengGao,Research Instructor: Ph.D. LAB MEMBERS tion 2017. PMID:28384222 doi: 10.1371/journal.pone.0175212. eCollec- BR. PLoSOne. 2017 Apr 6;12(4):e0175212. KL, Bourgogne A, MurrayBE, van Hoof A, Harvey its roleinvirulenceandfitness. Gao P, Pinkston Functional studiesofE. faecalisRNaseJ2and PMID: 27914083 BR. MethodsMolBiol. 2017;1535:229-241. darinia A, MurrayBE, EM, Sevick-Muraca Harvey Endocarditis. PinkstonKL, GaoP, SinghKV, Azh- Antibody GuidedMolecularImagingofInfective 29049345 pone. 0186706. eCollection2017. PMID: 19;12(10):e0186706. doi: 10.1371/journal. MC, MurrayBE, BR. Harvey PLoSOne. 2017Oct Stinemetz EK, GaoP, PinkstonKL, Montealegre AtlA septallocalizationandcellseparation. AtlA, bythezinc-metalloprotease, GelE, impacts Processing ofthemajorautolysinE. faecalis, KEY PUBLICATIONS factorregulationgoverning bacterial •Virulence formolecular •Antibody agentdevelopment RESEARCH PROJECTS diagnosis orsurgeonsintheremovalofdisease. plications tohelpguidephysicianswithclinical equipment, theseagentshavedirectclinicalap- terial infection. Combinedwithmodernimaging inimagingcancerandbac- with focusedefforts antibody-basedimaging agents also develops inantibodyengineering,achievements ourlab tage ofantibodyspecificityandtechnological the factorsthatgoverninfection. Taking advan- and our understandingofbacterialphysiology have beeninstrumentalinhelpingtoimprove research tools, antibodiesdesignedbyourlab both basicandtranslationalresearch. As basic target disease, antibodiesarepowerfultoolsfor infection imaging ofcancerandinfectiousdisease Engineered torecognizeandspecifically Live animal imaging to diagnose enterococcal endocarditis infection within the heart valveofarat. Liveanimalimaging todiagnoseenterococcalendocarditisinfection withintheheart septum ofdividingbacterialchains incellwallcleavage.important Beads(designatedbyredarrows)denote AtlA localizationatpolesand Bacterial chaining, thoughttoencouragecolonizationandinfectionisregulatedby AtlA, anenzyme infectious disease ofdiagnosticimagingagentsforcancerand Development Assistant Professor Barrett RowlandHarvey, Ph.D. CENTER FORMOLECULARIMAGING response to AD pharmacologicalintervention. target forpredicting onset, progression, and the headandneckcan beusedasadiagnostic to showthatperipherallymphaticsoutflow from McGovern MedicalSchool. This isthefirsttime the MitchellCenterfor Alzheimer’s diseaseat Claudia SotoandInesMoreno-Gonzalezin plaque accumulationincollaborationwith Drs. function atearlyonsetofamyloidbeta(A β) (AD) isimpairedandmayimpactglymphatic transgenic mousemodelsof Alzheimer’s disease showed thattheperipherallymphaticsystemof vasculature withintheheadandneckarea. We ultimately drainintotheperipherallymphatic (ISF) areexchangedthrough “glymphatics” that spinal fluid(CSF)andbraininterstitial the untreatedcohort. tumor shrinkageoccurredwhencomparedto successfully deliveredviathelymphaticsystem, lymphocyte–associated antigen4(CTLA-4)was that whentherepeateddoseofanti-cytotoxic T- systemic anti-tumorimmunity. We demonstrated stimulation totheregionallymphaticsfor LNs andreducetoxicitybylocalizingimmune to maximizedrugexposuretumor-draining forimmunotherapy lymphatic infusiondevice ratmodel.arthritis Recently, weusedthenovel to conventionalroutesofadministrationinan local andsystemicresponseswhencompared into thelymphaticscanresultinimproved infusion ofanimmune-attenuatingbiologic metastasis, hypertension, andinflammation. age patternsinanimalmodelsoflymphnode showed abnormallymphaticfunctionanddrain- and disease. Usingthisnoveltechnique, we ing andquantifyinglymphaticfunctioninhealth fluorescence (NIRF)imagingmethodsforimag- non-invasive,developed dynamicnear-infrared ticularly inthelymphaticsystem. Recently, I’ve movement offluidandmacromolecules, par- focuses oninvestigatingthemicrocirculatory biological questions. Mymainresearchinterest small animalimagingtechniquestoaddress Recent evidence demonstratesthatcerebral Recent evidence Recently, CMIdemonstratedthatdirect andapplicationof I leadthedevelopment liver. Asterisk, upperhardpalate in themouth. Double arrows, lymphaticvessels. Arrowhead, to theMLNs(arrow) after intrathecalinjection. circle), endoturbinate(openarrow), andfinally cisterna magnaandsupracerebellarcistern (red ofamouseshowingCSFdrainagetothe view in theventral, rightandleftlateral, anddorsal White light, NIRfluorescent, andmergeimages Develop PET/CT methodology forquantifying PET/CTmethodology •Develop methodsforassessingCSF new •Develop •Assess brainlymphaticsandcerebrospinal •Non-invasive functionallymphaticimaging RESEARCH PROJECTS myocardial infarctinmice. lymph nodemetastasis, inflammation, and conducting molecularimagingofcancerand and translatingimagingagents. Iamcurrently The CenterforMolecularImagingisdeveloping volve aroundmulti-modalitymolecularimaging. lymphovascular disorders Functional lymphaticimaginginanimalmodelsof Assistant Professor Sun KukKwon, Ph.D. dial infarctinmice infarct sizeingenetherapytrialsofmyocar- delivery route with intrathecaldeliveryofdrugsasaviable drainage inmiceandswinefortranslation mice phatics in Alzheimer disease(AD)transgenic fluid (CSF)outflowintotheperipherallym- metastasis andautoimmunedisorders lymph nodesofanimalmodelsnode to showtheefficacyofdrugdeliveryinto Other directionsofmyscientificinterestsre- with 20Gy. lymphatic abnormality ascomparedtomice only micetreatedwith 40Gyshowedpersistent days afterradiationwith20and40Gy;however, changes oflymphaticvasculatureinmice 21 tion. Ourpreliminarydatashowedsignificant the footofmice7, 21, and42daysafterradia- Magnified near-infraredfluorescentimages in Immunological Methods, 2017. cerebrospinal fluidinmice,” Inpress, Journal of “Fluorescence imagingoflymphaticoutflow Christian Velasquez, andEvaM. Sevick-Muraca, Sunkuk Kwon, ChristopherF. Janssen, Fred Therapy, 19;116, 2017. collagen-induced arthritis,” Research & Arthritis cept viananotopographyimprovesresponseto Sevick-Muraca. “Lymphatic deliveryofetaner- Russell F. Ross, CarolineE. Fife, andE. M. R. Harvey, Wenyaw Chan, JohnC. Rasmussen, Kwon, Ali Azhdarinia, KennethPinkston, Barrett Melissa B. Aldrich, FredC. Velasquez, Sunkuk pages, 2017. doi:10.1155/2017/7659242 Imaging, vol. 2017, ID7659242,Article 7 rescence Imaging,” Contrast Media&Molecular and Diet-InducedFluorescenceonInVivoFluo- fects ofDepilation-InducedSkinPigmentation Sunkuk KwonandEvaM. Sevick-Muraca, “Ef- of Anesthesia,Journal 30;1091-1094, 2016. lymphatic contractileactivityinmicevivo,” of lidocainewithandwithoutepinephrineon Sunkuk KwonandEvaM. Sevick-Muraca. “Effect 1100-1115, 2016. node inmice,” BiomedicalOpticsExpress, 7; tion aftersurgicalremovalofanaxillarylymph of internodalcollectinglymphaticvesselfunc- Sunkuk KwonandPrice, R.E. “Characterization KEY PUBLICATIONS

45 • PACT REPORT PACT IMM 46 • IMMPACT REPORT CENTER FORMOLECULARIMAGING of therolelymphatics inearlyvascular of thevasculardisease. A betterunderstanding are aresultoforcontribute tothedevelopment appear, andwhethertheselymphaticschanges the abnormallymphaticanatomyandfunction disease todetermineatwhatstageof additional subjectswithearlyvenousorarterial of venousdisease. We arecurrently imaging legs, includingthosewithnoexternalsign lymphatic abnormalitiesinallthecontralateral was mostsurprisingthatwealsoobserved the legswithadvanceddisease. However, what and demonstratedabnormallymphaticsinall a groupofpatientswithactivevenouslegulcers and poorimmuneresponse. We recentlyimaged of chronicswellingwithfibrotictissuechanges lymphedema,ease oftenco-develop acondition that patientswithadvancedchronicvenousdis- tory system. Ithasbeenknown formanyyears between thelymphaticsandbloodcircula- fluorescent contrastagent. and diseaseusingmicrodoseamountsofa quantify theircontractilefunctioninhealth image andcharacterizehumanlymphatics (NIRF) opticalimagingasawaytononinvasively and translationofnear-infraredfluorescence my researchhasfocuseduponthedevelopment MRI, andultrasound. years, Overthepastfew imaging modalitiessuchasscintigraphy, X-ray, orusingtraditionalclinical either withoureyes to distinguishfromthesurroundingtissues, clearfluids,primarily transport aredifficult they because thelymphaticsaretypicallysmalland absorption ofnutrientsfromthegut. However, tion, thelymphaticsprovideapathwayfor and isultimatelyreturnedtotheveins. Inaddi- lymphatics, processedforimmuneresponse, etc. The resultingsolutionistaken upbythe up cellwaste, foreigncontaminants, proteins, the watermovesthroughtissues, itpicks the smallgapsbetweentissuecells. As veins, waterleaksfromthevesselsentering system. and As bloodflowsthroughthe arteries understood, componentofthecirculatory One ofourprimaryfocusesistherelationship The lymphaticsystemisavital, yetpoorly and clinicalquestionsnotaddressedbyother biological the ultimategoalofansweringnew lymphatic imageprocessingandanalysiswith analyticaltoolsto facilitate and developing automating differentaspectsofthehardware, ogy, sensitivity, including improvingdevice in aslittletwoweeks. reduction intheextentofabnormallymphatics and necklymphedemahaveobserveda specificallyforsubjects withhead developed advancedpneumaticcompression device new imaging toassessthelymphaticresponsea radiation treatment. We arecurrentlyusingNIRF increasing withtime(months)aftertheendof the extentofabnormallymphaticsgenerally abnormal lymphaticsinthispopulation, with of Our imaginghasshownthedevelopment lymphedema.cancer survivorswilldevelop that75%ofheadandneck has beenreported in theheadandneckcancerpopulationasit cancer treatment. We interested areparticularly recovery, orlackthereof, ofthelymphaticsafter malformations. tionship betweenthelymphaticsandvascular disease. Additional studiesfocusontherela- and/or functioninallsubjectswithearly have observedabnormallymphaticanatomy efficacious therapeuticapproaches. To date, we ofmore disease mayenablethedevelopment translationforlymphaticimaging Device Assistant Professor John Rasmussen, Ph.D. We continue the development ofthistechnol- We continuethedevelopment We arealsousingNIRFimagingtoassess the e20154456, 2017(PMID:28356336). congenital lymphedema,” Pediatrics, 139(4): rescence lymphaticimagingofatoddlerwith E.M., andRasmussen, J.C., “Near-infrared fluo- Greives, M.R., Aldrich, M.B., Sevick-Muraca, 5(4):533-537, 2017(PMID:28623992). cular surgery: Venous andlymphaticdisorders, of Klippel-Trénaunay syndrome,” Journal ofvas- “Near-infrared fluorescencelymphaticimaging M.B., Greives, M.R., andSevick-Muraca, E.M., Rasmussen, J.C.,*Zvavanjanja, R.C.,* Aldrich, 39(6):1177-1188, 2017(PMID:28263428). to surgeryandradiation,” HeadandNeck, of theheadandnecklymphaticsinresponse Sevick-Muraca, E.M., “Longitudinal monitoring M.B., Maus, E.A., Blanco, A.I., Karni, R.J., and Rasmussen, J.C.,* Tan, I.,*Naqvi, S., Aldrich, KEY PUBLICATIONS •Translation oflymphaticimagingtothe ofcancer-related •Assessing thedevelopment •Understanding theroleoflymphaticsin RESEARCH PROJECTS technologies. pediatric population lymphedema anditsresponsetointervention rial disease - ofperipheralvenousandarte development 2017). et al., HeadandNeck, duced fromRasmussen, neck lymphedema(Repro- a patientwithheadand compression therapyin of advancepneumatic lymphatics aftertwoweeks Reduction ofabnormal (right) stage C4venousdisease phatics inapatientwith (left) andabnormallym- in ahealthyvolunteer Normal lymphaticvessels preclinical studies. While molecularimagingof a methodtoassesstransgene expressionin of far-redfluorescent protein(iRFP)represents vector deliveryandtransgeneexpression. Use quantitatively associatingMMPactivitywith dependsupon targeting andinfectionstrategy of tissueremodeling. Validation ofinvivoAAV by extracellularproteasesthatarebiomarkers ated virus(AAV)vectorsthatcouldbeactivated University wasthefirsttocreateadeno-associ- ofBiomedicalEngineeringatRice Department number ofcardiacdiseases. The Suhlabatthe Gene therapyhasthepotentialtotreata close to5million failure.Americans haveheart the leadingcauseofdeathinU.S. and diseaseis myocardial infarction(MI):Heart protease activatedvirusinanimalmodelof in childrenwithbraintumors. algorithm toassessbrainnetworkdysfunction with acomputationally-efficienttomographic Optical Tomography (fNIRS-DOT) imagingsystem Functional NearInfraredSpectroscopy-Diffuse high resolution, multi-pixeltechniquebased side effects. Currently, a wearedeveloping ment andminimizingbrainnetworkdysfunction are madeonbasisofoptimizingtumortreat- toms; chemotherapyandradiationtreatment present withbothcommonandunusualsymp- biologically diversegroupofneoplasmsthatcan brain tumorsrepresentananatomicallyand of cognitivedeclineaftertreatment. Childhood cancers, suchasleukemia, alsohaveevidence have influenceonbrainfunctioning. Childhood coids andanti-epilepticdrugs, whichbothalso ment,- gliomapatientsoftenreceiveglucocorti negative way. fromtheanti-tumortreat- Apart functioning, bothinapositiveaswell apy, andchemotherapyallmayaffectcerebral directed againstthetumor. Surgery, radiother- the tumoritselfandalsofromtreatment patients. These disturbancesmay resultfrom tions, arefrequentlydisturbedinbraintumor Brainfunctions,tumor: including cognitivefunc- brain functionalmappinginchildrenwith CENTER FORMOLECULARIMAGING Assessment ofdeliveryandinfection High-resolution diffuseopticaltomographyfor model ofmyocardialinfarction delivery andinfectionofproteaseactivatedvirusinanimal andassessmentof imaging inchildrenwithbraintumor; High-resolution diffuseopticaltomographyforbrainfunctional Assistant Professor Banghe Zhu, Ph.D. G., West, J., Sevick, E. M., Davis, A., OlmstedD. Alvarez-Urena, P., Zhu, B., Sonnet, C., Henslee, KEY PUBLICATIONS inheadandnecksurgeryclinical •Participate inperipheralvasculardisease •Participate •Assess thegenetherapyusingiRFP 3-Dtomographyalgorithm forbrain •Develop high-resolutiondiffuseoptical imag- •Develop RESEARCH PROJECTS protease-activatable virus(PAV) targeting. International (SI)unitsforassessingaccuracyof rescence measurementscalibratedinStandard MMP activityandexvivoiRFPNIRFfluo- mice withinducedmyocardialinfarcttodetect imaging.use We non-invasive, invivomicroPET/CTandNIRF targeting activegelatinases(MMP-2/-9)for adual-labeledimagingagent we developed inhumans.location oftheheart Previously, based techniquesarenotsuitedforthedeep employ MMPtargetedtherapies, fluorescence- diagnostic inclinicalgenetherapytrialsthat companion MMP couldprovideanimportant studies clinical studies reporter imaging ing system oxygen saturationof the children’s brain. 3-D (a), Coronal(b), Transverse (c), and Sagittal(d)cross-sectionsofthereconstructed in vivomicroPET/CTimagingof Express. 5(2):562-572(2014). ICCD andlaserdiode,” BiomedicalOptics ambient lightconditionsusingamodulated M., “Non-invasive fluorescenceimaging under Zhu, B., Rasmussen, J.C., andSevick-Muraca, E. 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 35(3),Medical Imaging, 802-811(2016). with SIUnitsofRadiance”, IEEETransactionson Using TraceableStandards Working Devices Performance ofFluorescenceMolecularImaging Sevick-Muraca, E. M., “Determining the Zhu, B., Rasmussen, J. C., Litorja, M., and tion”. J. Cell. Biochem.9999:1–8, 2017. Approach toSelectively Turn offBoneForma- E., ofaCell-basedGene “Development Therapy

47 • PACT REPORT PACT IMM IMMPACT REPORT • • • • 49 CRC Press, Taylor & Taylor CRC Press, KEY PUBLICATIONS KEY PUBLICATIONS V, Cristini WH, Driessen AS, Dobroff Hosoya H, D’Angelo Cardo-Vila M, Staquicini FI, Brinker LM, Dunphy DR, YS, Lin Proneth B, F, Ferrara S, K, Miyazono Stafford RJ, MP, Melancon Dogra P, Sidman RL, Brinker CJ, Kataoka K, Gelovani JG, Integrated nanotechnology Pasqualini R. W, Arap platform for tumor-targeted multimodal imaging Acad and therapeutic cargo release. Proc Natl 2016 A 113:1877-82, Sci U S A, Sreekumar Landua J, A, Chauviere Paine I, A Geometrically- Lewis MT. Rosen J, V, Cristini Constrained Mathematical Model of Mammary Gland Ductal Elongation Reveals Novel Cellular PLoS End Bud. Terminal Dynamics within the 2016 Comput Biol 12:e1004839, An Wang. Zhihui Koay, Eugene J. Vittorio Cristini, Introduction to Physical Oncology: How Mecha- nistic Mathematical Modeling Can Improve Cancer Therapy Outcomes. Francis Group. July 2017 Group. Francis LAB MEMBERS Ph.D. Post-doctoral Fellows: Joseph Butner, Bro- Terisse Graduate Students: Prashant Dogra, Oluwadara Tito, Jorge Naomi Hasegawa, cato, Coker Ph.D. Research Scientists: Satyanarayan Nandi, A mass transport model of pancreatic ductal adenocarcinoma (PDAC) A mass transport model of pancreatic ductal adenocarcinoma (PDAC) reveals that gemcitabine incorporation in the tumors is highly variable after accounting for and correlates inversely with the amount of stroma, hENT1 levels. Systems pharmacology and drug and invasion and drug response in local and metastatic cancers immunotherapy functionally linking biological behaviors at different scales) Vittorio Cristini, Ph.D. Vittorio Cristini, Director Professor and Max LevitRochelle and in the Neurosciences Chair biomedicine Translational are developingWe complex biological systems. challenge in methods to address a significant bridging the gaps be- i.e., multiscale modeling, and between tween different modeling methods from the molecular, models at different scales, based on to the cellular and tissue scales, a technique theory,” “dynamic density functional implemented in the physical sciences. pharmacokinetic-pharmacodynamic (PKPD) modeling. Many PKPD models based on ordinary differential equations (ODEs) 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 •Mechanistic modeling of tumor response to •Upscaling and downscaling framework (i.e., Our research program focuses on developing Physi- oncology. physical Translational Multiscale modeling. Biological PRECISION BIOMEDICINE FOR PRECISION CENTER mechanistic biophysical models for predicting tumor response to various treatment methods nano- targeted therapy, chemotherapy, (e.g., individual and immunotherapy) in medicine, patients using standard clinical diagnostic CT, such as histopathology, measurements, a multidisciplinary have taken We and MRI. 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. cal 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 com- plex normal and pathologic biological tissue. clinical translation of the mathematical Toward we have been investigating the effects 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 mecha- nisms 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. a book (monograph) in this just published We CRC area: An Introduction to Physical Oncology, Press. processes can occur across physical time and forming a complex system with space scales, multiple feedback and feed-forward loops. Advanced multiscale methods are therefore needed to simulate and predict the behavior of provide novel ways to investigate the structures the structures investigate ways to novel provide also We colitis. of diseased bones and ulcerative high resolution large-scale, multi-color, have state-of-the-art printers for both fast 3D models level and finished production prototypes of new surgical tools and instruments or patient- multiscale We provide specific organ models. to mathematical and computational modeling clinical trials focusing prospective aid current on understanding drug penetration barriers and tumor response in tumors and improving patient outcomes. the Center Collaboration with of Research Hubs include: Core Proteomics Translational • Clinical and Laboratory Service Center • Nanochemistry/3D-printing Computational Cancer in • NCI Programs and Nanomedicine Biology Facility Core Proteomics • UT System-wide Network Translational Clinical and • UTHealth/MDACC Technologies Center for Translational Ph.D. Cristini, Vittorio for Precision Center and Director, Professor Medicine for Molecular Institute Biomedicine, Levit Chair in the Neurosciences and Max Rochelle Fellow STAR System Texas of University

he Center for Precision Biomedicine Biomedicine he Center for Precision the mathematical, focuses on developing experimental, and analytical technologies

These efforts connect us with collaborators

that will deliver precise medication to the correct to the correct medication precise that will deliver by understanding tissues. This is accomplished the the underlying physiological problems, of indicative and genomic biomarkers proteomic the targeting of tissues or toxins, disease, selective use of targeted nanoparticles, and mechanistic to mathematical models of tissue and vasculature biological barriers to tissue and overcome predict penetration. Texas within the institutions UTHealth; across Methodist such as Baylor, Center, Medical through Texas and MD Anderson; and across for interactions with the centers/programs Science, nanomedicine Translational Clinical and and faculties studying disease researchers, genomics, and mechanisms using proteomics, state-of- have the IMM, we bioinformatics. At in-depth the-art providing mass spectrometers, analysis of cells, tissues, or biological proteomic targets of novel fluids, leading to the discovery for early for drug and biomarkers development medicine. precision detection and personalized new aptamers and multi- develop also We functional nanoparticle therapeutics for targeting Using pathological tissues, such as cancer. we custom-built NIRS/Ramen spectrometers,

PRECISION BIOMEDICINE FOR PRECISION CENTER T

IMM PACT REPORT PACT • • • • 48 50 • IMMPACT REPORT this problem, wehaveusedcomputationalap- not respondtotraditionaltherapies. To address metastasis, becausemetastaticcellsdo inpart that upto90%ofcancerdeathsaredue broad andcomplementaryareasofemphasis: cal samples. including cellculture, mousemodels, andclini- cell biology, and biochemistry;andusemodels range ofapproachesencompassinggenomics, repositioning ofdrugs. To dothis, weusea ofgenomicbiomarkers and the development translate thesefindingsintotheclinicthrough differentiation processes. Ourultimategoalisto whether throughacquiredmutationsortrans- lead tumorcellstoacquireametastaticstate, sion. that We wishtounderstandtheevents signaling programsunderlyingcancerprogres- CENTER FORPRECISIONBIOMEDICINE (bottom leftpanel). Inamousemodel, knockoutof ABCA1 leadstoasignificantreduction oflungmetastasesinabreastcancer model(rightpanel). tumors, highexpressionof ABCA1 isassociatedwith100%chance ofmetastasis, ofthepatientswithnormal whileonlyabout aquarter ABCA1 metastasize levels inmetastaticbreastcancercelllines (top leftpanel).We haveidentified acholesteroleffluxchannelthatisexpressed athigh In humanbreastcancer 1. Breastcancermetastasis. Itisestimated Our researchprogramencompassestwo Our labisfocusedonunderstandingthe •Intelligent computationalpipelinesforbioin- •Heterogeneity andprogressionofmetastatic •The roleofcholesteroltraffickingincancer RESEARCH PROJECTS of commonbioinformaticsalgorithms. ages aknowledgebasecontainingdescriptions - systemthatlever a backwards-chainingexpert tasks, presentinguswithfinishedresults. Itis that canautomaticallyplanandexecutethese intelligence, anartificial have developed BETSY, amplify ourabilitytodobioinformatics, we hypotheses,develop oranalyzeresults. To with bioinformaticstominepublicdatasets, Many ofourprojectsrequirestheintegration metastasis inpreclinicalmodels. pathways thathaveshowntheabilitytoinhibit pounds andFDA-approved drugstargeting novel We haveidentifiedaselectionofnatural com- exhibit phenotypesthatpromotemetastasis. proaches torepositiondrugstargetcellsthat metastasis Deciphering thesignalingprogramsunderlyingcancer CPRIT ScholarinCancerResearch Associate Professor Chang,Jeffrey Ph.D. formatic analysis cancers metastasis to-mesenchymal transition, andcancer stem celldifferentiation, theepithelial- 2. intelligenceforgenomicanalysis.Artificial Visiting Scientists: Visiting Luo,Ying JuhuaDan Baidya Research Assistant:Aurnab Zhu Graduate Student:Kevin Prjic, Ph.D.. HuanQiu, Ph.D. Post-doctoral Fellows: Weina Zhao, Ph.D., Sarah LAB MEMBERS ics 33(8), 2017. system.workflows usinganexpert Bioinformat- Chen XandChangJ.T.: Planningbioinformatics Oncotarget, 2016. Transition requiredestabilizationoflipidrafts. ties inducedbytheEpithelial-Mesenchymal Chang J.T.*: Motilityandstemcellproper- M.J.,Tisza SjolJ.S., ChenX., I.*, Levental and 76(7):2037-49, 2016. reducing membranefluidity. CancerResearch metastatic capacityofbreastcancercellsby Candidate anti-metastasisdrugssuppressthe Tan Z., ChenX., ManiS.A., andChangJ.T.: Zhao W., PrijicS., UrbanB., M.J.,Tisza LiL., KEY PUBLICATIONS * Co-Corresponding Authors * CENTER FORPRECISIONBIOMEDICINE fluid inuptofmscale. SERS methodscandetectbiomarkersinbody By combiningRSandnanotechnology, such surface enhancedRamanspectroscopy(SERS). targeted imagingandbiosensingmethods using in bonequalitywithdiabetesandaging. lational studiesinclinics, investigatingchanges Currently thissystemisbeingappliedfortrans- bonequalityinvivo. transcutaneously evaluate to in vivoRamansystemhasbeendeveloped of metastaticbonetotreatment. A noninvasive theresponse sess bonequalityandtoevaluate alterations. These markerscanbeusedtoas- to breastandprostatecancersinducedbone RSspectralmarkersthatarerelated developed stantly studiedinthelab. Inaddition, wehave andmechanicalfunctioniscon- properties variations anddiseaseonbonecompositional of fractureresistance. The effectofgenetic bone quality, determinants whichareimportant our laboratory. cal marginassessmentisalsobeingexploredin application ofRSincancerdiagnosisandsurgi- to improvediagnosisaccuracyinsitu. Further incorporation ofRStocolonoscopyisexpected of IBD:ulcerativecolitisandCrohn’s colitis. The the twodistinctyetoftenindeterminateforms shown over99.7%accuracyindifferentiating ics. InvitroRSstudiesoncolonbiopsieshave with inflammatoryboweldisease(IBD)inclin- using anRSfiberopticsystemtotestpatients changes intissueenvironment. We arecurrently disease- andaging-associatedbiochemical composition oftissueandissensitiveto response evaluation, andguidanceofsurgery. such asearlydiseasediagnosis, therapy Raman spectroscopy(RS), forclinicalproblems, opticaltools,focuses ondeveloping especially noninvasive insitudiagnosis. Ourresearch have demonstratedgreatpotentialinproviding Another area of research involves developing Another areaofresearchinvolvesdeveloping We haveextensiveexperienceinquantifying RS exploitssubtlechangesinthemolecular Optical spectroscopyandimagingtechniques increasing bonesignals fromdepths. risk assessment. D)Representativespectracollected concurrentlyfromonemeasurement, showing of bonecomposition, whichisindicativeofbone quality. C) Ongoingclinicalapplicationforfracture Ramansystemfortranscutaneousinvivodetection The scheme(A)andthepictureofportable cal factors on the biochemical properties of cal factorsonthebiochemicalproperties Guha, M. Younes, Y. Ye, X. Bi, Effectofphysiologi- H. Ding, A. W. Dupont, S. Singhal, L. D. Scott, S. Express, 2017, 8(7):3426-3439. Raman spectroscopy, 2017, BiomedicalOptics activity inulcerativecolitisusingendoscopic histologicaldisease of mucosallipidsreveals Scott, S. Guha, M. Younes, X. Bi, Invivoanalysis H. Ding, A. W. Dupont, P. Wang, S. Singhal, L. D. KEY PUBLICATIONS •Cancer-targeted imagingusingultrasensi- andapplyinvivoRamantechniques •Develop •Evaluate changesinbonequalityandme- andapplynoninvasiveendoscopic •Develop RESEARCH PROJECTS Optical spectroscopyandimagingformedicine Assistant Professor Xiaohong Bi, Ph.D. nanoparticles nanoparticles tive SERSimagingtechniqueandtargeted predict thefracturerisk bonequalityinpatients andto to evaluate in preclinicalmodelsandpatients diabetes, andotherbonemetabolicdisorders chanical functionswithaging, geneticdefect, other diseasesingastrointestinaltract colorectal cancer, Barrett’s esophagus, and information forinflammatoryboweldisease, Raman spectroscopytoprovidediagnostic Research Assistant: GuijinLu LAB MEMBERS Optics, 19(11):111606(2014) Bone inBreastCancer, Journal ofBiomedical Raman SpectralMarkersto Assess Metastatic A. Mahadevan-Jasen, andX. Bi, of Development H. Ding, J.S. Nyman, J.A. Sterling, D.S. Perrien, 6(28):26041-51 (2015) in pancreaticcancer, 2015, Oncotarget, 2015, X. Bi, M. Li, ZIP4silencingimprovesboneloss Ding, C. Houchen, RGPostier, CG Ambrose, Z. Li, Q. Zhang, X. Sun, J. Yang, H. Ding, D. Lebrun, K. eral Research, e-pubonlineSeptember2016 anti-TGF-β treatment,Journal ofBoneandMin- Osteogenesis Imperfectaandtheresponseto in mousemodelsofdominantandrecessive andbonecompositionparameters properties Correlations betweenbonemechanical M.M. Jiang, B. Dawson, B. Lee, C.G. Ambrose, X. Bi, I. Grafe, H. Ding, R. Flores, E. Munivez, 48(7): 902-909 study, Journal ofRamanspectroscopy, 2017, colon tissue– An invivoRamanspectroscopy

51 • PACT REPORT PACT IMM 52 • IMMPACT REPORT CENTER FORPRECISIONBIOMEDICINE diseases. andbioinformatics Systemsbiology diseases, GI-tractmicrobiome, HIVandother of chronicinflammation, neurodegenerative proteomicsinvestigation our labalsosupports eases. Inaddition, throughcollaborativeefforts, are pancreaticcancerandotherGI-tract dis- cal applications. Currently, our diseasefocuses innovation ofproteomicstechnologiesfor clini- association withcancerandotherdiseases; 4) characterization ofproteinglycationadductsin rodegenerative diseases;3)massspectrometric in malignances, chronicinflammation andneu- Investigation ofproteinglycosylationalterations early malignantsignalsinPDAC precursors;2) especially proteomechangesassociatedwith in pancreaticductaladenocarcinoma(PDAC), covery ofproteinbiomarkersanddrugtargets Our currentresearchinterestsinclude:1)dis- isolated cellsfromvariousclinicalspecimens. tissues, bloodand otherbodilyfluids, aswell disease andcontrolspecimens, includingtumor The samplesanalyzedincludeavarietyof to improvediagnosisortherapeutictreatment. identify cancerassociatedproteinbiomarkers PTM statusassociatedwithdiseases, andto ing tumorigenesis, toinvestigatechangesin understand themolecularmechanismsunderly- carried outwithvariousgoals, aimingtobetter cancer andotherdiseases. These studies are technologies areappliedtostudypancreatic our lab, massspectrometrybasedproteomics lead tomalfunctionofcellularprocesses. In protein-protein interactions, whichmayall post-translational modifications(PTMs), and may includechangesinproteinexpression, alterations thatareassociatedwithdiseases alterations atafunctionallevel. Proteome to investigatedisease-associatedmolecular clinical research, providingauniqueavenue ing asapowerfultoolintranslationaland quantitative proteomics, hasbeenemerg- detection ofdiseases. Proteomics, especially andearly targets fordrugdevelopment physiologic activitiesandhavebeenimportant ofcellular ecules thatareinvolvedinallsorts Proteins areessentialfunctionalbiomol- Development ofquantitativeproteomics and •Development biomarker •Quantitative proteomicstosupport •Characterization ofproteomealterationsin •Investigation ofthemacroheterogeneity •Characterization ofproteinglycationend- ofproteomicsignaturesforearly •Development RESEARCH PROJECTS selection andtherapeutictargets. marker discoveryforearlydetection, treatment other diseasemechanismstofacilitatebio- regulatory pathwaysassociatedwithcancerand analysis ofproteininteractionnetworksand componentsinourstudyfor are important with diseases Proteomics indecipheringproteomealterationsassociated Associate Professor Sheng Pan, Ph.D. bioinformatics fordigitalbiobanking colorectal cancer forulcerativecolitis-associated development ment responseinpancreaticcancer plasma/serum associatedwithdrugtreat- glycosylation pathways malignancy associatedalterationsinN- of proteinN-glycosylationtoelucidate in diabetics products (AGEs)linkedtopancreaticcancer detection ofpancreaticcancerindiabetics Proteomics inclinical andtranslationalstudies Stevanovic, Azhar Khandekar Stevanovic, Azhar Undergraduate ResearchInternship:Kristina Scientist:Lei Visiting Wang, M.D./Ph.D. Research Scientist: Yumi Sullivan Post-doctoral Fellow:HongPeng, Ph.D. LAB MEMBERS Research. 2017, 16(2):665-676. sition inplasmaanalysis”, Journal ofProteome based onoptimizeddata-independent-acqui- Brentnall TA, PanS, “Quantitative proteomics Chari ST, vonHallerPD, EngJK, FengZ, Yan Q, Nigjeh EN, ChenR, BrandRE, PetersenGM, cancer”,Clin Appl.201711(9-10). Proteomics ing galectin-3bindingproteininpancreatic glycopeptide analysis-detectionofcirculat Brentnall TA, PanS, “Spectral library-based Nigjeh EN, ChenR, Allen-Tamura Y, BrandRE, cancer”, 2017, ScientificReports, ;7(1):7950. to sensitizegemcitabine-resistantpancreatic S, “Disrupting glutaminemetabolicpathways dell J, JungL, Pillarisetty VG, Brentnall1 TA, Pan Chen R, LaiLA, Sullivan Y, Wong M, Wang L, Rid- KEY PUBLICATIONS CENTER FORPRECISIONBIOMEDICINE the principlesofphysics. 2)Canwefigureout try tofindoutsuchbiologicalmechanismusing pathways ofescape. The planofmyresearch to such biomarkers:geneticmutationpatternand intense research/studiesaregoingontoout begins? Inthetraditionalbiologicalapproach, from thenon-respondersbeforetreatment determine awaytoseparatetheresponders average. Two majorquestionsare:1)Canwe drugresistanceinabout a yearonthe develop the patientsresponds, andthoseresponding of thesetreatments, onlyaboutathirdof the lifeofmanypatients. However, forboth extended cancersandcertainly treating several past 5years, havebeenquitesuccessfulin and morerecentlytheimmunotherapyin The targetedtherapyinthepast12years, and immunotherapyforcancersusingthelawsofphysics Understanding universaldrugresistancefortargetedtherapy Adjunct Professor Satyanarayan Nandi, Ph.D. assumes thedominantcomponentof deficiency ofthemodelisittwocomponents: survival, andtheuniversaldrugresistance. The give theprogressionfreesurvival, durationof well asmakepredictions. The modeldoes a, bandctosetofbiologicalvariables, as to validatethemodel, correlatetheparameters with theoncologisttogetredactedpatientdata to thetherapy. We are nowtryingtocollaborate term representintrinsicresistanceofthetumor represent theeffectoftherapy, and3rd mor intheabsenceofanytherapy, the2ndterm the normalizedtumorvolumeattime be writtenasdy/dt=ay–by^2+ct. Here, yis differential equationdescribingthemodelcan resistance usingphysicsideas. The ordinary describe boththeresponseanduniversaldrug amathematicalmodelwhich have developed Sofar,develop? togetherwithDr. V. Cristini, I quired andadaptiveresistancethecancercells What aretheintrinsicresistance, andtheac- forthepatientswhodorespond? that develops the mechanismforuniversaldrugresistance The firsttermrepresentsthegrowthoftu- t =0. •Understanding universaldrugresistance RESEARCH PROJECTS models. the cancermetastasis, andbuildquantitative another project, Iamalsotryingtounderstand be continuedandassociatedriskfactors. In it willletusknowhowlongthetherapyshould has betterprobabilitytogetaresponse. Also, some guidancetotheoncologistwhichtherapy ful, thiswillgreatlybenefitpatients. Itwillgive complex situations. Ifthisapproachissuccess- plan toextendthemodelincludemore resistance tothetherapy. Oncevalidated, we much smallercomponent, whichisintrinsically mutations responsetothedrug, butthereisa •Understanding cancermetastasisusingphysi- Post-doctoral Fellow:JosephButner, Ph.D. LAB MEMBERS cancer treatments to targetedtherapyandimmunotherapyin cal andbiologicallaws:physicalbiology

53 • PACT REPORT PACT IMM 54 • IMMPACT REPORT CENTER FORPRECISIONBIOMEDICINE can reducecanceralone andenhancesanti- • effects inovariancancer. vascular maturationtoenhanceanti-tumor aptamer directeddeliveryofsiRNAimproves • increased survivalrates. breast cancermetastasisandleadingto cancer siRNAtothebonemarrow, reducing • following. recent cancer-relatedresearchhasshownthe kinetic andpharmacodynamicproperties. Our is safeandeffective, with desirablepharmaco- cancer. Furthermore, wehaveshownourmethod and moreimportantly, thespreadofmetastatic aptamer targetedapproachreducestumorsize drugs likePaclitaxel. We haveshownthatour chemotherapeutics agentssuchassiRNAor when usedincombinationtherapytogether However, moreeffective aptamersareeven using theE-selectinaptamercalledESTA1. recently showedintwobreastcancercelllines cancer inadose-dependentmanner, aswe aptamers cangreatlyreducetheburdenof E-selectin,CD44, AnnexinA2,and AXL.Such cells, includingthosetargetingtheproteins aptamers targetingbreastandovariancancer anumber of and oncologistshasdeveloped our translationalteamofchemists, biologists, nanoparticles.release ofthesmart tocreateaslowsustained silicon particles can beusedaloneorloadedintolarge agent fortheparticles. particles Suchsmart taining anti-canceragentstoactasatargeting con- used aloneorcanbecoupledtoparticles attachments (X-aptamers). X-aptamerscanbe which arecoupledwithdrug-likeorprotein-like ous forpatients. We usemodifiedDNAhybrids, create harshsideeffects, whicharedeleteri- Current treatmentsareoftenineffectiveor or infectiousorganisms, suchastuberculosis. tack specificdiseasedtissues, mainlycancers, particles,targeting agentsandsmart whichat- Our AXL aptamer(Kanlikiliceretal., 2017) Our Annexin A2 (Mangalaetal., 2016) ESTA1 directedanti- multistageparticles -Inrecentyears Aptamer Development The majorfocusofmylabistodevelop •Software toanalyzehowLymedisease X-aptamerstargetinginfectious •Developing toattack particles ofsmart •Development RESEARCH PROJECTS infection deficits. of long-termLymediseaseinfectionandpost- Antigenic variationisthoughttobethecause to escapefromthehost’s immunesystem. the bacteria’s proteinsurface, whichleads changes inthebacteria’s vlsElocus, andthus called antigenicvariationleadstolarge-scale of Lymedisease. A poorlyunderstoodprocess time duringB. burgdorfori infections, thecause tion) inthevslEsurfaceproteinasafunctionof significant DNAsequencechanges(recombina- of specialtysoftwarefortheanalysis single NGSdatafile. sis ofmanyX-aptamerprojectscontainedina and statisticalanalysispackagesfortheanaly- noise-reduction filters, DNAlengtherrorfilters, with easy-to-usegraphicaluserinterfaces, et al. 2014), acompletelyautomated program trivial. We thereforedeveloped Aptaligner (Lu sequences, andtheanalysisofthemisnot data filesoftencontaintensofmillionsDNA of next-generationsequencing(NGS)data. NGS novelsoftwarefor theanalysis and todevelop of thelabistoprovidebioinformaticssupport toxins andcancercheckpointproteins. targeting ofBruton’s Tyrosine Kinase, C. diff burden. Otherongoingworkfocusesonthe system andreducingtheM. tuberculosis (Tb) M. tuberculosis, therebyenhancingtheimmune tomacrophagesinfectedwith silicon particles ESTA1 andCD44aptamersdelivermesoporous recently (Leonardetal. 2017)showedthatour influenza andMycobacteriumtuberculosis. We targetingClostridiumdifficile(C.particles Diff), fectious diseasesincludesaptamersandsmart ing thedengue2virus, andcurrentworkonin- marrow. targeting theendotheliumoflymphomainbone • tumor effectsincombinatorialtherapy. Targeting cancerwithX-aptamersandnanoparticles Assistant Professor David Volk,Ph.D. escapes hostimmune systems diseases breast andovariancancers A recentprojectinthelabiscreation - Software Development Another focus Previously, anaptamertarget- wedeveloped aptamers(Liuetal.Developed 2018) Visiting Students:SindhuGangula,Visiting SommerLuu Brenda Saucedo(MSIII) Medical Students: Andrea Costello(MSIII), Research Associates: XinLi, M.S. Maria Zaske, Ph.D. Research Scientists:LokeshRao, Ph.D., Ana LAB MEMBERS al. Theranostics (2018)8:31-44. Cells and Tumor Cells. Liu, H., etal., Volk, D.E. et Polymorphonuclear Myeloid-derivedSuppressor A NovelDNA Aptamer forDual Targeting of al., Volk, D.E., etal, Molecules(2017)22:1355. rium Tuberculosis BurdeninMice. Leonard, F., et Increase SelfImmunityandReduceMycobacte- Thioaptamer Targeted DiscoidalMicroparticles Nucleic Acids(2017)9:251-262. Therapy Kanlikilicer, P., etal. Volk, D.E., etal. Molecular toneal MetastasisinOvarianCancerModels. Kinase Inhibits Tumor GrowthandIntraperi- Receptor Tyrosine of AXL Therapeutic Targeting KEY PUBLICATIONS cancer reduction. ESTA1 aptamerprovidesdose-dependent attackingcancer. slowreleaseparticles Smart CENTER FORPRECISIONBIOMEDICINE therapeutic. research, diagnosticassays, andpotentiallyasa synthetic alternativetoantibodiesforuse in tumor tissue. These X-aptamersprovidea binding toPD-L1expressiononpancreatic shown thesamecapacityasPD-L1antibody similar cellbindingintensity. XA-PDL1has antibodies, XA-PD1andXA-PDL1demonstrated X-aptamers. Compared toPD-1andPD-L1 / programmeddeath-ligand1(PD-L1)specific protein programmedcelldeath1(PD-1) successfully identifiedimmunecheckpoint the bead-basedX-aptamerlibrary, wehave tory antibodieswithnoimmunogenicity. Using are attractivealternateforimmunomodula- inflammatory pathologies. Modifiedaptamers withimmunomodulatoryantibodiesare events cancer growth. However, themajoradverse restore latentanti-tumorimmunityandcontrol as highlypromisingtherapeuticagentsto to atargetbiomoleculeinsolution. enabling rapidselectionofXAaffinityreagents chemically-modified DNAbasesandbackbones bead-based X-aptamer(XA)librariescontaining resistance andbinding. We developed further ester backbone(TA) forenhancednuclease thiophosphate substitutionsofthephosphate tamer librarythathasbeenmodifiedwith cation forvariousapplications. low cost, non-immunogenicity, andfacilemodifi- monoclonal antibodiesintermsoffeasibility, and offermanysignificantadvantagesover alternatives toantibodiesontargetedtherapy, and specificity. DNAaptamersareattractive can bindtoaspecifictargetwithhighaffinity oligonucleic acidorpeptidemoleculesthat for precisioncancertreatment. Aptamers are and targetedtherapyareattractiveapproaches therapy. Aptamer mediatedbiomarkerdiscovery prognosis, andpredictionofresponseto biomarkers areessentialforclinicaldiagnosis, ofprecisionmedicine.impotent parts Validated Beyond selection forsingletargetprotein,Beyond we Immune checkpointantibodiesareemerging We createdacombinatorialDNAap- Biomarker discoveryandtargetedtherapyare Cancer biomarker discoveryandtargetedtherapy Assistant Professor Hongyu Wang, M.D., Ph.D. Develop immune-checkpointblockade X- •Develop •Proteomics biomarkerdiscovery •Targeted cancertherapywithaptamermedi- RESEARCH PROJECTS extended tomanyothercancersordiseases. ery, imagingandtargetedtherapy, andcanbe platformforbiomarker discov- build upanew inthoseprojects will technologies developed applied inclinicaldiagnosisandprognosis. The specifically bindbiomarkersoncancercellsand targetedtreatmentforcancer.new of offer arapidandhighlyspecificdevelopment assembly aptamer selectionandnanoparticle and anti-tumoreffects. These methodsof A2 siRNAresultedinprofoundanti-angiogenesis tumor vasculature. Targeted deliveryof Annexin selective deliveryoftherapeuticagentstothe Annexin A2-specific aptamerthatallowsfor Sequence Endo28wascharacterizedasan that bindspecificallytothosetargetcells. or tumorcellsandsuccessfullyidentified TAs (TAs) againstpatient-derivedendothelialcells processes toidentifyapanelofthioaptamers based(Morph-X-Select)aptamerselection ogy used acell-basedSELEXandtissuemorphol- (Kd) (B)andincomparison toappropriateantibodies(C). L1 expressioncellsandtheirbindingaffinity wereanalyzedbyflowcytometry(A), dissociation constant Binding affinityofselectedXAs. SelectedXA-PD1-78andXA-PDL1-82 wereincubatedwithPD-1orPD- aptamers forcancerimmunotherapy delivery ated nanoparticle–drug The identifiedthioaptamer/X-aptamercan Selection and Validation.Selection Ward N, GorensteinDG, Volk DE. X-Aptamer Lokesh GL, Wang H, LamCH, Thiviyanathan V, print]; PMID:28912094 10.1016/j.biochi.2017.09.006. [Epubaheadof Sep 11. pii:S0300-9084(17)30230-4. doi: tion ofPD1/PD-L1X-Aptamers. Biochimie. 2017 Wang H, LamCH, LiX, West DL, Yang X. Selec- PMCID: PMC5070952. (*equalcontributions) insight.87754. PubMedPMID:27777972; 2016;1(17):e87754. doi:10.1172/jci. antitumor effectofchemotherapy. JCIInsight. maturation usingnoncodingRNAsincreases G, GorensteinDG, Sood AK. Improvingvascular PL, CooperLJ, RosenblattKP, Lopez-Berestein Zhang X, MaitiSN, N, Bulayeva ChoiHJ, Dorniak Gharpure KM, McGuireMH, Thiviyanathan V, E, BayraktarR, LiL, Tanaka T, Hu W, IvanC, Aguayo C, Nagaraja AS, RupaimooleR, Bayraktar Pradeep S, Yang X, HaemmerleM, Rodriguez- Somasunderam A, Volk DE, LokeshGL, LiX, *Mangala LS, *Wang H, *JiangD, Wu SY, KEY PUBLICATIONS Research Associate: LiLi, M.S. Research Associate: XinLi LAB MEMBERS 4939-7138-1_10. PubMedPMID:28730438. 2017;1632:151-174. doi:10.1007/978-1- Methods MolBiol.

55 • PACT REPORT PACT IMM IMMPACT REPORT • • • • 57 For patients presenting with genetically patients presenting For evidence for the is increasing there Finally, in principle, may be specifically guided into the in principle, may be specifically guided the within cell types and tissues present various seeking within the center are Faculty human body. robust methodologies efficient and to develop cells/tissues of to convert iPSCs into various including neural, blood, therapeutic interest to lung, muscle, and cartilage as how – as well maintain such cells/tissues for and best deliver therapeutic benefit. utilizing inherited disease, center faculty are technologies to gene editing developed recently the disease-causing mutations in either correct stem cells or iPSCs. The goal of tissue-resident of therapies that these studies is development in a patient’s the mutations include correcting either the cells, then delivering stem own from stem cells or cells/tissues derived corrected this them back into the same patient. Presently, within the center is being investigated approach lung (Cystic for muscle (muscular dystrophies), and blood (immune deficiencies). Fibrosis), within cancers of cells having specific presence properties typically associated with stem cells. of such the role interrogating Center faculty are cells in the initiation and maintenance of cancers lymphoma. cancer and such as ovarian Ph.D. R. Davis, Brian Director and Center Associate Professor Wallace Distinguished and Lorine G. Harold The C. Chair University he faculty, research research he faculty, trainees staff, and for of the Center There are at least two distinct classes of stem are There Stem Cell and Regenerative Cell and Regenerative Stem (CSCRM) are Medicine focused on experimental studies of the biological properties of stem cells in The both health and disease. in healthy (or normal) interest by stem cells is motivated in both their essential role – from normal development the fertilized egg to fully organism – as developed as in maintenance of well tissues and organs throughout of the hopes of life. One medicine regenerative is that this fundamental understanding of stem cells may be effectively stem translated into therapies in which healthy to can be employed cells, or their derivatives, cells and tissues lost as a consequence replace or disease. This of normal aging, injury, already therapeutic potential of stem cells has decades in which been demonstrated for several either bone blood stem cells (obtained from peripheral blood, or umbilical mobilized marrow, to regenerate been utilized blood) have cord patients the complete blood system in cancer is There undergoing chemotherapy for cancer. accumulating evidence that certain stem cells, facilitate such as mesenchymal stem cells, may inflammation. reducing of tissue damage by repair will find several you pages following the In examples of center faculty exploring the potential tissues of stem cells for repairing therapeutic value brain, muscle, cartilage, lung, such as spinal cord, and blood. within the center investigation cells under active for such therapeutic applications. The first of stem cells; such cells tissue-resident these are as organs such life in various throughout present lung involved intestine, and are bone marrow, of cells and tissues lost regeneration in active or aging, injury, due to normal cell turn-over, disease. A second class of stem cells of significant is investigators to center therapeutic interest induced pluripotent stem cells (iPSCs). iPSCs are patient-specific stem cells that can be generated any individual and easily obtained cells from from STEM CELL AND REGENERATIVE MEDICINE REGENERATIVE AND CELL FOR STEM CENTER T 407:259-270, 407:259-270, CRC Press, Taylor & Taylor CRC Press, Simulation examples of a multiscale hybrid model for simulating ductal carcinoma in situ (DCIS) growth. KEY PUBLICATIONS KEY PUBLICATIONS An Wang. Zhihui Koay, J. Eugene Vittorio Cristini, Introduction to Physical Oncology: How Mecha- Can Improve nistic Mathematical Modeling Cancer Therapy Outcomes. July 2017 Group. Francis Butner Dogra P, YL, Chuang Kerketta R, Z, Wang Simbawa Shen H, Xu R, A, Day TA, Brocato JD, Ferrari Curley SA, Mahmoud SR, AS, Al-Fhaid E, Theory and Experimental V. Cristini Koay EJ, M, of a Spatio-temporal Model of Che- Validation Cell Kill. Tumor to Enhance Transport motherapy 2016 PLoS Comput Biol 12:e1004969, Al-Fhaid Simbawa E, YL, Chuang Butner JD, A hybrid Z. Wang V, Cristini Mahmoud SR, AS, agent-based model of the developing mammary J Theor Biol terminal end bud. 2016 LAB MEMBERS Ph.D. Post-doctoral Fellows: Joseph Butner, Terisse Graduate Students: Prashant Dogra, Tito Jorge Naomi Hasegawa, Brocato, A hybrid multiscale modeling approach to study normal mammary gland development and breast cancer initiation Development of dynamic molecular target se- and validation methods identification, lection, with multiscale modeling Predictive modeling of cancer treatment • • Quantitative biologyQuantitative and medicine on biophysical erful) mathematical tools based propertiestheories to correlate physical of drug transport progression and treatment with tumor with experimental/clinical in- Together outcome. we use ODE- and PDE-based models vestigators, for each indi- to predict treatment outcome Since vidual patient prior to actual treatment. these tools are derived based on fundamental they are broadly principles of mass transport, The concept applicable to the clinical sciences. of this approach is also likely to be useful beyond context of cancer in any case where the drug delivery relies on local diffusion properties, demonstrating the general applicability and broader impact of his modeling method. RESEARCH PROJECTS • Zhihui (Bill) Wang, Ph.D. Wang, Zhihui (Bill) Associate Professor Our research program focuses on integrating Multiscale cancer modeling. Cancer Most Cross-scale drug target discovery. cancer modeling. We are Translational

and statistical methods physical, mathematical, with experimental investigations and patient data analysis to quantitatively study tumor are working to We progression and invasion. use our models to help biologists and medical scientists to simulate experimental procedures, optimize and predict clinical therapies and biological/ and test and refine their outcomes, three specific have We medical hypotheses. research areas. integrated phenomenon growth is an emergent, that spans multiple spatial and temporal biological scales resulting from dynamic and interactions between individual cells, between cells and their constantly changing address a Our research projects environment. challenging part of of current systems modeling and linking, cancer: bridging the gaps between, and tissue multicellular, cellular, the molecular, integrated a also have successfully We scales. combination of in vitro and in vivo experiments paired with patient data analysis with the These models examine mathematical models. how changes occurring at the molecular level percolate across and affect tumor growth behaviors at the tissue and tumor scales. mathematical models used in identifying cancer drug targets to date focus on the molecular and large-scale proteins, on genes, level (i.e., selection and However, signaling networks). identification of drug targets that account for and tumor-scale multicellular-, molecular-, behaviors is potentially more realistic and hence more powerful than focusing only on are developingWe cross-scale cell signaling. drug target evaluation methods for identifying potential drug targets and multi-target thera- peutics (high in both efficacy and safety while based minimizing unintended adverse effects), on single- and multiple-parameter perturbation algorithms. developing practical (relatively simple yet pow- PRECISION BIOMEDICINE FOR PRECISION CENTER

IMM PACT REPORT PACT • • • • 56 58 • IMMPACT REPORT CENTER FORSTEMCELL AND REGENERATIVE MEDICINE blood cellsbearinginherited geneticmutations that iswhenspontaneous mutationsarisingin project focuseson “natural genecorrection,” mutation.of therelevant The third laboratory original stemcellsbythegeneticcorrection lung orbloodstemcells, onlydifferingfromthe is thedeliverybacktopatientsoftheirown the CFand WAS projects, theultimateobjective to bloodstemcellsfortransplantation. Inboth – oriPScellswithsubsequentdifferentiation mutations inpatient-specificbloodstemcells we areseekingtocorrectthedisease-causing (WAS), aprimary immunedeficiency. Again, disorders suchasthe Syndrome Wiskott-Aldrich mutations responsibleforinheritedblood focuses onthesite-specificcorrectionofgene The secondmajorprojectinthelaboratory stem cellsdirectlyobtainedfromCFpatients. to correcttheCFmutationsintissue-specific fore-mentioned genecorrectionmethodologies approach. We arealsopresentlyutilizingthe order tofacilitateapersonalizedtherapeutic testing sensitivitytospecificCFdrugs–in specific iPScell-derivedlungepitheliumfor of ourobjectivesistoemployCFpatient- molecular andfunctionalcharacterization. One isolate earlylungprogenitorsforpurposesof ers intoiPScellsandutilizedtospecifically - introduction oflung-specificfluorescentreport corrected iPScells. We haverecentlyreported in lungepithelialcellsderivedfromthese demonstrated geneticandfunctionalcorrection –andhave from patientswithCysticFibrosis genetic mutationsiniPScelllinesderived directed repairtocorrectthemostcommon sequence-specific nuclease-mediatedhomology therapeutic approaches. We haveutilizedDNA stem/progenitor cell-based goal ofdeveloping system. This isbeingpursuedwith theultimate inherited disordersaffectingthelungorblood specific stemcellsderivedfrompatientswith pluripotent stem(iPS)cellsand/ortissue- mutations inthechromosomalDNAofinduced the sequence-specificgeneticcorrectionof My laboratoryhasasitsprimaryobjective laboratory (incollaboration withD. Kotton and F. Hawkins). can usesuchGFP-expressing cellstogeneratelung-specificorganoids andbronchospheresinthe andcorrectediPScells.lung cellsderivedfrom CysticFibrosis As shown intherighthandpanels, we Line.NKX2.1 GFPReporter We tospecificallyidentifyandisolate areutilizinganNKX2.1GFPreporter defects. StemCellReports. 2016, 7:139-148. and functional the lymphoiddevelopmental Syndromepatientnormalizes Wiskott-Aldrich tion ofinducedpluripotentstemcellsfroma B. Vandekerckhove, B.R. Davis:Genetic correc- Paschon, E.J. Rebar, M.C. Holmes, D. Kaufman, Liao, M. Gonzalez-Garay, J.C. Segovia, D.E. C. Ma, Z. Ni, Z. Garate, A.M. Crane, X.S. Li, W. T.J. Laskowski, Y.V. Caeneghem, R. Pourebrahim, KEY PUBLICATIONS •Characterization ofspontaneousgene •Correction ofiPSandbloodstemcellsfrom •Correction ofiPSandairwaystemcellsfrom RESEARCH PROJECTS patients withthe Syndrome. Wiskott-Aldrich asitoccursinvivo correction particularly We arepresentlyexaminingthisnatural gene somaticmosaicism.phenomenon ofrevertant correctedcells.revertant This givesrisetothe gene, followedbyinvivoselectionforthe result infunctionalrestorationofthedefective blood andlungdiseases Genetically correctedstemcellsfortreatmentofinherited C. HaroldandLorineG. Wallace DistinguishedUniversityChair Director oftheCenterforStemCellandRegenerativeMedicine Associate Professor Brian Davis, Ph.D. Wiskott-Aldrich Syndromedefects Wiskott-Aldrich mutations resultingincorrectionofinherited Syndromepatients Wiskott-Aldrich patients Cystic Fibrosis Matthias Research Staff:Dr. Ana M. Crane, Dr. Nadine Medical Students:NicholasKing Students: Varada Anirudhan Graduate Post-doctoral Fellows:Dr. LeilaRouhigharabaei LAB MEMBERS 21:1-17. alveolar epithelialcells. CellStem2017, man pluripotentstemcellsintofunctionallung M.F. Beers, D.N. Kotton. Differentiationofhu- Whitsett, F.S. Cole, E.E. Morrisey, S.H. Guttentag, A. Crane, B.R. Davis, F.V. White, J. Wambach, J.A. W. Zacharias, M. Ochs, K. Traber, L.J. Quinton, Vedaie, K. Hurley, A. Hinds, S.J. Russo, S. Kook, J.C. Jean, H. Heins, C-LNa, T.E. Weaver, M. A. Jacob, M. Morley, F. Hawkins, K.B. McCauley, 2017, 127:2277-2294. Stem Cells. Journal ofClinicalInvestigation man LungProgenitorsDerivedfromPluripotent Prospective IsolationofNKX2.1-expressingHu- J.R. Rock, J.M. Shannon, B.R. Davis, D.N. Kotton. Wong, A.S. Khalil, S.X.L. Huang, S. Guttentag, A. Kurmann, A. N. Hollenberg, S. Nguyen, B.G. Thomas, K.A. Benson, N. Skvir, A.M. Crane, A. F. Hawkins, P. Kramer, A. Jacob, I. Driver, D. CENTER FORSTEMCELL AND REGENERATIVE MEDICINE central nervoussystem. However, thenumberof for thetreatmentoftraumatic injuriestothe offer apotentiallyautologous cellsources Human inducedpluripotentstemcells(hiPSC) cellsourcesalsoneedtobeoptimized.relevant expansion anddifferentiationofclinically widespread clinicaluse, protocolsforthe injury. cavities insubacuteratmodelssospinalcord of axonsfromthehostacrossspinalcordlesion formulating matricestofacilitatetheextension matrices. Preliminary studieshavefocusedon farbetterthantraditional tissue development native tissuemicroenvironmentandsupport these hybridmatricescanbegintoemulatethe bioactive signalingandcontrolledarchitecture, respectively. When combinedwithadditional andcellularresponse,in scaffoldproperties signaling andbatchtoinconsistency their disadvantages, namely lackofbioactive bioactive signaling)polymers, whilemitigating and cellularresponse)natural(native both synthetic(consistencyinfabrication These matricesmaximizetheadvantagesof integrated intoadvancedhybridmatrices. using combinatorialmethods, willbe they peptide concentrationshavebeenidentified mechanical properties, andbioactivesignaling (porosity, featuresize, fiberalignment, etc.), tissue. Onceoptimalcomposition, architecture and integrationwiththesurroundinghost extracellular matrixfacilitatetissuerepair seek tounderstandwhichaspectsofthenative use instemcelltherapy. cellsources for expansion ofclinicallyrelevant seeks tooptimizescaffolddesignandthe Utilizing engineeringapproaches, thelaboratory cell biology, chemistry, andmaterialscience. techniques fromcell, molecular, andstem tory usesaninterdisciplinaryapproachinvolving traumatic braininjury, andstroke. The labora- toward clinicaltreatmentsforspinalcordinjury, tissueengineeringapproaches developing In ordertoadvancetissueengineering By examiningcell-materialinteractions, we The researchinmylaboratoryfocuseson staining isblue. Scalebars=50µm. network organization inresponsetothechangestiffnessof the hydrogelculturesubstrate. Nuclear induced pluripotentstem cellderivedneuralstemcytoskeletal organization(red)andneural andresponseofhuman Schematic ofhydrogelwithacontinuous gradient inmechanicalproperties MC,Mosley LimHJ, ChenJ, Yang Y-H, LiS, Liu KEY PUBLICATIONS • • RESEARCH PROJECTS onhiPSClineagechoice.properties gradient tostudytheeffectofmechanical of hydrogelswithcontaininga Young’s Modulus the covalenttetheringofproteinstosurface from hiPSC. Preliminarystudieshavefocusedon stem cellsandoligodendrocyteprogenitor cell culturesurfacesfordifferentiationofneural culture systemstooptimizetwo-dimensional to extendourknowledgeofthreedimensional to hiPSCdifferentiation. The laboratoryseeks have notbeenstudiedsignificantlyinrespect significantly affectcellulardifferentiationbut of thecellculturesurfacehavebeenshownto Both biochemicalandmechanicalproperties rent differentiationprotocolsisextremelylow. viable cellsfortransplantproducedfromcur- injuries engineeringapproachesforthetreatmentofCNS Tissue Assistant Professor Laura A. SmithCallahan, Ph.D. combinatorial approaches using concentration ormechanicalproperties suchasbioactivesignalingmoiety properties matrix Identification ofoptimalartificial progenitor cells cells toneuralstemandoligodendrocyte entiation ofhumaninducedpluripotentstem Optimization ofculturesurfacesforthediffer- Post-doctoral Fellows: T. HiranPerera;XiLu LAB MEMBERS Science &Engineering. 3(5):776-781, 2017. peptide His-Ala-Val-Asp-Ile. ACS Biomaterials concentration gradientofn-cadherinderived acrylate hydrogelscontainingacontinuous differentiation onpolyethyleneglycoldimeth- cell derivedneuralstemsurvivaland Callahan LA. Humaninducedpluripotentstem Kurosu YE, RavivarapuK, MC, Mosley Smith Lim HJ, KhanZ, Wilems TS, LiX, Perera TH, 153-160, 2017. His-Ala-Val-Asp-Lle. ActaBiomaterialia.56: tion gradientofn-cadherinderivedpeptide hydrogels possessingacontinuousconcentra- cultured onpolyethyleneglycoldimethacrylate differentiation ofmurineembryonicstemcells Concentration dependentsurvivalandneural Lim HJ, MC, Mosley Kurosu Y, SmithCallahanLA. 824-833, 2017. Biomedical MaterialsResearch: Part A. 105(3): continuous Young’s Modulusgradient. Journal of on apolyethyleneglycolhydrogelscontaining pluripotent stemcellderivedneuralcells neuronal differentiationofhumaninduced Y, SmithCallahanLA. Neuriteextensionand

59 • PACT REPORT PACT IMM 60 • IMMPACT REPORT CENTER FORSTEMCELL AND REGENERATIVE MEDICINE reprogramming active astrocytesintoneuronal major inhibitorforaxonal regeneration.Insitu stroke brainintoneurons. Astroglial scararethe astroglial cellsintheinjuredspinalcordor testing whetherwecandirectlyreprogram the cell graftforSCIandstroke. Recently, we are or glialprecursorcellstoidentifytheoptimal ficacy andlong-termsafetyofNSCs, neuronal Currently, wearetestingthetherapeuticef- preserve andrestoreneurologicalfunctions. hiPSC-derived NSCisaneffectivetherapyto These studiessuggestthattransplantationof spinal cordandpromotefunctionalrecovery. glias aftertransplantationintothecontused survive anddifferentiateintobothneurons drocytes. Importantly, hiPSC-derived NSCscan functional neurons, astrocytes, andoligoden- time hiPSC-derived NSCscanproliferateoverlong cursor cellsfromhiPSCs. Our resultsshowthat purify NSC, neuronal precursorcellsorglialpre- protocoltodifferentiateand We havedeveloped grafts withouttheneedofimmunosuppression. use, andespeciallytheabilitytogenerateiso- material withoutethicalconcernsofembryo advantages intermsofavailabilitysource with ESCs, hiPSCsoffersignificantadditional pluripotency transcriptionfactors. Compared cells byover-expressionoffourdevelopmental/ ESC-like cells reprogrammed from adult somatic remarkablepluripotent,are recentlydeveloped Human inducedpluripotentstemcells(hiPSCs) have additionalriskofteratomaformation. and graftrejection. CellsderivedfromhESCs cells areassociatedwithethicalcontroversy clinical trialsforSCIpatients. However, these cells (hESCs)-derivedhavebeenapprovedfor cells derivedfromhumanembryonicstem fetal cadaverbraintissueandneuralprogenitor central nervoussystemstemcellisolatedfrom which NSCresourceisoptimalforSCI. Human stroke. However, thereisnoconsensusasto diseases, includingspinalcordinjury(SCI)and promote functionalrecoveryafterneurological is provedapromisedtherapeuticapproachto Transplantation ofneuralstemcells(NSCs) in vitroandbeinducedtodifferentiateinto processes extendingtothesparedhostspinal cord(D). injury side(A, B). ofneurons(C)withlong GraftedhiPSC-NSCsdifferentiatedintocellswithmorphology The graftedNSCsfilledthecavityandmainly remainedinareasaroundtheinjuryepicenterat NSCs wasobservedinallanimalsreceived hiPSC-NSCgraftat2monthsaftertransplantation(A, B). week aftermoderateC5unilateralcontusion SCI(IH180kdyne)inruderats. Robust survivalofgrafted Survival ofgraftedhiPSC-derivedneuralstem cells. HumaniPSC-derivedNSCs weretransplantedat2 QL (2017). Humanneuralprogenitorsderived roeder GW, Wu JQ, Zhang YY, KimDH, andCao Liu Y, Zheng YY, LiSL, XueHP, SchmittK, Hergen- KEY PUBLICATIONS •Human iPSC-derivedneuralstemorprecursor •Treating neuropathicpainbyinvivorepro- •In vivoreprogrammingofreactiveastrocyte RESEARCH PROJECTS stroke. stem cell-basedtherapiestotreathumanSCIor or stroke. novel Ourlong-termgoalistodevelop to promotegreatfunctionalrecoveryafterSCI These twomechanismsmayworksynergistically replace thelostneuronsafterSCIorstroke. reprogrammed neuronalprecursorcellscould to promoteaxonalregeneration. The newly precursor cellswilldecreaseastrocyteinhibition Stem cellsforneurologicaldiseases Associate Professor Qi LinCao, M.D. cells forspinalcordinjuryandstroke gramming ofastrocytesafterSCI and chemogeneticapproachforSCIrepair Research Assistants: JunLi;ChrystineGallegos Post-doctoral Fellows: Zheng;XiuquanHe Yiyan LAB MEMBERS 299(Pt A):97-108. Kim DH, CaoQ.,. Experimentalneurology 2018; injury, ChengX, Zheng Y, BuP, QiX, FanC, LiF, roprotection targetaftertraumaticspinalcord Apolipoprotein Easanoveltherapeuticneu- co-corresponding authors). 7:41008. doi:10.1038. PMID:28106101 (* chronic stagesofspinalcordinjury. SciRep. long non-codingRNAsinthesub-chronicand (2017). The systematicanalysisofcodingand XF, GrillR, KimDH, CaoQL*and Wu JQ* DuranR,Cuevas-Diaz Yan H, Zheng YY, Huang 28073086 Stem CellRes.9:55-64. doi:10.1016. PMID: from integration-freeiPSCsforSCItherapy. cell productionforuseinourclinicalprotocols. treatments. These facilitiesallow clinicalgrade 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 significant engraftmentinthebrainlong cellular therapieshavebeensuccessfulwithout of theinnateimmuneresponseto TBI andhow TBI. We havebeeninterestedinthemodulation injuries, principallytraumaticbraininjury, or the useofcellulartherapiesforneurological CENTER FORSTEMCELL AND REGENERATIVE MEDICINE Our teamdirectstheGriffinStemCellLabora- Our currentresearchprogramfocuseson Hasen Xue, M.D.-Research Associate innate immunity Naama Toledo-Furman, Ph.D.-Flow Cytometry/ Amit Srivastava, Ph.D.-Assistant Professor Supinder Bedi, Ph.D.-Assistant Professor Cecilia Martin, Ph.D.-Research Associate Prabhakara-SrResearch Karthik Assistant Katherine Ruppert, Ph.D.-Sr Research Associate Scott Olson, Ph.D. - Assistant Professor Therapy Mitchell George, M.D.-TBI-Clinical andCell Louis Carrillo, M.D.-TBI-Clinical andCell Therapy Therapy Akshita ‘Jade’ Kumar, M.D.-TBI-Clinical andCell Cai-Programmer Analyst Yidao Joiya Arrington, MSN, RN-TBI Clinical Kosmach,Steven MSN, RN, CCRC-TBI Clinical LAB MEMBERS 2016. PMID:27800660, PMCID:PMC5367945 mononuclear cells. StemCells 35:1065-1079, adult traumaticbraininjuryusingbonemarrow Holcomb JB, KitagawaR. Treatment ofsevere Pedroza C, LeeDA, Worth L, Aisiku I, ChoiHA, Romanowska-Pawliczek A, Triolo F, DashPK, Cobbs L, JuranekJ, SavitzSI, JacksonML, Cox CS, HetzRA, LiaoGP, BM,Aertker Ewing- DMPK. 3(3):182-189, 2015. in traumaticbraininjuryresearch. ADMET& CS. Assessing bloodbrainbarrier permeability Smith PA, HetzRA, XueH, BediS, OlsonSD, Cox Liao GP, BA,Aertker KotaDJ, PrabhakaraKS, PMID: 28624058 a Meta-Analysis.JSurg Res214:38-48, 2017. progenitor celltherapyintraumaticbraininjury: Jackson ML, Srivastava A, CoxCS. Pre-clinical KEY PUBLICATIONS •Imaging ofmicroglialactivationinvivo •Novel deliverysystemsforstemcellsin •Amniotic fluidderivedMSCsforthetreatment •IND-enabling studiesusing APCs fortrau­ ofPhase1and2Clinical •Development Trials RESEARCH PROJECTS Cellular therapiesforneurologicalinjury George andCynthiaMitchellDistinguishedUniversityChair Professor Charles Cox, Jr., M.D. neurological injury bypass/hypothermic circulatoryarrest diseaseandcardiopulmonary genital heart of neurologicalinjuryassociatedwithcon- brain injury traumatic braininjury using non-ESCstem/progenitorcellsfor matic StephanieBaca-ProgramManager Christina Manager Willingham-Program MaxSkibber-Research Assistant Tushar Sharma-ScientificProgrammer Kevin Aroom-Scientific Programmer MatteoCostantini-Research Assistant DeepaBhattarai-SrResearch Associate Coordinator Romina Gipson-Love-QualityImprovement Sufira Kiran, GMP-QADirector Fabio Triolo, Ph.D.-GMP CenterDirector cell production. ofanovelbio-reactorforstem Development

61 • PACT REPORT PACT IMM 62 • IMMPACT REPORT genes. formyogenic iPS cellsusingknock-inreporters to studymyogenicdifferentiationofhumanES/ and MusculoskeletalSkinDiseases(NIAMS) grant awardfromNationalInstituteof Arthritis survival andengraftment. ing theeffectoflocaltissueperfusionincell celldelivery;aswellexplor- systemic/arterial involved incellhomingintothemuscleafter delivery andengraftment;studymechanisms vitro genecorrectionofiPScells;optimizingcell of iPScellsfrommusculardystrophypatients;in dystrophies andmusclemassinjuries. of thesecellsinmicemodelsmuscular ofinvivoregenerationpotential and evaluation inducers ofmyogenesisinhumanstemcells throughput screening(HTS)toidentifyimportant Other majorgoalsofthelabincludeusinghigh progenitors forfuturecellbasedtherapies. a crucialsteptoidentifyandisolatemyogenic myogenic progenitorsfromhumanES/iPScells; This willallowstudyingtheemergenceofearly early myogenicgenessuchasPAX7 andMYFF5. celllinesfor knock-in humanES/iPSreporter system) ourlabhassuccessfullygenerated editing technologies(suchasCRISPR/Cas9 for differenttypesofmusculardystrophies. cells (iPScells)forcelltherapyinmicemodels (ES cells)aswellinducedpluripotentstem methods forusinghumanembryonicstemcells years,ing thelastfew novel wehavedeveloped stem cellsforskeletalmuscleregeneration. Dur- CENTER FORSTEMCELL AND REGENERATIVE MEDICINE Our labiscurrentlyfundedbyaNIH(RO-1) Our researchteamalsoworksonderivation Here atIMM, byusingcutting-edgegene My lab’s maininterestisusingpluripotent transplantation. LacZ+donor-derivedfibers(blue) replacethefibrosisonemonthafter transplantation. B. musclemasslossinjury withmassivefibrosis(left)isrepaired usingbio-scaffold/stem cell Severe A. HydrogelseededwithLacZlabeled stemcellsdemonstratesgoodcellsurvival withinthescaffold. Muscle injuryrepairusingbio-scaffoldseeded withstemcellstorepairmusclemasslossinjury Jun 3;111(22):8275-80. Academy ofSciencestheUSA(PNAS).2014 myocytes invitro.ProceedingsoftheNational tent andembryonicstemcell-derivedskeletal structural featuresofhumaninducedpluripo- Perlingeiro R, Tabti N. Physiologicalandultra- Skoglund G, LainéJ, DarabiR, FournierE, KEY PUBLICATIONS • celldeliveryapproachesfor •Systemic/arterial •High throughputscreening(HTS)formyogenic •Gene correctionofmusculardystrophyiPS •Generation ofknock-inhumanES/iPS RESEARCH PROJECTS Skeletal muscleregenerationusinghumanstemcells Assistant Professor Radbod DarabiM.D., Ph.D. models formusclemasslossinjuries Using bio-scaffoldsforcelldeliveryinmice cell therapyinmusculardystrophies cells induction ofhumaniPSreporter cells usingCRISPR/Cas9system (PAX3, PAX7, MYF5) celllinesforearlymyogenicgenes reporter Jonathan Lo Undergraduate Student(RiceUniversity): Master Student(GSBS):JoseL. Ortiz-Vitali Research Associate: NadineMatthias Post-doctoral Fellow:Jianbo Wu LAB MEMBERS 16(2):220-8. Stem CellResearch. 2016Mar; Human iPSCellsUsingCRISPR/Cas9 Technology. tion and Validation ofPAX7 Linesfrom Reporter Wu J, HuntSD, XueH, Liu Y, DarabiR. Genera- srep18759. Reports. 2016Jan5;6:18759. doi:10.1038/ ated HomologousRecombination.Scientific Human iPSCellLineUsingCRISPR/Cas9Medi- tion andCharacterizationofaMYF5Reporter Wu J, HuntSD, XueH, Liu Y, DarabiR. Genera- CENTER FORSTEMCELL AND REGENERATIVE MEDICINE to structures including the periaqueductal grey to structuresincluding theperiaqueductalgrey the centralnucleusof amygdala(CeA)project for mediatingfearresponses. The neuronsof sential fortheformationoffearmemories and this associationarelargelyunknown. and neurochemicalmechanism(s)thatunderlie traumatic stressdisorder(PTSD), thecellular and stress-relateddisorders, especiallypost- comorbiditybetween studies havereported TBI through extinction. Although epidemiological in agradualreductionofthefearresponse absence oftheunconditionedstimulusresults exposure totheconditionedstimulusin to astheconditionedresponse. Repeated the trigger), itcausesafear responsereferred the conditionedstimulusalone(referredtoas When thepersonissubsequentlyexposedto isrobustandcanbelong-lasting.matic event is learnedrapidlyandthememoryfortrau- the unconditionedstimulus). This association occurred, (referredtoas andtheharmfulevent a smelland/orthecontextinwhichevent as theconditionedstimulus)suchasound, tween anotherwiseinnocuousstimuli(referred event, thesubjectcanformanassociationbe- ders. When apersonexperiences atraumatic lies atthecoreofmanystress-relateddisor- to extinguishthememoryofatraumaticevent and otherstress-relateddisorders. An inability logical consequencesofbraininjuryisanxiety andlong-lastingconsequences.short-term concussion isaprogressivediseasewithboth incident. indicatesthata Mountingevidence of concussionarerestrictedtothetraumatic lenged theperceptionthatconsequences sified asconcussion. Recentstudieshavechal- physician contacts), 87%ofwhichcanbeclas- (i.e., hospitalization, emergencyroomorother sustain adocumentedbraininjuryeachyear Control, approximately2.6million Americans sexes. According totheCentersfor Disease health problemforpersonsofallagesand brain injury, mTBI)hasemergedasamajor The amygdala is a key structurethatises- The amygdalaisakey One ofthemostcommonlong-termpsycho- Concussion (alsoknownasmildtraumatic 2017 [Epubaheadof print] tion ofContextualFear Memory. JNeurotrauma. andImpairsExtinc- the MedialPrefrontalCortex Reduces SpineDensityofProjectionNeurons in Moore AN, DashPK. Mild Traumatic BrainInjury Zhao J, HuynhJ, HylinMJ, JJ, O’Malley Perez A, KEY PUBLICATIONS •To investigatetheconsequencesofmito- •To investigateneurovascularfunctionafter •To identifyhowconcussionaltersneural RESEARCH PROJECTS disorders. to reduceamygdalaoutputandstress-related stage ofinjurymayenhancetheabilityIL gies toincreasespinedensityinthechronic disorders afterconcussion. Alternatively, strate- ofstress-related thedevelopment may prevent molecular strategiestoblockthelossofspines is reduced. Further, theseresultssuggestthat activity ofthecentralnucleusamygdala gest thattheabilityofILneuronstoinhibit for neuronalcommunication, theseresultssug- pyramidal neurons. As spinesare potentialsites reduction inthespinedensityoflayerII/III fication ofdendriticspinesshowedasignificant a lossofneuronsintheIL. Interestingly, quanti- thatthisdysfunctionisnot causedby revealed the conditionedstimulus. Ourcellularanalysis fearful memoryinspiteofrepeatedexposureto in theabilityofinjuredanimalstoextinguisha single mTBIcancauseprolongedimpairments and theformationofextinctionmemory. tion ofILneuronsfacilitatesextinctionlearning of extinctionisimpaired. Incontrast, stimula- and unconditionalstimuli, however, thememory learn theassociationbetweenconditioned activity ofILneuronsisblocked, animalscan 1). (Figure medial prefrontalcortex When the ofthe neurons withintheinfralimbic(IL)cortex controlled bytheprojectionsfrompyramidal The outputfromthecentralnucleus, inturn, is mus (mediatesreleaseofstresshormones). the paraventricularnucleusofhypothala- hypothalamus (increasesbloodpressure)and (controls thefreezingbehavior), thelateral Pramod Dash, Ph.D. Concussion andstress-relateddisorders Nina andMichaelZilkhaDistinguishedChair, NeurodegenerativeDiseaseResearch Professor metabolism afterconcussion chondrial plasticityandalteredbrainenergy concussion communication Using arodentmodel, wehavefoundthata block stress-relateddisorders. inhibitamygdala outputand infralimbic cortex Schematic drawingshowing howneuronsinthe Diagnostic andBiomedicalSciences Dr. assistantprofessorof CameronJeter: MedicineInstitute Ironman Sports pedic Surgery;Director, ConcussionProgramat Dr.- SummerOtt:associateprofessorofOrtho group director,ogy; DementiaandMemoryDisorders Dr. PaulSchulz:associateprofessorofNeurol- at MemorialHermannHospital-TMC Medicine; medicaldirector, EmergencyCenter Dr. chairofEmergency JamesMcCarthy: COLLABORATORS 10:29, 2016. Pathophysiology. and TBI MN, DashPK. Altered MitochondrialDynamics Fischer TD, HylinMJ, ZhaoJ, Moore AN, Waxham 18, 2016. Traumatic BrainInjury. JNeurosci.36(9):2809- Barrier PermeabilityfollowingExperimental linergic NicotinicReceptorsReduceBlood-Brain Hood KN, Moore AN. Activation of Alpha 7Cho- Dash PK, ZhaoJ, KoboriN, RedellJB, HylinMJ, Front SystNeurosci.

63 • PACT REPORT PACT IMM 64 • IMMPACT REPORT CENTER FORSTEMCELL AND REGENERATIVE MEDICINE development ordiseasestudiesbyusand development the generatedcellshave beenappliedforlung 2014, NatProtoc2015). As aproofofprinciple, at largequantities(Huangetal. NatBiotechnol lower (alveoli)respiratorylungepithelialcells become differenttypesofupper(airway) and that directshumanpluripotentstemcells to astep-wisedifferentiationstrategy developed critical stepinthisdirection, wehave previously of clinicallyapplicablecelltypes. As afirst, diseases reliesonthesuccessfulgeneration same donor). obtaining airwayandalveolarcellsfromthe subjects (includingtheadditionaldifficultyof limited availabilityoflungtissuefromhealthy duetothe cells areinadequateinlargepart approaches utilizingprimaryhumanadultlung human studies. In addition, experimental indemonstratingconcordancewith fall short existing animalexperimentalmodels, which a systemstemsfromthelimitationsof studies ofhumandiseases. The needfor such model hasemergedasanovelsystemfor organs. the extremelylimitednumberofavailabledonor is lungtransplantation, whichishamperedby definitive treatmentforend-stagelungdisease successful therapeuticapproaches. The only diseases, andconsequentlythereisalackof pathogenic mechanismsresponsibleforthese we stillhaveverylimitedunderstandingofthe wide. Despitethehugelungdiseaseburden, approximately 9milliondeathsannuallyworld- disease andlungcancertogetheraccountfor infections, chronicobstructivepulmonary causes ofdeathglobally. Lowerrespiratory diseases. Lungdiseasesareamongtheleading and treatmentofcurrentlynotcurablelung of theacquiredknowledgeintoprevention conditions. The long-termgoalistranslation the contextofbothnormalandpathological mechanisms oflungcellfatespecificationin pluripotent stemcellstostudythemolecular Realization ofstemcelltherapyinlung Recently, humanpluripotentstemcell-based My laboratoryisinterestedinapplyinghuman •Understanding thebasicmechanisms •Mapping thepathogenrecognitionpatterns •Use patienthiPSCdifferentiatedlungepithe- RESEARCH PROJECTS the subtype. both theairwayandalveolicellsdependingon the lung;andlungcancersthatcanarisein syndrome thataffectsthelowerrespiratoryof infectionandacuterespiratory destress severe the airway;influenzavirusinfectioninduced cystic fibrosis–ageneticdiseaseaffects in bothairwayandalveolar. Examplesinclude platform forstudyinglungdiseasesoriginate airway- andalveolar-fatedcellsprovidesavalid cells. The availabilityofeachtheseenriched either airwayepithelialcellsordistalalveolar progenitors towardsanenrichedpopulationof human pluripotentstemcell-derivedearlylung ing oncultureconditionsthatcandirectthe other researchgroups. Currently, wearework- disease modeling Human pluripotentstemcellsforlungregenerationand Assistant Professor Sarah XuelianHuang, M.B.B.S., Ph.D. the lungepithelialcellstopathogeninfection and thecellularmolecularresponsesof induced byH1N1virus influenzainfection as causativeforsevere immunity andtestraremutationsin TLR3 lial cellstoexamine TLR3 mediatedintrinsic lung epithelialcellsfor modelingairwayandlungdiseases. Schematic illustration ofhumanpluripotentstemcells-derived airwayand JCI89950. Epub2017May2. 2017 Jun1;127(6):2277-2294. doi:10.1172/ rived frompluripotentstemcells. JClinInvest. NKX2-1+/CD47+ humanlungprogenitorsde- Prospective isolationandsinglecellprofilingof Shannon, BrianR. Davis, andDarrellN. Kotton. Susan Guttentag, JasonR. Rock, JohnM. G. Wong, Ahmad S. Khalil, SarahX.L. Huang, Anthony N. Hollenberg, SineadNguyen, Brandon Nicholas Skvir, Ana M. Crane, Anita A. Kurmann, Driver, DylanC. Thomas, KatherineB. McCauley, Hawkins,Finn PhilippKramer, Anjali Jacob, Ian Aug 30;3(8):e1700521 grafts forlungbioengineering. SciAdv. 2017 Vunjak-Novakovic. Functionalvascularizedlung Snoeck, Bacchetta, Matthew andGordana Holly M. Wobma, SarahX.L. Huang, Hans-Willem Ya-Wen Chen, MauerBiscotti, Theresa Swayne, D. O’Neill, JinhoKim, KatherineCunningham, N. Valerio Dorrello, Brandon A. Guenthart, John KEY PUBLICATIONS NKX2.1 of lunglineagespecificationfrom molecular, genetic andepigeneticapproaches human lungandairwayprogenitorsusing + SOX2 + SOX9 + NKX2.1 + SOX2 + P63 +

and SubarachnoidHemorrhage. Stroke.2016 in thePathogenesisof Intracranial Aneurysm Type 1DomainContainingProtein1)Mutation S, DePalmaS, etal. THSD1 (Thrombospondin Santiago-Sim T, FangX, Hennessy M, Nalbach KEY PUBLICATIONS •Clinical trials •Genetic aneurysmresearch •Stem celltherapyforspinalcordinjury RESEARCH PROJECTS •Chiari malformations •Trigeminal neuralgia •Skull basetumorsandmeningiomas •Brain tumors, benignand malignant •Intracranial aneurysms and MemorialSloanKetteringCancerCenter. versity MedicalCollege, The New York Hospital the Dana-FarberCancerInstitute, CornellUni- Medical School, Brighamand Women’s Hospital, coming to Texas, IheldpositionsatHarvard at Harvard, thenneurosurgeryatUCSF. Priorto Medicine, Icompletedgeneralsurgerytraining of California, SanFrancisco(UCSF)Schoolof Stroke Association. National InstitutesofHealthandthe American geons. Iamtherecipientofgrantsfrom ’sReport Top 1%Doctors, and America’s Top Sur- and spinalcord. neural stemcellsforimplantationintothebrain in familialpatients. We alsoworktodevelop defect proventocauseintracranialaneurysms Our grouprecentlyidentifiedthefirstgene velopment, andtreatmentofbrainaneurysms. includes over100facultyandresidents/fellows. Healthcare System. Currently, ourgroup fortheMemorialHermann neuroscience efforts ence Institute(MNI), whereIleadtheclinical I alsoamdirectoroftheMischerNeurosci- Neurosurgery atMcGovernMedicalSchool. CENTER FORSTEMCELL AND REGENERATIVE MEDICINE I specializeinthefollowingdiseases: A graduateofStanfordandtheUniversity I wasnamedtotheUSNewsand World Our researchhasfocusedontheorigin, de- of Professor andChairoftheDepartment 2017 Jan5;19:55-64. iPSCs forSCItherapy. 2016;StemCellRes. neural progenitorsderivedfromintegration-free anyuan Zhang, DongH. Kim, QilinCao:Human Xue,Georgene W.Hergenroeder,Jiaqian Wu, Yu- Liu,Ying Zheng,Yiyan ShenglanLi, Haipeng Dec;47(12):3005-3013. Epub2016Nov15. and SubarachnoidHemorrhage.Stroke. 2016 in thePathogenesisofIntracranial Aneurysm Type 1DomainContainingProtein1)Mutation S, DePalmaS, etal. THSD1 (Thrombospondin Santiago-Sim T, FangX, HennessyM, Nalbach Dec;47(12):3005-3013. Epub2016Nov15. Variants. Identification ofthe THSD1R450XMutationin LargeFamilywithIAandtheSpectrum of THSD1Rare Advancing thefieldofneuroscience Memorial Hermann–TMC Director, MischerNeuroscienceInstitute L.Vivian ofNeurosurgery SmithDepartment Professor andChair Dong Kim, M.D. spinal cordinjury. Neurosurgery. 24May2017. stem cellsinchroniccervicalandthoracic intramedullary transplantationofhumanneural Huhn S, Curt A, GuzmanR. Emergingsafetyof Fehlings M, Anderson KD, Gage A, HsiehJ, S, Parr A, Ganju A, Aarabi B, KimD, CashaS, Levi AD, OkonkwoD, ParkP, Jenkins A, Kurpad 2017Jan20;7:41008.Scientific Reports. chronic andstagesofspinalcordinjury. coding andlongnon-codingRNAsinthesub- DH, CaoQ, Wu JQ. The systematicanalysisof Duran RC, Yan H, Zheng Y, HuangX, GrillR, Kim

65 • PACT REPORT PACT IMM 66 • IMMPACT REPORT CENTER FORSTEMCELL AND REGENERATIVE MEDICINE help ustounderstand themechanismofHSC lating abilitybysingle-cell RNAsequencing. through whichpre-HSCsgainefficientrepopu - are tryingtoclarifythemolecularmechanisms its mechanismhasyettobeelucidated. We adult-repopulating abilitywithinonedayand regionoftheembryo.aortic Pre-HSCsacquire repopulating HSCsareproducedfromthepara- produced intheembryo, pre-HSC andadult various lineagetracingmousemodels. contribution topostnatalB-1cellpool, utilizing itsreal dent B-1progenitorcellsandevaluate is toidentifythemainsourceofHSC-indepen- cell poolthatpersiststopostnatallife. Ouraim mouse embryoandcontributetoproducingB-1 inthe progenitors areproducedsomewhere fetal liver, suggestingthatHSC-independentB-1 highly purifiedHSCsinadultbonemarrowand publication showedlackofB-1cellpotentialin in HSC-deficientembryos. Ourdataandothers’ presence ofHSC-independentB-1progenitors tion assays. We the haverecentlyreported HSCs, basedontheresultsoftransplanta- progenitor cells, notfromadultbonemarrow decades thatB-1cellsarederivedfromfetal and autoimmunity. Ithasbeenpostulatedfor against variousinfections, atherosclerosis, rolesinthefirstlineofdefense ing important natural antibodieswithout T cellhelp, display - the peritonealandpleuralcavitiessecrete adoptive Bcells(B-2cells). B-1cellslocalizein cells thataredistinguishedfromconventional development. HSCs inalimitedtimewindowofembryonic how pre-HSCsmatureintoadult-repopulating HSCs andpersistintopost-natallife, and2) progenitors areproducedindependentfrom we areaskingquestions1)innateB-1cell are producedinthemouseembryo. Specifically, before thefirsthematopoieticstemcells(HSCs) produced from “hemogenic endothelialcells” waves ofvarioushematopoieticcellsare Knowledge obtained from aboveprojectswill At thesametimewhenB-1progenitorsare B-1 cellsareuniquemurineinnateimmune My labistryingtounderstandhowmultiple Biol. 94(8):719-728, 2016. potential regulatoryfunctions. ImmunolCell is expressedbyasubsetofhumanBcellswith S, MorseIiiHC. Plasmacellalloantigen ENPP1 YoonJ,H,Kim YC, Wang YoshimotoM, Abbasi KEY PUBLICATIONS moleculesforHSCmatura- •Identify important •Understanding themultiplewavesofhema- •Elucidating cellintrinsicandextrinsic •Lineage tracingforHSC-independentand/or RESEARCH PROJECTS deficient patients. for hematologicaldisordersandimmune in vitro, whichmightopenapathofcelltherapy from human iPS cells those human counterparts and B-1cellproductioninvivo, andtoproduce peritoneal B-1cellsafter birth. (AnnNY Acad Sci2015) and expand. FetalliverB-1progenitor cellsderivedfrom YS hemogenic ECandHSCmatureintothe liver. Around atE11, thefirst HSCsemergefromhemogenicECsat AGMregionandalsoseedfetalliver pleura (P-Sp)produceB-1progenitorcells priortoHSCemergence. These progenitorcellsseedfetal peritoneal cavity. splanchno- Hemogenicendothelialcells(ECs)intheyolksac(YS)andpara-aortic embryo andcontributestoB-1progenitors inthefetalliverandpostnatalB-1acellpooladult Hypothesis: fromyolksachemogenicendothelialcellsinthemouse The firstwaveofB-1celldevelops hematopoiesis inthemouseembryo Elucidating themechanismsofmultiplewaves Associate Professor Momoko Yoshimoto Kobayashi, M.D., Ph.D. RNA-sequencing tion inthemouseembryoutilizingsingle-cell embryo topoiesis andHSCproductioninthemouse ability renewal mechanisms formaintainingB-1acellself- embryos toadults from HSC-dependent B-1celldevelopment Ph.D. MichihiroKobayashiM.D.,Assistant Professor: LAB MEMBERS 8:1563-1572, 2017. Hematopoietic StemCells. StemCellReports. for B-1aandB-2LymphocytesinClonalPre- Yoshimoto M, BernsteinID. A CommonOrigin DJ, PoulosMG, ButlerJM, RafiiS, Yoder MC, Hadland BK, B,Varnum-Finney MandalPK, Rossi Cells.35:1053-1064, 2017. and KitSignalsinEarly T CellProgenitors. Stem Tyrosine PhosphatasePRL2MediatesNotch Daniel LacorazzaH, ZhangZY, Liu Y. Protein Carlesso N, Yoder MC, KapurR, KaplanMH, S,Rodriguez Yashiro-Ohtani Y,Pear WS, M, GaoR, ChenS, Yao C, Dong Y, ZhangL, Kobayashi M, NabingerSC, Bai Y, Yoshimoto continue applyingpatient-specific iPSCsand date uniquegenefunction intumoretiology. We therefore providesa powerful systemtoeluci- gene alteration. The patient-specificiPSCmodel of osteosarcomamustbeduetothesingle formed fibroblasts, anyrecapitulatedfeatures these iPSCsweregeneratedfromnon-trans - mutation mediateddiseasepathogenesis. Since “disease inadish” platformforelucidatingp53 that ourestablishedLFSdiseasemodelisa ferentiation andtumorigenicability, suggesting features includingdefectiveosteoblasticdif- derived osteoblastsrecapitulateosteosarcoma Gingold, etal, Trends Cancer2016). LFSiPSC- p53 inosteosarcoma(Lee, etal, Cell2015; the pathologicalmechanismscausedbymutant using LFSpatient-specificiPSCstodelineate Li-Fraumeni syndrome(LFS)diseasemodelby ESCs. We haveestablishedthefirsthuman patient-specific iPSCsand/orengineered cancer pathologicalmechanismsbyapplying transplantation-based therapies. ing, cell personalizedhealthcareandeventually discovery anddevelopment, screen- toxicology to elucidatediseasepathogenesis, fordrug generate differentiatedcellsthatcanbeused a powerfulandunlimitedsourceofcellsto like embryonicstem(ES)cells, iPSCsprovide ferentiate intoallcelllineagesofanorganism indefinitelyand dif- their abilitytoself-renew Shinya Yamanaka in2006. Characterizedby byDr.cell (iPSC)methodologiesdeveloped become feasiblewithinducedpluripotentstem perfect modelsystemtostudyosteosarcoma. osteosarcoma development, whichprovidesa suppressor gene, haveincreasedincidenceof caused bygermlinemutationsinthep53tumor inherited autosomaldominantcancerdisorder with Li-Fraumenisyndrome(LFS), agenetically with osteosarcomadevelopment. Patients and RB1deletion)arestronglyassociated children. Geneticalterations(e.g., p53mutation among leading causeofcancermortality CENTER FORSTEMCELL AND REGENERATIVE MEDICINE Our researchisdedicatedtounderstand Modeling humangeneticdiseasehasrecently After leukemia, osteosarcomaisthesecond Familial cancersyndromesinadish Assistant Professor Dung-Fang Lee, Ph.D. new therapeutic compounds. new ties fortestingexisting both WT andmutantp53 -associated pathway related drugsanddiscovering - Integration of3Dorganoid andorgans-on-chipwithLFSiPSCdisease modeloffersexcitingopportuni PSCs withdifferentp53mutations, whichprovidesvaluableresourceforprecision cancermedicine. clinical trialsinadish. Precisegenomeeditingtechniquesmakeit possibletoexpandthebankof model andprimarycelllines, andholdspotentialsinmodelingLFS associatedcancersandfacilitating LFS iPSCmodelovercomesthelimitations ofcurrentLFSdiseasemodelslikemousemodel, zebrafish forLFSandp53mutation-associatedtumors.The applicationofLFSiPSCmodeltodrug development Ewing sarcoma. CancerRepRev. 1(2):1, 2017. Jianan Shen, Dung-FangLee. The histogenesisof Jian Tu, ZijunHuo, JulianGingold, RuingZhao, KEY PUBLICATIONS •Model familialcancersyndromewithpredis- • analysesandcharacterization •Systems-level RESEARCH PROJECTS illuminate cancerpathologicalmechanisms. TALEN/CRISPR geneticallyengineeredhESCsto iPSC approaches position toosteosarcomabypatient-specific ment - during LFS-associatedosteosarcomadevelop Systematic analysesofgenomealterations coma of mutantp53inLFS-associatedosteosar- Visiting Scholars:Jian Visiting Tu, Donghui Wang Liu Technicians: Ying Students: RuojiZhou, BrittanyEJewell Zhu Post-doctoral Fellows: An Xu, MoLiu, Dandan LAB MEMBERS 38(10):908-927, 2017. targeting oncogenicp53. Trends PharmacolSci. precisioncancertherapy platform todevelop Li-Fraumeni syndromediseasemodel: A C Strong, RuiyingZhao, Dung-FangLee. Ruoji Zhou, An Xu A, JulianGingold, Louise Med. 23(8):737-755, 2017. pathogenesis andiPSCmodeling. Trends Mol Dung-Fang Lee. Osteosarcoma:Molecular Linchao Lu, RuiyingZhao, LisaL Wang, Yu-Hsuan Lin, BrittanyEJewell, JulianGingold,

67 • PACT REPORT PACT IMM 68 • IMMPACT REPORT faithfully reflectsthe endogenousNEUROG2expressionalong thedifferentiationpathway(B, C). bodies (EBs)whichglow redunderthefluorescencemicroscope (A). NEUROG2antibodystaining(green) confirmsthatmCherry(red, nativesignal)expression celllinemadeusingtheCRISPR/Cas9 system.A Neurogenin2knockin humaniPSCreporter NEUROG2-mCherry humaniPSCclonesareinduced asembryoid We haveoptimizedthemoreclinicallyrelevant, retically omittheneedforimmunesuppression. autologous materialsforpatients, whichtheo- stem cells(iPSCs). Mostcritically, iPSCsprovide reprogrammed tohumaninducedpluripotent keratinocytes, andbloodcells, canbe somatic cellssuchasdermalfibroblasts, tion factors, OCT4, SOX2, KLF4andC-MYC, targets forthetreatmentofCNSdiseases. Our long-termgoalistoidentifytherapeutic pathogenesis inspinalcordinjuryandstroke. pathwaysandthe corresponding developmental vivo models. We focusondissectingtheneural biological approachesappliedtoinvitroand combined geneticandmolecularcellular and CNSinjury. Ourresearchentailstheuseof (ii) pathogenesisofneurodegenerativedisease andregenerativemedicine,cell biology and al researchinthefollowingtwoareas:(i)stem CENTER FORSTEMCELL AND REGENERATIVE MEDICINE By transientoverexpressionoffourtranscrip- We havebeenpursuingbasicandtranslation - Assistant Professor Liu,Ying Ph.D. for CNSinjury Human pluripotentstemcellsincell-basedtherapy Hergenroeder, G.W., Wu, J., Zhang, Y., Kim, D.H., Liu, Y.*, Zheng, Y., Li, S., Xue, H., Schmitt, K., KEY PUBLICATIONS •Molecular changesingeneexpressionregula- •Down syndromediseasemodelingusing •Identification ofoptimalneurallineage •Generation ofpatient-specific, integration-free RESEARCH PROJECTS in diseaseanddevelopment. applied toin-depthstudyofsignaltransduction iPSCs. These neurallineagespecific cellsare andgenecorrectionsofpatient eage reporters CRISPR/Cas9 systemincreationofneurallin- adapted thehighlyefficientgenomeeditingtool drug screeningandtesting. Recentlywehave studies, neuralregenerationandrepair, and types fordiseasemodeling, transplantation and glialprogenitors, aswellmaturecell specific iPSCsintoneuralstemcells, neuronal performed directeddifferentiationofpatient- integration-free hiPSCgenerationprotocoland tory networksinglioblastoma patient derivediPSCsandneuralpopulations cord injury progenitors forcell-basedtherapyinspinal iPSCs Research Associate: HaipengXue GraduateStudent:DiJia Visiting Post-doctoral Fellow:ShenglanLi LAB MEMBERS of print] org/10.1155/2017/9182748 [Epubahead 8, 2017. ID9182748,Article https://doi. pluripotent stemcells. Neural Plasticity. Oct neural progenitorsderivedfromhuman-induced Y. (2017). Effectsofpropofoltreatmentin Long, B., Li, S., Xue, H., Sun, L., Kim, D. H., Liu, PMCID:PMC5485764 omtn.2017.06.007. PMID:28918057 8:64–76, doi. http://dx.doi.org/10.1016/j. Nucleic Acids.15September2017.Mol Ther CRISPR/Cas9 activationofmultiplegenes. transposon-based One-step piggyBac Li, S., Zhang, A., Xue, H., Li, D., Liu, Y. (2017) PMID:28073086 ahead ofprint](*correspondingauthors) 64. doi:10.1016/j.scr.2017.01.004. [Epub therapy. StemCellRes.2017 Jan5;19:55- derived fromintegration-freeiPSCsforSCI Cao, Q*. (2017)Humanneuralprogenitors microenvironment is underway. of quiescentcellsand theirinteractionwith their survivalinbone marrow. Characterization to understandthemolecularmechanisms of and increasedtumorformation, itisimportant ment andpotentiallyledtodrugresistance cells canescapethechemotherapeutictreat - and causesosteolyticlesions. Sincequiescent nancy thatproliferatesprimarilyinbonemarrow microenvironment. MMis aplasmacellmalig- and theirinteractionwiththebonemarrow ing rolesofthequiescentmultiplemyeloma applications. in cancercellswillhavedirecttranslational pounds thattargetdrugresistancepopulations cells arehighlydrugresistant, discovery ofcom- of PAX5 silencedcells. GiventhatPAX5 silenced select compoundstargetthesurvivalpathways and prestwicklibraries. We discoveredthat compounds ofNCIoncology, customclinical, screening usinglibrariescomprisedof3991 the bonemarrow. silencing ledtoincreasedcancercellsurvivalin potential tumorsuppressor. Moreover, PAX5 in xenograftmice, indicatingthatPAX5 isa lymphoma leadstoincreasedtumorformation discovered thatPAX5 silencinginmantlecell nant ofnormalBcelllineagedevelopment. We those factorsispairedbox5(PAX5), adetermi- with bonemarrowmicroenvironment. Oneof lignant Bcellsanddelineatingtheirinteraction selective growthandsurvivaladvantagestoma- to deciphermolecularpathwaysthatconfer chemotherapeutic agents. for dormantclones, whichareoftenresistantto bone marrownichesprovidea “hiding place” tion fromchemotherapy-inducedapoptosis. The multiple myelomacellgrowthaswellprotec- niche, whichprovidesanenvironmentfor malignant cellsandtheirlocalbonemarrow also requirescomplexrelationshipsbetween dependent ontheirgenomicabnormalitiesbut CENTER FORSTEMCELL AND REGENERATIVE MEDICINE We arealsoconductingaresearchdelineat- We haveconductedhighthroughputdrug The majorgoalsofmyresearchprogramare The behaviorofcancercellsisnotonly taken by an inverted microscope(x10).taken byaninverted were seededandafter21days, cellgroupingsof>40cells werecountedasacolony. The pictureswere TRIM44 over-expressedcells(TRIM44 the bonemicroenvironment Deciphering mechanismsofhumancancercellsurvivalwithin Jerold B. KatzDistinguishedProfessorshipinStemCellResearch Associate Professor Nami McCarty, Ph.D. •Delineating transcriptionfactornetworkson ofsmallmoleculeinhibitors •Development •Survival mechanismsofdormantmultiple RESEARCH PROJECTS ization inthecelldeterminedrugresistance determine whetherBACH2sub-cellularlocal- closely workwithcollaboratorsatcliniciansto works inmantlecelllymphomas. We willalso address rolesforPAX5-BACH2 signalingnet- drug resistantlymphomas: We willcontinueto test theirefficaciesinthepatients. models forpre-clinicalstudiesandplanto andtestthesecompounds inanimal develop drugresistance.that develop We willfurther selectively targetmantlecelllymphomacells screening toidentifythecompoundsthat have conductedhighthroughputchemical to targetdrug-resistantlymphomas: We chemotherapies.cells evade delineate howdormantmultiplemyeloma with bonemarrowmicroenvironmentto genes inthemultiplemyelomainteraction continue tocharacterizefunctionsofthese different nichesofthebonemarrow. We will quiescent multiplemyelomacellsfromthe ray analysestoidentifygenesexpressedin the bonemarrow: We conductedmicroar- myeloma cellsandtheirmicroenvironmentin OE ) increasedcolonyformationinmethylcellulosemedium. Cells Research Assistants: ZhengChen, M.S. Graduate Students:FarahLadha Liu, Ph.D., Quach Tu Hue, Ph.D. Post-doctoral Fellows: Tsung ChinLin, Ph.D., Lyn LAB MEMBERS revision. HIF-1αstabilization. Leukemia, 2017, Under and survivalintheosteoblasticnichevia quiescent multiplemyelomacell2occupancy McNiece, IandMcCarty, N. TRIM44 promotes Chen, Z., Lin, T-C., Bi, X., Lu, G., Dawson, B.C., Blood” asanEditor’s pick. 2017. wasfeaturedin This article “this weekin dispersal duringhypoxia. Blood130:763-776. coordinates mantlecelllymphomasurvivaland L.J., andMcCarty, N. BifurcatedBACH2control Zhang, H., Chen, Z., Miranda, R.N., Medeiros, 13:627-628, 2017. regulatorynetwork.IL6-autophagy Autophagy cancer chemoresistancebytargetingthe TGM2- Zhang, H., andMcCarty, N. Tampering with KEY PUBLICATIONS outcome andpatientsurvival.

69 • PACT REPORT PACT IMM 70 • IMMPACT REPORT CENTER FORSTEMCELL AND REGENERATIVE MEDICINE to be stably maintained even aftertransplanta- to bestablymaintained even allowthe culture-madecartilage ing thatthey joint progenitorfrom hPSCs, and2)demonstrat- following twogoals;1)generatingtheembryonic permanent cartilage), the weaim toachieve after transplantation(i.e., jointcartilage-like permanently thatstaysascartilage cartilage mice. Inordertoestablishmethodsgenerate when transplantedintoimmunocompromised of themaremineralizedandturnedintobone inculture.hyaline-like cartilage However, most chondroprogenitors arecapableofgivingriseto precursors (chondroprogenitors). All such differentiate intocorrespondingchondrocyte areabletoexpandand demonstrated thatthey three embryonicoriginsofchondrocytes, and purified fromhPSCs, progeny representingthe and tilage?: developed We havepreviously regenerative medicine. PSCs apromisingsourceofembryoniccellsfor that mimicembryogenesis, makinghuman(h) of humanbodyinculturethroughprocesses are expectedtodifferentiateintoanycell-types from anembryo, orinducedfromadultcells, Pluripotent stemcells(PSCs), whetherderived best fortheregenerationofadultjointcartilage. formation: i.e. jointprogenitorwouldbethe the embryoniccell-typesresponsibleforjoint embryogenesis. Therefore, wehypothesizethat distinct populationsofprecursorcellsduring from aredeveloped growth-plate cartilage cartilage, destinedtoformbone. The jointand inthelimbandvertebra,cartilage is a transient permanent cartilage. Incontrast, growth-plate afterexpansion.cartilage isa Jointcartilage tendency toyieldunsuitableand/orunstable the problemsoflowyieldcellsandtheir cells suchasmesenchymalstromalface therapies usingchondrocytesandadultstem however,damaged cartilage; currentcell application ofstemcellstotherepair tilage ofjointsisnotspontaneously been considerableinterestintheclinical repaired afterinjuryinhumans. There has prog What isthemolecularbasisofstem/ The car enitor cells to form permanent car- permanent enitor cellstoform Annie &BobGrahamDistinguishedChairinStemCellBiology Associate Professor Naoki Nakayama, Ph.D. Pluripotent stemcelldifferentiationandlineagespecification Joint articular cartilage repairwith cartilage •Joint articular •Comparative omicsanalysestoelucidate definingtheprocessofchondrogene- •Further RESEARCH PRO in future. expansion culturemethodforadultstemcells capacity, whichmaybeappliedtoimprovethe in elucidatingthemechanisticbasisofsuch withadultstemcells.achieve We areinterested sion ofchondrogenicactivityiscurrentlyhardto capacity.cartilage-forming Suchstableexpan- extended periodoftime, withoutlossoftheir the hPSC-derivedchondroprogenitorsforan conditions thatmaintainedandexpanded genitors: establishedculture We previously of PSC-derivedhumanchondropro- expansion cells;long-term forming formation ofpermanentcartilage. tion ofadditionalsignalingmechanismsforthe capacity,cartilage-forming andontheelucida- to elucidatemolecularbasisoftheirpermanent characterization ofthejointprogenitor-likecells Therefore, wearecurrentlyfocusingbothonthe plantation (i.e. pseudo-permanentcartilage). limited boneformingcapacityaftertrans- the standardchondroprogenitorsdisplayingvery madefrom cAMP signalingleadstocartilage In addition, wehavedeterminethatcontrolling inmice.maintained asunmineralizedcartilage with thejointprogenitor-likecellsisinfact made have recentlydiscoveredthatcartilage progenitor markergeneloci. Furthermore, we carry fluorescencemarkergenesinthejoint like cells, wehavegeneratedhiPSClinesthat characterizethejointprogenitor- and further joint progenitor-likecellsfromhPSCs. To purify tively generate, andtoalimitedextent, expand cells. discoveredawaytoselec- We previously repairtocurrentlyavailable cartilage articular tion, showsuperiorcapacityof andthatthey Larg animal models progenitor-like cellsinimmunocompromised hPSC-derived chondroprogenitorsandjoint joint progenitor-likecells formationfromthehPSC-derived cartilage the molecularbasisofpermanent, hyaline and jointprogenitors sis fromthehPSC-derivedchondroprogenitors e quantity of articular cartilage- e quantityofarticular JECTS from NSGmiceafter8weeks. from jointprogenitor-likecells(2)recovered progenitors (1), andpermanent-like cartilage fromstandardchondro- Bone-forming cartilage ist: NadineMatthias Senior Research Associate and Animal Special- Pothiawala Research Assistants:Azim LAB MEMBERS ,Stem CellReports inrevision. human pluripotentstemcell-derivedcartilage” control ofthepost-transplantationalfate Nakayama, N. (2017) “Pre-transplantational Yan, Q., Pigeot, S., Martin, I., Huard, J. and Lee, J.Y., Matthias, N., Pothiawala, A., Ang, B.K., 3:16065. genic potential” Mol. Ther. MethodsClin. Dev., is Attributed toHighCellSurvivalandChondro- Derived StemCellsfor Repair Cartilage Articular “The SuperiorRegenerativePotentialofMuscle N., Poddar, M., Hogan, M., andHuard, J. (2016) Li, H., Lu, A., Tang, Y., Beckman, S., Nakayama, Inc., Rijeka, Croatia. In PluripotentStemCells,pp. 385-425. InTech, chondroprogenitors: Promiseandchallenges” regeneration usingpluripotentstemcell-derived K., Qing, Y., andHuard, J. (2016) “Cartilage Nakayama, N., Lee, J.Y., Matthias, N., Umeda, KEY PUBLICATIONS patients affectedbycancer. treatmentoptionsfor we hopetoidentifynew proaches tostudythetumormicroenvironment, tumor. Byapplicationofbioengineeringap- the systemicspreadofcancerfromprimary tion ofoncogenicpathwaysthatcontributeto of fluidforceoninvasivepotentialandactiva- theimpact rienced bycancercellsandevaluate vasculature, wemodulate theshearstressexpe- microchips designedtomodelthelymphatic potential ofcancercells. Using biomimetic stresses mayplayinregulatingmetastatic unclear whatrolelymphaticorvascularshear been implicatedintumorbiolog transplantation assays. fluidics, pharmacology, mousegenetics, and stemcellfunction,evaluate includingmicro- We employvarioustoolsandapproachesto and immunomodulatoryfunctionofstemcells. of fluidforcethatpotentiatestemcellpotential mediating thesignalingcascadedownstream players are currentlyunderinvestigationaskey A numberofgeneticandbiochemicalpathways hematologic disordersandneurologicaltrauma. and othercellulartherapiesfortreatmentof improved sourcesofhematopoieticstemcells this informationinthelaboratorytoexpand are especiallyinterestedinhowwemightuse and inthecontextofdiseaseinjury. We function duringembryogenesis, address howfrictionalforcepromotesstemcell potential andbehavior and lymphflowinthelymphaticsimpactscell generated bybloodflowinthevasculature CENTER FORSTEMCELL AND REGENERATIVE MEDICINE Fluid flowandhydrostaticpressurealsohave One armofourresearchisdesignedto Our labstudieshowbiomechanicalforce . intheadult, y, butitremains represent asignificant reductionincancercellmotilitybytreatment with YTIPunderfluidflow. with asmallpeptide YTIP thatinterrupts YAP1 function, aproto-oncogene, inthecancercells. Asterisks site tosecondarymetastatic sitesinthebody. The increased migrationcanbeblockedbytreatment cancercellmotility,Flow elevates formovementofcellsfromtheprimarytumor aprocessimportant through themicrochanneltomimiclymphaticflow. is treatedwithcollagenmatrix, followed byintroductionofcancercells. Culturemediumispushed through thelymphaticvasculature. A microchannelembeddedinthecenterofmicrofluidicsdevice monitoring ofgeneactivityunderconditionsthatacancercellmayexperienceduring metastasis We engineerbiomimeticmicrochipsthatpermitreal-timevisualizationofcancercellmigrationand metastasis Biomechanical forceregulatescellpotentialandcancer Assistant Professor Pamela Wenzel,Ph.D. promote cancercellmotility. NatureCommuni- Wenzel, P.L. Fluidshearstressactivates YAP1 to Zhang, S., Sevick-Muraca, E.M., Hagan, J.P., & Lee, H.J., Diaz, M.F., Price, K.M., Ozuna, J.A., 35:1259-1272, 2017. mesenchymal stromalcells. StemCells. immune regulatoryfunctionofbonemarrow Wenzel, P.L. Biomechanicalforcespromote Omichi, R., Bedi, S., Olson, S.D., Cox, C.S., & J.A., Liao, G.P., Aroom, K.R., Xue, H., Gu, L., Aertker, B.M., Alexander, A.J., Price, K.M., Ozuna, Diaz, M.F., Vaidya, A.B., Evans, S.M., Lee, H.J., KEY PUBLICATIONS •Fluid flowininitiationofmetastasis ofhematopoieticand •Mechanobiology RESEARCH PROJECTS mesenchymal stemcells Research Associate:MiguelDiaz LAB MEMBERS 10.1016/j. biomechanical force. marrow mesenchymalstromalcellsinducedby regulates anti-inflammatoryfunc-tionofbone & Lee, as beingofspecialsignificanceinitsfield. af8e-8d975d8fe8e6. Recommended inF1000Prime media/story.htm?id=e2ea8aec-975e-4ee1- news storyatUTHealth:https://www.uth.edu/ cations. Wenzel, H.J., Diaz,M.F.,Ewere, 8:14122, P .L. Focal adhesionkinasesignaling cellsig.2017.06.012, 2017. Cellular Signalling. This workwasfeaturedina A., Olson, S.D.,Cox, 2017. 38:1-9. doi: C.S.,

71 • PACT REPORT PACT IMM 72 • IMMPACT REPORT Pinpoint key transcription factorsandregula- •Pinpoint key •Characterize molecularsignaturesandidentify •Investigate geneexpressionandregulatory RESEARCH PROJECTS poietic precursorandstemcells. efficient expandingandmanipulatinghemato- differentiation ingeneralandprovideinsightfor and model fortheanalysisofcellself-renewal and differentiation. This studycanserveasa stages ofhematopoieticprecursorself-renewal oftheregulationearly comprehensive view global interactionnetworkandanovel in thisprocess. Currently we aregeneratinga together withRUNX1, regulators are important control theswitch. We haveidentifiedthat TCF7, componentsthat approaches toidentifykey are usingintegratedgenomicandproteomic and lymphocytic)cellasamodelsystem. We using multipotentEML(erythroid, myeloid, anddifferentiation precursor cellself-renewal of theregulatorynetworkshematopoietic area ofherresearchinterestliesinthestudies and toincreasetransplantationsafety. The other into desiredneuralcelltypesmoreefficiently, targets todirectthedifferentiationofstemcells regulatorsastherapeutic tify andmodulatekey and itsclinicalimplications. The goalisto iden- mechanism ofstemcellneuraldifferentiation are crucialinunderstandingthemolecular ation sequencingtechnologies. These studies factors andregulatoryRNAsusingnext-gener- expression andtheregulationoftranscription neurological diseases. We arestudyinggene and safetreatmentforspinalcordinjury effective neural differentiationanddeveloping focus ofthegroupisinvestigatingstemcell governing stemcelldifferentiation. Onemajor gene transcriptionandregulatorymechanisms bioinformatics andfunctionalassaystounravel systems-based approachesinvolvinggenomics, CENTER FORSTEMCELL AND REGENERATIVE MEDICINE tory RNAs, regulators to andmodulatekey neurological diseases therapeutic targetsforspinalcordinjuryand mechanisms duringstemcelldifferentiation and My laboratorycombinesstemcellbiology tubulin (TujIII red)labelsbothimmatureandmature neurons. Nuclei(blue)arestained byDAPI. Immunofluorescence labelingofneuronsderivedfromH1human embryonicstemcells(hESCs). beta- Associate Professor Jiaqian Wu,Ph.D. and neuralinjuries Gene transcriptionandregulationofstemcelldifferentiation Zheng, Y., Li, S., Xue, H., Schmitt, K., Hergenro - Integration-Free iPSCsforSCItherapy. Liu, Y., Human NeuralProgenitorsDerivedfrom Jun 8. 10.1186/s40169-017-0150-9. Epub2017 Clin TransMed (2017)6:20. Dec;6(1):20. doi: Duran,Cuevas-Diaz R., Wei, H.C., and Wu, J.Q. Single-cell RNA-sequencinginthebrain. 20;7:41008. doi:10.1038/srep41008. 2017 R., Kim, D.H., Cao, Q., and Wu, JQ. SciRep. Jan Duran, R., Yan, H., Zheng, Y., Huang, X., Grill, chronic stagesofspinalcordinjury. Cuevas-Diaz non-coding RNAsinthesub-chronicand The Systematicanalysisofcodingandlong 2016. Gene Regulation. Springer PublisherDordrecht Wu, J. Q. (ed.)(2016). Transcriptomics and KEY PUBLICATIONS and improveefficiency steer thedirectionofstemcelldifferentiation Undergraduate Student: Vy Hong Diaz Duran, Haichao Wei, XiaominDong Post-doctoral Fellows: Yanan You, RaquelCuevas LAB MEMBERS PMID: 28727410 2017 Jul27. doi:10.1021/acsnano.7b01087. Chen, M., Zhou, Y-H., andMao, J.Nano. ACS Chen, K., Coelho, P.G., Tovar, N.M., Kim, S.H., Zhang, Y.,S.,Zhou, Wang, Y.,Sheu, T.,J., Wu, Xiang, L., He, L., Yang, G., Zheng, J., Wang, C., Crosstalk inOrganDevelopment. Jiang, N., Exosomes MediateEpithelium−Mesenchyme Jun 8. 10.1186/s40169-017-0150-9. Epub2017 Clin TransMed Duran,Cuevas-Diaz R., Wei, H.C., and Wu, J.Q. Single-cell RNA-sequencinginthebrain. scr.2017.01.004. 2017 Cao, Q. StemCellResearch. doi:10.1016/j. eder, G.W., Wu, JQ., Zhang, Y., Kim, D.H., and (2017) 6:20. Dec;6(1):20. doi: drugs thatexploittheiruniqueproperties. in thetumorandhowwecantargetthemwith is tounderstandthisminorpopulationofcells determining thefutureoftumor. Ourgoal “cancer stemcell,” whereastherestcannot 1:2000 tumorcellsofagivencanceristrue pancreas, andesophagus. Remarkably, about lethal cancerssuchasthoseoftheovaries, to identifyandcapturestemcellsofhighly tory istouseourstemcellcloningtechnology drug discoverytomitigatetheseconditions. diseasemodelsand ofrelevant development such, patient-derivedstemcellswilldrivefuture the stemcellpopulationinthesepatients. As COPD, aredrivenbypermanentchangesin disease, lungdiseases, suchasasthmaand of theseorgans, suchasinflammatorybowel of themajorchronicinflammatorydiseases derived. Importantly, wearefindingthatmany were them toformtheorganfromwhichthey as immaturecellsandyetoncommandtrigger as wecangrowthemforyearsinatesttube kidney. These stemcellsarequiteremarkable lung, andorganssuchasliver, pancreas, and tissues suchasthegastrointestinaltract, the capturing thestemcellsofhighlyregenerative CENTER FORSTEMCELL AND REGENERATIVE MEDICINE A secondbutrelateddirectionofthelabora- for technology new My laboratorydevelops sampled forexpansion andCNVanalysis(right). Derivation ofpatient-specific cancerstemcell(CSC)libraryof clonesfromhigh-gradeovariancancercase (left). FourCSCclones Stem cellsofregenerativeandmalignantepithelia CPRIT Scholar Assistant Professor Wa Xian, Ph.D. •Intratumor heterogeneity amongstem •Cloning andtargeting stemcells •Intestinal stemcellvariation- underly RESEARCH PROJECTS to sufferrelapses6-24monthslatertypically acompleteresponseonly patients achieving ably sensitivetochemotherapywithmany High-grade ovariancancer(HGOC)isremark- cells ofhigh-grade ovarian cancer intodysplasiaandmalignant cancer.evolve haveachanceto esophagus beforethey that eliminatethestemcellsofBarrett’s cancer, wearefocusedonidentifyingdrugs Given thepoor5-yearsurvivalratesforthis esophageal adenocarcinomaby30-100-fold. of that increasestheriskfordevelopment Barrett’s esophagusisaprecancerouslesion esophagus of precancerouslesions:Barrett’s resolving thisdisease. therapeutictargetsfor and identifyingnew andchronicfeaturesofIBD ing thepathology these patients, forunderstand- whichmaykey identify unusualstemcellsinthecolonsof to wedeveloped the stemcelltechnology intestinal epithelialbarrier. We haveapplied microbiome ofthegut, andtheintervening interplay betweentheimmunesystem, the by environmentalfactorsconsistentwithan genetic componentbutseemsdominated 1.6 million Americans. hasa The etiology and difficulttomanageconditionsaffecting Crohn’s andulcerativecolitis, remainserious Inflammatory boweldisease, including boweldisease ing inflammatory Senior ResearchScientist:Brian Wang, M.D. Post-doctoral Fellow: Wei Rao, Ph.D. LAB MEMBERS Cell MolGastroenterolHepatol. 4, 161-164. as aUniqueand Targetable Entity. Xian W, McKeonF. (2017). Barrett’s StemCells 2016 Jan19;7:10380 sion ofaprecancerouslesion. NatureComm. sells suggestspathstoinitiationandprogres- Xian W. MutationalspectrumofBarrett’s stem B, CrumCP, NagarajanN, HoKY, McKeon F, and CC, L,Wilson BlaszykH, RolshudD, LiuJJ, Howitt T, Hu YY, DelubaM, SrivastavaS, Ming T, Khor Yamamoto Y, Wang X, D, Bertrand KernF, Zhang intestinal stemcells. Nature522, 173-178. W. (2015). Cloningandvariation of groundstate Lacy, DB, Ho, KY, Crum, CP, McKeon, F, andXian, Sylvester,,FA, Hyams, JS, Devers, T, Bronson, R, CC, B, Chevalier D, Bertrand Nagarajan, N., B, FarrowMA, KernF, NingG, Hong, Y, Khor X,Wang Yamamoto Y,LH,Zhang T, Howitt Wilson KEY PUBLICATIONS these differencesrelatetoresistantclones. examine intratumorheterogeneityandhow ies ofcancerstemcellsfromeachpatientto phenomenon, wearegeneratinglargelibrar- cellular andmolecularbasisofthisresistance with resistantproperties. To understandthe

73 • PACT REPORT PACT IMM 74 • IMMPACT REPORT T This andthe exponentially increased incidence of increases the riskofmorbidityandmortality. to stress, andasaconsequence, dramatically which leadstoanimpaired ability torespond during theprogressive depletionofstemcells, aging isthelossoftissueregenerative potential membrane damage.Auniversal characteristicof mutations, andcumulative DNA,protein or nuclear dysmorphology, accumulationofgenetic mitochondrial damage,telomere attrition, changes remains ill-defined,butmayinclude that isresponsible foraging-related degenerative faces today. Theprecise nature ofthedamage challenges thatthemedicalresearch community which represent fundamentalandpressing for preventing ordelayingage-associated diseases, molecular basisofaginganddeveloping strategies major focusofthecenteristounderstand diabetes, andcancerincreases withage.The sensorineural defects,cardiovascular disease, neurodegeneration, osteoporosis, sarcopenia, of numerous debilitatingdiseases,including organ systems.Asaconsequence,theincidence ofall of homeostasisandfunctionalreserve system. Agingresults intheprogressive attrition demandsonourhealthcareextraordinary TISSUE ENGINEERING AND AGING RESEARCH ENGINEERING ANDAGING CENTER FORTISSUE increasing elderlypopulation,placing population are shiftingtoward an he demographicsoftheAmerican research. on tissueengineeringandaging our efforts in thearea ofagingresearch sowe cansynergize We are alsoestablishingnumerous investigators stem cells,toextendhuman healthandlifespan. rejuvenating factors,derived from functional of novel approaches fortheuse ofstemcellsor of thisresearch willresult inthedevelopment functional stemcells.Thesuccessfulcompletion identify thetherapeuticfactorssecreted by young, In addition,we are proteomics to performing lifespan inmousemodelsofacceleratedaging. not aged,stemcellshave onhealthspanand systemic injectionoffunctional,young, but followingdramatic therapeuticeffectsobserved determine themechanism(s)underlying mechanisms. In to addition,we are trying cells dysfunctionthrough noncellautonomous blood vessel) orsystemicfactorsindrivingstem of themicroenvironment (muscle,vascularity, autonomous mechanisms,aswell astheeffects We are currently examiningtheintrinsic,cell stem cellsbecomedysfunctionalwithage. on determiningthepathway(s)through which tissue rejuvenation. for by theloss offunctionalstemcellsnecessary led tothehypothesisthatagingiscaused,inpart, numerous degenerative diseasesintheelderlyhas Our research program iscurrently focusing TISSUE ENGINEERING AND AGING RESEARCH ENGINEERING ANDAGING CENTER FORTISSUE Canada andtheU.S. havevolunteeredforthis More than400womensufferingwithSUIin urinary incontinenceandmyocardialinfarction. in clinicaltrialsforthetreatmentofstress isolated bymyteamarecurrentlybeingutilized industry. Muscle-derivedcellsthathavebeen havebeenlicensed to that wehavedeveloped international recognition, andtechnologies processes. Myteamhasreceivednationaland resulting fromnaturalandacceleratedaging the musculoskeletalsystem, includingthose of diseases, conditions, andinjuriesthat affect plications ofMDSPCsfortreatmentavariety involveinvestigatingap- Our researchefforts repair andregenerationofavarietytissues. techniques fortranslationtotheclinicaidin tissue engineering, andapplicationofthese and moleculartechniquesforgeneediting areas ofinterestincludebasicstemcellbiology cells (MDSPCs). Myresearchteam’s primary the useofmuscle-derivedstem/progenitor regenerative medicineapplicationsbasedon of genetherapy, tissueengineering, and The focusofmyresearchisintheareas Muscle-Derived Stem/Progenitor Cells(MDSPCs)fromMurine and HumanSkeletalMuscle B, Weiss K, HuardJ. RhoA/ROCKinhibition X,Mu Tang Y,K,Chen W,LuA, Takayama Wang 2017 Oct24;8(1):1118. PMID:29061963. fibrosis. NatureCommunications. NatCommun. critically regulateskeletalandcardiacmuscle integrins onperivascularmesenchymal1cells H, HuardJ, PeaultB, HendersonNC. αv MA, Kendall TJ, DaiXJ, Li Y, IredaleJP, Simpson DW, RuminskiPG, GrayGA, SinghM, Campbell Greenhalgh SN, Thompson AI, ConroyKP, Griggs J, DobieR, Wallace RJ, Mackinnon AC, SmithJR, Murray IR, GonzalezGonzálezGalofreZN, Baily KEY PUBLICATIONS •Bone abnormalitiesandhealingdefectsin ofbiologicalapproachesto •Development •Muscle stemcellsreprogrammedthroughge- RESEARCH PROJECTS stem celltherapy(PhaseIIIClinical Trial). Muscle-derived stem/progenitorcellapplications Director, IMMCenterfor Engineeringand Tissue Aging Research Surgery ofOrthopedic Department Professor & Research ChairforOrthopedic Vice Johnny Huard, Ph.D. cardiac repair vate deliveryofmusclestemcellstoimprove muscular dystrophy, andbiomimeticcoacer- ment syndromeinjury - improve functionalrecoveryaftercompart defects withaging and non-autonomousmechanismsofstem anti-fibrotic therapy, andcellautonomous nome engineeringforautonomouslyregulated Barbara Lipari Michelle Ramirez, MaryHall, M.B.A., Ph.D., Ling (Jeannie)Zhong, Pena, Martha Sarah Amra, Research Team: HaiyingPan, HaiziCheng, M.D., (Amy)Lin Chih-Yi Students:Zhenhan(Stephen) Deng,Visiting Scientist:Fan Visiting Yang, Ph.D. M.B.A., Ph.D., Yan Cui, M.D. Senior ResearchScientists:PolinaMatre, Sun, Ph.D., SealyHambright,William Ph.D. len) Gao, Ph.D., Chieh(Judy) Tseng, Ph.D., Xuying Post-doctoral ResearchFellows;Shanshan(El- Ph.D., KrishnaSinha, Ph.D. M.D., XiaodongMu, Ph.D., XueqinGao, M.D., Assistant Professors:PingGuo, Ph.D., Aiping Lu, LAB MEMBERS 1382. Epub2016Sep22. PMID:27572850. ated aging. Res.2017Jul;35(7):1375- JOrthop cells isolatedfromamurinemodelofacceler- observed inmuscle-derivedstem/progenitor signaling playsacriticalroleinthedefects FH, RobbinsPD, NiedernhoferLJ, HuardJ. mTOR Cummins JH, Yurube T, KurodaR, KurosakaM, Fu Takayama K, Kawakami Y, LavasaniM, MuX, 1;26(15):2813-2824. PMID:28549178. stem celldepletion.2017 Aug HumMolGenet. treatment indystrophicmuscle:implicationsfor improves thebeneficialeffectsofglucocorticoid

75 • PACT REPORT PACT IMM 76 • IMMPACT REPORT TISSUE ENGINEERING AND AGING RESEARCH ENGINEERING ANDAGING CENTER FORTISSUE mimics theclinicalmanifestations ofhuman phin doubleknockout (dko)modelthatclosely found boneabnormalities inadystrophin/utro- bound intheirseconddecadeoflife. We have 3,000 boys. Patientsoftenbecomewheelchair- deadly musclediseasethatinflictsabout 1in phy model muscle andboneinamusculardystro- model. inamurine etate (MIA)-inducedosteoarthritis forthetreatmentofmonoiodoac - particularly repair,factors forhMDSC-mediatedcartilage vectors andbiomaterialstodelivergrowth translational. Inthisproject, weareusingviral repairareclinically defects orforosteoarthritis in murinemodels, fortreating osteochondral murine muscle-derivedstemcells(mMDSCs) thritis orosteoar- age-related cartilage injury mediated bonerepair. both donorhMDSCsandhostsonhMDSC- are alsoinvestigatingtheeffectsofage cell (hMDSC)-mediatedboneregeneration. We factors toenhancehumanmuscle-derivedstem therapy andusingbiomaterialstodelivergrowth and growthfactorstomediateexvivogene orthopaedics. vectors We areinvestigating new resection representmajorissuesinclinical fractures causedbytraumaticinjuryorcancer bone regeneration muscular dystrophy. inadiseasemodelof investigating bonebiology as osteoporosisandosteoarthritis. Iamalso conditions,age-related boneandcartilage such ment ofbonedefects, non-unionfractures, and translational studiestouseMDSCsforthetreat- repair.therapy forboneandcartilage Iconduct muscle-derived stemcells(MDSCs)andgene research team. Myresearchfocusesonusing Duchenne musculardystrophy(DMD)isa Investigation ofinteractions between The applicationofstemcells, including Human muscle-derivedstemcellsfor Large segmentalbonedefectsandnon-union Human muscle-derivedstemcellsfor I amamemberofDr. JohnnyHuard’s M, SunX, CumminsJH1, HuardJ. Cyclooxygen- X,Usas A,LuKozemchak Gao Tang Y,Poddar KEY PUBLICATIONS •Bone abnormalitiesinmusculardystrophy •Using humanmuscle-derivedstemcellsfor •Utilizing humanmuscle-derivedstemcellsand RESEARCH PROJECTS patients. strategies toimprovethelifequalityofDMD mechanisms whichmaybetranslatedintonew interact inthismousemodelordertounveil and defecthealing, andhowmusclebone muscular dystrophyaffectsthebonequality DMD. Forthis project, weareinvestigatinghow and repair regeneration Muscle-derived stemcellsforboneandcartilage Assistant Professor Xueqin Gao, M.D., Ph.D. bone defectmodel. Cox-2 -deficientMDSCBMP4/GFP-mediated boneregeneration isimpairedinvivousingacritical-size (NIH RO1awardedtoDr. Huard) vivo genetherapyandbiomaterialscaffolding. repairusingex andosteoarthritis cartilage gene therapyforbonetissuerepair PMID:24843069 effects. FASEB J. 2014 Aug;28(8):3792-809. differentiationvs.bone repair: paracrine host cellsinmuscle-derivedstemcell-mediated Proctor A, ChenCW, HuardJ. Roleofdonorand Gao X, Usas A, ProtoJD, Lu A, CumminsJH, 70. PMID:24856105 of BMP.2014 Aug;35(25):6859- Biomaterials. muscle-derived stemcellsandthecriticalrole tion withhumanmesenchymalstemcellsand RS, HuardJ. A comparisonofboneregenera- J, Kozemchak AM, Wang B, CumminsJH, Tuan X,Usas A,Gao Tang Y,Lu A,J,Schneppendahl Tan [Epub aheadofprint], PMID:27354351 Hum MolGenet. 2016Jun27. pii:ddw172. autonomous andnon-autonomousmechanisms. cell-mediated boneregenerationviacellular ase-2 deficiencyimpairsmuscle-derivedstem TISSUE ENGINEERING AND AGING RESEARCH ENGINEERING ANDAGING CENTER FORTISSUE important growthfactors thatmayaccelerate important to obtainandmanipulate. PRPcontainsmany of autologousgrowth factorsandisbotheasy ment modalitiessinceitcontainsanabundance advantages overotherpotentialbiologictreat - tissue repair. let richplasmaformusculoskeletal improve thebeneficialeffectofplate - The useofbiomimeticmicrospheresto tors. through thereleaseofcytokinesorgrowthfac- but alsosynergisticallyenhanceangiogenesis cardiac regenerativepotentialoftheMDSCs, not onlypromotecellretention, survival, andthe areaofresearchthatcould represents anew vate, asanoveldelivery vehiclefortheMDSCs, The useofcytokines, orgrowth factor-coacer- cardiac regenerativepotentialoftheMDSCs. MDSCs lowcardiomyogenicpotentiallimitsthe to limitedcellretentionandsurvival. Moreover, intramyocardial injectioninPBS)whichleads poor approachesfordeliveringthecells(direct CCM. However, limitations, several suchasa represent aviabletherapeuticcellsourcefor & myocardialinfarction, confirmingthatMDSCs arena forthetreatmentofbladderdysfuntion human MDSCshavealreadyenteredtheclinical numbersinculture.relevant Moreimportantly, muscle derivedstemcells(MDSCs), toclinically function. We havesuccessfullyexpandedhuman repair injuredmyocardiumandimprovecardiac into theheart, isapromisingapproachto involves thetransplantationofexogenouscells and regeneration. muscle stemcellsforcardiacrepair of delivery Biomimetic coacervate RESEARCH PROJECTS of research. diseases. Currently, wehavethreemajorareas related cell therapyfortreatinghumansport ofgenemodificationandstem development Platelet-rich plasma(PRP)hassignificant Cellular cardiomyoplasty(CCM), which Our labisfocusingonthediscoveryand of insulin. Stemcellsholdtremendouspotential beta-cells failtoproducesufficientquantities levels. whenthepancreas Diabetesdevelops system andacriticalcontrollerofbloodsugar tive tract organs. Stem cellsandregeneration indiges- formation. subsequent fibrocartilage injuries viathesuppressionofangiogenesisand the beneficialeffectsofPRPonchondral hypothesize thatinhibitionof VEGF willenhance degeneration.vessel formationandcartilage We healing buthavebeenshowntostimulateblood growth factorsarebeneficialforskeletalmuscle other typesoftissues. Forexample, angiogenic type oftissuebutproducedetrimentaleffectsin promote tissuehealingandregenerationinone cating thisissue, growthfactorsmay certain subsequent tissuedamage. compli- Further inflammationand are knowntopromotefurther contains largequantitiesofsubstancesthat TGF-β1, amongmanyothers). However, PRPalso tissue healing(includingPDGF, VEGF, IGF, and sports relateddiseases sports Stem cell/Genetherapyandtissueengineeringfortreating Assistant Professor Ping Guo, Ph.D. scar formation(bluein Trichrome staining)whenco-injected with TGFβ1 siRNA(bottompanel). andreduced previously preserved thenormaltissuestructurein infarctedzoneaswereported with TGFβ1siRNA.MDSCs Fourweeksafterinjection, MDSCsreducedtheinfarctedareaandpartially SCs (withRFPtag)wasdetectedbyanti-RFPantibody(toppanel). We alsohavetestedco-injectionof Transplantation ofmusclederivedstemcellsintothemyocardialinfarcted heart. EngraftmentofMD- The pancreas is a vital part ofthedigestive The pancreasisavitalpart Gonzalez. Research Assistants: ElizabethMorris, Sabrina Graduate Students:IsaacCastillo Post-doctoral Fellows:ShanshanGao LAB MEMBERS 2016, 202:227-236. Organs, ontheHorizon!CellsTissues New Guo, Yong Li, MuscleInjuriesandRepair: What’s Johnny Huard, Aiping Lu, XiaodongMu, Ping logia, 2017, Sept. 7, publishedonline. Manipulation inPancreatic Alpha Cells. Diabeto - GcgCreERT2 KnockinMiceas A Toll forGenetic Joseph Fusco, XiangweiXiao, GeorgeGittes. Chiyo Shiota, KrishnaPrasadan, PingGuo, KEY PUBLICATIONS “mini organ” thatmimicsthenativepancreas. placed inaBiohub, whichisabioengineered source ofinsulin-producingcellsthatcouldbe anykindofcellandhencecouldbea virtually havethepotentialtobecome because they

77 • PACT REPORT PACT IMM 78 • IMMPACT REPORT •To analyzethecell non-autonomouseffects RESEARCH PROJECTS histopathologies inmuscledystrophy. stemcelldysfunctionandimprovethe alleviate therapiesto novelandclinicalrelevant develop host. The resultsobtainedfromthisstudywill for thefunctionofmusclestemcellsin from donor, butalsohaveabeneficialeffect hance theoutcomeofstemcelltransplantation improve thisenvironment, thiswillnotonlyen- young/or pregnant WT and dystrophicmicewill We hypothesizedbloodexchangebetween roenvironment inthemuscleofDMDpatients. unsatisfied outcomeduetotheharmfulmac- in DMDhasbeenwidelyinvestigatedbutwith genitor cellsfromhealthydonorsfortreatment cure forDMD. Transplantation ofmusclepro- genetic diseaseandthereisstillnoeffective Duchenne musculardystrophy(DMD)islethal for treatmentofmusculardystrophy. come by improvingmicroenvironment their myogenicpotential. pregnancy maystimulateMPCsandimprove suggest thatcirculatingfactorspresentduring serum obtainedfrompregnantmice. Ourresults genic capacitywhenculturedinthepresenceof non-pregnant micedisplayedenhancedmyo- non-pregnant mice. Inaddition, MPCsfrom the significantly improvedwhencomparedtothatof ity ofMPCsisolatedfrompregnantmicewas also foundthatmyogenicdifferentiationcapac- compared tonon-pregnantcontrolmice. We healing followingcardiotoxininjurywhen that pregnantmiceexhibitacceleratedmuscle the motherandfetus. We havedemonstrated model ofasharedcirculatorysystembetween Pregnancy representsauniquebiological Myogenic Progenitor Cells(MPCs). ing andMyogenic Differentiationof ate aging. of muscleinjury, musculardisease, andacceler- peutic musclederivedstemcellsfortreatment RESEARCH ENGINEERING ANDAGING CENTER FORTISSUE Enhance stemcelltherapeutic out- Pregnancy ImprovesMuscleHeal- Our groupfocusesonidentificationofthera- Mu X,Mu Tang Y,K,Chen W,LuA, Takayama Wang Jun 22;12(6):e0179270. PD, NiedernhoferLJ, HuardJ. PLoSOne.2017 Proto JD, Lu A, Dorronsoro A, Scibetta A, Robbins KEY PUBLICATIONS •Parabotic pairingbetweendystrophicmice • • Assistant Professor Aiping Lu, M.D. treatment ofmusculardiseaseandaccelerateaging Identification oftherapeuticmusclederivedstemcellsfor histopathology and WT miceforimprovingmuscleandheart wound healing The roleofpregnancyinmuscleinjuryand tween myogeniccellsandnon-myogenic To investigatethecell-cellinteractionbe- inactivation ofERCC1(NIH, PO1) heterochronic parabiosisandtissue-specific induced byagingandstressonMDSPCsusing Aug; 100(2):142-6. Huard J, MuX, Lu A.Pharmacol Ther. Clin Genet. 2016 Aug 1;25(15):3216-3231. dar M, SunX, CumminsJH, HuardJ. HumMol X,Usas A,LuKozemchak Gao Tang Y,Pod- Organs. 2016;202(3-4):227-236. Huard J, Lu A, MuX, GuoP, Li Y. CellsTissues 2016. Clin Dev. 2016Nov30;3:16065. eCollection dar M, HoganMV, HuardJ. Mol Ther Methods Li H, Lu A, Tang Y, Beckman S, Nakayama N, Pod- 1;26 (15):2813-2824. B, Weiss K, HuardJ.2017Aug HumMolGenet. mice. serum frompregnant after stimulationwith differentiation capacity hibit improvedmyogenic non-pregnant miceex- (MPCs) isolatedfrom Muscle progenitorcells pregnant mice. healing afterinjuryin Improvement ofmuscle 2016 transplantation. Notch signalingpathways, andmusclestemcell methods includetheinhibitionof Wnt, ALDH, or cells andreducemuscleatrophy. Potential ways toimprovethefunctionofmusclestem osteosarcoma mousemodel, andexpecttofind stem cellsinmediatingmuscleatrophy an treatments. We arestudyingtheroleofmuscle in reducedenduranceofpatientstoclinical associated withcancercachexiaandresults cells onreducingmuscleatrophy. cachexia, andpotential effectsofmusclestem atrophy inosteosarcoma-inducedcancer regulators oftissueagingandcellsenescence. ing pathwayshavebeenshowntobeimportant regulators: Notch, Wnt, RhoA, andmTOR signal- cular dystrophicdisease. stemcell Some key such asthatresultingfromprogeriaandmus- been observedindiseasedskeletalmuscle, of regenerativepotentialstemcellshave regenerating muscles. toreducefibrosisandimprovefunctionof effort defects indiseasedandagedmusclesan regulatory mechanismsofmusclestemcell following areas: ossification, andosteosarcoma). cachexia-related muscleatrophy, heterotopic (i.e., DuchenneMuscularDystrophy, cancer ous musculoskeletaldisordersandconditions premature aging, andthemechanismsofvari- the mechanismsofstemcellsenescenceand rently, weareespeciallyinterestedinstudying aging, (osteosarcoma). andcancerbiology Cur- prevention, musculardystrophy, premature stem cellbiology, woundregeneration, fibrosis Surgery,Orthopaedic andourstudiesinvolve and of Aging ResearchandtheDepartment group intheCenterfor Engineering Tissue ofDr.team ispart JohnnyHuard’s research RESEARCH ENGINEERING ANDAGING CENTER FORTISSUE 3) Improvementofwound healingbyreducing Skeletal muscleatrophyisfrequently 2) Investigatingthemechanismsofmuscle Accelerated exhaustion, senescence, and loss 1) Understandingthecellularandmolecular Currently, Iamfocusingmyresearchinthe Our “Stem Cellsin Aging andCancer” research •Investigating themechanismsofmuscleatro- •Understanding ofthecellularandmolecular RESEARCH PROJECTS stem cells. application ofrelaxin, MMPs, andmultipotent regeneration ofdiseasedsofttissueswiththe fibrosis, weareinvestigatinghowtoenhance muscle) isusuallyaccompaniedbyexcessive ing ofdiseasedskeletalmuscle(i.e., dystrophic skeletal muscleoramputateddigits. Sinceheal- of fibrosisduringthehealingprocessinjured hormone relaxinandMMPsintheprevention fibrosis orapplicationofstemcells. were maintainedinmyogenic differentiationmedium. Treatment ofZ24 muscle phenotypesinprogeroidmice tors delaysmusclestemcellsenescenceandrescuesdefective Inhibition ofNF-Bsignalingwithnovelsmallmoleculeinhibi- Assistant Professor Xiaodong Mu, Ph.D. increased myotubeformation (MyosinHeavyChain/MHC+)after NF-κBinhibitionwhenZ24 adecreaseinSA-β-gal+cells;themyogenesis assayalsorevealed the cellsenescenceassay revealed the myogenesispotential. Z24 reducing muscleatrophy and potentialeffectsofmusclestemcellson phy inosteosarcoma-inducedcancercachexia aging, aswellrolesofcancerstemcells of acceleratedagingincontrasttonormal signaling inregulatingstemcellsenescence including therolesofNF-κB, Notch, and Wnt cell defectsindiseasedandagedmuscles, regulatory mechanismsofmusclestem theeffectsof reported We previously -/- MPCswithNF-κBinhibitordecreasedtheratioofsenescent cellsandincreased -/- MPCsweretreatedwithnovelNF-κBinhibitor(50µM)for4days, and •Improvement ofsofttissuewoundhealingby accelerated aging. Res.2016 JOrthop Aug 30 genitor cellsisolatedfromamurinemodelof defects observedinmuscle-derivedstem/pro- J. mTOR signalingplaysacriticalroleinthe M, FuFH, RobbinsPD, NiedernhoferLJ, Huard Cummins JH, Yurube T, KurodaR, Kurosaka Takayama K, Kawakami Y, LavasaniM, MuX, 2016. Cachexia CausedbyOsteosarcoma. Sarcoma. Mediates SkeletalMuscle Atrophy inCancer C, Tebbets J, HuardJ, Weiss K. NotchSignaling Mu X, Agarwal R, MarchD, Rothenberg A, Voigt 1;26(15):2813-2824. PMID:28549178. stem celldepletion.2017 Aug HumMolGenet. treatment indystrophicmuscle:implicationsfor improves thebeneficialeffectsofglucocorticoid B, Weiss K, HuardJ. RhoA/ROCKinhibition X,Mu Tang Y,K,Chen W,LuA, Takayama Wang KEY PUBLICATIONS reducing fibrosisorapplicationofstemcells -/- MPCs

79 • PACT REPORT PACT IMM IMMPACT REPORT • • • • 81 In addition to basic and translational addition In synthetic small molecules that modulate the synthetic small molecules that modulate molecules activity of these targets as potential lead for drug and 3) characterization of discovery; animals and humans in response antibodies from to viral infections and experimental vaccines. TTI has from the translational side, research On in the establishment of two UT-spin resulted for off biotech companies and granted licenses multiple biologic therapeutics. TTI two major is building programs, research drugTherapeutic platforms: 1) the discovery and Antibody Lead Optimization Monoclonal and 2) the Natural Platform Development Discovery Drug Molecular and Small Products The drug platforms not Platform. discovery TTI they also internal projects, only support with other scientists support projects collaborative based institutions, and other Texas at IMM, the countryinstitutions around the and around world. Ph.D. Zhiqiang An, Director & Center Professor Chair in Distinguished University Welch Robert A. Chemistry exas Therapeutics Institute at The Brown Brown Institute at The exas Therapeutics of Molecular Institute Foundation (TTI) was established in Medicine TTI investigators have brought in significant brought TTI have investigators TTI include: 1) activities at research Current TEXAS THERAPEUTICS THERAPEUTICS TEXAS INSTITUTE 2010 with funding from the Texas Emerging Emerging Texas the 2010 with funding from Texas of University The Fund, Technology Health Texas of University and The System, for the discovery, Science Center at Houston of and commercialization development, therapeutic agents and diagnostic tools. conducted at the center focuses on the Research validation of drugidentification and targets, for and establishment of proof-of-principle therapeutics. pharmaceutical and biotechnology funding from and & Johnson, companies, including Johnson of Institutes the National and from Merck, and Research the Cancer Prevention Health, and the Department of Energy. Texas, of Institute scientific made significant have TTI researchers fungal of cancer biology, in the areas discoveries and cancer antibody drugnatural products, development. and enzymes signaling mechanisms of receptors in human diseases; 2) critical roles that have and of biologics, natural products, discovery T and develop the stem cell therapy to inhibit development of PCa in bone capacity of MRL/ responsible for high healing to under- MpJ (super-healer) mouse strain regeneration stand the mechanisms of tissue The long-term goal of our research program is Proteomic approach to identify factors •Proteomic approach to identify to understand the role of epigenetic regulators and develop therapies for better bone health in age-related osteoporosis and other bone disorders. KEY PUBLICATIONS The Polycomb PHF19 (2012). Brien et al. binds H3K36me3 and recruits PRC2 and the NO66 demethylase to hitherto active genes during embryonic stem cell differentiation. 2012 Nature Structure and Molecular Biology, Dec;19(12):1273-81. Dandan Zhang, Xing Zhou, Wu, Minhao Tao, Yue Krishna M Sinha, Benoit deCrombrugghe, into the Structural Insights Jianye Zang (2013). Interactions of Histone Demethylase NO66 with of Journal Transcription. Osterix to Repress Gene 16430-16437. 288, Biological Chemistry 2013, Zhou Xin Yasuda, Hideyo Krishna M Sinha, Osterix and Benoit deCrombrugghe (2013). and NO66 histone demethylase control the chromatin architecture of Osterix target genes J Bone Mineral during osteoblast differentiation. Epub 2013 Sep 23. Research. R Krahe, Yasuda, H J Deng, K Sinha, Q Chen, R Behringer and B de Crombrugghe (2015). Mesenchymal Deletion of Histone Demethylase J NO66 in Mice Promotes Bone Formation. Epub 2015 Feb 26. Research, Bone Mineral in PCa progression and bone metastasis, in PCa progression and bone metastasis, Osterix by lysine methylation in differentiation of skeletal muscle stem cells into osteoblasts and bone formation renewal and depletion of skeletal muscle- and in the repair of derived stem cells, musculoskeletal tissues using next-gen sequencing approaches The histone demethylase NO66 has NO66 has The histone demethylase Bone is the most susceptible organ for metas- Musculoskeletal tissue regeneration using adult stem cell adult stem cell regeneration using tissue Musculoskeletal disorders and non-malignant skeletal therapy in malignant Krishna Sinha, Ph.D. Krishna Sinha, Assistant Professor To investigate the oncogenic function of NO66 •To To study post-translational modification of •To modified for expression of osteoblast specific modified for expression of improve Osterix (Osx) to transcription factor, Osx plays a their bone regenerative capacity. of osteo- critical role in turning on expression (Figure).blast genes during differentiation role in PCa and bone an oncogenic metastasis. particularly tases by nearly all types of cancer, and skeletal prostate and breast cancers, metastases lead to severe defects in bone NO66 is upregulated architectures during aging. have found that NO66 levelsWe in lung cancer. are elevated in prostate cancer patient samples indicate that Our data and xenografted samples. NO66 overexpression promotes the prolifera- In tion and invasion of prostate cancer cells. mice femurs of male SCID xenograft studies, implanted with NO66-overexpressing PC3 cells have significant bone loss compared with mice NO66 suggesting that with control PC3 cells, plays an oncogenic role in PCa progression we are also In addition, and bone metastasis. exploring research avenues to test the potential of muscle stem cell therapy in treating PCa and skeletal lesions. RESEARCH PROJECTS study epigenetic mechanisms of self- •To muscle derived progenitor/stem cells My research interests are focused on control in multipotent Epigenetic

Osterix serves as molecular switch for gene activation in osteoblasts. epigenetic control of cell fate determination, specifically on critical factors responsible for bone formation and bone homeostasis as well as prostate cancer-induced skeletal metastases. Our long-term research plan is to identify and study gene function in age-related bone information in order to provide pivotal disorders, that will inform the development of preventive measures and treatments for bone disorders. I have research group, Huard’s Since joining Dr. broadened my research interests to include gene therapy approaches that use multipotent skeletal muscle-derived stem cells in the regeneration of musculoskeletal tissues during aging and disease. muscle-derived potentials of skeletal stem cells. Skeletal (MPCs) are highly regenerative and multipotent that have ability to differentiate into various cell car- bone and types including skeletal muscle, important Epigenetic mechanisms play tilage. role in self-renewal capacity of adult stem cells through transcription control of genes that are My current involved in stemness of stem cells. research focuses on the epigenetic regulation by histone modification in control of those genes which are involved in the multipotent abilities We of MPCs and their regenerative capacity. are using chemical compound inhibitors and CRISPR/Cas9 gene editing tools to ablate the MPCs are function of epigenetic regulators. TISSUE FOR TISSUE CENTER AGING AND ENGINEERING RESEARCH

IMM PACT REPORT PACT • • • • 80 82 • IMMPACT REPORT •Cancer therapeutic monoclonalanti- •Antibodies responsetoviralinfections •HER3 mediatedcellsignalingand resistance •Cancer antibodydrug RESEARCH PROJECTS have fourmajorresearchareas. cancer andinfectiousdiseases. Currently, we antibiotics againsthumandiseasesincluding oftherapeuticantibodiesand development INSTITUTE TEXAS THERAPEUTICS a comprehensiveantibody drugdiscovery discovery.body drug Ourgrouphasbuilt and humans. the experimentalvaccinesinrhesusmonkeys vaccines byprofilingantibodyresponseto project toaidthedesignofHCMVanddengue effective viralvaccine. We haveanongoing isolates isoneapproachtodeveloping antibodies againstabroadrangeofclinical munogenic vaccinesthatinduceneutralizing and vaccination.Designofhighlyim- their modeofactions. HER3 targetingantibodiesandunderstanding EGFR antibodytherapies;and3)generationof plays inresistancetocurrentanti-HER2and mediated cellsignaling;2)theroleHER3 therapies. Ourgroup isworkingon:1)HER3 the resistancetocurrentEGFRandHER2 limited andHER3hasbeenimplicatedin ing ofthedrugresistancemechanismsis resistance iswidespread. Currentunderstand- treatment ofsomecancertypes, butdrug and HER2exhibitedclinicalbenefitsforthe in variouscancertypes. Agents targeting EGFR family receptorsignalinghasbeenimplicated therapy. Ablated regulationintheHER/ErbB HER3 targetingantibodiesforcancer the tumormicroenvironment. function mediatedbyproteolyticenzymesin immunity byimpairmentofantibodyeffector mechanismofcancersuppression of a new munity. Ourrecentstudieshavedemonstrated is largelybasedonstudiescellularim- nized asahallmarkofcancerandthisnotion mechanisms. Immunesuppressionisrecog- Our groupfocusesonthediscoveryand antibiotics oftherapeuticantibodiesand Discovery anddevelopment Robert A. Welch DistinguishedUniversityChairinChemistry Professor andCo-Directorofthe Texas Therapeutics Institute Zhiqiang An,Ph.D. memory B-cellsspecifictoviralgH/gL/ Lin Xiaetal. 2017 of Active evolution AAC.01719-17. Antimicrob. Agents Chemother. doi:10.1128/ Bispecific Antibody.an Anti-CD3/Anti-gB Using megalovirus-Infected CellsbyRedirecting T Cells Weixu Mengetal, 2017. Targeting HumanCyto- KEY PUBLICATIONS and D’. Antimicrob. Agents Chemother. doi:10.1128/AAC.01719-17 with uninfected ARPE-19 cells (imageD’). Magnificationis10Xfor A, B, C, DandE, 40Xfor A’, B’, C’, that T cellsmigrated towardstheHCMV-infected ARPE-19 cells. T-cells dispersedinthewells areevenly seeded with ARPE-19 cellswithorwithoutHCMVinfectionasindicatedbyGFP. Images A’ andB’ shows ence ofbispecificantibody. Tcellsandbispecificantibodywereadded toa6-wellplatethatwas Light microscopyof T cells incubatedwith ARPE-19 cellswithorwithoutHCMVinfectioninthepres- gray, andgLiscoloredyellow. Journalof 2017,91(7)e02033-16. Virology surface renderingwas generatedusingtheChimeravisualization package. GlycoproteingHiscolored vidual structures. The randomconicaltilt(RCT)methodwasused toreconstructthe3Dstructures. The Fab 1-85, orFab1-103andtheiroverlay, whichwasrotated180°horizontallywithrespecttotheindi- EM 3Dreconstructionofpentamerbound byFabs. Structuresofthepentamer boundwithFab2-15, targeting variouscancertypes. collaborative antibodydrugdiscoveryprojects humanization. Currently, wehavemultiple expressing Bcells, affinitymaturation, and encoding genesfromindividualantibody phage display, deepsequencingofantibody optimization technologiessuchasantibody platform withafocusonantibodylead Research Scientist: Ahmad S. Salameh Georgina Research Coordinator: T. Salazar Graduate Students: Yuanzhi Chen YanhongXu,Xiaohua Ye Wang, Yixiang Ku, Leike(Simon)Li, Wenxin Luo, Qihui Wang, Post-doctoral Fellows:Xun(Mark)Gui, Zhiqiang LAB MEMBERS e02033-16. Pentameric Complex. Journal of 91(7) Virology gH/gL/pUL128-131 Antibodies Targeting Viral Human CytomegalovirusStrainsConferredby Sha Haetal. 2017. NeutralizationofDiverse 73654–73669. megalovirus infection. Oncotarget. 8(43): healthy subjectswithsilenthumancyto- pUL128/130/131 pentamericcomplexin including pneumocandinB for thefamilyofechinocandinantifungaldrugs, characterized biosynthetic pathways responsible tion inhumandiseases. For example, wehave forpharmaceuticalinterven- families relevant as modelorganisms, especially biosynthetic of microorganismsusingfilamentousfungi predict geneticallyencodedchemicaldiversity intervention ininfectiousdiseasesandcancers. bioactivechemistryuseful in to generatenew synthetic mechanismsthatcanbeharnessed harbor biosyntheticgeneclustersandnovelbio- that naturalproduct-producingmicroorganisms products. We seektotestvarioushypotheses ing biosyntheticpathwaysofbioactivenatural medicinal chemistrysynthesis, andelucidat- products throughfermentationtosupport for therapeuticapplications, makingnatural involves testingmicrobialnaturalproducts lecular biology, andmicrobiology. Ourresearch sciences, organicchemistry, biochemistry, mo- diverse backgrounds, includingpharmaceutical members ofourlabandcollaboratorsfrom Institute ofMolecularMedicinebringstogether in the Texas Therapeutics Instituteandthe Multidisciplinary microbialbiomedicalresearch molecules forinventioninhumandisease. molecules continuetoyieldbreakthrough and geneticcapacityforsynthesisoforganic drugs.most important Their hyper-biodiversity INSTITUTE TEXAS THERAPEUTICS coccus neoformans, thecausalagentof Crypto- growth ofhumanpathogens, includingCrypto- bioactive naturalproductsthatinhibit new molecules. derivativesandoverproducedrug-precursor new synthesisof these naturalproductstosupport organisms toaidinsupplyinglargequantities of and genomicmanipulationintheproducing potency. Inparallel, weusepathwaygenetics analogueswithincreased purity andnew strainswithimprovedproduct to producenew re-programmed pneumocandinbiosynthesis ecule for the antifungal drug CANCIDAS. We have Our labemploysgenomicstointerpretand Microorganisms haveproducedmanyofour We alsocarryoutfundamentaldiscoveryof 0 , mol- thestarting encoding cyclosporine Cand A. independent. C. ChemicalstructureofcyclosporinC. D. Comparisonofbiosyntheticgeneclusters at 37°C, butnotatambienttemperature (25°C). Antifungal activityofamphotericinB is temperature (12 ml). B. ExtractsofA. felinacontainingcyclosporinCdisruptgrowthofCryptoccocusneoformans ture. A.felina, Cylcosporin C-producingfungus,Amphichordagrownonwheatfermentationmedium Cylcosporin CdisruptsthegrowthofCryptococcus meningitis pathogenathumanbodytempera- metabolites forinfectiousdiseasesandcancertherapies Genome mining, biosynthesisanddiscoveryofmicrobial Kay and Ben Fortson DistinguishedChairinNeurodegenerativeDiseaseResearch Kay andBenFortson Professor Gerald Bills, Ph.D. •Discovery of new antifungalsandother ••Discovery ofnew ofmethodsforreprogramming ••Development •Biosynthesis ofnaturalproductsandpathway RESEARCH PROJECTS with otheracademicandindustriallaboratories. extracts areavailablethroughcollaborations to theactivity-causingnaturalproducts. These preparative HPLCandbioautography, guideus More refinedchromatographictechniques, e.g., our bioassaysagainstthetargetpathogen. fractions oftheextractsareidentifiedthrough such asflashorcolumnchromatography, active pathogens. After preliminarychromatography, directed atfindingmoleculesthataffecthuman biological effectsusinganensembleofassays foruseful of fermentedfungiareevaluated coccus meningitisandcryptococcosis. Extracts therapeutic agents useful fortreatinghumandiseases to discoveroroverproducenaturalproducts transcription ofbiosyntheticgenesfungi engineering forimprovedantifungals Academy ofSciences, InstituteofMicrobiology). Prof. Professor: MeichunXiang(Chinese Visiting Post-doctoral Fellow:Dr. NanLan. LAB MEMBERS FUNK-0009-2016. Spectrum4:doi:10.1128/microbiolspec. ology secondary metabolitesfromtheFungi. Microbi- Bills, G.F. &J.B. Gloer. 2016. Biologicallyactive Journal ofNatural Products.80:2101–2109. . pseudauxarthron rapeptides fromAuxarthron Anti-Cryptococcus phenalenonesandcyclictet- tholomeusz, Z. An, J. Gloer&G. Bills. 2017. Li, Y., Q. Yue, J. Dinith, D. Swenson, G. Bar- doi.org/10.1016/j.bmcl.2017.11.001. organic &MedicinalChemistryLettershttps:// derivatization andanticancerevaluation. Bio- of sphaeropsidin A: Isolation, semi-synthetic recognizedsource lus candidusisanewly A.V. Kornienko&G.F. Bills. 2017. Aspergil- Li, Y., R. Scott, A.R. Hooper, G.A. Bartholomeusz, KEY PUBLICATIONS

83 • PACT REPORT PACT IMM 84 • IMMPACT REPORT tumor cells which express high levels oftarget tumor cellswhichexpress highlevels is onlyreleasedonce the ADC bindsandenters tached toacytotoxic chemical “warhead” that of ahighlyspecificmonoclonalantibody at- similar toguidedmissiles. ADCs arecomprised therapies. ofCSC-based targetforthedevelopment new role incancerandcouldserveasapromising findings suggestthatLGR5playsanimportant adhesion incoloncancercells. Altogether, these small GproteinRac1andincreasecell-cell ing GTPase Activating Protein1)torecruitthe the scaffoldproteinIQGAP1(IQMotifContain- recently, wehavefoundthatLGR5interactswith in stemcellsurvivalandtumorigenesis. More tentiate Wnt signaling, regulatorypathway akey the secretedgrowthfactorsR-spondinstopo- discovery thatLGR5functionsasareceptorof Dr. Qingyun’s (Jim)Liu’s laboratoryledtothe is stillrelativelyunknown. workin Myprevious the functionandmechanismofLGR5inCSCs liver, gastric, andovariancarcinomas. However, othermajortumortypes,in several including has beenshowntobesignificantlyelevated and metastasis. Inaddition, LGR5expression colon CSCsarecapableofdrivingtumorgrowth colon tumorsandthatonlyLGR5-positive pled Receptor5)ishighlyexpressedinprimary destroy CSCs. novel therapiesthatcanultimatelytargetand of cancer. Therefore, itisessentialtodevelop a majorimpedimentfortheeffectivetreatment sistance, metastasis, andrelapse, makingthem growth. CSCshavebeenimplicatedindrugre- havethecapacitytofueltumor cells sincethey as cancerstemcells(CSCs)ortumor-initiating normal stemcells. These cellsarereferred to a subpopulationofcancercellsthatbehavelike eral differentmalignanttumortypesthereexists INSTITUTE TEXAS THERAPEUTICS will targetanddestroycolontumors CSCs, innovative antibody-drugconjugates(ADCs) that (Leucine-rich repeat-containing lished thattheadultstemcellmarkerLGR5 My research is focused on the development of My researchisfocusedonthedevelopment Recent studieshaveunequivocallyestab- hasshownthatwithinse Emerging evidence , Gprotein-cou- v-

Treatment withanti-LGR5 ADCs eradicatestumorsin axenograftmodelofcoloncancer. colon cancer. target CSCsforthetreatmentanderadicationof and generateinnovativetherapeuticleadsto the functionandmechanismofLGR5inCSCs of CSCs. Ourworkwillleadtotheelucidationof in thecontrolofstemnessanddrugresistance the signalingmechanism(s)ofLGR5anditsrole Furthermore, wearecontinuingtoinvestigate in micepriortoantibody-drugconjugation. measure tumoruptakeandbiodistribution ourantibodiesasdiagnostics and to evaluate Azhdarinia’s group, weareusingPET/CTimaging ADC development. IncollaborationwithDr. Ali cancertargetsforfuture and characterizingnew LGR5-targeting ADCs andarealsoidentifying to modifyandimprovetheefficacyofour grafts. We arecurrentlytakingnovelapproaches coulddestroycoloncancercellsandxeno- they monomethyl-auristatin E(MMAE)andshowed targeted ADCs thatincorporatethecytotoxin antigen. We havesuccessfullygeneratedLGR5- tumors andcancerstemcells Therapeutic targetingandmechanismsofLGR5incolon Assistant Professor Kendra S. Carmon, Ph.D. (LAMP1, green)where drugisreleasedtodestroythecancercells(ADC/LAMP1co-localization, yellow). Anti-LGR5 ADC (red) bindsLoVo coloncancercells, internalizes, tothelysosome andistransported Cancer Ther. recurrence,testinal tumorsandprevents Mol. antibody-drug conjugateeradicatesgastroin- An, Z., Liu, Q., andCarmon, K.S. LGR5-targeted Gong, X., Azhdarinia, A., Ghosh, S.C., Xiong, W., 292(36):14989-15001, 2017. cells viatheIQGAP1-Rac1pathway. JBiolChem. cell adhesioninstemcellsandcoloncancer nov K.A., LiuQ.J. LGR5receptorpromotes cell- Moore C.M., MasuhoI., D.J.,Timson Martemya- Carmon K.S., GongX., J.,Yi Wu L., Thomas A., KEY PUBLICATIONS •Investigation oftheLGR5signalingmecha- ofantibody-drugconjugatesto •Development RESEARCH PROJECTS drug resistance nism anditsroleincancerstemcells target colontumorsandcancerstemcells 15(7):1580-90, 2016. associated withsecondmessengercAMP, • • • RESEARCH PROJECTS failure. and heart human diseases, including cancer, chronicpain, and inexploringtheirpotentialusesvarious isoform specificEPAC inhibitorsandagonists secondgeneration engaged indeveloping of fatalrickettsioses. Currently, weareactively EPAC andtreatment inhibitorsintheprevention Recently, wehaveidentifiedapotentialuseof signalingmolecules.of thisfamilyimportant physiological functionsanddiseasesrelevance and EPAC knockoutmousemodelstostudythe first-in-classEPACdeveloped selectiveinhibitors for thetreatmentofhumandiseases. We have be exploitedandcontrolledpharmaceutically signaling moleculessothattheirfunctionscan way specificmodulatorsfortheseimportant intricacies ofEPAC proteinsandtodesignpath- (EPAC). Ourgoalsaretounravelthesignaling exchange proteinsdirectlyactivatedbycAMP and functionofafamilycAMPsensors: chemical biology, tounderstandthestructure and withpharmacology ics andcellbiology approaches, couplingbiochemistry, biophys- of humandiseases. We applymultidisciplinary forthedevelopment jor stresssignalimportant INSTITUTE TEXAS THERAPEUTICS

human diseases, Examine therolesofEPAC proteinsinmajor candidates targetingEP ofnoveldrug Preclinical development (EPAC), exchange proteinsdirectlyactivatedbycAMP Structural andfunctionalanalysesofthe Our laboratorystudiesintracellularsignaling models andpharmacologicalinhibitors failure,and heart usingEPAC knockoutmouse bacteria Rickettsia, funded byNIH of microbialinfectionscausedbytick-borne fundedbyNIH suchascancer, chronicpain, AC forthetreatment ama- Loss ofEPAC1 increasesleptinsensitivityandprotects micefromhigh-fatdietinducedobesity 36:2440-1250, 2016. white adiposetissue. regulation ofleptinexpressionandsecretionin homeostasis: by cAMPisoform1inenergy X. Roleofexchangeproteindirectlyactivated Wang Hu, Y KEY PUBLICATIONS cAMP -mediatedcellsignalinganddrugdiscovery Walter andMaryMischerDistinguishedProfessorinMolecularMedicine Professor Xiaodong Cheng, Ph.D. of Epac1suppressesmitochondrialfissionand Inhibition X. Cheng, and J. Chen, G., Du, M., Cai, C., F. Mei, Z., Wang, G., W. Robichaux, H., Wang, PMC5021373/ http s ., :// , H.,Tong Robichaux, W.G.,Kim,E.R.,Mei,F www.ncbi.nlm.nih.gov/pmc/articles/ and Cheng, , Q.,Xu,M.,Chen,J.,andCheng, Molecular CellularBiology. . C., Research Assistants: PeiLuo Post-doctoral Fellows: Robichaux William Li Yue Instructor: Scientist: Wenli YangResearch Research FangMei Assistant Professor: LAB MEMBERS 7:6200,Reports. 2017. proteins directlyactivatedbycAMP. Scientific titative andisoform-specificassayforexchange Zhu, Y., Mei, F. andCheng, X. A cell-based, quan- Scientific Reports. 6:36552, 2016. neointimainducedbyvascularinjury.prevents

85 • PACT REPORT PACT IMM 86 • IMMPACT REPORT crine prostatecancers. Similarly, afterbecoming become AR-low/negative aggressiveneuroendo - AR-positive prostate adenocarcinoma cancers the acquisitionofresistanceto ARPIs, some of thesetwomajorsolidcancertypes. Upon cess incellularplasticityanddrugresistance which isincreasinglyacceptedasacritical pro- called neuroendocrinedifferentiation(NED), to betterunderstandandtargetaprocess inhibitors. The commonthemeinthistopicis cancers (NSCLC)becomeresistanttoEGFR inhibitors (ARPIs), andhow non-smallcelllung generationofandrogenreceptorpathway new how prostatecancersbecomeresistanceto and drugresistance. Inparticular, westudy tigate themechanismsofcancercellplasticity step ofcancermetastasis. tounderstandcolonization,key therate-limiting genes formetastaticcancercellsmaybethe fibrosis, andstem cellphenotypes. Essential in cancermetastasis, drugresistance, organ opmental process, role toplayakey isbelieved cancer cells(i.e. essentialgenes). EMT,- adevel survival andproliferationofhighlymetastatic (EMT), whilesomeothersareessentialfor tors controlepithelial-mesenchymaltransition for cancermetastasis. Someofthenovelregula- unknownregulators eral crucialbutpreviously functional screens,- ourlabhasidentifiedsev metastasis. Through genomics, RNAiandcDNA therapiesagainstcancer new and todevelop need tobetterunderstandcancermetastasis Therefore, thereisahugeunmetmedical metastatic cancersaremostlyunsuccessful. ortreathuman current approachestoprevent metastasis isstillpoorlyunderstoodandthe for over90%ofcancerdeath. However, cancer ofpatient’sother parts body, isresponsible cancers. ofbettertherapeutics forhuman development biomarkersanddrug targetsforthe identify new drug resistanceofhumancancercells, (2)to knowledgeofcancermetastasisand cal new INSTITUTE TEXAS THERAPEUTICS Another researchtopicsinthelabistoinves- Cancer metastasis, thespreadoftumorto My researchprogramsare(1)toobtaincriti- tissue microarray). breast cancersissignificantlyassociatedwithworseoutcomes(D-TCGA data, andE-breastcancer cell invasionincultureandbonemetastasismousexenografts(A-C). ofZAKinhuman Higherlevel criticalmetastatic-promotingproteincallZAKkinase.We discoveredanew ZAKiscrucialforcancer •Precision oncology, basedonmolecularpro- epigeneticregula - •Studying andinhibitingkey •Targeting metastasiskinasesthatwe new RESEARCH PROJECTS NED. orovercomedrugresistancerelatedto prevent and currentlythereisnoeffectivetreatmentsto very aggressive. NEDisstillpoorlyunderstood cancer, which isneuroendocrineinnatureand demonstrates phenotypesofsmallcelllung resistant toEGFRinhibitors, someNSCLC Studying andtargetingcancermetastasisdrugresistance Assistant Professor Wenliang Li, Ph.D. and patient-derived-xenograftsinmice files anddrugsensitivitiesofpatienttumors tors toovercomedrugresistanceincancer and breastcancers discovered toinhibitprogressionofprostate Visiting Scholar: Zhou Scholar: Ting Visiting Liu Ying Post-doctoral Fellows:Zheng Wang, NingSu, LAB MEMBERS Cell Death&Disease(inpress). epithelial-mesenchymal transitionincancer. S., Li, W*. MixedlineagekinaseZAKpromotes Li, L., Su, N., Zhou, T., Zheng, D., Wang, Z., Yuan, 1536, 2017Mar. of thrombospondin-1. Oncogene 36(11):1525- gression throughHDAC2-mediated suppression tumor angiogenesisandprostatecancerpro- Li, W*. Beta-adrenergicsignalingpromotes Hulsurkar, M., Li, Z., Li, X., Zhang, Y., Zheng, D., KEY PUBLICATIONS TEXAS THERAPEUTICS INSTITUTE TEXAS THERAPEUTICS current efforts are focused on further optimizing arefocusedonfurther current efforts models ofcolorectal and ovariancancer. Our displayed robustanti-tumor activityinanimal their drugconjugates. The modifiedantibodies series ofanti-LGR4antibodiesandgenerated We alsohaveidentifiedandcharacterizeda efficacy inpreclinicalmodelsofcoloncancer. LGR5 antibodiesshowedexcellentanti-tumor recently, weshowedthatdrugconjugatesofant- adenocarcinomas andcolorectalcancer. Most has amajorroleintheaggressivenessoflung In particular, weuncoveredthatRSPO3-LGR4 together toregulatecellgrowthandmigration. have nowelucidatedhowRSPOsandLGRswork for thesurvivalandgrowthofstemcells. We called R-spondins(RSPOs)thatareessential as receptorsofagroupstemcellfactors Previously, wediscoveredthatLGR4-6function survival ofnormalstemcellsandtumorcells. LGR6 (LGR4-6)thatplaycriticalrolesinthe surface receptorscalledLGR4, LGR5, and 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 inwhichonlyasubpopulationof 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 of cancertherapeutics Investigation ofnormalandcancerstemcellsforthediscovery Janice DavisGordonChairforBowelCancerResearch Professor (Jim)Liu,Qingyun Ph.D. internalized intothe lysosome (Green)incancercells. Internalization andlocalization ofanti-LGR4antibodyintoovarian cancercells. LGR4antibody(Red)is ProcNatl Wnt/beta-catenin signaling. Acad Sci orphan receptorsLGR4andLGR5toregulate Liu, Q. R-spondinsfunctionasligands ofthe Carmon, K.S., Gong, X, Lin, Q., Thomas, A., and KEY PUBLICATIONS •Optimization ofantibody-drugconjugatestar- •Identification ofleadmoleculestargeting •Investigation oftherolesaberrantexpres- •Determination ofthefunctionandmechanism •Delineation ofsignalingmechanismsstem RESEARCH PROJECTS other typesofmalignancies. as potentialtreatmentforcolorectalcancerand these drugleadstargetingtheRSPO-LGRsystem of colorectalandother geting theRSPO-LGRsystemfortreatment therapeutics the RSPO-LGRsystemasnovelanticancer metastasis oflungandcoloncancer sion oftheRSPOsincontroltumor cancer cellgrowth of thereceptorsincontrolnormaland cell receptors Wu Research Associates:Wangsheng AliceYu,Ling Post-doctoral Fellow:SoohyunPark, Jianghua Tu LAB MEMBERS Biol Chem. 292:14989-15001(2017). cancer cellsviatheIQGAP1-Rac1pathway. J cell-cell adhesioninstemcellsandcolon myanov, K.A., Liu, Q. J.:LGR5receptorpromotes Moore, C.M., Masuho, I., Timson, D.J.,- Marte Carmon, K.S., Gong, X., Yi, J., Wu, L., Thomas, A., Cancer Ther. recurrence.testinal tumorsandprevents Mol antibody-drug conjugateeradicatesgastroin- Z., Liu, Q., Carmon, K.S. (2016). LGR5-targeted X.,Gong, Azhdarinia, A.,Ghosh,S.,Xiong, W., An, E1221-E1229 (2014). Wnt signaling. ProcNatl Acad SciUS A. 111: RSPO-LGR4 functionsviaIQGAP1topotentiate Carmon, K. S., Gong, X., Yi, J., Thomas, A., Liu, Q. S A,U 108:11452-11457 (2011). 15:1580-1590 (2016).

87 • PACT REPORT PACT IMM 88 • IMMPACT REPORT drug conjugate(ADC) showsenhancedcellkillingpotencyagainst humancancercellscomparedto ADCs constructedusing traditionallinearlinkers. Enzymatic conjugation ofacancer-specificantibody, in ourgroup, abranchedlinkerdeveloped and thehighlycytotoxicdrugMMAF. Theresultingantibody- lack ofefficientandversatilemethods. Recently, more drugsremainsunexploredbecauseofthe potential of ADC linkers thatcanloadtwoor load onlysinglecytotoxicdrugs. The clinical technology, most todate ADC linkersdeveloped toimprove Despite extensiveefforts ADC linker medicinal chemistry, andchemicalbiology. advantage ofthepowerorganicchemistry, ment ofnovelchemical ADC linkersbytaking based chemotherapy. forsuccessfulimplementation of is akey ADC- tumor sites. Thus, theuseofproper ADC linkers release ofthehighlyactivedrugonlyat bridge, enablingselectivedeliveryandprecise and thehighlyactivedrugprovidesaspecific propriate chemicallinkerbetweentheantibody too toxictouseforcancertreatment. An ap- the useofhighlyactivedrugsthathavebeen tumors whileavoidinghealthytissues, enabling deliver potentanticancerdrugsselectivelyto effectivecancertherapeutics.developing ADCs molecularplatformfor advancethisnew further ADCs haveinspiredscientistsandcliniciansto 2017), successfultreatmentoutcomesusing than 60promising ADCs inclinicaltrials(asof Kadcyla®, Besponsa®, and Mylotarg®) and more with the4FDA-approved ADCs (Adcetris®, of anticancertherapeutics. As demonstrated a rapidlygrowingandextensivelypotentclass INSTITUTE TEXAS THERAPEUTICS - Our primaryinterestisinthedevelop Antibody-Drug Conjugates(ADCs)represent therapeutics antibody-drug conjugates(ADCs)towardinnovativecancer Branched linker forconstructingefficacious technology Assistant Professor Kyoji Tsuchikama,Ph.D. Tsuchikama,K.*,Shimamoto, Y., Anami, Y. DOI:10.1007/s13238-016-0323-0. tion andlinkerchemistries,Protein&Cell, conjugates: recentadvancesinconjuga- Tsuchikama, K.*, An, Z. (2016) Antibody-drug KEY PUBLICATIONS •Quorum sensing-guideddrugdelivery •Structural optimizationof ADC linkersforhigh •Design, synthesis, ofnovel andevaluation RESEARCH PROJECTS candidates forthefutureclinicalstudies. valuable additionstothecurrentlistofdrug a novel ADC platformprovidinganumberof overcoming suchunsolvedissuesandestablish willprovideaninnovativeapproachfor ogy envisage thatourmulti-loading ADC technol- medication andrecurrenceofmalignancy. We chemotherapy leadingtodiscontinuationof heterogeneity areunsolvedissuesincancer ity issues. The drugresistanceand tumor the cancerdrugresistanceandheterogene- to producenext-generation ADCs forcombating currently advancingthemulti-loadingstrategy loading linkers. Basedonthissuccess, weare than thoseconstructedwithtraditionalsingle- are moreeffectiveincancercellkillingtests thatourdual-loading We haverevealed ADCs installation oftwomoleculesontoanantibody. loading ADC linkersenablingsimpleandeasy anefficientmethodusingdual- we developed plasma stabilityandrapiddrugrelease (ADCs) multi-loading antibody-drugconjugates branched chemicallinkersforconstructing pharmacology. organic chemistry, medicinalchemistry, and/or for PostdoctoralFellowwithexperiencein The Tsuchikama labhasanopenposition Chisato Tsuchikama,Ph.D. Fellows: YasuakiPost-doctoral Anami,Ph.D., LAB MEMBERS lecular Chemistry, 15, 5635–5642. Conjugates withHighPotency. Organic &Biomo- for ConstructingHomogeneous Antibody–Drug Enzymatic ConjugationUsingBranchedLinkers C. C., Zhang, N., An, Z., Tsuchikama, K.*(2017) Anami, Y., Xiong, W., Gui, X., Deng, M., Zhang, 406–410. coccus aureus. ACS InfectiousDiseases, 3(6), gates SelectivelyRecognizeandKillStaphylo- Conju- Peptide (2017) Truncated Autoinducing TEXAS THERAPEUTICS INSTITUTE TEXAS THERAPEUTICS studies suchashigh content fluorescenceimag- technologiesare usedinour State-of-the-art by proteolyticimpairment ofantibodyhinge. triggered nisms ofcancerimmuneevasion proteolytic impairmentandtoidentifymecha - cancer patientstodeterminefactorsinfluencing models, andstudieswithclinicalsamplesfrom vitro 2Dand3Dcellco-cultures, mousetumor array ofexperimentalapproachesincluding in ment. To testourhypothesis, weemployawide to animmunesuppressivetumormicroenviron- antibody anticancerimmunitybutalsoleads hinge cleavageofantibodiesnotonlyweakens metalloproteinases (MMPs). Suchproteolytic impairment throughahingecleavagebymatrix microenvironment aresusceptibletoproteolytic tumor associatedantigens(TAA) inthetumor we hypothesizethatantibodiesrecognizing byothers,on ourrecentfindingsandreports and reducedantitumorefficacyinvivo. Based effector functionagainstcancercellsinvitro hinge cleavageshowedalossofimmune microenvironment. Trastuzumab withasingle lytic impairmentofantibodyIgGinthetumor ofproteo- demonstrated theprevalence as ahallmarkofcancer. Ourresearchhas immunity forimprovementofcancertreatment. therapeutic strategiestomodulateanticancer of sion ofantibodyimmunityanddevelopment - centered onbetterunderstandingoftumoreva patients. Ourcurrentresearchprogramsare targeted therapiestobenefitmorecancer forimprovementofthesecancer importance tance totherapeuticantibodiesisofparamount Understanding themechanismofcancerresis- therapeutic antibodiesarewidelyreported. both innateandacquiredresistancetothese treatment ofmanytypescancer. However, factor VEGF, havebeensuccessfullyusedfor targetingtumor angiogenesis and bevacizumab antibodies, suchastrastuzumabagainstHER2 types ofcancer. Tumor targetingmonoclonal shown clinicalsuccessfortreatmentofvarious drug modalityforcancertreatmentandhave Cancer immune evasion isrecognized Cancer immuneevasion Monoclonal antibodiesarebecomingamajor McLelian, HuaZhu, P. Stuart Adler, Michael Pete A. Dephillips, Zhang, Ningyan JasonS. Nickle, RichardR. Rustaandi, MelissaHamm, Dai Wang, I-Ming Wang, Josef Vlassak, DavidC. Daniel C. Freed, Aiming Tang, John W. Loughney, Sha Ha, FengshengLi, C. Matrhew Troutman, KEY PUBLICATIONS of •Understand mechanismsofcancerevasion •Determine theroleofproteolytichinge RESEARCH PROJECTS anticancer immunotherapies. ofeffective molecular targetsfordevelopment tumor microenvironmentandtoidentifykey between cancercellsandhostimmunityinthe the roleofantibodiesindynamicinteraction long-term goalofourresearchistounderstand models. usinginvitroandvivoThe evaluation select novelanticancerantibodiesforfunctional todiscoverand antibody platformtechnology antibodies. We haveestablishedamonoclonal (FACS),cell sorting andsingle-cellcloningof ing, massspectrometry, fluorescenceactivated CCR-15-1057. immunohistochemistry method(IHC), seeClinicalCancerResearch, 2015;doi:10.1158/1078-0432. Detection ofproteolytichingecleavageantibodiesintumortissuesfrombreast cancer patientsby modulation ofanticancerimmunity Cancer resistancemechanismstotherapeuticantibodiesand Associate Professor Zhang,Ningyan Ph.D. antibody immunotherapeutics. suppression. cleavage ofantibodiesincancerimmune Post-doctoral Fellows: YanhongPh.D.Post-doctoral Wang, Xuejun Fan, M.D., Ph.D., Wei Xiong, Ph.D. Research Associate/Scientists: HuiDeng, M.S., LAB MEMBERS Chemotherapy. doi:10.1128/AAC.01719-17 Bispecific Antibody. Redirecting T CellsUsingan Anti-CD3/Anti-gB Human Cytomegalovirus-InfectedCellsby Tong-Ming Fu, Zhiqiang An. (2017) Targeting Freed, ShaHa, Dr. Dai Wang, Zhang, Ningyan Leike Li, XuejunFan, RobbieD. Schultz, Daniel Weixu Meng, Aimin Tang, Xiaohua Ye, XunGui, try. DOI:10.1039/c7ob01027c. High Potency. Organic &BiomolecularChemis- Homogeneous Antibody-Drug Conjugateswith tion UsingBranchedLinkersforConstructing Tsuchikama, Kyoji. (2017)EnzymaticConjuga- Zhang, Cheng, Zhang, Ningyan, An, Zhiqiang, Anami, Yasuaki, Xiong, Wei, Gui, Xun, Deng, Mi, of Virolog. JournalVol 91, e02033-16. viral gH/gL/pUL128-131pentamericcomplex. virus strainsconferredbyantibodiestargeting Neutralization ofdiversehumancytomegalo- A. MaVoy, Zhiqiang An, Tong-Ming Fu(2017) Antimicrobial Agents and Antimicrobial Agents

89 • PACT REPORT PACT IMM 90 • IMMPACT REPORT T E IMM isacriticalcomponentofthiscommitment. centersatUTHealth-establishment ofkeyservice customers accesstocutting–edgetechnology. The infrastructure andtoprovide toourfacultyand improvea systematicprocess tofurther our forourresearch capacity,services we have initiated providing highqualityandeffective support offered. staffed by top inthetechnologies research experts cost-effective manner. IMM’s Centersare Service colleagues andcustomerswhileoperatingina toour technology andthehighestqualityservices centergoalistoprovideservice thelatest protein technologiesandanimalmodels.Our external funding tocontinuedevelopment of strengthen the collaborators’ abilityto attract Results center will generatedfrom theservice cloning, antibodyexpression andpurification. toantibodyidentification,molecular services centeristoprovide and service technicalsupport discovery communities.Theobjective ofthe lead optimizationfortheresearch anddrug early discovery ofmonoclonal antibodiesand in fill thegapofmuch-neededexpertise centerwill engineering andexpression service and otherrelated keytechnologies. Our antibody infrastructure required forantibodyengineering and by thelackofaccesstoexpertise academic research laboratoriesisoften hindered antibodies intheearlystageofdiscovery from However, advancement ofsomethepromising in discovering candidates. antibodydrug new academic researchers are increasingly engaged discovery anddevelopment,stage ofdrug and biotechnologyindustriesare inthecenter including cancer. Even thoughthepharmaceutical breakthrough incombatinghumandiseases A IMM Service Centers xpression ntibody To accomplishIMM’s strategicgoalof Antibody therapeuticsrepresents amajor and molecularlevels usingDNAand preventing diseaseatthegenetic,cellular he IMMisfocusedonstudyingand E S ngineering ervice C entr

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C therapies. ultimate goaloftranslatingbasicresearch tonovel the optimized therapeuticantibodieswiththe translational imaging studiesshouldbeacquainted are orwish to beinvolved insmall animal/ a facilitythatallresearchers atUTHealth who C analysis, andbioinformaticsdataprocessing. including samplepreparation, massspectrometric to provide anintegrated proteomics analysis, andtrainedpersonnel edge instrumentation centercontainsthecutting- profiling. Theservice assays, andmetaproteomics formicrobiome verification, development oftargetedproteomics suchasbiomarker discoverysupports, and and/or otherbiofluids. We alsoprovide advanced such ascellandtissueextracts,plasma/serum, purified protein samplestocomplexmixtures in abroad rangeofresearch specimensfrom identify andquantitateproteins andtheirPTMs community, andotherexternalorganizations. at Houston, Texas Medical Centerresearch totheentire UTHealthservices ScienceCenter center provides proteomics state-of-the-art proteins and otherbiomolecules.Our service functions, aswell asprotein interactionwith post-translational modifications(PTMs),protein changes inprotein compositionandexpression, state. Such proteome alterationsmayinclude associated withadiseasestatusorbiological proteome alterations or perturbation-induced research andclinicalspecimenstoidentifysteady These studiesoften require extensive analysesof and theevaluation oftherapeuticefficacy. and therapies,butalsofordiseasediagnosis only fortheunderstandingofdiseaseprocesses increasingly focusedontranslationalstudiesnot C P roteomics entr oll linical The IMMCenterfor Molecular Imaging is providedThe basicservices are designedto Current trends inbiomedicalresearch are abor

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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 disorders, gastrointestinal disorders, projects incancer, discovery, drug autoimmune researchers across thenationontranslational scientists, aswell asacademicandindustry tointeractwithclinicians,clinician- expertise “collaboration” centerutilizes thisexisting technologies intoclinicaltrials. imaging and pioneeringtranslationofnew of targetedprobes, innovative algorithms, design,andchemistry instrumentation, new different aspectsofmolecularimaging,including faculty memberswhoseresearch focuseson and ledby seven engineeringandbasicscience with. Thecenterisdirected by Dr. Eva Sevick organizations. investigators from UTHealth andexternal chargetoallresearchavailable onafee-for-services are These instruments analysis andcellsorting. forboth andtechnical expertise instrumentation, centerprovides Theservice training, be sorted. cells andadditionallyisolatecellpopulationsto rare populations of gather informationonvery timeframeand large numberofsamplesinashort allowThese instruments scientiststoevaluate a as theypassthrough liquidinfront ofthelasers. type. cells suchasrelative size, complexity, andcell be measured of individual todetermineproperties wavelengths. The fluorescencevarying canbethen labelled andthenpassedinfront oflasers variety of cellular componentsare fluorescently the characteristicsofcellsinfluid. Typically a F low The newly formed The newly Molecular Imaging Thousands ofcellscanbeanalyzed persecond Flow isatechniqueusedtoanalyze cytometry C ytometry S ervice C entr C medical imagingfilessuchasMRIdata. 3D printerscanusestandard STLfilesand andacademics.The products forbothindustry of producing bothprototype modelsandfinal resolution (14microns) 3Dprinterscapable Recently, we have added twolarge-scale,high- production, andchemicalconjugationservices. other diseases.Thecenteralsooffersnanoparticle we alsodevelop aptamersforthemodulationof such asovarian, breast, andpancreatic cancers, regulation oftheprotein. ofchemotherapeuticagentsorthedown-delivery RNA forspecifictargetingofproteins forthe purification ofDNAaptamers,X-Aptamers, and Building. floorsofthe and fourth Fayez S. Sarofim Research established in2012andislocatedonthethird N T research. of microsurgery, cellculture, andstemcells indifferentintellectual/technical support aspects gene targeting,derivation celllines,and ofnew ofthefacilityalsoinclude rats, theservices derivation ofgenetically-engineered miceand andre-to theproduction, cryopreservation, for celldifferentiation studies. In addition generation ofknock-out/knock-inmiceand are highlyeffective forthe in thelaboratory basis. organizations onafee-for-service investigators from UTHealth andexternal knock-out mouseanimalmodelsforallresearch It hasgeneratedover transgenicand 800new center,service whichwasestablishedin1998. Center operatesa Transgenic andStem Cells ransgenic entr anochemistry While mostofourprojects targetcancers, It specializes inthediscovery, synthesis,and The Centerwas Service Nanochemistry The stemcelllinesthathave beenderived Our Immunology andAutoimmune Diseases

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91 • PACT REPORT PACT IMM IMMPACT REPORT • • • • 93 Gift Report Gift * IMM advisory council member ** deceased Zhiqiang An, Ph.D. and Alan Baden *Betty Foundation Jr. The Harry E. Bovay, Cahill Robert Deleon Jose Mary Errera and Robert JD PhD, Cindee R. Ewell, M. Frois Theodore *Louana H. and Inc Foundation Family Frois L. Gordon *Steven Foundation Hamman and MaryJosephine George L. Jurek Roberta Wilson Kilroy Jeanie Knight Amy and Edward Mackel *Amber and John McDonald and John *Patricia McWilliams *Laura and Bradley **Ms. Kathryn G. Neuhaus O’Donnell Gerald and Helene and Charles Parker *Marsha Poyner F. Estate of Herbert MBA, PCM Schreiber, Roberts *Shavonnah The Will Under of EmoryKate Thompson, Trust C. Thompson Foundation Private Tidwell and Weaver Thank you to all of our supporters! New Gifts and Bequests Fiscal Year 2017 and Bequests Fiscal New Gifts 2017 2016 2015 Federal Government State Government Foundations Industry Service Centers Gifts/Endowments 2014 Note - ARRA funds Excludes all Sponsored Projects based on award received Service Centers and Endowments/Gifts based on expenses 2013 0 40% Total Funds Supporting Research Supporting Funds Total Endowment/Gifts Service Center Sponsored Projects $7,500,000 $5,000,000 $2,500,000 $25,000,000 $22,500,000 $20,000,000 $17,500,000 $15,000,000 $12,500,000 $10,000,000 FY 17 59 27% 20% 57 FY 16 Total Expenses Supporting Research Supporting Expenses Total FY 15 56 4% 6% 3% 52 FY 14 Number of Faculty Number 53 FY 13

IMM By the Numbers By IMM

IMM PACT REPORT PACT • • • • 92 94 • IMMPACT REPORT The Jerry Rubenstein andMaury Foundation Distinguished Professorship in Jerold B.KatzDistinguished Professorship inStem CellResearch Janice Davis Gordon Chairfor Bowel CancerResearch James T. Willerson Distinguished ChairinCardiovascular Medicine E.Bovay,Harry Jr. Distinguished University ChairinMetabolic Disease Research George Josephine &Mary Hamman Foundation Distinguished Professorship in Dr. Edward Randall,Jr. Memorial Fund D. Dudley andJudy White Oldham Research Fund Cullen ChairinMolecular Medicine The Carolyn Frost Keenan Professor inCardiovascular Disease Research C. Harold andLorineG. Wallace Distinguished University Chair Becker Family Foundation Professorship inDiabetes Research Annie andBobGraham Distinguished ChairinStem CellBiology Medicine EndowmentsInstitute of Molecular Heart Disease Research Cardiovascular Research What's theplan? on thewo goals foryourselfandyourlo 713.500.3200 Contact TheOfficeofEstateand and letushelpyoudevelopaplantosupporttheIMM. rl d

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George andCynthia Mitchell Distinguished ChairinNeurosciences William S.Kilroy, Sr. Distinguished University ChairinPulmonary Disease Walter Mischer &Mary Distinguished Professorship inMolecular Medicine The Welch Foundation Endowment andRelated Sciences inChemistry Rodney J.Sands New Initiatives Stem CellResearch Endowment Rochelle andMax LevitChairintheNeurosciences Robert A. Welch Distinguished University ChairinChemistry Kozmetsky Family Chair inHuman Genetics IMM General Endowment E.BovayHarry Lecture Series inMolecular Medicine Marjorie B.Poyner Endowment for Medical Research intheInstitute of Pierce Runnells Memorial Research Fund Research Nina andMichael Zilkha Distinguished ChairinNeurodegenerative Disease Nancy andRichKinderDistinguished ChairinCardiovascular Disease Research Hans J.Muller-Eberhard andIrma Gigli Distinguished ChairinImmunology Hans J.Muller-Eberhard Lecture Series ElizabethMary Holdsworth Distinguished University ChairinMetabolic and Disorders Linda andRonny Finger Foundation Distinguished ChairinNeuroimmunologic The Laurence andJohanna Favrot Distinguished Professorship inCardiology Kay andBen Fortson Distinguished ChairinNeurodegenerative Disease Research John S.Dunn, Sr. Research Scholars Molecular Medicine forPrevention ofHuman Diseases Inflammatory Disease Research who

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95 • PACT REPORT PACT IMM 96 • 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 Michael Blackburn, Ph.D. Executive Vice President, Chief Academic Officer Barbara J. Stoll, M.D. Dean, McGovern Medical School

IMM Senior Administrator John F. Hancock, M.A., M.B., BChir., Ph.D., Sc.D. Executive Director

IMM Advisory Council Members Mr. John E. McDonald, Chair Mr. Alan P. Baden Regent David J. Beck Mr. John Barney Beckworth Mrs. Louana H. Frois Mr. Theodore M. Frois Irma Gigli, M.D. Mr. Steven L. Gordon Dr. John F. Hancock Mr. John Mackel III Mr. Rodney H. Margolis Mr. D. Bradley McWilliams Mr. D. Dudley Oldham Mr. Charles R. Parker Ms. Beth Robertson Mr. Clive Runnells Shavonnah Roberts Schreiber, M.B.A., P.C.M. Mr. Ralph B. Thomas

The University of Texas System Board of Regents Sarah Martinez Tucker, Chair Paul L. Foster, Vice Chair Jeffery D. Hildebrand, Vice Chair Ernest Aliseda David J. Beck Jaciel Castro Kevin P. Eltife R. Steven Hicks Janiece Longoria James C. “Rad” Weaver