division of Immunogenetics division of Immunogenetics

Mission The Division of Immunogenetics is committed to providing basic science support aimed at better understanding the etio-pathogenesis of pediatric diseases to more efficiently and correctly diagnose, prevent, and possibly 136 better treat them. 137

DEPARTMENT OF PEDIATRICS 2012 ANNUAL REPORT DEPARTMENT OF PEDIATRICS 2012 ANNUAL REPORT division of Immunogenetics division of Immunogenetics

faculty and staff To block auto-, without the use of immunosuppressant, Nick Giannoukakis and Massimo Trucco have success- fully completed a National Institutes of Health (NIH)-funded, U.S. Food and Drug Administration (FDA)-approved Massimo Trucco, MD Yong Fan, PhD Steven Ringquist, PhD phase I clinical trial that was designed to test the safety of a treatment in which autologous DC—rendered functionally Hillman Professor of Research Assistant Professor Research Assistant Professor immature by exposing them to CD40, CD80, and CD86 antisense oligodeoxyribonucleotides (AS-ODN) to reduce Pediatric costimulatory molecule levels at their surface—were administered to T1D patients with established disease. Leukocytes Nick Giannoukakis, PhD William Rudert, MD, PhD of the patient were obtained by apheresis, and DCs were then isolated and expanded. DC engineered in GMP facilities Suzanne Bertera, PhD Associate Professor Research Associate Professor to express low levels of CD40, CD80, and CD86 were then injected into the patient by intradermal administration at an Research Assistant Professor Jing He, MD, PhD Tatyana Votyakova, MD, PhD anatomical site proximal to the pancreas. DCs then migrate to the nearest (i.e., pancreatic) lymph nodes, where they are Rita Bottino, PhD Research Assistant Professor Research Assistant Professor able to interrupt the vicious circle that maintains islet-specific (i.e., insulitis). In the pancreas, DCs acquire Research Associate Professor new beta cell-specific from apoptotic cells, leading to the eventual display of these antigens to naïve T cells in Jonathan Piganelli, PhD the pancreas-draining lymph nodes. The lack of costimulatory molecules will result in an anergizing signal to the T cells, H. Henry Dong, PhD Associate Professor induce regulatory immune T and B cells, and limit the T-cell mediated anti-beta cell epitope-spreading phenomenon. In Associate Professor particular, a role for tolerogenic dendritic cell-induced B-regulatory cells in T1D was formally ascertained. The abroga- tion of the autoimmune diabetogenic insult should be sufficient to promote the rescue of any remaining insulin-producing beta cells and/or neogenesis of other insulin-producing cells in the host endocrine pancreas, even after the onset of the 138 disease. This possibility waits to be tested in a phase II (efficacy) trial involving new-onset diabetic patients. This thera- 139 peutic approach should, in fact, be more successful when DC injections start close to the clinical onset of the disease or even before the clinical onset in patients at high risk. To help promote islet transplantation as a more successful and easily OVERVIEW OF DIVISION approachable alternative to insulin administrations, making available a virtually endless source of functional islets, Bottino and Bertera tested the possibility of using pigs as islet donors. An alpha 1-3 galactosyltransferase knockout pig was used he Division of Immunogenetics is committed to providing basic science support aimed at better understanding as the recipient of additional transgenes useful to reduce complement activation and blood coagulation in the recipient. the etio-pathogenesis of pediatric diseases to more efficiently and correctly diagnose, prevent, and possibly treat Our ability to isolate still functional islets for performing auto-transplants in chronic pancreatitis patients who underwent them. In particular, the Pediatric Research Section of the Diabetes Institute of the University of Pittsburgh, total pancreatectomy to reduce their excruciating pains helped to successfully test the pig to nonhuman primate recipient whichT is administratively considered a component of the division, is the forum created to foster studies on islet xenotranplants. The extremely promising results were recently reported. Also, Yong Fan and William Rudert’s efforts (T1D), with the specific goal of finding new avenues for more efficiently predicting who will eventually convert to the were focused on characterizing factors able to break immune central and peripheral tolerance, thus promoting autoimmu- disease, for preventing the disease in genetically predisposed children, and for finding therapeutic approaches more nity, and in conceptualizing clinically useful contra-measures. Under the supervision of Steven Ringquist, next-generation efficient than daily injections of recombinant insulin. Toward these aims, members of the division are currently exploring sequencing (using the Roche/454 instrument) genotyping of human leukocytle (HLA) loci, critical to define sus- different aspects of diabetes etio-pathogenesis. ceptibility to T1D and determinant to achieving long-term survival of allogeneic transplants, was compared with more classical typing approaches, like the ones which use sequence specific oligonucleotide (SSO) probes, or the T1D is a disease in which mechanisms common to the majority of autoimmune diseases contribute to its pathological imputation of genotypes from high-density SNP analysis. Jon Piganelli continued to pursue the goal of better defining the process. What seems certain is that, at some point postnatally, environmental triggers activate the of effects of inflammation and ROS production on the demise of pancreatic beta cells to provide new approaches to alleviate an individual who is genetically predisposed to . Once silent, auto-reactive T cells, which have somehow their effects not only in the animal model but also in humans. Tatyana Votyakova is determining the role of mitochondrial escaped thymic censorship, are then activated and become able to reach and kill their targets (i.e., the insulin-producing performance and level of mitochondrial reactive oxygen species in islet viability prior to their transplantation, focusing beta cells of the pancreas) by recognizing self-markers on them. Resident antigen presenting cells, like the dendritic cells on this aspect of potential injury from hypoxia and reperfusion inevitably arising in the course of pancreas harvesting and (DCs), respond to the micro-environmental anomaly (e.g., cell death) and initiate the autoimmune process by phagocy- islet isolation. All these aspects have the major target of better defining the outcome of transplantable isolated islets of the tizing dead beta cell debris and then migrating out of the tissue and into the proximal lymph nodes. Through presenta- pancreas. Jing He supports the other faculty efforts by characterizing the morphological changes in immune cells and dif- tion of the newly acquired self-antigens, the DCs trigger the activation and proliferation of new naïve T lymphocytes. ferent organs to provide structural evidences. Finally, Henry Dong was analyzing the new phenomenon of type 2 diabetes The recurrent activation of additional T cells against once “ignored” self-antigens, or other epitopes of the same antigen, (T2D) in children, focusing on the role of factors, like FoxO1 and O6, as the possibly acting genes in this pathology. constitutes a vicious circle, known as the “epitope spreading” phenomenon. In fact, once activated, new T cells leave the and extravasate into the target tissues, where they destroy additional target cells recognizing new antigens. Once a large enough portion of insulin-producing cells is lost, the clinical onset of the disease presents.

To date, there are no cures for autoimmune disorders in general and for T1D in particular. In T1D, significant tissue destruction has already occurred at the moment of the clinical onset, and there is very little that can be done to restore function of the target organ other than hormone replacement or transplantation of the lost cells, which in itself is immunologically challenging. It would be far easier to manipulate either the immune system of the recipient patients or transplant the cells. Even better would be to intervene in genetically at-risk individuals, in a manner by which tolerance to tissue-restricted antigens of the self could be restored well in advance of disease onset.

DEPARTMENT OF PEDIATRICS 2012 ANNUAL REPORT DEPARTMENT OF PEDIATRICS 2012 ANNUAL REPORT division of Immunogenetics division of Immunogenetics

RESEARCH AND OTHER SCHOLARLY ACTIVITIES

Massimo Trucco, MD This study suggested the reconstructed , the team successfully reconsti- • Reviewer, Journal of Clinical Endocrinology and Metabolism that in a monkey, like tuted mature and functional T and B cells in nude mice. • Reviewer, Journal of Immunology Research in the mouse, a drastic Utilizing this approach, the researchers should become • Reviewer (on reviewers panel), Medical Science Currently, the most actively pursued therapeutic reduction of expres- able to not only create autoimmunity in the monkey, Monitor International approaches to avoid the daunting complications of sion of self-antigens but also generate the basis for an efficient, safe, new • Reviewer, Transplantation T1D are islet allo-transplantation, stem cell-based islet (e.g., insulin) in the therapeutic approach to cure T1D. replacement, beta cell regeneration attempts, and viral mTECs might favor Major Lectureships and Seminars vector-based gene therapies. Although significant advances • “Practical Ways to Achieve Targets in Diabetes Care,” the generation of an Study Sections have been made in all of these areas, and an enormous • Member, TrialNet Operations Strategic Review Group University of Colorado School of Medicine Office of autoimmune reaction number of therapeutic approaches to cure diabetes has • Speaker, Health Research Advisory Committee meeting, Continuing Medical Education and Children’s Diabetes specifically directed been successfully tested in the NOD mouse—the geneti- Commonwealth Universal Enhancement (CURE) Foundation, Denver, Colo., July 2011 against the insulin- cally diabetes-prone non-obese diabetic mouse strain, Program, Harrisburg, Pa., September 2011 • “Can We Really Prevent and/or Cure Diabetes?: producing cells of the whose etiopathogenesis is widely held to parallel the one • Member, Scientific Advisory Board, Diabetes Cellular Therapy for Diabetes Prevention,” Keystone, pancreas, eventually 140 that occurs in humans—the majority of them simply did Massimo Trucco, MD Research Institute, University of Miami, Miami, Fla., Colo., July 2011 141 bringing the animal to Division Chief, Immunogenetics not work in humans. The gap between mice and humans November 2011 • Giornate Diabetologiche Di Palazzo Ducale, Genova, clinically overt T1D. seems to be too large to justify the translation of therapies • Member, Special Emphasis Panel for the review of R43 Palazzo Ducale, November 2011 efficacious in mice directly to human individuals. But, On this basis, the team intended to engineer autoim- applications submitted in response to RFA-DK-11-024, • “Tavola Rotonda, I protagonisti della ricerca scientifica: even if the nonhuman primate (NHP) seem to be the best mune diabetes in monkeys by safely removing the thymus “Small Business Innovation Research to Develop New Testimonianze di ricercatori italiani che lavorano animal model available for testing new therapeutics on the from Cynomolgus monkeys (i.e., Macaca Fascicularis), Method and Technologies Able to Identify Individuals all’Estero e quelli rimasti in Italia,” November 2011 basis of the phylogenetic similarities between monkeys transducing it in vitro with a lentivirus expressing monkey- at Risk of Developing T1D,” National Institute of • Diabete … Un’Iniezione Di Fiducia: “Combattere il and humans, it has to be considered that NHP do not insulin-specific short-hairpin RNA (shRNA) to knock Diabetes and Digestive and Kidney Diseases (NIDDK), diabete, con mezzi fisiologici,” A.G.D. Sicilia Convegno spontaneously develop autoimmune diabetes. down insulin expression, and then to return the transduced Bethesda, Md., March 2012 Socio/Scientifico, Catania, Italy, May 2012 thymus back into the donor monkey (whose mature B and • T1D TrialNet Steering Committee Meeting, Reston, • “Cellular Treatment of Autoimmunity in T1D,” Given these considerations, Massimo Trucco’s research T cells had been concomitantly depleted), to facilitate the Va., April 2012 “Microsphere-based treatment of autoimmunity in team thought it useful to find the means to promote onset of autoimmunity against pancreatic beta cells. Even • Beta-cell Consortium (BCBC) Investigator T1D,” Sanofi Advisory Board Meeting, Bridgewater, autoimmune diabetes in NHP. Toward this objective, the if the might not have transfected all the cells of the Retreat 2012, Chantilly, Va., May 2012 N.J., June 2012 team was motivated by the evidence proving that insulin thymic medulla, a drastic reduction in insulin expression/ • Sanofi T1D Mellitus Advisory Board Meeting, • “Cellular Therapy for Diabetes Treatment and expression in the thymus can regulate the negative selection presentation was expected to be sufficient to favor the Bridgewater, N.J., June 2012 Prevention,” Third Annual Sanford T1D Symposium— of auto-reactive T cells, being the first self-antigen whose development of a specific autoimmune reaction. Following • Takeda T1D Advisory Board Meeting, Philadelphia, Pa., Recent Advance in the Immunology of T1D, Sioux lack of expression in the thymus is sufficient to break central depletion of all immuno-competent cells in the thymec- June 2012 Falls, S.D., July 2012 immune tolerance toward pancreatic beta cells. To gain tomized monkey and transplantation of the transduced the functional insights of this ectopic insulin expression, thymus back to it, the team observed a successful repopula- Advisory Committee Memberships Professional Affiliations/Society Memberships the team took advantage of the Cre-lox system to knockout • Research Advisory Committee, Children’s Hospital • Italian Society of Medical tion of its immune cells (mature and functional T and B the mouse Ins2 gene specifically inAire -expressing medul- of Pittsburgh of UPMC • Italian Society of Immunology and Immunopathology lymphocytes) after roughly three months from transplan- lary thymic epithelial cells (mTECs), without affecting • Cochrane-Weber Endowed Fund Advisory Committee • American Society for Histocompatibility and tation. However, no more than ~5% of insulin reduction its production in the pancreatic beta cells. These ID-TEC • Rangos Research Center Steering Committee, Immunogenetics in the transplanted thymus was achieved, by the process (i.e., carrying insulin-deficient mTECs) mice—previously Children’s Hospital • American Association for the Advancement of Science of transducing ex vivo the sliced thymic tissue, exposed also crossed to an Ins1 knockout background—spontane- • American Diabetes Association, Inc. in vitro to the monkey insulin-specific shRNA expressing ously developed diabetes around three weeks after birth, Editorships • New York Academy of Science vector. This reduction was insufficient to break central even in a strain’s background expressing diabetes-resistant • Editor, Review of Diabetic Studies • American Society of Clinical Investigation tolerance and failed to promote clinically overt diabetes in H-2b MHC molecules. Beta cell-specific, autoimmune • Associate editor, Pediatric Diabetes • Society for Pediatric Research the recipient monkey. It was then concluded that the only destruction was observed and documented by the presence • Reviewer, Advances in Experimental Biology and Medicine • Società Italiana di Diabetologia way to achieve better transduction with the vector was in the islets of effector T cells, directed specifically against • Reviewer, Biotechniques • American Association of Immunologists to decompose the thymic tissue so to obtain a single cell insulin epitopes. Even more pertinent to the new aim were • Reviewer, Diabetologia population, which could be more efficiently exposed to the the results obtained from ID-TEC-thymus transplantation • Reviewer, Diabetes shRNA-carrying vector. The problem remained then how experiments in a nude (thymus-deprived) mouse, which • Reviewer, Diabetes Care to reconstruct the thymus in order to transplant it back proved how the Ins2-depleted thymus was sufficient, to: • Reviewer, Endocrinology to the donor monkey. In the mouse, this final problem 1) successfully reconstitute the animal’s repertoire, • Reviewer, Human Immunology was solved utilizing a process that successfully repopulates 2) break its central tolerance, and 3) induce anti-insulin • Reviewer, Journal of Autoimmunity a thymic scaffold with enriched mTECs. Transplanting autoimmunity.

DEPARTMENT OF PEDIATRICS 2012 ANNUAL REPORT DEPARTMENT OF PEDIATRICS 2012 ANNUAL REPORT division of Immunogenetics division of Immunogenetics

Suzanne Bertera, PhD Editorships The islet lab directed by Bottino is also a reference center • “Autoimmune Diabetes in Nonhuman Primates: • Editorial Board, ISRN Endocrinology for isolating human islets from deceased donors for the A Model,” Division of Immunogenetics Seminars, Research • Reviewer, purpose of distribution to international investigators carry- February 2012 Suzanne Bertera focuses her research on genetic immuno- • Reviewer, Transplant Immunology ing out diabetes research. • “The Formation of a Collaborative Consortium modulation to improve allo- and xeno-islet transplantation. • Reviewer, Diabetes to Address Islet Encapsulation,” Division of She is testing several therapeutic genes to prolong function • Reviewer, ISRN Endocrinology Editorships Immunogenetics Seminars, May 2012 of allogeneic and xenogeneic islets in an autoimmune • Managing editor, Journal of Pancreas • “Porcine Islet Xenotransplantation Using Genetically mouse model. Other projects include quality-control test- Professional Affiliations/Society Memberships • Guest editor, Anatomy Research International Engineered Islets–An Update,” Thomas E. Starzl ing of the isolated islets in vivo with immunocompromised • International Pancreas and Islet Transplant Association • Reviewer, Diabetes Transplantation Institute, Pittsburgh, Pa., May 2012 mouse models. In a related project, Bertera is studying • International Xenotransplantation Association • Reviewer, Diabetologia • “Pancreatic Islet Transplantation. Clinical Experience,” the differences in the immunologic responses of mice to • American Association of Laboratory Animal Science • Reviewer, Cell Transplantation Cell Therapy Course (A. Soto-Gutierrez), University genetically altered pig islets. She also provides isolated • American Diabetes Association • Reviewer, Transplantation of Pittsburgh, July 2012 mouse islets for the Diabetes Institute and other related • Reviewer, American Journal of Transplantation researchers. • Reviewer, Pediatric Diabetes Professional Affiliations/Society Memberships Rita Bottino, PhD • Reviewer, Endocrine • American Diabetes Association She also is investigating the biological properties of Research • Reviewer, Hepatobiliary and Pancreatic Diseases International • Cell Transplantation Society 142 C-peptide, a small peptide generated in the pancreatic beta 143 Rita Bottino is responsible for the isolation and provision • Reviewer, Xenotransplantation • Transplantation Society cells as part of the normal insulin production and secreted of pancreatic islets obtained from adult and pediatric • Reviewer, Journal of Transplantation • International Pancreas and Islet Transplant Association in the bloodstream in healthy individuals. This study patients affected by chronic pancreatitis and undergoing • Reviewer, World Journal of Gastroenterology • International Xenotransplantation Society combines C-peptide with insulin therapy in diabetic mice total or partial pancreatectomy at UPMC and the Cleve- • Reviewer, PLoS One on a high-fat diet to reduce the amount of atherosclerosis land Clinic. Following surgical removal of the pancreas, formed compared to insulin therapy alone. a necessary treatment for untreatable pain, autologous Major Lectureships and Seminars Henry Dong, PhD islets transplantation is a safe and unique way to prevent • “Pig to Monkey Islet Xenotransplantation,” grand Bertera is a member of the Islet Isolation Core team with Research or reduce the onset of surgical diabetes. This procedure rounds, Thomas E. Starzl Transplantation Institute, Henry Dong focuses on studies of the molecular basis that Rita Bottino. She participates in pancreatic islet isolation is highly specialized and is carried out in a very limited Pittsburgh, Pa., December 2011 links insulin resistance to diabetic dyslipidemia in subjects from human, porcine, and nonhuman primates (NHP) number of centers worldwide. Stemming from the experi- • “Outcome of the First of 4-5 with morbid obesity and T2D. Insulin resistance is defined donors. The human islets are distributed to JDRF-funded ence of this successful program, Bottino, in collaboration GE Pig Islets in Diabetic Monkey Recipients,” as inept responsiveness of tissues to insulin. To overcome investigators (supported by the recently concluded fund- with the Department of Surgery, is working on the imple- Transplantation Institute, Pittsburgh, Pa., insulin resistance in peripheral tissues, the pancreatic beta ing, JDRF #31-2008-381). The porcine and NHP islets mentation of a program of allogeneic islet transplantation. January 2012 cells are called upon to produce more insulin via a com- are used in basic research within and outside the division This cell therapeutic approach involves the purification of pensatory mechanism, which over time can lead to beta- as well as for primate xenotransplantation studies. Bertera viable human islets from deceased organ donors and their cell failure and overt diabetes. Despite tremendous efforts also is a member of the autotransplant islet isolation transplantation in patients affected by T1D. to elucidate the molecular basis of insulin resistance, team, which is vital for patients with chronic pancreatitis a unifying idea to account for the pathophysiology of requiring surgical removal of the pancreas. The islets are Bottino’s research interests focus on developing preclinical insulin resistance in obesity and T2D is still lacking. Using isolated from the excised pancreas and then infused back to experimental models of diabetes and islet xenogeneic transgenic, gene knockout, and gene transfer approaches, the patient as an autologous cell transplant. This is being transplantation. One of the main goals is to develop a Dong’s lab focuses on the characterization of forkhead done in collaboration with the Department of Surgery model of autoimmune diabetes, similar to human T1D, transcription factors in glucose and lipid metabolism to of the Cleveland Clinic and more recently with UPMC in Cynomolgous monkeys. The approach involves ex vivo understand how insulin resistance perturbs carbohydrate in Pittsburgh. To date, there have been more than 40 knocking down of key genes in the thymus and thymus metabolism, contributing to the development of diabetic patients, including pediatric cases, receiving this therapy, transplantation. The project stems from a successful dyslipidemia. His research team identified the forkhead and more are being scheduled. murine model developed by Massimo Trucco and Yong Fan. transcription factor FoxO1 as an important regulator of She is continuing to collaborate with several members lipid metabolism, providing a scientific rationale that tar- Genetic manipulation of pig donor tissue is also the basis of the Department of Surgery and Thomas E. Starzl geted FoxO1 inhibition by small-molecule drugs would for rendering porcine cells more compatible to humans Transplantation Institute as head of the Rodent Islet Core, improve lipid metabolism for treating diabetic dyslip- and monkeys in transplantation settings. In collaboration a subdivision of the Islet Isolation Core, providing mouse idemia. Dong published these studies in the Journal of with the Thomas E. Starzl Transplantation Institute and islets, transplanting mice with islets under the kidney Clinical Investigation, Diabetes, and Endocrinology. The Revivicor, Bottino is testing the effect of porcine islets capsule, and helping to plan these projects and analyze significance of his studies was reported byScience expressing xenogeneic molecules aimed at reducing islet the generated data. Daily, University Times (University of Pittsburgh), loss following transplantation in diabetic monkey and Scientific American. recipient. The long-term goal is to identify an appropriate porcine source of islets as an alternative to cadaveric donor islets for clinical transplantation.

DEPARTMENT OF PEDIATRICS 2012 ANNUAL REPORT DEPARTMENT OF PEDIATRICS 2012 ANNUAL REPORT division of Immunogenetics division of Immunogenetics

Dong also focuses on the development of insulin replace- • Reviewer, International Liver Aire+ medullary thymic epithelial cells (mTECs), and such and adenoviral gene vectors exhibit prolonged survival ment therapy for T1D. T1D is caused by the lack of • Reviewer, Metabolic Syndrome and Related Disorders thymic ectopic expression is essential to establish central after transplantation. Intact murine islets are transduced insulin production in the pancreas. Insulin gene therapy • Reviewer, International Journal of Obesity self-tolerance toward pancreatic beta cells. In addition, in vitro with fourth-generation lentiviral and adenoviral is being developed as an alternative insulin replacement • Reviewer, European Journal of Pharmacology the team identified a population of insulin-expressing, gene vectors encoding a variety of soluble immunoregula- therapy because it offers great potential for achieving Aire+MHCII+CD11c+ antigen-presenting cells of bone tory proteins and cytokines and transplanted into diabetic long-term blood glucose control without eliciting immune Major Lectureships and Seminars marrow origin in the , which might play essential and autoimmune diabetic allogeneic murine recipients. rejection. The insulin gene is delivered through a gene • “FoxO1 in Lipid Metabolism,” University of Alabama maintenance roles in regulating peripheral tolerance of Transplants expressing inhibitors of CD8 cytotoxic vehicle to the liver, such that insulin will be produced Birmingham, Department of Medicine, Comprehensive beta cells under defective central selection conditions. To T-cell-mediated cytolysis and cytotoxicity exhibit pro- in liver cells and released into the blood stream. His Diabetes Center, Birmingham, Ala., September 2011 further investigate the roles of Aire+, self-antigen-express- longed survival. Improvements in vector delivery as well research has provided proof-of-principle that sustained • “FoxO1 in Insulin Action and Lipid Metabolism,” ing cells in tolerance induction, Fan’s team has developed as transplant site conditioning improve the outcome of insulin production at a basal level in the liver is sufficient Cincinnati Diabetes and Obesity Center, University a number of animal models in which it was able to achieve survival. This approach will be translated to facilitate to prevent urine ketone and relieve diabetes symptoms in of Cincinnati College of Medicine, Cincinnati, Ohio, either targeted deletion of specific islet autoantigens or the transplantation and survival of porcine xenogeneic diabetic animals. To improve this procedure, his research May 2011 depletion of Aire-expressing cells. Autoimmunity affecting islets in nonhuman primates prior to human safety trials. team has designed an autoregulated system to control multiple organs and tissues was observed in these animal insulin production to achieve insulin release in a glucose- Professional Affiliations/Society Memberships models, implying that defective TSA expression in immune Molecular Mechanisms and Pathways of Immunoregulation • American Heart Association 144 dependent manner. In this autoregulated system, insulin organs might contribute to etiology of multiple autoim- in Regulatory Leukocyte Populations Induced by the Diabetes- 145 • American Diabetes Association production will be stimulated when blood sugar is high mune diseases. therapeutic Dendritic Cells and Particles. The objective of • New York Academy of Sciences and suppressed when blood sugar is low. His research is in this study is to identify immune cells with immunoregula- preclinical studies to validate this highly regulated system Editorships tory/immunosuppressive activity that are upregulated in Honors for improving blood sugar control without the need of • Editorial Board, World Journal of Biological Chemistry successfully treated diabetic recipients of the autologous • Career Development Award, American daily insulin injection in animal models of T1D. Dong has • Reviewer, Pediatrics Diabetes dendritic cells and microparticle vaccines. FACS, micro- Diabetes Association received research grants and a career development award • Reviewer, Cellular Immunology scopic, whole-body imaging, conventional biochemistry, • Patent Pending, Ref No.: 01812, “Targeting FoxO1 from the American Diabetes Association for this study. • Reviewer, PLosOne and a variety of immune assays are used to identify the for Treating Hypertriglyceridemia,” Innovator(s): leukocyte subpopulations that are upregulated as well as H. Henry Dong Study Sections Professional Affiliations/Society Memberships to ascertain the possible intracellular mechanisms by which • Ad hoc member, NIH National Institute of Diabetes and • American Diabetes Association the nucleic acids in the cell and particle vaccines activate Digestive and Kidney Diseases (NIDDK), Integrative Yong Fan, PhD immunoregulation pathways. TLR-mediated signaling Physiology of Obesity and Diabetes Study Section, 2011 appears to be involved in the mechanism by which some Research Nick Giannoukakis, PhD • Ad hoc member, NIH NIDDK, Cellular Aspects of of the nucleic acids induce suppressive capacity in dendritic The major focus of Yong Fan’s group is to understand Diabetes and Obesity Study Section, 2011 and 2012 Research cells. Successfully treated recipients of the dendritic cells the roles of tissue-specific autoantigen (TSA) expres- Cell and Particle Vaccines to Prevent T1D. The objective of and the particles exhibit increased prevalence of Foxp3+ sion in both primary and secondary lymphoid organs in Editorships this study is to develop immunoregulatory dendritic cells regulatory T cells as well as a novel population of B cells establishing and maintaining immune tolerance. The goals • Editorial Board, World Journal of Gastrointestinal and microparticle carriers of dendritic cell-modifying with immunosuppressive activity. Further identification are to develop better therapeutics for the prevention and Pathophysiology agents toward prevention and reversal of T1D. Autologous of the upregulated cell populations may offer an additional treatment of autoimmune diseases, such as T1D. • Editorial Board, Immunology, Endocrinology, dendritic cells are engineered ex vivo with that cell type(s) that can be harnessed as alternative T1D and Metabolic Agents downregulate co-stimulation. Microparticle carriers of cell therapeutics. The success of adaptive immunity depends on its capa- • Editorial Board, Journal of Geriatric Cardiology such nucleotides are used in vivo to achieve downregulation bility to effectively distinguish self from nonself and • Editorial Board, Open Endocrinology Journal of co-stimulation. Both methods successfully prevent T1D Study Sections take actions accordingly. Both central and peripheral • Reviewer, Journal of Clinical Investigation in mouse models and reverse new-onset disease. A phase • Biological and Chemical Technologies, SBIR Study tolerogenic mechanisms are essential to maintain such • Reviewer, Cell Metabolism I trial demonstrated that the dendritic cells were safe and section, National Science Foundation, 2012–sent immune tolerance. Developing with high • Reviewer, American Journal of Transplantation well tolerated, and intriguing data suggested that they may • Reviewer, NIDDK: F30, F31 and F32 NRSA affinities to autoantigens of specific tissues and organs are • Reviewer, Journal of Applied Physiology be beneficial in new-onset diabetic patients. The micropar- Fellowship Grants, 2009–present negatively selected within the thymic medulla, whereas • Reviewer, Diabetes ticles are in the preclinical phase of safety testing and are • Reviewer, NIDDK: ZDK-GRB-R-J2, K and T32 T cells escaped the central selection and are inactivated • Reviewer, Hepatology poised to surmount critical studies enabling eventual phase Awards, 2011 by various peripheral tolerogenic mechanisms, includ- • Reviewer, Diabetologia I trials. • Grant reviewer, Regular and Innovative Grants ing induced anergy, ignorance, and active suppression • Reviewer, Diabetes Care Study Section, JDFR mediated by T regulatory cells. Recent studies highlighted • Reviewer, Metabolism Replication-defective as Vehicles with Which Insulin • Grant reviewer, Regular Grants Study Section, the importance of TSA ectopic expression in the stroma • Reviewer, Proteomics Cell Transplants Can Be Engineered to Resist Immune Rejection American Diabetes Association of immune organs in regulating self-tolerance. Using • Reviewer, Trends in Endocrinology in Diabetic Recipients. The objective of this study is to • Grant reviewer, Regular Grants Study Section, gene targeting and transgenic technique, Fan’s research • Reviewer, Medical Science Monitor determine whether intact allogeneic islets of Langerhans Canadian Diabetes Association team previously demonstrated that insulin, the primary • Reviewer, Journal of Molecular Medicine engineered to express immunosuppressive soluble proteins • Grant reviewer, Regular Grants Study Section, islet autoantigen in T1D etiopathogenesis, is expressed in • Reviewer, Molecular Therapy following transduction with fourth-generation lentiviral Diabetes UK

DEPARTMENT OF PEDIATRICS 2012 ANNUAL REPORT DEPARTMENT OF PEDIATRICS 2012 ANNUAL REPORT division of Immunogenetics division of Immunogenetics

• Reviewer, Thrasher Foundation, 2010–present been involved with numerous projects. Collaborating with LAG-3, or lymphocyte activation gene 3, is upregulated on The goal will be attained through an interdisciplinary • Program reviewer, Government of Belgium, Belgian Rita Bottino, He has tested the regeneration of trans- CD4 T cells early after antigen recognition, binding with approach involving development of molecular and Science Policy Office, Interuniversity Attraction Poles planted porcine islets obtained from genetically manipu- high affinity to MHC class II on APCs to temper the clonal statistical tools to identify perturbations of expression (IAP) Phase VII lated pigs in diabetic nonhuman primates. Collaborating burst and control aberrant immune responses. For an gene networks, as well as the integration of levels of • Reviewer, Framework Consortium Projects, with Yong Fan, He has studied immunocytochemical efficient T-cell response to occur, LAG-3 must be extracel- analyses ranging from mathematics, cellular, molecular, Government of Chile (CONFICYT), 2009–present changes in different organs (thymus, spleen, lymph nodes, lularly cleaved by the redox-dependent metalloproteinase in silico and in vitro interactions, and pathway modeling • Reviewer, Hellenic Republic (Government of pancreas, and liver). These organs were from different TACE, allowing successful CD4-class II engagement and to understand molecular networks. Greece Post-graduate State Scholarships; Medicine genetic background mice. In order to demonstrate the progression toward effector function. In T1D, reducing Committee), 2011–present balance between autoimmunity and self-tolerance in LAG-3 shedding to impede CD4 T-cell responses may Professional Affiliations/Society Memberships T1D pivots on the antigenic presentation of peptides show promise in restraining autoreactive T-cell activation/ • American Association for the Advancement of Science Editorships derived from the thymus epithelial expression of a single expansion and hindering disease progression. The lab • Editor, Clinical and Developmental Immunology, tissue-specific gene, He studied the infiltration and has previously shown that a manganese metalloporphyrin William A. Rudert, MD, PhD special issue on T1D Immunological Tolerance location of lymphocytes of both CD4+ and CD8+ subsets catalytic antioxidant (MnP) can scavenge reactive oxygen and Immunotherapy, 2012 in the mouse different organs. He also is involved in the species, decrease proinflammatory cytokine production, Research • Reviewer, Gene Therapy morphological study of reconstructed thymus transplanted prevent diabetes transfer and impair TH1 cell function in William Rudert’s experience in HLA molecular typing is 146 • Reviewer, Cell Transplantation to mouse kidney. Collaborating with Suzanne Bertera, diabetogenic models. However, the mechanism of MnP- facilitating the high-throughput molecular typing to 147 • Reviewer, Immunology He has studied the immunocytochemistry on the differ- mediated TH1 hyporesponsiveness is unknown. Therefore, characterize T1D patients relative to susceptibility to the • Reviewer, Diabetes entiation of umbilical cord blood–derived stem cells into redox modulation during diabetogenic TH1 cell activation disease or its complications. • Reviewer, Molecular Therapy islet cell. Collaborating with Jon Piganelli, He is involved can decrease LAG-3 cleavage by reducing enzymatic • Reviewer, Clinical and Experimental Immunology in the study of immune reaction on islets transplanted to activity of the redox-dependent TACE and, consequently, • Reviewer, Journal of Immunology mouse kidneys. impair the transition to autoreactive CD4 T cell effector • Ad hoc reviewer, Transplantation function. Furthermore, redox-modulated LAG-3+ T cells • Ad hoc reviewer, Nature Genetics Professional Affiliations/Society Memberships may obtain a regulatory phenotype, inhibiting autoreactive • Reviewer, American Journal of Transplantation • American Diabetes Association responses and transfer of diabetes. This hypothesis is being • Reviewer, Pharmacoeconomics tested by trying to further determine the direct relationship • Reviewer, Human Gene Therapy of redox-modulated, TACE-dependent LAG-3 shedding Jon Piganelli, PhD • Reviewer, Pediatric Diabetes and TH1 effector function, and if redox-modulated • Reviewer, Journal of Leukocyte Biology Research CD4+LAG-3+ T cells can inhibit diabetic spleen and • Reviewer, Diabetes Technology and Therapeutics Jon Piganelli’s research team has identified Chromogranin diabetogenic CD4+ T cell transfer, as well as provide A as the antigen for the well-characterized diabetogenic peripheral regulation of autoimmune diabetes. Professional Affiliations/Society Memberships T cell clone BDC-2.5, which the team has studied for a • American Society of Gene Therapy number of years. Team members are now poised to assess Study Sections • American Diabetes Association the interaction of endoplasmic reticulum (ER)-stress • Grant reviewer, American Diabetes Association • European Society of Gene and Cell Therapy induced posttranslational modification of Chromogranin • Federation of American Societies for A in beta cells to determine if ER-stress facilitates the Advisory Committee Memberships Experimental Biology interaction and the exposure of this protein to the immune • Chair, Recruiting Committee for the Interdisciplinary system. Also, they are assessing whether the ER stress Biomedical Sciences Graduate Program leads to a situation of posttranslational modification of the • Representative, Interdisciplinary Biomedical Sciences Jing He, MD, PhD protein as a result of the fluctuations in calcium contact in Graduate Program, University of Pittsburgh to the Research the cytoplasm. The researchers also are expressing CHgA Annual Biomedical Research Conference for Minority Jing He’s expertise is in morphology of tissue and in prokaryotic and eukaryotic expression systems to deter- Students, Berkeley, Calif. organs. She is interested in reorganizing and analyzing mine whether PTM is necessary for antigenicity. They are three-dimensional images, using both conventional and interested in these studies as they help to better understand Steven Ringquist, PhD fluorescent confocal microscopy. Her three-dimensional the question of end organ autoimmunity and why certain image study on the mouse islet vascular structure has been organs are more prone to self-reactive recognition. Research published. He is focused on projects related to structures T1D is among the most common chronic diseases of and functions in various species islets and the morphological Work also continues on the role of redox-dependent childhood. Steven Ringquist’s research goal is to elucidate changes of NOD mouse islets, including the support of signaling in immune system activation. It is known that molecular networks affecting T1D susceptibility that are capillaries, different kinds of immune antigens, and the ectodomain shedding of key transmembrane proteins is directly influenced by inherited genetic variants. Increased communication between islet cells. He also works as an involved in regulating inflammation, a primary mechanism understanding of the gene networks underlying disease advisor on morphological and immuno-histochemical of chronic immune-mediated diseases, such as T1D. risk will aid in the development of accurate screening tools studies in the Division of Immunogenetics, and she has as well as creation of new therapeutic treatments.

DEPARTMENT OF PEDIATRICS 2012 ANNUAL REPORT DEPARTMENT OF PEDIATRICS 2012 ANNUAL REPORT division of Immunogenetics division of Immunogenetics

Rudert is assisting Yong Fan in the validation of his Tatyana Votyakova, PhD Study of Mitochondrial Functions in Development of Kidney Study of Mechanism of Oxidative Stress Related to Metabolic diabetic model based on impaired thymic expression of Complications. About 30% of diabetic patients develop Derangements of Diabetes. Increased production of reactive insulin. The consistent and rapid onset of diabetes in this Research kidney complications at some point of the disease. The aim oxygen species (ROS) in mitochondria underlies major Tatyana Votyakova’s expertise is in energy metabolism, model makes it an ideal system for studying the T cells that of this project is to investigate the role of oxidative stress systemic diseases such as diabetes and pathological side mechanisms of oxidative stress, and mitochondrial have critical roles in the disease process. Recently devel- and bioenergetic factors in the development of diabetic effects accompanying acute conditions such as tissue physiology. She utilizes her expertise working on the oped methods for massive sequencing of the mRNAs that nephropathy using a rodent model. Two diabetic mice transplantation and ischemia. On the model of isolated projects connected to islet biology and diabetes-related encode the T-cell receptors are being used to characterize strains with the same (the Akita mutation), but mitochondria, Votyakova’s team directly demonstrated that pathophysiology. the T cells present at the onset of islet destruction. with different severity of complications, are used in this after temporal hypoxia, resuming O2 supply leads to an project. The Akita mutant mice on the DBA background increase in mitochondrial ROS generation. Using combi- The Role of Mitochondrial Functions in Beta-cell Physiology A parallel diabetes model system based on reduced thymic exhibit prominent features of diabetic nephropathy, nation of experimental model of isolated mitochondria and in the Scope of a Better Preservation of Donor Islets and Their insulin expression is being developed in Cynomolgus whereas in C57BL/6 (B6) background, kidney functions computational approaches, new insights have been gained Transplantation into a Diabetic Patient. The well-being of monkeys. This is expected to be a genuine autoimmune are less changed. At a young age, both types of diabetics into mechanisms of ROS production in mitochondria, transplanted beta cells is critical to the success of blood model of diabetes in subhuman primates. If successful, demonstrate compensation in their kidney mitochondria which are related to hypoxia-induced increase of ROS glucose normalization in recipient diabetic patients. The this primate model will be a valuable proving ground for activity. With aging, B6-Akitas that did not develop and different substrate supply relevant to diabetic condi- production of insulin, its storage, and timely release to potential human therapies for T1D. The need to focus complications stay in par with their normal age-matched tions. This project was performed in collaboration with maintain glucose homeostasis is a high-energy, demanding 148 on eventual human applications has also led to a project counterparts, whereas diabetic DBA-Akita mice, which computational biologists from the University of Alabama 149 function. This notion implies an important role of to develop minicircle DNA technology as the means of develop complications, lose compensatory features of and University of Barcelona (Spain). mitochondria in beta cells. The aim of this project is to safely and permanently modifying cells able to prevent or their kidney mitochondria. Knowledge on mechanisms elucidate mitochondrial mechanisms linked to beta-cell suppress the autoimmune attack that leads to diabetes. underlying development of diabetic nephropathy will be Editorships performance in vitro and in vivo with practical issues in instrumental in finding new therapeutic approaches for • Reviewer, Metabolism mind of better islet preservation and assessment of their Rudert also is investigating the molecular mechanisms that preventive treatment in patients with diabetic kidney. • Reviewer, Current Medicinal Chemistry mediate C-peptide effects on vascular endothelial cells, potential to survive and function well in a recipient’s body. In particular, it was established that exposure of which may be protective against the vascular complications Study of Protective Mechanisms of Catalytic Antioxidant in Major Lectureships and Seminars islets to xeno plasma in vitro resulted in decrease of their of diabetes. Efforts are being made to identify a receptor Development of Autoimmune Diabetes. Votyakova is assisting • “Islets from Mitochondrial Point of View: Islet respiratory activity, indicating once again importance specific for C-peptide. Jon Piganelli and Meghan Delmastro in their investigation Respiration as a Measure of Their Well-being,” invited of mitochondrial performance in islets survival. Tatyana of protective mechanisms of catalytic antioxidant against speaker, Thomas E. Starzl Transplantation Institute, Votyakova has continued the project in collaboration with development of autoimmune diabetes in mice. The data Faculty Research Meetings, Pittsburgh, Pa., May 2012 Rita Bottino and Suzanne Bertera. have been collected, indicating involvement of mitochon- • “The Ways Mitochondria Produce Free Radicals,” drial mechanisms in this process; in particular, there is invited speaker, Mitochondria, Metabolism, and evidence that catalytic antioxidant render protection via Disease,” across-campus seminar, June 2012 modulation of respiratory activity of mice splenocyte. In this project, Votyakova helps with experimental design, Professional Affiliations/Society Memberships • American Diabetes Association practical issues, and interpretation of acquired data. • Biophysical Society

TEACHING ACTIVITIES

Medical School Lectures and Courses • Pancreatic Islet Transplantation: Basic Mechanisms, Cell Therapy Course, Department of Pathology (Trucco) • Pancreatic Islet Transplantation: Clinical Experience, Cell Therapy Course, Department of Pathology (Bottino) • Instruction for visiting scholars in rodent islet isolation techniques and murine microsurgery (Bertera) • Training and laboratory orientation of graduate and undergraduate students rotating through the laboratory in small- animal handling and surgical techniques, separation of mononucleocytes from human blood samples, and molecular techniques including DNA extraction and PCR analysis, Department of Human Genetics, Graduate School of Public Health (Bertera) • Chapter of Diabetic Dyslipidemia, Molecular Pathology Course (Dong) • Introduction to Immunobiotherapeutics, course director, Department of Immunology, School of Medicine (Giannoukakis) • and Malignancy, MSBMG 3510: Advanced Topics in Gene Expression, , and Biochemistry, School of Medicine, graduate students in the Molecular Biochemistry Program (Giannoukakis)

DEPARTMENT OF PEDIATRICS 2012 ANNUAL REPORT DEPARTMENT OF PEDIATRICS 2012 ANNUAL REPORT division of Immunogenetics division of Immunogenetics

• The Thyroid, MSCMP 2730, Department of Pathology, School of Medicine, graduate students in the Molecular THREE-YEAR BIBLIOGRAPHY Pathology Program (Giannoukakis) • Stem Cells: Endocrine Pancreas, Department of Pathology, School of Medicine (Giannoukakis) 2010 Schulte BM, Kramer M, Ansems Mavalli MD, Digirolamo DJ, Fan Y, • Neuroendocrine-immune System Interactions MSIMM 3230: Immunology and Human Disease, graduate students of M, Lanke KHW, van Doremalen N, Riddle RC, Campbell KS, van Groen Al-Masri M, Krishnamurthy M, Li J, Piganelli JD, Bottino R, Trucco T, Frank, SJ, Sperling MA, Esser KA, the Immunology Program, Department of Immunology, University of Pittsburgh School of Medicine (Giannoukakis) Bamman MM, Clemens TL. Distinct Fellows GF, Dong HH, Goodyer CG, M, Galama JMD, Adema GJ, van • Histology and immunohistochemistry background and techniques for graduate students and fellows; mentoring them Wang R. Effect of forkhead box O1 Kuppeveld FJM. Phagocytosis of growth hormone receptor signaling in microscope technology (He) (FOXO1) on beta cell development enterovirus-infected pancreatic beta- modes regulate skeletal muscle cells triggers innate immune responses development and insulin sensitivity in • Molecular pathobiology, Module 3 section leader (Piganelli) in the human fetal pancreas. Diabetologia, 53:699, 2010. in human dendritic cells. Diabetes, mice. J Clin Invest, 120(11):4007, 2010. • Introduction to Immunobiotherapeutics (Piganelli) 59:1182, 2010. Phillips BE, . Drug • Methods in Logic and Medicine (Piganelli) Casu A, Echeverri GJ, Bottino R, Giannoukakis N Sklavos, M, , Tse H, delivery technologies for autoimmune • Immunobiotherapeutics course, Department of Immunology, University of Pittsburgh School of Medicine (Votyakova) van der Windt DJ, He J, Ekser B, Ball Bertera S Bottino S, Ayares D, Cooper DKC. Insulin R, He J, Beilke J, Coulombe M, Gill disease. Expert Opin Drug Deliv, secretion and glucose metabolism in R, Crapo J, Trucco M, Piganelli JD. 7(11):1279, 2010. Research Mentorship alpha 1,3-galactosyltransferase knock- Redox modulation protects islets from • Kritika Kachapati, PhD student, Department of Rheumatology and Clinical Immunology (Trucco) out pigs compared to wild-type pigs. transplant-related injury. Diabetes, Zahid M, Phillips BE, Albers SM, , Watkins SC, Robbins • Terri C. Thayer, PhD student, Department of Immunology (Trucco) Xenotransplantation, 17:131, 2010. 59:1731, 2010. Giannoukakis N PD. Identification of a cardiac specific • Maria Grupillo, MS, research fellow, ISMETT, Italy, project title: Regulation of Immune Self-tolerance Toward protein transduction domain by 150 Coudriet GM, He J, Trucco M, Trucco M. Beta-cell regeneration: From in vivo 151 biopanning using a M13 phage peptide Pancreatic Beta Cells Via Islet Autoantigen Expression in Immune Systems (Trucco and Fan) Mars WM, Piganelli JD. Hepatocyte science fiction to challenging reality. • Antonina Coppola, MS, research fellow, University of Palermo, Italy, project title: Immune Tolerogenic Roles growth factor modulates interleukin-6 Pediatric Diabetes,11:292, 2010. display library in mice. PLoS ONE, 5(8): production in bone marrow derived e12252, 2010. of Aire-expressing Cells (Fan) macrophages: Implications for Tse HM, Thayer TC, Steele C, Cuda • Giulio Gualtierotti, MS, research fellow, University of Pittsburgh, Department of Pediatrics, Division of inflammatory mediated diseases. CM, Morel L, Piganelli JD, Mathews Batinic-Haberle I, Spasojevic I, Tse CE. NADPH oxidase-deficiency HM, Tovmasyan A, Rajic Z, Clair Immunogenetics, project title: Role of Autoimmunity against Islet Autoantigen 69 (ICA69) in T1D Development (Fan) PLoSONE, 5(11):e15384, 2010. regulates T helper lineage commitment DK, Vujaskovic Z, Dewhirst MW, • Asako Tajima-Otsubo, MD, visiting scholar, Jikei University School of Medicine, Japan, project title: Islet Autoantigen Kamagate A, Kim DH, Zhang T, Slusher and modulates autoimmunity. J Piganelli JD. Erratum to: Design of Mn porphyrins for treating oxidative 69 (ICA69) in Etiopathogenesis of T1D (Fan) S, Gramignoli R, Strom SC, Bertera S, Immunology, 185:5247, 2010. stress injuries and their redox-based • Valentina Di Caro, PhD, international research fellow (RiMed), project Ringquist S, Dong HH. Forkhead box O1 links insulin action to endoplasmic van der Windt DJ, Smetanka C, regulation of cellular transcriptional title: Interactions of Foxp3 with Gene Promoters Driving Interleukin Macedo C, He J, , reticulum stress. Endocrinology, Lakomy R Bottino activities. Amino Acids, December 24, Expression (Trucco and Giannoukakis) 151:3521, 2010. R, Ekser B, Echeverri GH, Metes D, 2010. [Epub ahead of print] • Xuehui Geng, PhD, research associate, project title: Ijzermans JNM, Trucco M, Cooper DKC, Lakkis FG. Investigation of Chen J, Gusdon AM, Piganelli Characterization of C-peptide Receptor (Rudert) Morran MP, Casu A, Arena VC, Pietropaolo S, Zhang YJ, Satin LS, lymphocyte depletion and repopulation JD, Leiter EH, Mathews CE. mt- • Kristia Hamilton, MSc candidate, rotation student (Giannoukakis) Nd2(a) modifies resistance against Trucco M, Becker DJ, Pietropaolo using alemtuzumab (Campath-1H) • Angela Pardee, Department of Immunology, PhD Thesis M. Humoral autoimmunity against in cynomolgous monkeys. Am J autoimmune type 1 diabetes in NOD , 10:773, 2010. mice at the level of the pancreatic -cell. Committee member (Giannoukakis) the extracellular domain of the Transplantation neuroendocrine autoantigen IA-2 Diabetes, 60(1):355-9, 2011. Epub • Meghan Delmastro, Department of Immunology, heightens the risk of type 1 diabetes. Wu J, Devlin B, Ringquist S, Trucco October 27, 2010. PhD Thesis Committee member Endocrinology, 151:2528, 2010. M, Roeder K. Screen and clean: A tool for identifying interactions in - Tse HM, Thayer TC, Steele C, Cuda CM, (Giannoukakis) Perdomo G, Kim DH, Zhang T, Qu S, wide association studies. Genetic Morel L, Piganelli JD, Mathews CE. • Meghan Delmastro, Department Thomas EA, Toledo FG, Slusher S, Epidemiology, 34:275, 2010. NADPH oxidase deficiency regulates of Immunology, PhD Thesis Fan Y, Kelley DE, Dong HH. A role Th lineage commitment and modulates Crossett A, Kent BP, Klei L, Committee chair (Piganelli) of apolipoprotein D in triglyceride Ringquist autoimmunity. J Immunol, 185(9):5247- , , Roeder K, Devlin B. metabolism. J Lipid Res, 51:1298, 2010. S Trucco M 58, 2010. Epub September 29, 2010. • Gaia Bellone, Thesis Committee, Using ancestry matching to combine Department of Statistics, Carnegie Potter KJ, Abedini A, Marek P, Klimek family-based and unrelated samples Batinic-Haberle I, Spasojevic I, Tse Mellon University (Trucco) AM, Butterworth MP, Driscoll M, for genome-wide association HM, Tovmasyan A, Rajic Z, St Clair DK, Baker R, Nilsson MR, Warnock GL, studies. Statistics in Medicine, 2010. Vujaskovic Z, Dewhirst MW, Piganelli • Rachel Tobin, summer Oberholzer J, Bertera S, Trucco M, doi:10.1002/sim.4057 JD. Design of Mn porphyrins for student (Bottino) Korbutt GS, Fraser PE, Raleigh DP, treating oxidative stress injuries and • Richard Peluso, summer Verchere CB. Islet amyloid deposition Coudriet GM, He J, Trucco M, their redox-based regulation of cellular limits the viability of human islet grafts Mars WM, Piganelli JD. Hepatocyte transcriptional activities. Amino Acids, student (Piganelli) growth factor modulates interleukin-6 but not porcine islet grafts. PNAS, May 16, 2010. [Epub ahead of print] • Adam Attaar, summer 107:4305, 2010. production in bone marrow derived student (Votyakova) macrophages: Implications for inflam- Stadinski BD, Delong T, Reisdorph N, matory mediated diseases. PLoS ONE, Reisdorph R, Powell RL, Armstrong 5(11): e15384, 2010. M, Piganelli JD, Barbour G, Bradley B, Crawford F, Marrack P, Mahata SK, Lu C, Kumar PA, Fan Y, Sperling MA, Kappler JW, Haskins K. Chromogranin Menon R. A novel effect of GH on A is an autoantigen in type 1 diabetes. macrophage modulates macrophage- Nat Immunol, 11(3):225-31, 2010. Epub dependent adipocyte differentiation. February 7, 2010. Endocrinology, 151(5): 2189, 2010.

DEPARTMENT OF PEDIATRICS 2012 ANNUAL REPORT DEPARTMENT OF PEDIATRICS 2012 ANNUAL REPORT division of Immunogenetics division of Immunogenetics

Thayer TC, Delano M, Liu C, Chen Wherrett DK, Bundy B, Becker DJ, Giannoukakis N, Phillips B, Finegold Ekser B, Ezzelarab M, Hara H, van Cifarelli V, Lee S, Kim DH, Zhang T, 2011 J, Padgett LE, Tse HM, Annamali M, DiMeglio LA, Gitelman SE, Goland R, D, Harnaha J, Trucco M. A phase I der Windt DJ, Wijkstrom M, Bottino Kamagate A, Slusher S, Bertera S, Gottlieb PA, Greenbaum CJ, Herold , Moldawer LL, Mathews Luppi P, , . FOXO1 Crossett A, Kent BP, Klei L, Ringquist Piganelli JD (safety) study of autologous tolerogenic R, Trucco M, Cooper DK. Clinical Trucco M Dong HH KC, Marks JB, Monzavi R, Moran CE. Superoxide production by macro- mediates the autocrine effect of S, Trucco M, Roeder K, Devlin B. dendritic cells in type 1 diabetic xenotransplantation–The next great Using ancestry matching to combine A, Orban T, Palmer JP, Raskin P, phages and t cells is critical for the patients. Diabetes Care, 34:2026, 2011. medical revolution? Lancet, 379: endothelin-1 on endothelial cell survival. family-based and unrelated samples Rodriguez H, Schatz D, Wilson DM, induction of autoreactivity and type 1 672, 2012. Mol Endocrinol, 26:1213, 2012. for genome-wide association studies. Krischer JP, Skyler JS, Type 1 Diabetes diabetes. Diabetes, 60:2144, 2011. Marigliano M, Bertera S, Grupillo TrialNet GAD Study Group (including , , Grupillo M, , . A role Statistics in Medicine, 29:2932, 2011. M, Trucco M, Bottino R. Pig-to- Bottino R Bertera S Giannoukakis N Trucco M Trucco M). Antigen-based therapy with Sheng H, Spasojevic I, Tse HM, nonhuman primates pancreatic islet Melvin PR, Humar A, Mazariegos G, for tolerogenic dendritic cell-induced TRIGR Study Group (Akerblom HK, glutamic acid decarboxylase (GAD) Jung JY, Hong J, Zhang Z, Piganelli xenotransplantation: Overview. Curr Moser J, Walsh M, Fung J, Gelrud B-regulatory cells in type 1 diabetes Krischer J, Virtanen SM, Berseth C, vaccine in patients with recent-onset JD, Batinic-Haberle I, Warner DS. Diab Rep, 11:402, 2011. A, Slivka A, Soltys K, Wijkstrom, mellitus. Curr Opin Endocrinol Neuroprotective efficacy from , 19:279, 2012. Becker D, Dupre J, Ilonen J, Trucco type 1 diabetes: A randomized double- Trucco M. Isolation of human islets Diabetes a lipophilic redox-modulating M, Savilahti E, Koski K, Pajakkala E, blind trial. Lancet, 378:319, 2011. Phillips BE, Trucco M. Immunotherapy- for autologous islet transplantation in Franciscus M, Lough G, Bradley B, mnporphyrin, mntnhex-2-pyp: based strategies for the treatment children and adolescents with chronic Goodyer WR, Gu X, Liu Y, Bottino R, Koski M, Knip M). The trial to reduce Orban T, Bundy B, Becker DJ, Dimeglio Rodent models of ischemic stroke of autoimmune diabetes. Current pancreatitis. J Transplantation, 2012. Crabtree GR, Kim SK. Neonatal-cell IDDM in the genetically at risk (TRIGR) LA, Gitelman SE, Goland R, Gottlieb and subarachnoid hemorrhage. J Pharmaceutical Design, 17:3217, 2011. doi:10.1155/2012/642787, 2012 development in mice and humans is study: Recruitment, intervention and PA, Greenbaum CJ, Marks JB, Monzavi Pharmacol Exp Ther, 338:906, 2011. regulated by calcineurin/NFAT. Dev R, Moran A, Raskin P, Rodriguez H, Grupillo M, Lakomy R, Geng X, Styche Grupillo M, Gualtierotti G, He J, , 23(1):21-34, 2012. follow-up. Diabetologia, 54(3):627, 2011. Cell Russell WE: Schatz D, Wherrett D, Delmastro MM, Piganelli JD. Oxidative A, Rudert WA, Trucco M, Fan Y. An Sisino G, Bottino R, Rudert WA, Di Caro V, D’Anneo A, Phillips B, Wilson DM, Krischer JP, Skyler JS, stress and redox modulation potential improved intracellular staining protocol Trucco M, Fan Y. Essential roles of Walsh RM, Aguilar Saavedra JR, Lentz 152 153 Engman C, Harnaha J, Lakomy R, Type 1 Diabetes TrialNet Abatacept in type 1 diabetes. Clin Dev Immunol, for efficient detection of nuclear insulin expression in Aire+ tolerogenic G, Guerron AD, Schemen J, Stevens 2011:593863, 2011. Epub May 18, 2011. T, , , Hatipoglu B. Styche A, Trucco M, Giannoukakis Study Group (including Trucco proteins in YFP-expressing cells. dendritic cells in maintaining peripheral Trucco M Bottino R Improved quality of life following after N. IL-7 matures suppressive CD127+ M). Costimulation modulation with BioTechniques, 51:417, 2011. self-tolerance of islet b-cells. Cellular Manni ML, Tomai LP, Norris CA, Foxp3+ T cells into CD127-CD25HIGH abatacept in patients with recent-onset Immunology, 273:115, 2012. total pancreatectomy and autoislet Thomas LM, Kelley EE, Salter RD, Achkar J-P, Klei L, de Bakker PIW, transplantation for chronic pancreatitis. Foxp3+ Tregs. Clin Exp Immunol, type 1 diabetes: A randomized, double- Crapo JD, Chang LY, Watkins SC, March 7, 2011. doi:10.1111/j.1365- blind, placebo-controlled trial. Lancet, Bellone G, Rebert N, Scott R, Lu Y, Delmastro MM, Styche AJ, Trucco M, J Gastrointest Surg,16:1469, 2012. , Oury TD. Extracellular 2249.2011.04334.x 378:412, 2011. Piganelli JD Regueiro M, Brzezinski A, Kamboh Workman CJ, Vignali DAA, Piganelli superoxide dismutase in macro- MI, Fiocchi C, Devlin B, Trucco M, J. Modulation of redox balance leaves Mao W, You T, Ye B, Li X, Dong HH, phages augments bacterial killing by Hill JA, Li F, Xu H. Reactive oxygen Cifarelli V, Trucco M, Luppi P. Anti- Cifarelli V, Geng X, Styche A, Lakomy Ringquist S, Roeder K, Duerr RH. murine diabetogenic TH1 T cells ‘LAG- inflammatory effects of C-peptide R, Trucco M, Luppi P. C-peptide promoting phagocytosis. Am J Pathol, Amino acid position 11 of HLA-DRb1 3-ing’ behind. Diabetes, 61:1760, 2012. species suppress cardiac NaV1.5 prevent endothelial dysfunction in type reduces high glucose-induced 178(6):2752, 2011. is a major determinant of expression through Foxo1. PLoS One, Bonner SM, Pietropaolo SL, Fan Y, 2012;7(2):e32738. Epub 2012 Feb 29. 1 diabetes. Immun Endoc and Metab apoptosis of endothelial cells 6p association with ulcerative colitis. Ramsgaard L, Englert JM, Manni ML, Chang Y, Sethupathy P, Morran MP, PMID: 22400069 Agents in Med Chem, 11:59, 2011. and decreases NAD(P)H-oxidase Genes and Immunity, 2011. doi: reactive oxygen species generation. Milutinovic PS, Gefter J, Tobolewski 10.1038/gene.2011.79 [Advance Beems M, Giannoukakis N, Trucco G, J, Crum L, Coudriet GM, , Sklavos MM, Coudriet GM, Delmastro Selivanov VA, Votyakova TV, Diabetologia, 2011. doi: 10.1007/ Piganelli JD online publication] Palumbo MO, Solimena M, Pugliese Pivtoraiko V, Zeak JA, Sukhomlin s00125-011-2251-0 Zamora R, Vodovotz Y, Enghild JJ, Oury A, Polychronakos C, Trucco M, M, Bertera S, Coneybeer JT, He J, TD. Lack of the receptor for advanced , . Administration T, Trucco M, Roca J, Cascante M. Peskovitz MD, Torgerson TR, Ochs Pietropaolo M. Sequence variation in Trucco M Piganelli JD Reactive oxygen species production Kim DH, Zhang T, Ringquist S, glycation end-products attenuates e. HD, Ocheltree E, McGee P, Krause- promoter of Ica1 gene, which encodes of a negative vaccination induces hypo- by forward and reverse electron fluxes Dong HH. Targeting FoxO1 for coli pneumonia in mice. PLoS One, Steinrauf H, Lachin JM, Canniff J, protein implicated in type 1 diabetes, responsiveness to islet allografts. Cell in the mitochondrial respiratory chain. hypertriglyceridemia. Curr Drug 6(5):e20132, 2011. Epub May 23, 2011. Greenbaum C, Herold KC, Skyler JS, causes transcription factor autoimmune Transplantation, 2012. PLoS Computational Biology, 7(3): Targets, 12(9):1245, 2011. Weinberg A, Type 1 Diabetes TrialNet regulator (AIRE) to increase its binding Hager EJ, , Tse HM, van der Windt DJ, , Kumar e1001115, 2011. Piganelli JD Study Group. Effect of rituximab and down-regulate expression. JBC, Bottino R Yang S, Xu H, Yu S, Cao H, Fan J, Ge Gibson KM. Aberrant expression of on human in vivo immune 287:17882, 2012. G, Wijkstrom M, Hara H, Ezzelarab M, costimulatory molecules in splenocytes Ekser B, Phelps C, Murase N, Casu A, Stefan M, Simmons RA, Bertera S, C, Fransceschi RT, Dong HH, Xiao responses. J Allergy Clin Immunol, of the mevalonate kinase-deficient Ayares D, Lakkis FG, , Cooper Trucco M, Esni F, Drain P, Nicholls G. Foxo1 mediates insulin-like growth 128:1295, 2011. van der Windt DJ, Marigliano M, Trucco M RD. Global deficits in development, factor 1 (IGF1)/insulin regulation of mouse model of human hyper-IgD He J, Votyakova TV, Echeverri GJ, DKC. Clinical islet xenotransplantation– function, and gene expression osteocalcin expression by antagonizing syndrome (HIDS). J Inherit Metab Dis, Kim DH, Perdomo G, Zhang T, Ekser B, Ayares D, Lakkis FG, Cooper How close are we? Diabetes, 2012. in the endocrine pancreas in a Runx2 in osteoblasts. J Biol Chem, May 24, 2011. [Epub ahead of print] Slusher S, Lee S, Phillip BE, Fan Y, DKC, Trucco M, Bottino R. Early , Dendritic deletion mouse model of Prader-Willi 286 ( 21):19149, 2011. Giannoukakis N, Gramignoli R, Strom islet damage after direct exposure Giannoukakis N Trucco M. Di Caro V, D’Anneo A, Phillips B, cell therapy for type 1 diabetes syndrome. Am J Physiol Endocrinol S, Ringquist S, Dong HH. FoxO6 of pig islets to blood–Has humoral Lo CM, Obici S, , Haas M, Engman C, Harnaha J, , suppression. Immunotherapy, 2012. Metab, 300:E909-22, 2011. Dong HH Trucco M integrates insulin signaling immunity been underestimated? Cell Lou D, Kim DH, Liu M, D’Alessio D, Giannoukakis N. Phosphatidylinositol- with gluconeogenesis in the liver. Transplantation, 2012. doi:10.3727/ 3-kinase activity during Selivanov VA, Cascante M, Friedman M, Marigliano M, Casu A, Bertera S, Woods SC, Tso P. Impaired insulin in vitro Diabetes, 60:2763-74, 2011. 096368912X6530 [Advance online dendritic cell generation determines Schumaker MF, Trucco M, Votyakova Trucco M, Bottino R. Hemoglobin secretion and enhanced insulin publication] . Multistationary and oscillatory A1C percentage in nonhuman sensitivity in cholecystokinin-deficient suppressive or stimulatory capacity. TV modes of free radicals generation by primates: A useful tool to monitor mice. Diabetes, 60(7):2000, 2011. Immunol Res, 50:130, 2011. Schulte BM, Lanke KHW, Piganelli JD, 2012 the mitochondrial resperiatory chain diabetes before and after porcine Kers-Rebel ED, Bottino R, Trucco M, Zheng Y, Wen Y, George AM, Steinbach Lin M, Tse HM, Delmastro MM, Bertera Huijbens RJF, Radstake TRDJ, Galama revealed by a bifurcation analysis. PLoS pancreatic islet xenotransplantation. Haidet J, Cifarelli V, Trucco M, Luppi AM, Phillips BE, , S, Wong CT, Lakomy R, He J, Sklavos Computational Biology, 2012. 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DEPARTMENT OF PEDIATRICS 2012 ANNUAL REPORT DEPARTMENT OF PEDIATRICS 2012 ANNUAL REPORT