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Principal Investigator: GEORGE K. GITTES
Organization: UNIVERSITY OF PITTSBURGH AT PITTSBURGH
Fiscal Year: 2019
Award: $374,150
Funding agency: National Institute of Diabetes and Digestive and Kidney Diseases
PROJECT SUMMARY AND RELEVANCE
An ideal solution to the treatment or cure of diabetes mellitus would be the formation of new functioning
β-cells from the patient’s own tissues, thereby avoiding the need for transplant immunosuppression.
Abundant recent data has suggested that α-cells are a likely source for endogenous transdifferentiation
into β-cells. Here, we describe a pancreatic intraductal viral delivery system in the mouse, where a single
infusion of an adeno-associated virus (AAV) carrying a pdx1/mafA expression vector in a diabetic mouse
can induce robust and durable α-cell transdifferentiation into β-cells through neogenesis, with recovery of
over 60% of the β-cell mass within 4 weeks and persistent, indefinite euglycemia. Serendipitously, when
this β-cell neogenesis was induced in NOD mice, the mice became euglycemic for 4 months or more,
without any additional therapy or immunosuppression. To our knowledge no clinically applicable β-cell
replacement therapy in NOD mice has been successful without immunosuppression. We suspect that the
neogenic β-cells may not be rejected because they are “imperfect” β-cells by RNA-seq analysis. Since
pancreatic duct injection is routinely performed in humans as a relatively simple, non-surgical procedure,
and since numerous viral gene therapy trials are currently ongoing for several diseases, we feel that our
approach may be rapidly translatable to humans with diabetes mellitus, potentially both type 1 and type 2.
In this proposal we will first better delineate the phenotype of these neogenic mouse β-cells derived from
α-cells in terms of function and resistance to stressors that normally can cause β-cell death. We will then
strive to better understand how they form, their proliferative capacity, RNA expression and gene
methylation profile. We will then study ways to optimize their formation through promoter work for the
virus construct, and to better understand ways that anti-AAV neutralizing antibodies may affect the
effectiveness of this gene therapy approach. Next, we will pursue the feasibility of a novel lipid
nanoparticle technology to replace the need for AAV in the induction of α-to-β-cell transdifferentiation.
Since glucagon has been shown to play an important role in α-to-β-cell transdifferentiation, we will study
its role in this system. We will study the potential therapeutic effect of such α-to-β-cell
transdifferentiation in models of type 2 diabetes mellitus. Lastly, we will investigate the function of
human islets that have undergone α-to-β-cell transdifferentiation, both in vitro and in vivo. In summary,
we feel that the proposed studies, if successful, should position us well in preparation for clinical trials in
humans with diabetes.
Terms: <Adeno-Associated Viruses><Adult-Onset Diabetes Mellitus><Affect><Alloxan><Alpha Cell><Antidiabetic Hormone><Apoptosis><Apoptosis Pathway><Assay><Autoimmune><Autoimmune Process><Beta Cell><Bioassay><Biologic Assays><Biological Assay><Biology><Blood Glucose><Blood Sugar><Body Tissues><Brittle Diabetes Mellitus><Capsules><Cell Body><Cell Communication and Signaling><Cell Death><Cell Signaling><Cells><Clinical Trials><D-Glucose><DNA Methylation><DNA Therapy><Data><Dependoparvovirus><Dependovirus><Dextrose><Diabetes Mellitus><Diabetic mouse><Disease><Disorder><Duct><Duct (organ) structure><Ductal><Dysfunction><ER stress><Effectiveness><Family><Functional disorder><Gene Transcription><Gene Transfer Clinical><Gene therapy trial><General Viruses><Genes><Genetic><Genetic Intervention><Genetic Transcription><Glare><Glucagon><Glucagon Cell><Glucagon Secreting Cell><Glucose><Glukagon><Goals><HG-Factor><Human><Hyperglycemia><Hyperglycemic-Glycogenolytic Factor><IDDM><Immunosuppressants><Immunosuppression><Immunosuppression Effect><Immunosuppressive Agents><Immunosuppressive Effect><In Situ><In Vitro><Inbred NOD Mice><Infusion><Infusion procedures><Injections><Insulin Cell><Insulin Secreting Cell><Insulin-Dependent Diabetes Mellitus><Intervention><Intervention Strategies><Intracellular Communication and Signaling><Islands of Langerhans Transplantation><Islands of Pancreas Transplantation><Islets of Langerhans Grafting><Islets of Langerhans Transplantation><Juvenile-Onset Diabetes Mellitus><Ketosis-Prone Diabetes Mellitus><Ketosis-Resistant Diabetes Mellitus><Kidney><Kidney Urinary System><Location><Maturity-Onset Diabetes Mellitus><Metabolic><Methylation><Mice><Mice Mammals><Modeling><Modern Man><Murine><Mus><NIDDM><NOD Mouse><Non-Insulin Dependent Diabetes><Non-Insulin-Dependent Diabetes Mellitus><Non-Obese Diabetic Mice><Noninsulin Dependent Diabetes><Noninsulin Dependent Diabetes Mellitus><Nonobese Diabetic Mouse><Pancreas><Pancreatic><Pancreatic Islets Transplantation><Pancreatic duct><Patients><Phenotype><Physiopathology><Play><Position><Positioning Attribute><Preparation><Procedures><Process><Programmed Cell Death><RNA Expression><RNA Seq><RNA sequencing><RNAseq><Recovery><Recurrence><Recurrent><Replacement Therapy><Resistance><Role><SCID Mice><Severe Combined Immunodeficient Mice><Signal Transduction><Signal Transduction Systems><Signaling><Slow-Onset Diabetes Mellitus><Source><Stable Diabetes Mellitus><Stress><Study Section><Sudden-Onset Diabetes Mellitus><System><T1 DM><T1 diabetes><T1D><T1DM><T2 DM><T2D><T2DM><Technology><Therapeutic><Therapeutic Effect><Tissues><Transcription><Transgenic Mice><Transplantation><Type 1 Diabetes Mellitus><Type 1 diabetes><Type 2 Diabetes Mellitus><Type 2 diabetes><Type I Diabetes Mellitus><Type II Diabetes Mellitus><Type II diabetes><Universities><Viral><Viral Genes><Virus><Wirsung canal><Work><adeno associated virus group><adult onset diabetes><beta cell replacement><biological signal transduction><capsule><cell transformation><clinical applicability><clinical application><cytokine><diabetes><diabetes mouse model><diabetic><endoplasmic reticulum stress><euglycemia><experience><experiment><experimental research><experimental study><expression vector><gene delivery system><gene therapy><gene transfer trial><gene-based therapy><genetic therapy><genomic therapy><hyperglycemic><immune suppression><immunosuppressive><in vivo><insulin dependent diabetes><interventional strategy><islet><islet beta cell transplantation><islet cell transplant><islet cell transplantation><islet transplantation><juvenile diabetes><juvenile diabetes mellitus><ketosis prone diabetes><ketosis resistant diabetes><lipid nanoparticle><maturity onset diabetes><necrocytosis><neutralizing antibody><non-obese diabetic (NOD) mice><non-viral gene delivery><nonobese diabetic (NOD) mice><nonviral gene delivery><novel><pathophysiology><process optimization><promoter><promotor><renal><resistant><social role><stressor><theories><transcriptome sequencing><transdifferentiation><transformed cells><transplant><type 2 DM><type I diabetes><type II DM><type one diabetes><type two diabetes><α-cell><β-Cell replacement><β-cell><β-cells><βCell>