Joslin Diabetes Center Inc

DIABETES AND ENDOCRINOLOGY RESEARCH CENTER

PERL: A MULTICENTER CLINICAL TRIAL OF ALLOPURINOL TO PREVENT GFR LOSS IN T1D

TRAINING GRANT IN DIABETES AND METABOLISM

DEVELOPMENTAL GENES AND THE ORIGIN OF FAT

DIABETES CARE AND TREATMENT PROJECT

INSULIN RECEPTOR STRUCTURE AND TURNOVER

EXERCISE REGULATION OF GLUCOSE HOMEOSTASIS ADMINISTRATIVE SUPPLEMENT

ROLE OF IGF-1 AND INSULIN RECEPTORS BETA-CELL SURVIVAL

NOVEL MECHANISMS FOR EXERCISE TRAINING EFFECTS ON GLUCOSE HOMEOSTASIS

INSULIN RECEPTOR SUBSTRATES AND INSULIN ACTION

ROLE OF BMPS IN ADIPOGENESIS, MITOCHONDRIAL FUNCTION AND ENERGY HOMEOSTASIS

CELLULAR MECHANISMS THAT PROMOTE OR PREVENT LIPOTOXICITY

PATHWAYS TO BETTER HEALTH THROUGH A NEW HEALTH CARE WORKFORCE AND COMMUNITY ENGAGEMENT

TARGETING INFLAMMATION IN TYPE 2 DIABETES: CLINICAL TRIAL USING SALSALATE

PI 3-KINASE ISOFORMS IN INSULIN ACTION

POST-DPP FOLLOW-UP STUDY

SIGNALING MECHANISMS THAT DETECT STRESS AND MAINTAIN HOMEOSTASIS

MECHANISM OF ENDOCYTIC TRAFFICKING OF CHOLESTEROL

PATERNAL CONTRIBUTIONS TO METABOLIC DISEASE IN OFFSPRING: ENVIRONMENT, EPIGENETICS, AND SPERM

NATURAL HISTORY OF MICROALBUMINURIA IN TYPE 1 DIABETES

RIBOSOME HETEROGENEITY AS A MECHANISM FOR METABOLIC REGULATION

INSULIN RECEPTOR STRUCTURE AND TURNOVER

FIBROBLAST GROWTH FACTOR AND ENERGY METABOLISM

VALIDATION OF POTENTIAL PROTECTIVE FACTORS FROM DIABETIC COMPLICATIONS

EARLY MYOCARDIAL REMODELING AND PROGRESSIVE KIDNEY FUNCTION DECLINE IN TYPE 1 DIABETES - SUMMARY A LARGE PROPORTION OF THE EXCESS CVD MORBIDITY AND MORTALITY EXPERIENCED BY INDIVIDUALS WITH T1D OCCUR IN CONJUNCTION WITH DIABETIC KIDNEY DISEASE (DKD), WHICH IS ASSOCIATED WITH A STRIKING INCREASE IN THE RISK OF CORONARY ARTERY DISEASE (CAD) AND HEART FAILURE. THE LATTER IS FREQUENTLY DUE TO THE DEVELOPMENT OF DIABETIC CARDIOMYOPATHY – A DIABETES-SPECIFIC ALTERATION OF THE MYOCARDIUM. THE ETIOLOGIC LINKS BETWEEN DKD AND CARDIOMYOPATHY ARE NOT CLEAR, BUT PRELIMINARY DATA FROM OUR GROUP SUGGEST A PIVOTAL ROLE OF THE KIDNEY FUNCTION DECLINE COMPONENT OF DKD RATHER THAN ALBUMINURIA. SPECIFICALLY, USING AN MRI-DERIVED MARKER OF CARDIOMYOCYTE SIZE, WE HAVE OBSERVED THAT PATIENTS WITH T1D WHO ARE LOSING KIDNEY FUNCTION BUT STILL HAVE PRESERVED GFR HAVE SUBCLINICAL SIGNS OF MYOCARDIAL REMODELING, AS INDICATED BY A LARGER CARDIOMYOCYTE SIZE AND A REDUCTION OF MYOCARDIAL FIBER SHORTENING DURING SYSTOLE AS COMPARED TO T1D PATIENTS WITH STABLE KIDNEY FUNCTION. THE OVERALL GOAL OF THIS COLLABORATIVE PROPOSAL, WHICH IS IN RESPONSE TO RFA-HL-21-014, IS TO TAKE ADVANTAGE OF THE LATEST DEVELOPMENTS IN CARDIAC IMAGING AND BIOMARKER PLATFORMS TO CHARACTERIZE THE CARDIAC INVOLVEMENT IN PATIENTS WITH T1D AND DKD, FOCUSING ON THE INITIAL EVENTS IN THE DEVELOPMENT OF DIABETIC CARDIOMYOPATHY. “GFR DECLINERS” (GFR LOSS IN THE PREVIOUS 3-6 YEARS =3 ML/MIN/YEAR, N=100) AND “GFR NON- DECLINERS” (N=100) WITH T1D AND CKD STAGE 1-3A, ALONG WITH NON-DIABETIC CONTROLS (N=100) OF SIMILAR AGE AND CKD STAGE, WILL UNDERGO A GADOLINIUM-ENHANCED CARDIAC MAGNETIC RESONANCE (CMR) AND A GATED CARDIAC CT SCAN TO QUANTIFY CORONARY ARTERY CALCIUM (CAC). THROUGH THESE STUDIES, WE WILL ADDRESS THE FOLLOWING SPECIFIC AIMS: 1. TO EVALUATE THE PRESENCE AND SEVERITY OF MYOCARDIAL REMODELING AMONG T1D PATIENTS AND ASSESS ITS RELATIONSHIP WITH EARLY PROGRESSIVE KIDNEY FUNCTION DECLINE. CARDIOMYOCYTE SIZE (TIC) AND INTERSTITIAL FIBROSIS (MEASURED AS EXTRACELLULAR VOLUME [ECV]) WILL BE QUANTIFIED BY CMR AND COMPARED AMONG GFR DECLINERS, GFR NON-DECLINERS, AND NON-DIABETIC SUBJECTS, AND ALSO RELATED TO ALBUMINURIA AND PRESENCE AND SEVERITY OF CAD. 2. TO ASSESS THE RELATIVE CONTRIBUTION OF CARDIOMYOCYTE HYPERTROPHY AND INTERSTITIAL FIBROSIS TO IMPAIRED CARDIAC FUNCTION AMONG T1D PATIENTS. INDICES OF CARDIAC FUNCTION AND MYOCARDIAL STRAIN WILL BE DERIVED FROM THE CMR DATA AND EVALUATED FOR THEIR ASSOCIATION WITH CARDIOMYOCYTE SIZE (TIC) AND INTERSTITIAL FIBROSIS (ECV), IN RELATION TO THE SEVERITY OF CONCOMITANT CAD. 3. TO GAIN INSIGHTS INTO THE DISEASE PROCESSES INVOLVED IN THE ETIOLOGY OF MYOCARDIAL REMODELING AND ASSESS WHETHER THESE OVERLAP WITH THOSE INVOLVED IN THE PROGRESSIVE KIDNEY FUNCTION DECLINE. IN TARGETED STUDIES, WE WILL FOCUS ON SERUM PROTEINS IMPLICATED IN HEART FAILURE OR PREVIOUSLY ASSOCIATED WITH INCREASED RISK OF GFR LOSS. IN UNTARGETED STUDIES, WE WILL LEVERAGE THE LATEST DEVELOPMENTS IN MULTIPLEXED ASSAYS TO EVALUATE SERUM PROTEIN PROFILES IN A SYSTEMATIC FASHION. WITH THE INFORMATION GENERATED BY THIS STUDY ON HAND, WE WILL BE OPTIMALLY POSITIONED TO DEVELOP NEW STRATEGIES AND POSSIBLY NEW DRUGS TO PREVENT CVD IN T1D.

EPITRANSCRIPTOMICS IN HUMAN OBESITY AND TYPE 2 DIABETES - PROJECT SUMMARY DYSFUNCTION OF MULTIPLE METABOLIC CELLS CONTRIBUTE TO THE PATHOPHYSIOLOGY OF THE TWO MAJOR PATHOLOGICAL STATES, OBESITY AND TYPE 2 DIABETES, WHICH HAVE A HUGE ECONOMIC BURDEN WORLDWIDE. IDENTIFYING THE ETIOLOGY OF THE TWO LINKED PATHOLOGICAL STATES CONTINUES TO BE AN AREA OF INTENSE RESEARCH WITH A FOCUS ON IDENTIFYING NEW REGULATORY PATHWAYS WITHIN TISSUES THAT IMPACT METABOLIC FUNCTION AND CAUSE DYSFUNCTION. THERE IS A CONTINUING NEED TO DEVELOP A SYSTEMS BIOLOGY FRAMEWORK THAT INTEGRATES REAL PATIENT DATA TO INFORM SIGNALING WITHIN CELLS AND CROSSTALK BETWEEN TISSUES TO ENABLE IDENTIFICATION OF POTENTIAL NEW TARGETS FOR EFFECTIVE THERAPY. AN EMERGING AREA OF SIGNIFICANCE IN HUMAN OBESITY AND TYPE 2 DIABETES (T2D) IS EPIGENETICS THAT HAS PRIMARILY FOCUSED ON DNA METHYLATION AND PROTEIN MODIFICATIONS. A MISSING LINK IN THIS EPIGENETIC FRAMEWORK IS THE MODIFICATION(S) IN MRNA AND CHROMATIN-ASSOCIATED REGULATORY RNAS (CARRNAS) THAT HAVE BEEN SHOWN TO CONTRIBUTE TO GENE EXPRESSION REGULATION AT BOTH POSTTRANSCRIPTIONAL AND TRANSCRIPTIONAL LEVELS. CURRENT DATA INDICATE ALTERED EXPRESSION OF VARIOUS REGULATORS IN METABOLIC TISSUES (E.G. HUMAN ISLETS, ADIPOSE, LIVER, SKELETAL MUSCLE) IN OBESITY AND TYPE 2 DIABETES; HOWEVER, THE SIGNIFICANCE OF THESE ALTERATIONS AND THE IMPACT ON MRNA AND PROTEIN EXPRESSION IN REAL HUMAN TISSUES IS VIRTUALLY UNKNOWN. CONSISTENT WITH THE REQUIREMENT OF A RC2 MECHANISM, WE WILL DEVELOP A COMPREHENSIVE DATABASE OF FUNCTIONALLY IMPORTANT RNA MODIFICATIONS IN SELECTED HUMAN TISSUES. WE SEEK TO INTERROGATE ALTERATIONS IN THE MODIFICATIONS OF RNA, SPECIFICALLY, THE TWO MOST IMPORTANT MODIFICATIONS, N6-ADENOSINE METHYLATION (M6A) AND PSEUDOURIDYLATION IN KEY METABOLIC CELL TYPES THAT ARE RELEVANT FOR TYPE 2 DIABETES AND OBESITY. IN THIS PROPOSAL, WE WILL: 1) INTERROGATE THE MRNA AND CHROMATIN- ASSOCIATED RNA MODIFICATIONS, M6A-SEQUENCING AND BID-Ψ-SEQUENCING AT BASE RESOLUTION WITH STOICHIOMETRY INFORMATION IN KEY METABOLIC TISSUES FROM PATIENTS WITH T2D AND OBESITY; 2) PERFORM M6A AND Ψ QTL STUDIES IN TISSUES FROM PATIENTS WITH T2D AND OBESITY AND VALIDATE THE FUNCTIONAL RELEVANCE OF KEY MRNA AND CHROMATIN- ASSOCIATED RNA MODIFICATIONS IN IN VITRO STUDIES; 3) INTERROGATE COMPREHENSIVE TRANSCRIPTION AND ENHANCER ACTIVITY USING ATAC-SEQ IN THE METABOLIC TISSUES FROM PATIENTS WITH T2D AND OBESITY; 4) PERFORM EQTL STUDIES USING TRANSCRIPTIONAL ACTIVITY DATA OBTAINED FROM KAS-SEQ AND RNA MODIFICATION RESULTS ON CARRNAS IN T2D AND OBESE TISSUES, AND PERFORM VALIDATION IN IN VITRO STUDIES; AND 5) DEVELOP A LARGE-SCALE RESOURCE DATABASE ON MRNA AND CHROMATIN-ASSOCIATED RNA MODIFICATIONS AND TRANSCRIPTION ACTIVITY WITH CORRELATION OF COMMON GENOMIC FEATURES FOR THE LARGER SCIENTIFIC COMMUNITY. THE SUCCESS OF THIS PROJECT WILL LAY GROUND FOR THE BROADER COMMUNITY TO TAKE ADVANTAGE OF THESE DATA AND OUR ANALYSES FOR FUTURE BASIC AND TRANSLATIONAL INVESTIGATIONS AND THERAPEUTIC DEVELOPMENTS.

USE OF CONTINUOUS GLUCOSE MONITORING WITH AUTOMATED CLINICAL DECISION SUPPORT TO REDUCE HYPOGLYCEMIA IN OLDER ADULTS WITH TYPE 1 DIABETES

UNCOVERING CONSERVED LIPID HOMEOSTASIS MECHANISMS THAT MEDIATE LONGEVITY AFTER GERM CELL LOSS IN C. ELEGANS

ROLE OF THE KALLIKREIN-KININ SYSTEM IN DIABETIC RETINOPATHY

GENETIC MODIFIERS OF THE EFFECT OF INTENSIVE GLYCEMIC CONTROL ON CVD RISK

USING EX VIVO, IN VIVO MODELS AND PATIENT MUTATIONS TO INTERROGATE PANCREATIC EXOCRINE-ENDOCRINE CROSS TALK - PROJECT SUMMARY TYPE 1 DIABETES AND ITS METABOLIC CONSEQUENCES CONTINUE TO BE AMONG THE MOST SIGNIFICANT BIOMEDICAL CHALLENGES IN THE US AND WORLDWIDE TODAY. IN ADDITION TO MORBIDITIES SPECIFICALLY RELATED TO DIABETES THE DISEASE IS ASSOCIATED WITH COMPLICATIONS SUCH AS RENAL FAILURE, LIPID DISORDERS, CARDIOVASCULAR DISEASE AND CANCER AND IS A MAJOR CAUSE OF DEATH WORLDWIDE. THUS THERE IS AN URGENT NEED FOR A BETTER UNDERSTANDING OF THE PATHOGENESIS THAT PROMOTES THE LOSS OF INSULIN-SECRETING BETA CELLS TO PLAN BETTER THERAPEUTICS TO COMBAT THE DISEASE. SEVERAL STUDIES HAVE ARGUED THAT DIVERSE CELL TYPES THAT MAKE UP THE PANCREATIC NICHE CONTRIBUTE TO THE PATHOGENESIS OF THE DISEASE PROCESS. THUS A BETTER UNDERSTANDING OF THE INTER-CELLULAR COMMUNICATION BETWEEN MAJOR CELLS SUCH AS THE ACINAR AND DUCT CELLS AND ISLET CELLS ARE WARRANTED. WE HAVE STUDIED THE ROLE OF THE MUTANT CARBOXY ESTER LIPASE (CEL) GENE, EXPRESSED IN ACINAR CELLS, ON ITS ABILITY TO BE TAKEN UP BY BETA CELLS AND CAUSE DEFECTS IN ITS FUNCTION AND GROWTH. THE GOAL OF THIS PROPOSAL IS TO DISCOVER THE CHANGES THAT OCCUR WHEN SECRETOME FROM ACINAR VERSUS DUCT CELLS ARE INCUBATED WITH HUMAN ISLET AND SS-CELLS. THE DATA FROM THESE STUDIES WILL GENERATE NEW HYPOTHESIS FOR TESTING THAT WILL ALLOW A DEEPER INTERROGATION OF THE CROSS-TALK BETWEEN HUMAN ACINAR, DUCT AND ISLET CELLS. WE WILL ADDRESS THE FOLLOWING AIMS IN THIS PROPOSAL: AIM 1) DETERMINE THE ABILITY OF HUMAN ACINAR VERSUS HUMAN DUCT CELLS TO DIRECTLY IMPACT ISLET CELL FUNCTION. WE WILL ISOLATE AND CHARACTERIZE EVS, FROM HUMAN INDUCED PLURIPOTENT STEM (HIPS) CELL DERIVED ACINAR- VERSUS DUCT-LINEAGE COMMITTED ORGANOIDS, AND INCUBATE THEM WITH HUMAN ISLET/SS-CELLS AND HUMAN PANCREAS SLICES TO DIRECTLY EXAMINE THE CONSEQUENCES ON ISLET CELL BIOLOGY. WE WILL ALSO EXAMINE THE EV CARGO WITH A PARTICULAR FOCUS ON FRAGMENTS DERIVED FROM TRANSFER RNAS. AIM 2) WE WILL INTERROGATE THE ABILITY OF EVS, DERIVED FROM ACINAR-DERIVED ORGANOIDS GENERATED FROM THE HIPS CELLS FROM MODY8 PATIENTS, TO DIRECTLY REGULATE HUMAN ISLET-SS-CELL BIOLOGY. THE USE OF HUMAN ORGANOIDS AND HUMAN PANCREAS SLICES WILL PROVIDE TRANSLATIONAL RELEVANCE OF OUR STUDIES. THE RESULTS AND DATASETS OBTAINED FROM THESE EXPERIMENTS WILL COMPLEMENT THE EFFORTS OF THE HUMAN ISLET RESEARCH NETWORK AND PROVIDE NOVEL RESOURCES TO BE SHARED WITH THE LARGER SCIENTIFIC COMMUNITY.

SELECTIVE INSULIN RESISTANCE - DIABETIC VASCULAR DISEASE

SKN-1 REGULATION AND OXIDATIVE STRESS RESISTANCE IN C. ELEGANS

METABOLIC PATHWAYS OF INCREASED CARDIOVASCULAR RISK IN TYPE 2 DIABETES

CHARACTERIZATION OF CARDIOVASCULAR DISEASES (CVD) IN PEOPLE WITH LONG DURATION TYPE 1 DIABETES - ABSTRACT CARDIOVASCULAR DISEASE (CVD) IS THE MAJOR CAUSE OF MORTALITY IN TYPE 1 DIABETES (T1D). SOME CVD RISK FACTORS ARE SHARED BETWEEN PEOPLE WITH T1D AND TYPE 2 DIABETES (T2D). HOWEVER, DIFFERENCES IN DYSLIPIDEMIA, SEVERITY OF INSULIN RESISTANCE, DISEASE ONSET, RESPONSES TO GLYCEMIC CONTROL, AND PRESENCE OF AUTOIMMUNITY SUGGEST THAT THE PATHOGENESIS OF CVD MAY DIFFER BETWEEN T1D AND T2D. DIABETIC NEPHROPATHY (DN) IS A MAJOR RISK FACTOR FOR CVD IN T1D; BUT WHILE IT OCCURS IN 40% OF THOSE WITH T1D, CVD IS STILL THE MAJOR CAUSE OF MORTALITY IN THOSE WITHOUT DN. AUTOIMMUNITY MAY ALSO CONTRIBUTE TO CVD, SINCE THE RISKS OF ATHEROSCLEROSIS ARE ELEVATED IN SEVERAL AUTOIMMUNE DISEASES. UNDERSTANDING THE PATHOGENESIS OF CVD IN TID IS HAMPERED ALSO BY THE LACK OF COMPARATIVE PATHOLOGICAL STUDIES OF CORONARY VESSELS OF AGING PEOPLE WITH T1D VERSUS T2D AND NON-DIABETIC SUBJECTS. PRELIMINARY STUDIES FROM THE MEDALIST STUDY, A LARGE COHORT (N=1019) WITH >50 YEARS OF INSULIN-DEPENDENT T1D, SHOWED THAT WHILE ONLY 13% HAVE DN, 40% EXHIBITED SIGNIFICANT CVD HISTORY, WHICH CORRELATED WITH CORONARY ARTERY CALCIUM (CAC) SCORES BY CT AND MYOCARDIAL DYSFUNCTION BY CARDIAC MRI (CMR). WHILE >90% OF MEDALISTS POSSESS HIGH-RISK HLA ALLELES FOR CLASSIC AUTOIMMUNE T1D, 8% ALSO HAVE KNOWN GENES FOR MONOGENIC DIABETES. THE MEDALIST BIOBANK INCLUDES PLASMA/SERUM SAMPLES AND POSTMORTEM ORGAN SPECIMENS (HEARTS WITH CORONARY VESSELS, AORTAE, KIDNEY, PANCREASES AND OTHERS). PILOT STUDIES IN MEDALISTS PROVIDE THE FIRST EXTENSIVE CHARACTERIZATIONS OF INFLAMMATORY AND METABOLOMICS PROFILES OF T1D AND THEIR ASSOCIATIONS WITH CAC SCORES AND CMR PARAMETERS, WHICH MAY INDICATE DIFFERENCES WITH PUBLISHED DATA ON T2D. PILOT MORPHOLOGICAL STUDIES OF MEDALISTS’ CORONARY VESSELS CLEARLY IDENTIFIED IMMUNE CELL INFILTRATES, INCLUDING CD3+ T-CELLS. THE ROLE OF AUTOIMMUNITY IN EXACERBATING ATHEROSCLEROSIS IS CLEARLY DEMONSTRATED BY STUDIES USING NEWLY-CREATED APOE-/-/NOD MICE WITH AUTOIMMUNE DIABETES CLOSELY MIMICKING T1D, WHICH EXHIBITED SIGNIFICANTLY MORE ATHEROSCLEROTIC PLAQUES CONTAINING ELEVATED SUBSETS OF PRO-INFLAMMATORY T-CELLS AND LESS REGULATORY T-CELLS THAN IN NON-DIABETIC APOE-/-/CONGNOD CONTROL MICE. IN THIS PROPOSAL, WE AIM TO CHARACTERIZE CVD IN T1D BY CLINICAL, BIOCHEMICAL, IMAGING, METABOLOMIC AND PATHOLOGICAL STUDIES, AND THEIR ASSOCIATIONS WITH AUTOIMMUNITY, VIA COMPARATIVE STUDIES OF THE MEDALIST COHORT WITH T2D, MONOGENIC DIABETES AND NON-DIABETIC SUBJECTS. OUR SPECIFIC AIMS ARE AS FOLLOWS: SP. AIM 1: TO CHARACTERIZE ATHEROSCLEROSIS AND MYOCARDIAL STRUCTURE AND FUNCTION IN TYPE 1 DIABETES OF LONG DURATION (JOSLIN MEDALIST STUDY) BY CVD HISTORY AND IMAGING, PRESENCE OF AUTOIMMUNITY, BETA CELL FUNCTION, INFLAMMATORY AND METABOLOMICS MARKERS, METABOLIC CONTROL AND MICROVASCULAR DISEASES. SP. AIM 2: TO COMPARE THE PLAQUE CHARACTERISTICS AND COMPOSITION OF IMMUNE CELLS, INCLUDING MONOCYTES, MACROPHAGES, B-CELLS AND T-CELLS (CD3+, CD4+ AND CD8+ T CELLS) AND SUBTYPES OF T-CELLS (TREG, TFH, TH1 AND TH17) IN THE ATHEROSCLEROTIC PLAQUES OF THE CORONARY VESSELS AND PERIPHERAL ARTERIES FROM PATIENTS WITH T1D, T2D, MONOGENIC DIABETES AND NO DM.

IDENTIFICATION OF RETINOID-BINDING PROTEIN 3 (RBP3): A PROTECTIVE FACTOR AGAINST DIABETIC RETINOPATHY USING RETINA FROM PEOPLE WITH EXTREME DURATION

TRANSCRIPTIONAL AND EPIGENETIC REGULATION OF THERMOGENIC ADIPOCYTE PROGRAM - PROJECT SUMMARY/ABSTRACT OBESITY AND ITS METABOLIC SEQUELAE ARE RAPIDLY INCREASING IN THE UNITED STATES AND WORLDWIDE, LEADING TO HIGH MORBIDITY AND MORTALITY IN TYPE 2 DIABETES, CARDIOVASCULAR DISEASE, AND CERTAIN CANCER. UNDER THE CURRENT PANDEMIC, OBESITY HAS BEEN RECOGNIZED AS A KEY RISK FACTOR FOR SEVERE COVID-19. CENTRAL TO THESE PATHOLOGIES IS ADIPOSE TISSUE. THERE ARE FUNCTIONALLY DISTINCT TYPES OF ADIPOSE TISSUE. WHITE ADIPOSE TISSUE IS THE PRIMARY SITE OF THE TRIGLYCERIDE STOREHOUSE. IN CONTRAST, THERMOGENIC FAT, WHICH CONSISTS OF CLASSICAL BROWN ADIPOSE TISSUE (BAT) AND INDUCIBLE BEIGE/BRITE ADIPOCYTES, CONCENTRATES ON THERMOGENIC ENERGY EXPENDITURE. IT HAS BEEN RECENTLY REPORTED THAT PEOPLE WITH BAT HAVE A SIGNIFICANTLY LOWER PREVALENCE OF CARDIOMETABOLIC DISEASES, HIGHLIGHTING THE METABOLIC BENEFITS AND THERAPEUTIC POTENTIAL OF BAT IN HUMANS. TO MAKE THE THERAPEUTICS POSSIBLE, IMPROVED KNOWLEDGE OF THE REGULATION OF THERMOGENIC ADIPOCYTES IS URGENTLY NEEDED. THE THERMOGENIC FUNCTION OF BROWN AND BEIGE ADIPOCYTES ARE COORDINATELY REGULATED BY SPECIFIC TRANSCRIPTIONAL AND EPIGENETIC REGULATORS. WHILE TRANSCRIPTION OF THE THERMOGENIC GENE UNCOUPLING PROTEIN 1 (UCP1) IN RESPONSE TO BETA-ADRENERGIC STIMULATION HAS BEEN BROADLY STUDIED, LITTLE IS KNOWN ABOUT HOW HISTONE POSITIONING AND CHROMATIN FOLDING INFLUENCES THE EXPRESSION OF UCP1 AND OTHER THERMOGENIC GENES. USING AN UNBIASED CRISPR-BASED SCREEN, WE IDENTIFIED THE HISTONE VARIANT H2A.Z AND THE LIM DOMAINING-CONTAINING ZINC-FINGER PROTEIN CRIP2 AS TRANS-ACTING FACTORS RECRUITED TO THE UCP1 PROMOTER/ENHANCER REGION BY BETA3-ADRENERGIC RECEPTOR STIMULATION. IMPORTANTLY, DELETION OF H2A.Z OR CRIP2 IN MATURE BROWN ADIPOCYTES NOT ONLY IMPEDED UCP1 TRANSCRIPTION, BUT ALSO REDUCED THE EXPRESSION OF MULTIPLE THERMOGENIC GENES AND LED TO IMPAIRED CELLULAR THERMOGENESIS. THIS PROPOSAL WILL DETERMINE THE SIGNALING EVENTS MEDIATING THE ACTIVATION OF CRIP2 AND H2A.Z DEPOSITION AND THE IMPACT OF CRIP2 OR H2A.Z DEFICIENCY IN THE CELLULAR THERMOGENESIS AND BIOENERGETIC PROFILES OF THERMOGENIC ADIPOCYTES MURINE AND HUMAN ORIGINS. SINCE HISTONE VARIANTS PLAY AN IMPORTANT ROLE IN DETERMINING CHROMATIN REMODELING, WE WILL EXAMINE HOW CRIP2-H2A.Z INTERACTION INFLUENCES CHROMATIN ARCHITECTURE, THEREBY REGULATING THERMOGENIC TRANSCRIPTION AND CELLULAR RESPIRATION. TO ESTABLISH THE PHYSIOLOGICAL SIGNIFICANCE OF CRIP2 AND H2A.Z IN METABOLIC REGULATION, WE WILL GENERATE BROWN FAT-SPECIFIC CRIP2 OR H2A.Z KNOCKOUT MICE AND THOROUGHLY CHARACTERIZE THEIR METABOLIC PHENOTYPES. COMPLETING THE PROPOSED STUDIES WILL INCREASE FUNDAMENTAL KNOWLEDGE ON THE ROLE OF CHROMATIN REMODELING IN THE REGULATION OF THERMOGENIC PROGRAM AND PAVE WAYS TO ESTABLISH NEW THERAPEUTIC APPROACHES FOR COMBATING METABOLIC DISEASES.

ANIMAL STUDIES INVESTIGATING MOLECULAR TRANSDUCERS OF PHYSICAL ACTIVITY

GUT MICROBIAL FERMENTATION PRODUCTS OF MUSCLE-DERIVED LACTATE AS MEDIATORS OF EXERCISE AND METABOLISM - PROJECT SUMMARY/ABSTRACT THE GUT MICROBIOME MAKES SIGNIFICANT CONTRIBUTIONS TO WHOLE-BODY GLUCOSE METABOLISM AND INSULIN SENSITIVITY, IN PART, THROUGH PRODUCTION OF SHORT CHAIN FATTY ACIDS (SCFA). RECENT EVIDENCE SUGGESTS THAT MICROBIAL SCFA PRODUCTION MAY BE INCREASED BY EXERCISE TRAINING, AND THAT SCFA MAY BE IMPORTANT POSITIVE REGULATORS OF EXERCISE PERFORMANCE AND SKELETAL MUSCLE METABOLISM AND FUNCTION. HOWEVER, IT IS NOT KNOWN HOW GUT MICROBES COULD REGULATE SCFA PRODUCTION IN RESPONSE TO EXERCISE. OUR WORK IDENTIFIES LACTATE UTILIZING BACTERIA (LU-BAC), WHICH CAN CONVERT LACTATE TO SCFA, AS POTENTIAL SOURCES OF SCFA DURING EXERCISE. AS CIRCULATING LACTATE LEVELS INCREASE DURING MODERATE TO HIGH INTENSITY EXERCISE, COMBINING LU-BAC SUPPLEMENTATION WITH EXERCISE MAY RESULT IN HIGHER LEVELS OF CIRCULATING SCFA, THUS ENHANCING THE METABOLIC BENEFITS OF EXERCISE. INDIVIDUALS WITH IMPAIRED GLUCOSE TOLERANCE HAVE LOWER GUT LU-BAC CONTENT, AND BLUNTED METABOLIC AND AEROBIC IMPROVEMENTS IN RESPONSE TO EXERCISE TRAINING. THUS, IN ADDITION TO IMPROVING METABOLIC HEALTH, LU-BAC SUPPLEMENTATION MAY ENHANCE THE HEALTH BENEFITS OF EXERCISE BY AMELIORATING THE METABOLIC DEFECTS THAT LEAD TO IMPAIRED TRAINING RESPONSE. WE HYPOTHESIZE THAT LACTATE PRODUCED BY MUSCLE WITH REGULAR EXERCISE IS USED BY LU-BAC TO GENERATE SCFA, WHICH THEN ACT ON SKELETAL MUSCLE TO IMPROVE METABOLISM AND FUNCTION. WE PROPOSE THIS NOVEL GUT-MUSCLE AXIS LEADS TO IMPROVED METABOLIC HEALTH AND EXERCISE RESPONSE. ONE AIM OF THIS PROPOSAL IS TO TEST LU-BAC SUPPLEMENTATION AS A TREATMENT FOR IMPAIRED GLUCOSE TOLERANCE AND LOW RESPONSE TO EXERCISE IN MOUSE MODELS OF METABOLIC DISEASE THAT PARTIALLY REFLECT THE PATHOLOGIES OF TYPE 1 AND TYPE 2 DIABETES. A SECOND AIM IS TO DETERMINE THE SPECIFIC CONTRIBUTION OF LACTATE FERMENTATION BY LU-BAC TO CIRCULATING AND TISSUE SCFA LEVELS WITH EXERCISE, AND WHETHER LU-BAC-DERIVED SCFA CONTRIBUTE TO THE HEALTH BENEFITS OF EXERCISE TRAINING. OUR THIRD AIM IS TO DEFINE THE MECHANISMS IN SKELETAL MUSCLE BY WHICH SCFA LEAD TO ENHANCED EXERCISE PERFORMANCE AND METABOLIC HEALTH. SPECIFICALLY, WE WILL DETERMINE WHETHER SCFA RECEPTORS AND TRANSPORTERS IN MUSCLE ARE NECESSARY FOR THE POSITIVE EFFECTS OF SCFA ON MUSCLE METABOLISM AND FUNCTION. THIS WORK WILL HAVE A BROAD IMPACT ON THE FIELDS OF EXERCISE, METABOLISM, AND MICROBIOLOGY BY DETERMINING THE MECHANISMS BY WHICH THE MICROBIOME CAN ENHANCE MUSCLE METABOLISM AND ADAPTATION TO EXERCISE. WE ANTICIPATE OUR RESULTS WILL LEAD TO DEVELOPMENT OF A LIVE BIOTHERAPEUTIC TO IMPROVE EXERCISE RESPONSE AND METABOLIC HEALTH.

IMPROVING GLYCEMIA & REDUCING DIABETES DISTRESS IN ADOLESCENTS & YOUNG ADULTS WITH T1D - PROJECT SUMMARY/ABSTRACT ADOLESCENTS AND YOUNG ADULTS (AYA) WITH TYPE 1 DIABETES (T1D) ARE ESPECIALLY AT RISK FOR MISSED DIABETES VISITS AND SUBOPTIMAL SELF-CARE BEHAVIORS, AS EVIDENCED BY THEIR POOR GLYCEMIC CONTROL WITH VERY HIGH A1C LEVELS. FURTHER, DIABETES DISTRESS IS COMMON IN AYA WITH T1D AND CONTRIBUTES TO THEIR SUBOPTIMAL SELF-CARE BEHAVIORS AND OUTCOMES. THE AMERICAN DIABETES ASSOCIATION (ADA) RECOMMENDS ROUTINE SCREENING FOR DIABETES DISTRESS AND, WHEN IDENTIFIED, REFERRAL FOR DIABETES EDUCATION OR MENTAL HEALTH TREATMENT. PERSONS EXPERIENCING DIABETES DISTRESS ARE AT INCREASED RISK FOR MISSED MEDICAL APPOINTMENTS AND DEFICIENT DIABETES SELF-MANAGEMENT. THIS IS CONCERNING BECAUSE, WITHOUT INTERVENTION, DIABETES DISTRESS DOES NOT REMIT AND EITHER REMAINS STABLE OR INTENSIFIES OVER TIME. FURTHER, AYAS MAY NOT ADEQUATELY ACCESS OR USE ADVANCED DIABETES TECHNOLOGIES DUE TO PERCEPTIONS OF POSSIBLE INCREASES IN SELF-CARE BURDEN AND DIABETES DISTRESS WHEN, IN FACT, SUCH TECHNOLOGIES COULD AID IN THEIR EFFORTS TO ENHANCE SELF-CARE AND IMPROVE GLYCEMIC CONTROL. THUS, THERE IS AN URGENT NEED TO DEVELOP PRACTICAL AND WIDELY ACCESSIBLE INTERVENTIONS THAT TARGET DIABETES DISTRESS, SELF-CARE, AND GLYCEMIC CONTROL AND INCLUDE FACE-TO-FACE AND REMOTE VISITS TO FACILITATE UPTAKE. IN THE PROPOSED 2-YEAR RANDOMIZED CLINICAL TRIAL, WE WILL IMPLEMENT AND ASSESS THE IMPACT OF A BEHAVIORAL/PSYCHOEDUCATIONAL INTERVENTION TO REDUCE DIABETES DISTRESS, INCREASE ACCEPTANCE OF SELF-CARE BEHAVIORS (WITH OR WITHOUT DIABETES TECHNOLOGIES), AND IMPROVE GLYCEMIC OUTCOMES IN AYAS WITH T1D. AS A GROUP, AYAS WITH T1D REPRESENT THE SUBPOPULATION OF ALL PERSONS WITH T1D ACROSS THE LIFESPAN THAT HAS THE POOREST GLYCEMIC CONTROL, REACHING A PEAK MEAN A1C VALUE >9% AT AGE ~19. AYAS WITH T1D OFTEN EXPERIENCE DIABETES DISTRESS, WHICH IS LINKED TO POOR GLYCEMIC CONTROL, MAKING THEM IDEAL PARTICIPANTS FOR THE PROPOSED STUDY. WE WILL RECRUIT 180 AYAS (AGES 14-25) WITH T1D DURATION OF AT LEAST 1 YEAR AND SUBOPTIMAL DIABETES CONTROL (A1C 8-13%). PARTICIPANTS WILL BE RANDOMLY ASSIGNED TO A USUAL CARE CONTROL CONDITION OR THE INTERVENTION GROUP. THE INTERVENTION GROUP WILL PARTICIPATE IN MONTHLY INTERVENTION SESSIONS WITH A STUDY INTERVENTIONIST DURING THE FIRST YEAR. OF THE MONTHLY SESSIONS, 4 WILL FOCUS ON IMPROVING GLYCEMIC OUTCOMES IN FACE-TO-FACE SESSIONS AND 8 WILL FOCUS ON REDUCING DIABETES DISTRESS IN REMOTE, VIRTUAL SESSIONS TO LIMIT STUDY BURDEN (AND IN RECOGNITION OF PARADIGM CHANGES IN CARE DELIVERY DURING SARS- COV-2 PANDEMIC). TO ENSURE ADEQUATE RECRUITMENT AND RETENTION, THE CONTROL GROUP WILL RECEIVE A DELAYED INTERVENTION AND PARTICIPATE IN THE MONTHLY SESSIONS DURING THE SECOND YEAR OF THE STUDY. BOTH GROUPS WILL USE CONTINUOUS GLUCOSE MONITORING (CGM) FOR 14 DAYS EVERY 3 MONTHS AND COMPLETE SURVEYS EVERY 6 MONTHS. A1C WILL ALSO BE MEASURED CENTRALLY EVERY 6 MONTHS. WE WILL COMPARE THE TWO GROUPS ON THE PRIMARY GLYCEMIC OUTCOME OF PERCENT TIME-IN-RANGE (TIR) OF 70-180 MG/DL ASSESSED BY CGM AND ON THE SECONDARY OUTCOMES OF DIABETES DISTRESS AND A1C FROM BASELINE TO 1 YEAR. WE HYPOTHESIZE THAT THE INTERVENTION GROUP WILL HAVE AN IMPROVEMENT IN PERCENT TIR, DIABETES DISTRESS, AND A1C COMPARED WITH THE USUAL CARE CONTROL GROUP.

GENETICS OF CORONARY ARTERY DISEASE IN TYPE 2 DIABETES

ALTERATIONS IN POST-RECEPTOR INSULIN SIGNALING IN DIABETES AND INSULIN RESISTANCE - ABSTRACT: THIS IS A REVISED GRANT APPLICATION ENTITLED “ALTERATIONS IN POST-RECEPTOR INSULIN SIGNALING IN DIABETES AND INSULIN RESISTANCE.” INSULIN AND IGF-1 ACTING VIA THEIR COGNATE RECEPTORS (IR AND IGF1R) TO PRODUCE A WIDE RANGE OF METABOLIC AND GROWTH EFFECTS ON MOST CELLS IN THE BODY. OVER MANY YEARS, WORK FROM MY LAB HAS BEEN DEVOTED TO UNDERSTANDING THE INTERMEDIATE SIGNALS IN THIS PROCESS AND HOW THESE MAY BE ALTERED IN DISEASE. THUS, WE HAVE CHARACTERIZED EXTENSIVELY THE ROLES OF INSULIN RECEPTOR SUBSTRATE PROTEINS IN COUPLING IR AND IGF1R TO DOWNSTREAM EFFECTOR SYSTEMS, THE IMPORTANT ROLE OF PI-3 KINASE AND AKT IN THE METABOLIC ACTIONS OF INSULIN, AND EFFECTS OF MAP/MTOR/S6K KINASE PATHWAY IN GROWTH PROMOTION. THESE STUDIES HAVE LED TO DEVELOPMENT OF AN INTEGRATED MODEL OF THE INSULIN SIGNALING NETWORK IN WHICH THERE ARE CRITICAL NODES OF SIGNAL DIVERGENCE THAT PROVIDE COMPLEMENTARY INFORMATION TO DIFFERENT DOWNSTREAM ACTIONS OF INSULIN. THESE CRITICAL NODES ALSO PROVIDE IMPORTANT SITES OF POSITIVE AND NEGATIVE REGULATION THAT CAN LEAD TO ALTERATIONS OF INSULIN ACTION IN DISEASE. RECENTLY, WE HAVE BEGUN TO DISSECT THE FULL PHOSPHOPROTEOME DOWNSTREAM IR/IGF1R AND, THROUGH THIS, HAVE IDENTIFIED TWO NEW FORKHEAD TRANSCRIPTIONAL MEDIATORS OF INSULIN/IGF-1 SIGNALING, FOXK1 AND FOXK2. FROM A DISEASE PERSPECTIVE, WE HAVE ALSO SHOWN HOW DIFFERENT INSULIN RESISTANT STATES ALTER THE INSULIN SIGNALING NETWORK IN DIFFERENT TISSUES. WE HAVE ALSO DEVELOPED IPS CELL MODELS TO FOCUS ON IDENTIFICATION OF CELL AUTONOMOUS COMPONENTS OF INSULIN RESISTANCE IN HUMAN DISEASE. INDEED, AS SHOWN IN OUR PRELIMINARY DATA, MYOBLASTS DERIVED FROM T2D IPSCS DEMONSTRATE DEFECTS IN DOWNSTREAM SIGNALING AND METABOLIC FUNCTION IN VITRO MIRRORING THE DEFECTS FOUND IN VIVO. MORE IMPORTANTLY, THESE CELLS ALSO SHOW DYSREGULATION OF A MULTIDIMENSIONAL PHOSPHORYLATION NETWORK - BOTH INSIDE AND OUTSIDE THE CLASSICAL INSULIN SIGNALING CASCADE. IN THIS GRANT, WE WILL FOCUS ON TWO INTERRELATED SPECIFIC AIMS: 1) ELUCIDATE THE FUNDAMENTAL DIFFERENCES IN INSULIN SIGNALING IN T2D AND OTHER INSULIN RESISTANT STATES IN VITRO USING TARGETED AND GLOBAL PHOSPHOPROTEOMICS OF HUMAN IPS CELL-DERIVED MYOBLASTS FROM NORMAL INDIVIDUALS, T2D PATIENTS AND NON-DIABETIC INDIVIDUALS WITH INSULIN RESISTANCE. WE WILL ASSESS HOW THESE CHANGES AFFECT CELLULAR FUNCTION AND PARTICIPATE IN INSULIN RESISTANCE. 2) DEFINE THE ROLE OF TWO NEW DOWNSTREAM TRANSCRIPTIONAL REGULATORS IN INSULIN ACTION, FOXK1 AND FOXK2. WE WILL IDENTIFY THE GENES REGULATED BY FOXK1/2, DETERMINE HOW THEY COMPLEMENT OTHER TRANSCRIPTIONAL REGULATORS IN INSULIN REGULATION OF CELLULAR FUNCTION, AND HOW THEY ARE ALTERED IN DIABETES. WE WILL ALSO DEFINE FOXK REGULATED GENES USING CHIP-SEQ. FINALLY, WE WILL CREATE MICE WITH TISSUE SPECIFIC DELETION OF FOXK1, FOXK2 AND SELECTED COMBINATORIAL KNOCKOUTS TO DEFINE THEIR COMPLEMENTARY ROLES IN INSULIN-REGULATED GENE EXPRESSION AND INSULIN ACTION IN VIVO. TOGETHER THESE STUDIES WILL LEAD TO A NEW LEVEL OF UNDERSTANDING INSULIN SIGNALING AND ITS ALTERATIONS IN DIABETES, PROVIDE DEEPER UNDERSTANDING OF INSULIN REGULATION OF GENE EXPRESSION AND PROVIDE NEW POINTS FOR THERAPY OF TYPE 2 DIABETES AND OTHER INSULIN RESISTANT DISORDERS.

DISSECTING THE THERMOGENIC ADIPOSE NICHE - PROJECT SUMMARY/ABSTRACT ADIPOSE TISSUE PLAYS A CRITICAL ROLE IN THE REGULATION OF WHOLE-BODY ENERGY METABOLISM, AND ADIPOSE DYSFUNCTION DIRECTLY LINKS TO THE ETIOLOGY OF SEVERAL COMMONLY SEEN METABOLIC DISEASES, SUCH AS TYPE 2 DIABETES MELLITUS AND CARDIOVASCULAR DISEASES. THERE ARE SEVERAL DIFFERENT ADIPOSE DEPOTS DISPERSED THROUGHOUT THE BODY. WHITE ADIPOSE TISSUE (WAT) IS THE PRIMARY SITE OF THE TRIGLYCERIDE STOREHOUSE. IN CONTRAST, THERMOGENIC FAT, WHICH CONSISTS OF CLASSICAL BROWN ADIPOSE TISSUE (BAT) AND INDUCIBLE BEIGE/BRITE FAT, IS SPECIALIZED FOR THERMOGENIC ENERGY EXPENDITURE. IN HUMANS, INDIVIDUALS WITH DETECTABLE BAT HAVE A SIGNIFICANTLY LOWER PREVALENCE OF CARDIOMETABOLIC DISEASES, POINTING TO THE METABOLIC BENEFITS AND THERAPEUTIC PROMISE OF BAT. THERMOGENIC ADIPOSE TISSUE CAN RAPIDLY RESPOND TO ENVIRONMENTAL CHALLENGES BY MODULATING CELLULAR COMPOSITIONS AND CELL- TO-CELL INTERACTIONS. SUCH ADAPTATION IS KEY TO MAINTAINING METABOLIC HEALTH. INTERCELLULAR COMMUNICATIONS WITHIN THE THERMOGENIC ADIPOSE NICHE ORCHESTRATE ADIPOSE TISSUE DEVELOPMENT AND PLAY AN ESSENTIAL ROLE IN ADIPOSE TISSUE TURNOVER, EXPANSION, AND REMODELING IN RESPONSE TO EXTERNAL STIMULI. TO DISSECT THE COMPLEX CELLULAR MAKEUP OF THERMOGENIC FAT, WE PERFORMED SINGLE-CELL RNA-SEQUENCING ANALYSIS OF BAT FROM MICE HOUSED AT DIFFERENT TEMPERATURES. THE RESULTS REVEALED A HIGH DEGREE OF HETEROGENEITY OF THERMOGENIC ADIPOSE NICHE AND SHOWED COLD EXPOSURE INDUCED DYNAMIC CHANGES OF CELLULAR COMPOSITION IN BAT. IMPORTANTLY, WE IDENTIFIED A NOVEL POPULATION OF ADIPOSE PROGENITOR CELLS (APCS), WHICH WAS DERIVED FROM THE VASCULAR SMOOTH MUSCLE (VSM) LINEAGE AND UNIQUELY EXPRESSED TRPV1 (TRANSIENT RECEPTOR POTENTIAL CATION CHANNEL SUBFAMILY V MEMBER 1). LINEAGE TRACING STUDIES DEMONSTRATED THAT THE TRPV1-EXPRESSING VSM-APCS COULD PROLIFERATE AND DIFFERENTIATE INTO HIGHLY THERMOGENIC ADIPOCYTES IN RESPONSE TO COLD STIMULATION. WORK IN PROGRESS SHOWED THAT IMPAIRED ADIPOGENIC DIFFERENTIATION OF TRPV1-EXPRESSING APCS RESULTED IN REDUCED EXPRESSION OF THERMOGENIC GENES IN BAT AND WAT, SUGGESTING AN IMPORTANT ROLE OF THESE CELLS IN THERMOREGULATION. ADDITIONAL PRELIMINARY DATA SHOWED THAT THE TRPV1-EXPRESSING APCS COULD RESPOND TO LOCAL SIGNALS AND DIFFERENTIATE INTO THERMOGENIC ADIPOCYTES. THESE EXCITING FINDINGS HAVE LED US TO PROPOSE A MODEL INVOLVING INTERCELLULAR COMMUNICATIONS IN WHICH INTERPLAYS BETWEEN THE APCS AND INDUCTIVE SIGNALS SHAPES ADIPOCYTE DIFFERENTIATION AND FUNCTION. IN THIS PROPOSAL, WE WILL TEST THIS HYPOTHESIS BY DETERMINING THE PHYSIOLOGICAL ROLE OF THE TRPV1-EXPRESSING APCS IN METABOLIC REGULATION, IDENTIFYING THE ENDOGENOUS AND EXOGENOUS STIMULI OF TRPV1-POSITIVE APCS, AND MAPPING THE CELLULAR INTERACTOME OF THE THERMOGENIC ADIPOSE NICHE USING COMPUTATIONAL TOOLS AND SPATIAL TRANSCRIPTOMIC ANALYSIS. SUCCESSFUL COMPLETION OF THE PROPOSED STUDIES WILL PROVIDE NEW INSIGHT INTO THE ROLE OF INTERCELLULAR COMMUNICATIONS IN THE REGULATION OF ADIPOSE TISSUE DEVELOPMENT AND FUNCTION.

OPTIMIZING CGM USE AND METABOLIC OUTCOMES IN YOUTH WITH TYPE 1 DIABETES

CAUSAL CONNECTIONS BETWEEN AXON GUIDANCE PROTEINS AND EARLY PROGRESSIVE KIDNEY FUNCTION DECLINE IN DIABETES - PROJECT SUMMARY/ABSTRACT END STAGE KIDNEY DISEASE (ESKD) DUE TO DIABETES CONTINUES TO RISE DESPITE IMPROVEMENTS IN GLYCEMIC CONTROL AND WIDESPREAD TREATMENT WITH RENO-PROTECTIVE DRUGS. A MORE COMPLETE UNDERSTANDING OF THE MECHANISMS RESPONSIBLE FOR THE PROGRESSIVE DECLINE IN KIDNEY FUNCTION IS URGENTLY NEEDED TO IDENTIFY NEW, MORE EFFICACIOUS METHODS OF TREATMENT AND PREVENTION OF ESKD IN DIABETES. BY STUDYING DIABETIC PATIENTS WITH ADVANCED KIDNEY DISEASE WITH PROTEINURIA AND CKD=3, WE OBTAINED TWO NOVEL FINDINGS RELATED TO PROGRESSION TO ESKD IN BOTH T1D AND T2D. USING FOLLOW-UP DATA FROM TWO INDEPENDENT COHORTS AND GLOBAL MIRNAS PLATFORM, WE FOUND A SPECIFIC PROFILE OF PLASMA MIRNAS STRONGLY ASSOCIATED WITH PROGRESSION TO ESKD. THESE MIRNAS SPECIFICALLY TARGETED AXON GUIDANCE PATHWAY (AGP) PROTEINS. STRIKINGLY, CIRCULATING LEVELS OF 6 AGP PROTEINS (TWO LIGANDS EFNA4, EFN5 AND 4 RECEPTORS EPHA2, EPHB2, EPHB6, UNC5C) WERE VERY STRONGLY ASSOCIATED WITH THE EXTENT OF KIDNEY STRUCTURAL LESIONS IN THE T2D PIMA INDIAN COHORT AND PROGRESSION TO ESKD IN FOUR INDEPENDENT COHORTS. OUR FINDINGS ARE THE FIRST TO IMPLICATE CIRCULATING AGP PROTEINS IN THE PROGRESSION TO ESKD IN DIABETES. TO FOLLOW-UP THESE FINDINGS, WE PROPOSE A MULTI-DISCIPLINARY STUDY TO ASSESS THE ROLE OF THE AGP PROTEINS IN THE DEVELOPMENT OF EARLY PROGRESSIVE KIDNEY FUNCTIONAL DECLINE (PKFD) AND LEARN ABOUT MECHANISMS THROUGH WHICH CIRCULATING AGP MAY DAMAGE THE KIDNEY. THREE COMPLEMENTARY STUDIES ARE PROPOSED. DR. KROLEWSKI’S LAB WILL EXAMINE WHETHER ELEVATED LEVELS OF TWO EXEMPLARS OF AGP PROTEINS EFNA4 AND EPHA2 ARE ASSOCIATED/CAUSALLY RELATED TO THE DEVELOPMENT OF EARLY PKFD. DR. KRETZLER’S LAB WILL EXAMINE HOW CIRCULATING LEVELS OF THE AGP PROTEINS MAY IMPACT CANDIDATE ETIOLOGICAL PATHWAYS IN KIDNEY CELLS. DR. KORSTANJE’S LAB WILL EXAMINE WHETHER GENETIC MANIPULATION OF CIRCULATING LEVELS OF EFNA4 AND EPHA2 IN MICE MODELS OF DIABETES LEAD TO ONSET OF DIABETIC KIDNEY DISEASE. THE PROPOSED RESEARCH HAS THE FOLLOWING SPECIFIC AIMS: AIM #1: WE WILL EXAMINE THE EFFECT OF VARIATION OF CIRCULATING LEVELS OF THE CANDIDATE MIRNAS AND AGP PROTEINS EXAMINED AT BASELINE IN THE TWO NESTED CASES-CONTROL STUDIES OF T1D PATIENTS WITH NORMO-ALB (N=390) AND MICRO- ALB (N=430) ON RISK OF EARLY PKFD DURING 7-15 YEARS OF FOLLOW-UP. AIM #2: WE WILL EXAMINE THE LINK BETWEEN CIRCULATING LEVELS OF THE CANDIDATE AGP PROTEINS AND DYSREGULATION OF CANDIDATE ETIOLOGICAL PATHWAYS USING SINGLE CELL RNA SEQUENCING RESULTS OBTAINED FROM RESEARCH KIDNEY BIOPSIES FROM PIMA INDIANS WITH T2D. AIM #3: WE WILL TEST EPHA2 AND EFNA4 FOR THEIR ABILITY TO MODIFY RENAL PHENOTYPES IN A DIABETIC MOUSE MODEL AND TEST WHETHER MIR-328-5P AND MIR-339-5P, WHICH ARE BOTH DOWNREGULATED DURING PROGRESSION TO ESKD AND HAVE ORTHOLOGS IN THE MOUSE, CAN AFFECT THE IMPACT OF THESE AGP PROTEINS ON DIABETIC KIDNEY DISEASE. THE PROPOSED RESEARCH HAS A VERY HIGH PROBABILITY OF GENERATING NOVEL FINDINGS. ITS SUCCESS IS ASSURED BY THE FACT THAT THE CO-INVESTIGATORS HAVE A TRACK RECORD OF COLLABORATION, AND STUDY COHORTS AND BIOBANKS OF SPECIMENS FROM ALL MEMBERS OF THE JOSLIN KIDNEY STUDY AND PIMA INDIAN KIDNEY STUDY ARE AVAILABLE.

DEVELOPMENT OF PROGNOSTIC ALGORITHMS TO IDENTIFY SUBJECTS AT HIGH RISK OF ESKD IN TYPE 2 DIABETES - PROJECT SUMMARY/ABSTRACT WITH THE RISING PREVALENCE OF DIABETES IN THE US AND OTHER COUNTRIES, THERE IS AN ONGOING RESEARCH EFFORT TO FIND BIOMARKERS ALLOWING THE IDENTIFICATION OF PATIENTS WITH DIABETES AT HIGH RISK OF END STAGE KIDNEY DISEASE (ESKD). WITH SUPPORT FROM NIH AND JDRF, WE HAVE IDENTIFIED 21 SERUM PROTEINS THAT WERE SIGNIFICANTLY ASSOCIATED WITH INCREASED RISK OF KIDNEY FUNCTION LOSS AND ESKD IN THE JOSLIN KIDNEY STUDY, AND HAVE DEVELOPED AN AD HOC OLINK MULTIPLEX ASSAY (SO CALLED JOSLIN KIDNEY PANEL [JKP]) TO MEASURE THESE BIOMARKERS. PRELIMINARY DATA STRONGLY SUGGEST THAT A SUBSET OF THE JKP CAN SIGNIFICANTLY IMPROVE THE ABILITY TO PREDICT ESKD RISK IN SUBJECTS WITH TYPE 2 DIABETES (T2D) WHEN ADDED TO GFR AND ALLBUMINURIA. IN THIS PROPOSAL, WE AIM TO VALIDATE THESE PRELIMINARY FINDINGS IN OTHER SETTINGS, IN ORDER TO DEVELOP IMPROVED ALGORITHMS FOR ESKD RISK PREDICTION. WE INTEND TO ACCOMPLISH THESE GOALS USING EXISTING DATA AND SPECIMENS FROM INDIVIDUALS WITH AND WITHOUT T2D FROM 1. THE CHRONIC RENAL INSUFFICIENCY COHORT (CRIC) STUDY; AND 2. THE ACTION TO CONTROL CARDIOVASCULAR RISK IN DIABETES (ACCORD) TRIAL AND ITS FOLLOW-UP STUDY ACCORDION. OUR SPECIFIC AIMS ARE: 1: TO IDENTIFY THE MOST INFORMATIVE OF THE 21 BIOMARKERS IN THE JOSLIN KIDNEY PANEL AND EVALUATE THEIR PERFORMANCE, WHEN ADDED TO GFR AND ALBUMINURIA, IN PREDICTING ESKD RISK AMONG SUBJECTS WITH T2D AND CHRONIC KIDNEY DISEASE. WE WILL MEASURE THE 21 PROTEINS OF THE JKP IN BASELINE SERUM SPECIMENS FROM ~1,500 CRIC PARTICIPANTS WITH T2D, AND WILL USE THESE DATA TOGETHER WITH GFR AND ALBUMINURIA TO DEVELOP AND INTERNALLY VALIDATE MULTI-MARKER PROGNOSTIC ALGORITHMS PREDICTING THE RISK OF ESKD (PRIMARY OUTCOME) OR THE COMPOSITE OF ESKD AND/OR 50% LOSS OF KIDNEY FUNCTION (SECONDARY OUTCOME) DURING 10 YEARS OF FOLLOW-UP. 2: TO EVALUATE THE GENERALIZABILITY OF FINDINGS FROM CRIC TO T2D INDIVIDUALS WITH A BROADER SPECTRUM OF KIDNEY FUNCTION. WE WILL ASSAY THE JKP IN BASELINE SERUM SPECIMENS FROM A CASE- COHORT SAMPLE OF ~2,000 ACCORD/ACCORDION PARTICIPANTS AND WILL USE THESE DATA TO INVESTIGATE THE GENERALIZABILITY OF THE PREDICTIVE ALGORITHMS BUILT IN CRIC TO DIABETIC PATIENTS WITH DIFFERENT CHARACTERISTICS. THE PROGNOSTIC MODELS DEVELOPED IN AIM 1 AND EXTERNALLY VALIDATED IN AIM 2 WILL BE USED TO BUILD A WEB-BASED KIDNEY RISK CALCULATOR FOR THE ESTIMATION OF THE 10-YEAR RISK OF ESKD IN A CLINICAL SETTING. 3: TO EVALUATE THE TRANSFERABILITY OF THE KIDNEY RISK CALCULATOR FROM DIABETIC TO NON-DIABETIC KIDNEY DISEASE. WE WILL MEASURE THE 21 JKP BIOMARKERS IN BASELINE SERUM SAMPLES FROM ~1,700 NON-DIABETIC SUBJECTS FROM THE CRIC STUDY AND WILL ASSESS THE PERFORMANCE OF THE KIDNEY RISK CALCULATOR DEVELOPED IN AIM 2 IN PREDICTING THE RISK OF ESKD AND ESKD/50% KIDNEY FUNCTION LOSS IN PATIENTS WITH NON-DIABETIC KIDNEY DISEASE. THE PROPOSED RESEARCH HAS A HIGH LIKELIHOOD OF RESULTING IN THE DEVELOPMENT OF IMPROVED PROGNOSTIC TOOLS FOR THE STRATIFICATION OF PATIENTS WITH DIABETES ACCORDING TO THEIR RISK OF PROGRESSION TO ESKD. THIS WOULD BE A GREAT ADVANCEMENT FOR OPTIMIZING PATIENT CARE AND FOR IMPROVING THE EFFICIENCY OF CLINICAL TRIALS OF NEW ESKD-PREVENTING INTERVENTIONS.

INTERACTION BETWEEN GENES, ENVIRONMENT, THE MICROBIOME AND METABOLOME IN TYPE 2 DIABETES AND METABOLIC SYNDROME

OPTIMIZING SELF-MANAGEMENT ADHERENCE AND GLYCEMIC CONTROL IN OLDER TEENS WITH T1D

EMBRYONIC GENE EXPRESSION DURING DIABETIC EMBRYOPATHY

FUNCTIONAL VALIDATION OF GENE MODIFICATIONS THAT PROTECT BETA CELLS AGAINST AUTOIMMUNITY IDENTIFIED BY GENOME-WIDE CRISPR CAS9 SCREENING

ROLE OF SERPINB1 IN BETA CELL GROWTH

AGING AND REJUVENATION OF THE HEMATOPOIETIC STEM CELL NICHE

LIPOTOXICITY AND MAINTENANCE OF METABOLIC HEALTH - ABSTRACT LIPOTOXICITY IS AN IMPORTANT MECHANISM OF COMPLICATIONS IN TYPE 2 DIABETES, INCLUDING NON-ALCOHOLIC FATTY LIVER DISEASE (NAFLD). THROUGH A GENETIC SCREEN, WE DISCOVERED THAT LOSS OF SMALL NUCLEOLAR RNAS (SNORNAS) EMBEDDED WITHIN INTRONS OF THE RIBOSOMAL PROTEIN L13A (RPL13A) LOCUS CONFERS RESISTANCE TO LIPOTOXICITY IN A CELL CULTURE MODEL. THE FOUR RPL13A SNORNAS, SNORD32A, SNORD33, SNORD34, AND SNORD35A, GUIDE 2¢-O- METHYL NUCLEOTIDE MODIFICATIONS AT FIVE SITES ON NASCENT 18S AND 28S RIBOSOMAL RNAS THAT LIE IN CLOSE PROXIMITY TO KEY FUNCTIONAL CENTERS OF THE RIBOSOME STRUCTURE. THIS STUDY WILL TEST THE HYPOTHESIS THAT KNOCKOUT OF RPL13A SNORNAS PROTECTS FROM LIPOTOXIC TISSUE DAMAGE IN A MOUSE MODEL OF HIGH FAT DIET-INDUCED NAFLD. WE WILL DEFINE THE MECHANISTIC LINKS AMONG SNORNA ABUNDANCE, MRNA TRANSLATION, AND DIETARY NUTRIENT EXPOSURES. FINDINGS FROM THIS WORK WILL PROVIDE NEW INSIGHTS INTO THE PATHOPHYSIOLOGY OF NAFLD AND OTHER DISEASES OF LIPID OVERLOAD AND HAVE THE POTENTIAL TO IDENTIFY NEW THERAPEUTIC TARGETS FOR TREATMENT OF METABOLIC DISEASES.

SEARCH FOR BIOMARKERS TO MEASURE PROGRESSION OF DIABETIC NEPHROPATHY IN TYPE 1 DIABETES; APPLICATION OF PROTEOMICS TECHNOLOGIES

PYRUVATE KINASE M2 LEVELS AND ACTIVATION AS PROTECTIVE FACTORS FOR DIABETIC NEPHROPATHY

UNDERSTANDING THE ROLE OF THE COMPLEMENT PROTEOME IN PROGRESSIVE DIABETIC KIDNEY DISEASE

UNDERSTANDING, PREDICTING AND PREVENTING TYPE 2 DIABETES IN YOUTH, BOSTON CLINICAL CENTER (UPP STUDY) - PROJECT SUMMARY / ABSTRACT RISING OBESITY HAS LED TO AN UNPRECEDENTED INCREASE IN PRE-DIABETES (PRED) AND TYPE 2 DIABETES (T2D) INCIDENCE IN CHILDREN AND ADOLESCENTS, A WORRISOME TREND AMPLIFIED BY THE COVID 19 PANDEMIC. BASED ON CURRENT EVIDENCE, IT REMAINS DIFFICULT TO PREDICT WHETHER CHILDREN AND ADOLESCENTS WITH PRED WILL PROGRESS TO T2D. OUR PROPOSAL AIMS TO ADDRESS KEY QUESTIONS IN THE PATHOGENESIS OF T2D, FOCUSING ON MODIFIABLE RISK FACTORS. LEVERAGING A COLLABORATION BETWEEN BOSTON CHILDREN’S HOSPITAL, JOSLIN DIABETES CENTER, AND MASSACHUSETTS GENERAL HOSPITAL, IN PARTNERSHIP WITH COMMUNITY HEALTH CENTERS IN THE GREATER BOSTON AREA, WE PROPOSE TO RECRUIT A DIVERSE COHORT OF EARLY PUBERTAL YOUTH (AGES 7-15, N=300) WITH PRED, ELEVATED BMI (=95TH PERCENTILE), A POSITIVE FAMILY HISTORY OF DIABETES, AND ONE OR MORE ADDITIONAL RISK FACTORS. WE PROPOSE TO COMBINE RIGOROUS ANNUAL CLINICAL STUDIES WITH CONVENIENT REMOTE ASSESSMENTS, TO GAIN A GRANULAR UNDERSTANDING OF METABOLIC, HORMONAL AND ENVIRONMENTAL FACTORS CONTRIBUTING TO T2D PATHOGENESIS. IN AIM 1, WE WILL TEST WHETHER MEASURES OF GLUCOSE HOMEOSTASIS AND BETA CELL FUNCTION DIFFER BETWEEN YOUTH WITH PRED WHO PROGRESS TO T2D VERSUS THOSE WHO REVERT TO NORMOGLYCEMIA OR REMAIN PRED. WE PROPOSE TO ANALYZE GLYCEMIA AND BETA CELL FUNCTION USING ORAL GTT AND INCRETIN HORMONE LEVELS, AND ASSESSMENTS ADAPTABLE TO COMMUNITY SETTINGS, E.G., CONTINUOUS GLUCOSE MONITORS AND HOME A1C KITS. IN AIM 2, WE WILL TEST WHETHER FITNESS LEVEL AND AMOUNT OF PHYSICAL ACTIVITY DIFFER BETWEEN YOUTH WITH PRED WHO PROGRESS TO T2D VERSUS THOSE WHO REVERT TO NORMOGLYCEMIA OR REMAIN PRED. WE PROPOSE TO EVALUATE FITNESS LEVEL USING DETAILED CLINICAL ASSESSMENTS, INCLUDING VO2 MAX, ASSESSMENTS ADAPTABLE TO COMMUNITY SETTINGS INCLUDING GRIP STRENGTH, AND FREE-LIVING ASSESSMENTS USING WEARABLES AND APP-BASED ACTIVITY TRACKING. IN AIM 3, WE WILL TEST WHETHER EVOLUTION OF BODY COMPOSITION DURING GROWTH AND PUBERTY PREDICTS PROGRESSION TO T2D. WE PROPOSE TO ANALYZE BODY COMPOSITION (TOTAL AND VISCERAL FAT) USING DXA, AND HEPATIC FAT USING ECHOGRAPHY-BASED ASSESSMENTS, GONADAL HORMONES, ADRENAL ANDROGENS, AND MEDIATORS OF GROWTH HORMONE ACTION, AS WELL AS ASSESSMENTS ADAPTABLE TO COMMUNITY SETTINGS, E.G., BMI AND APP-BASED DIETARY SURVEYS. AS SECONDARY AIMS, WE PROPOSE TO CREATE A DATA REPOSITORY TO ALLOW EVALUATION OF SOCIAL AND ENVIRONMENTAL FACTORS CONTRIBUTING TO T2D ONSET AT A CONSORTIUM LEVEL, WITH MEASURES INCLUDING SOCIAL DETERMINANTS OF HEALTH, NEIGHBORHOOD AND GEOGRAPHIC CHARACTERISTICS (USING GEOCODING TECHNIQUES), AND THE ENVIRONMENTAL EXPOSOME. WE ALSO PROPOSE THE CREATION OF A BIOLOGICAL REPOSITORY TO ALLOW MULTI-OMICS STUDIES TO IDENTIFY GENOMIC, EPIGENETIC, AND/OR METABOLOMIC MARKERS FOR PROGRESSION FROM PRED TO T2D IN YOUTH AT A CONSORTIUM LEVEL. WE PROPOSE TO COLLECT A RICH BIOREPOSITORY OF LONGITUDINAL SAMPLES (I.E., PLASMA, PBMCS, URINE, STOOL, HAIR) FROM ALL PARTICIPANTS THAT WILL SET THE STAGE FOR FUTURE SYSTEMS BIOLOGY-DRIVEN STUDIES. TOGETHER, THESE STUDIES WILL PERMIT DEVELOPMENT OF A PREDICTIVE MODEL BASED ON VARIABLES EASILY MEASURABLE AT THE COMMUNITY LEVEL, WHICH CAN THEN BE APPLIED TO THE DETECTION AND TREATMENT OF YOUTH AT HIGHEST RISK OF T2D.

CONTROL OF HSC PROLIFERATION AND MIGRATION BY THE TRANSCRIPTION FACTOR EGR1

ROLE OF BROWN FAT-DERIVED SPECIALIZED PRO-RESOLVING LIPID MEDIATORS IN INFLAMMATION AND METABOLISM

SEARCH FOR NEW MODY GENES AND MOLECULAR STUDIES OF THEIR FUNCTION IN BETA CELLS

GLUCAGON PUMP THERAPY FOR POST-BARIATRIC HYPOGLYCEMIA: MERGING PHYSIOLOGY AND ENGINEERING - PROJECT SUMMARY/ABSTRACT BARIATRIC SURGERY IS A POTENT TREATMENT FOR OBESITY-RELATED METABOLIC DISEASE AND MARKEDLY IMPROVES GLUCOSE CONTROL IN T2D. HOWEVER, ALONG WITH THIS METABOLIC SUCCESS COMES AN INCREASED RISK OF SEVERE HYPOGLYCEMIA (TERMED POST-BARIATRIC HYPOGLYCEMIA, OR PBH), WITH ESTIMATED PREVALENCE OF UP TO 30% OF PATIENTS AFTER BOTH GASTRIC BYPASS AND SLEEVE GASTRECTOMY. CURRENT THERAPIES FOR PBH ARE LIMITED AND INCOMPLETELY EFFECTIVE, AND INDIVIDUALS WITH SEVERE HYPOGLYCEMIA AND UNAWARENESS LACK SUFFICIENT WARNING TO TREAT BEFORE RAPID ONSET OF NEUROGLYCOPENIA AND IMPAIRED SAFETY. PREVIOUS STUDIES OF PBH BY OUR GROUP AND OTHERS HAVE DEMONSTRATED POTENTIAL DRIVERS OF HYPOGLYCEMIA IN THIS CONDITION: (1) INCREASED INSULIN SECRETION, LINKED TO POSTPRANDIAL INCRETIN SECRETION (INCLUDING GLP1), (2) INCREASED INSULIN-INDEPENDENT GLUCOSE UPTAKE, (3) REDUCED SECRETION OF GLUCAGON AND OTHER COUNTERREGULATORY HORMONES, AND (4) ALTERED INTESTINAL METABOLISM. ONE PROMISING APPROACH TO PREVENT AND TREAT SEVERE HYPOGLYCEMIA IN PBH IS CLOSED-LOOP GLUCAGON THERAPY. IN PRIOR STUDIES, WE COLLABORATED WITH ENGINEERING COLLEAGUES TO DEVELOP AND TEST A CLOSED-LOOP SYSTEM WHICH UTILIZED CGM IN COMBINATION WITH A NOVEL CLOSED-LOOP ALGORITHM TO DETECT INCIPIENT MEAL-RELATED HYPOGLYCEMIA AND DELIVER BOLUSES OF GLUCAGON VIA A PATCH PUMP. USING A DOUBLE-BLIND, PLACEBO-CONTROLLED CROSSOVER PILOT STUDY IN THE CLINICAL RESEARCH UNIT, WE DEMONSTRATED EFFICACY AND SAFETY OF THIS CLOSED-LOOP GLUCAGON PUMP SYSTEM, WITH INCREASED NADIR GLUCOSE AFTER MIXED MEAL AND REDUCED NEED FOR RESCUE GLUCOSE. BUOYED BY THE SUCCESS OF THESE MEAL-FOCUSED STUDIES, WE NOW PROPOSE TO FURTHER DEFINE GLUCAGON SENSITIVITY, SECRETION, AND TURNOVER AND GLYCOGEN METABOLISM IN PBH, IN ORDER TO DEVELOP AN ENHANCED, MORE EFFECTIVE, CLOSED-LOOP GLUCAGON ALGORITHM AND CLINICAL THERAPEUTIC SYSTEM RESPONSIVE TO GLYCEMIC EXCURSIONS WHICH OCCUR NOT ONLY DURING THE POSTPRANDIAL PERIOD, BUT ALSO DURING OVERNIGHT HOURS AND DURING/AFTER ACTIVITY. TO ACHIEVE THIS GOAL, WE HAVE ASSEMBLED A MULTIDISCIPLINARY TEAM INCLUDING CLINICAL INVESTIGATORS WITH EXPERTISE IN PBH AND GLUCAGON AND GLUCOSE METABOLISM, AND BIOMEDICAL ENGINEERS WITH EXPERTISE AND AN ESTABLISHED TRACK RECORD IN CLOSED-LOOP PUMP SYSTEM DEVELOPMENT AND CLINICAL TESTING AND IMPLEMENTATION. THUS, THE PROPOSED STUDIES WILL ADDRESS AN IMPORTANT KNOWLEDGE AND THERAPEUTIC GAP IN PBH, AND PROVIDE FOUR INNOVATIVE DELIVERABLES TO ADVANCE OUR UNDERSTANDING OF AND THE CLINICAL CARE OF THIS CHALLENGING PATIENT POPULATION: (1) KEY QUANTITATIVE DATA FOR GLUCAGON SENSITIVITY, SECRETION AND TURNOVER AND GLYCOGEN METABOLISM AS CONTRIBUTORS TO HYPOGLYCEMIA DURING POSTPRANDIAL, POSTABSORPTIVE, AND POST-ACTIVITY STATES IN PBH, (2) DEVELOPMENT OF AN IN SILICO MODEL OF THE PBH PATIENT, ALLOWING IMPLEMENTATION AND TESTING OF A NOVEL CLG ALGORITHM, (3) IMPLEMENTATION OF THE CLG ALGORITHM IN A SYSTEM DRIVING ON-BODY PUMP DELIVERY OF MICRODOSES OF GLUCAGON GUIDED BY CGM DATA, AND (4) AFTER REGULATORY APPROVAL, COMPLETION OF A PILOT RANDOMIZED OUTPATIENT, HOTEL-BASED CLINICAL TRIAL OF THE CLG SYSTEM IN PATIENTS WITH PBH.

TRANSLATIONAL PROGRAMMING OF BETA-CELLS IN RESPONSE TO GLUCOSE TOXICITY - ABSTRACT CONTROL OF INSULIN BIOSYNTHESIS IN BETA-CELLS OCCURS PRIMARILY AT THE LEVEL OF TRANSLATION OF THE INSULIN MRNA. AN ACUTE RISE IN GLUCOSE LEVELS STIMULATES INSULIN TRANSLATION BY A PROCESS THAT INVOLVES BINDING OF PROTEINS TO THE 5' UNTRANSLATED REGION OF THE INSULIN MRNA. SIMULTANEOUS TRANSLATIONAL UPREGULATION OF OTHER SECRETORY GRANULE PROTEINS ALSO CONTRIBUTES TO GLUCOSE-STIMULATED INSULIN SECRETION AND PROVIDES EVIDENCE THAT BETA-CELLS COORDINATELY REGULATE TRANSLATION OF FUNCTIONALLY RELATED MRNAS. BY CONTRAST, EXPOSURE OF BETA-CELLS TO SUSTAINED HIGH GLUCOSE SUPPRESSES GLUCOSE-STIMULATED INSULIN TRANSLATION AND SECRETION. THE MECHANISMS RESPONSIBLE FOR DYSREGULATION OF INSULIN TRANSLATION AND WHETHER THE DELETERIOUS EFFECT OF CHRONIC HIGH GLUCOSE IMPACTS TRANSLATION OF OTHER PROTEINS ARE NOT KNOWN. THIS PROJECT WILL TEST THE HYPOTHESIS THAT GLUCOSE TOXICITY LEADS TO EARLY ALTERATIONS IN MRNA-SPECIFIC BETA-CELL TRANSLATION THAT ARE MEDIATED BY INTRINSIC PROPERTIES OF MRNAS AND BY DYNAMIC REMODELING OF THE RIBOSOME ITSELF. FURTHERMORE, WE PROPOSE THIS PROGRAMMATIC TRANSLATIONAL DYSREGULATION CONTRIBUTES TO IMPAIRED GLUCOSE-STIMULATED INSULIN SECRETION. KNOWLEDGE GAINED FROM THIS STUDY HAS THE POTENTIAL TO IDENTIFY NOVEL THERAPEUTIC TARGETS FOR PREVENTION OF PROGRESSIVE BETA-CELL FAILURE IN DIABETES AND FOR OPTIMIZING FUNCTIONALITY OF EX VIVO GENERATED BETA-CELLS FOR CELL REPLACEMENT THERAPY.

ROLE OF SLC2A2/GLUT2 IN EMBRYO AND STEM CELL METABOLISM, SELF-RENEWAL, AND PATHWAYS INVOLVED IN DIABETIC EMBRYOPATHY

ELUCIDATING AND TARGETING BETA-CELL SENESCENCE AND ITS SASP - ABSTRACT THIS PROPOSAL SEEKS TO ELUCIDATE THE MECHANISMS OF SS-CELL SENESCENCE, AN AGING HALLMARK, AS A CONTRIBUTOR TO TYPE 2 DIABETES (T2D) AND IDENTIFY OPTIMAL THERAPEUTIC TARGETS. PANCREATIC INSULIN SECRETING SS-CELLS, CRUCIAL TO GLUCOSE HOMEOSTASIS, ARE HEAVILY SECRETORY CELLS, EQUIPPED TO RESPOND TO SMALL CHANGES IN BLOOD GLUCOSE LEVELS AND HIGHLY SUSCEPTIBLE TO STRESS BY NUTRIENT OVERLOAD. MY WORK HAS IDENTIFIED THAT MOUSE AND HUMAN SS- CELLS UNDERGO SENESCENCE IN RESPONSE TO INSULIN RESISTANCE (IR), LEADING TO LOSS OF CELLULAR IDENTITY, IMPAIRED FUNCTION AND SECRETION OF A UNIQUE SENESCENCE-ASSOCIATED SECRETORY PHENOTYPE (SASP). ADDITIONALLY, I SHOWED THAT SENOLYSIS IMPROVED BLOOD GLUCOSE LEVELS AND RECOVERY OF SS-CELL FUNCTION AND IDENTITY. I HYPOTHESIZE THAT CELLULAR SENESCENCE AND ITS SASP ARE TARGETABLE DRIVERS OF SS-CELL DYSFUNCTION AND LOSS. MY GOALS ARE TO UNDERSTAND THE MECHANISMS BEHIND SS-CELL SENESCENCE AND IDENTIFY THE OPTIMAL THERAPEUTIC STRATEGY. AIM 1. IDENTIFY THE CELL AUTONOMOUS DRIVER(S) OF SS-CELL SENESCENCE AND ITS FUNCTIONAL EFFECTS. BASED ON OUR MODELS OF IR AND DNA DAMAGE, WE HYPOTHESIZE THAT CYCLIN-DEPENDENT KINASE INHIBITOR P21CIP1 IS UPREGULATED EARLY IN SS-CELL SENESCENCE AND IS FOLLOWED BY P16INK4A. GENETIC GAIN- AND LOSS-OF-FUNCTION STRATEGIES WILL BE USED TO COMPARE THE EFFECTS OF P21CPI1 AND P16INK4A ON MOUSE AND HUMAN SS-CELL FUNCTION, IDENTITY AND SASP. ADDITIONALLY, THE FUNCTIONAL CHANGES OF SENESCENT CELLS WILL BE PINPOINTED. THIS AIM WILL DEFINE THE CELL AUTONOMOUS MOLECULAR MECHANISM(S) THAT DRIVE SS-CELL SENESCENCE AND ITS FUNCTIONAL CONSEQUENCES. AIM 2. ELUCIDATE THE NON-CELL AUTONOMOUS EFFECTS OF THE SS-CELL SASP. THE HYPOTHESIS IS THAT SS-CELL SENESCENCE CAN BE DRIVEN BY A NON-CELL AUTONOMOUS MECHANISM THROUGH SASP FACTORS, CAPABLE OF IMPAIRING THE FUNCTION AND GENE IDENTITY OF NEIGHBORING CELLS AND PRECIPITATING THEIR ENTRY INTO SENESCENCE. TO EVALUATE THE EFFECTS OF SASP UPON NEIGHBORING SS-CELLS, WE WILL TEST THE EFFECTS OF THE OVERALL AND SPECIFIC SELECTED FACTORS ON INSULIN SECRETION, SENESCENCE STATUS AND GENE EXPRESSION. ADDITIONALLY, WE WILL TEST THE TEMPORO-SPATIAL EFFECTS OF SASP ON NEIGHBORING CELLS USING OUR P21CIP1-DTOMATO RED MIP:GFP REPORTER MICE. THIS AIM WILL TEST THE EFFECTS OF SS-CELL SASP ON NON-SENESCENT CELLS. AIM 3. COMPARE THE EFFECTS OF SENOLYTIC AND SENOMORPHIC DRUGS IN THE RECOVERY OF SS-CELL FUNCTION AND IDENTITY. IN OUR PREVIOUS STUDIES, SENOLYSIS EFFECTIVELY RESTORED SS-CELL FUNCTION AND IDENTITY BUT A DECREASE IN THE NUMBER OF SENESCENT CELLS (SENOLYSIS) MAY BE DETRIMENTAL TO AN ALREADY INADEQUATE BETA CELL MASS, SO PERHAPS ONLY INHIBITING THEIR SASP (SENOMORPHIC EFFECT) WOULD RENDER SIMILAR BENEFICIAL RESULTS. THE HYPOTHESIS IS THAT SENORMORPHIC DRUGS WILL RESTORE SS-CELL FUNCTION AND IDENTITY WITHOUT IMPACTING CELL MASS. THIS AIM WILL COMPARE THE EFFECTS OF SENOLYTIC AND SENOMORPHIC DRUGS ON ISLETS OF HUMAN DONORS THAT HAVE ONE OR MORE OF THE FOLLOWING CHARACTERISTICS IMPAIRING GLUCOSE METABOLISM: OLDER AGE, IR AND T2D. THIS AIM SEEKS TO IDENTIFY THE OPTIMAL PHARMACOLOGICAL MECHANISMS TO RECOVER THE FUNCTION AND CELLULAR IDENTITY WITHOUT MASS IMPAIRMENT.

KIDNEY TRANSPLANT OUTCOMES AND APOL1

CHARACTERIZATION OF RISK FACTORS FOR EXCESSIVE CARDIOVASCULAR DISEASES (CVD) FROM CIRCULATING AND CARDIOVASCULAR TISSUES OF PEOPLE WITH WELL-CONTROLLED CHRONIC TYPE 1 DIABETES - ABSTRACT PEOPLE WITH TYPE 1 DIABETES (T1D) HAVE EXCESSIVE RISKS FOR CARDIOVASCULAR DISEASES (CVD), EVEN WHEN THEY HAVE WELL-CONTROLLED TRADITIONAL CVD RISK FACTORS, SUCH AS HYPERTENSION, HYPERLIPIDEMIA, AND HYPERGLYCEMIA, WITHOUT NEPHROPATHY (DN) OR SIGNIFICANT SYSTEMIC INSULIN RESISTANCE (IR). THE PRESENCE OF CHRONIC AUTOIMMUNITY IN T1D MAY PLAY A ROLE SINCE MANY AUTOIMMUNE DISEASES ARE ASSOCIATED WITH INCREASED CVD RISK. THE LACK OF KNOWLEDGE REGARDING THE PATHOLOGIES OF CVD IN T1D IS PARTIALLY DUE TO THE DIFFICULTY OF OBTAINING CARDIOVASCULAR TISSUES FROM PEOPLE WITH T1D AND THE LACK OF TRUE MICE MODELS OF T1D AND ATHEROSCLEROSIS. HOWEVER, THE AVAILABILITY OF ARTERIES FROM T1D, T2D AND NON-DIABETIC SUBJECTS FROM THE CARE-T1D AND MEDALISTS’ (T1D FOR >50 YEARS) REPOSITORIES HAVE MADE IT POSSIBLE TO HAVE DETAILED COMPARATIVE ANALYSIS OF POTENTIAL DIFFERENCES AMONGST THEM. CVD STUDIES OF MEDALISTS BY HISTORY AND CORONARY ARTERY CALCIUM (CAC) SCORE SHOWED ACCELERATED ATHEROSCLEROSIS DESPITE WELL- MANAGED DIABETES AND ABSENCE OF DN. ADVANCED GLYCATION END-PRODUCTS (AGES) AND INFLAMMATORY CYTOKINES, IL1Β AND INTERFERON-Γ CORRELATED TO CVD HISTORY NOVEL AUTOANTIBODIES (AABS) TO NFKBIB AND LOSS OF INSULIN SIGNALING WERE FOUND IN ARTERIES IN BOTH PEOPLE AND MICE (APOE-/-/NOD) WITH T1D. PLASMA METABOLOMICS SHOWED DIFFERENT AND OPPOSITE CHANGES IN T1D AND T2D, IN PATHWAYS RELATED TO ADAPTIVE IMMUNITY AND CVD. SINGLE CELL SEQUENCING (SCRNA) OF AORTA FROM DIABETIC MOUSE MODELS OF ATHEROSCLEROSIS DUE TO DELETION OF INSULIN RECEPTORS SHOWED ELEVATED INFLAMMATORY VASCULAR SMOOTH MUSCLE CELLS. WE POSTULATED THE NOVEL IDEA THAT CHRONIC AUTOIMMUNITY IN T1D CAN PROMOTE INFLAMMATORY PROCESSES IN THE ARTERIAL WALL, CAUSING SELECTIVE INSULIN RESISTANCE ONLY IN THE ARTERIES TO ACCELERATE ATHEROSCLEROSIS. TO TEST THIS HYPOTHESIS, WE PROPOSE: SP. AIM 1:TO COMPARE THE PATHOLOGIES, CELLULAR COMPOSITION (USING SPATIAL TRANSCRIPTOMICS) AND INSULIN SIGNALING OF THE ATHEROSCLEROTIC PLAQUES FROM PEOPLE T1D, VS T2D AND NON-DM USING TISSUES FROM THE CARE-T1D. SP. AIM 2: TO CHARACTERIZE DIFFERENCES IN CIRCULATORY NOVEL ANTI-VASCULAR AUTOANTIBODIES, INFLAMMATORY CYTOKINES AND METABOLITES OF PEOPLE WITH T1D, MONOGENIC DM, T2D AND NON-DM WHICH CAN BE CORRELATED TO SEVERITY OF CAC OR CVD HISTORY. SP. AIM 3: TO TEST DEFINITELY WHETHER ENHANCING INSULIN ACTIONS TARGETED TO ENDOTHELIAL CELLS (EC) CAN DECREASE INNATE AND ADAPTIVE IMMUNITY INDUCED INFLAMMATION AT THE ARTERIAL WALL AND ATHEROSCLEROSIS, USING T1D MOUSE, ECIRS1-/-/APOE-/-/NOD MICE.

FEEDING DRIVES HSF1 TRANSCRIPTIONAL PROGRAMS REQUIRED FOR GLOBAL PROTEIN SYNTHESIS

REGULTION OF SERINE/THREONINE KINASES IN SKELETAL MUSCLE

IDENTIFICATION OF NOVEL LIPID MEDIATORS WITH ANTI-DIABETIC EFFECT

INTERROGATION OF DYNAMIC RNA MODIFICATIONS IN BETA CELLS IN TYPE 1 DIABETES

SEARCH FOR THE MOLECULAR CAUSES OF DIABETIC EMBRYOPATHY

PROTECTIVE FACTORS AGAINST THE DEVELOPMENT OF MICROVASCULAR COMPLICATIONS

ROLE OF HYPERGLYCEMIA IN INTRACEREBRAL HEMORRHAGE

PHYSIOLOGICAL FACTORS DRIVING MATURATION OF NEONATAL BETA CELLS

MAPPING GENES FOR PROTEINURIA IN TYPE II DIABETES

EXERCISE AND SKELETAL MUSCLE SIGNALING MECHANISMS

EXERCISE REGULATION OF GLUCOSE METABOLISM IN SKELETAL MUSCLE

ORGANIZATION, TIME MANAGEMENT, AND DIABETES SELF-CARE

IN VIVO GENE EDITING OF HEMATOPOIETIC STEM AND PROGENITOR CELLS

AGING OF THE BETA CELL AND TYPE 2 DIABETES

INTRINSIC ANTI-INFLAMMATORY ACTIONS OF AMPK ACTIVATION RENDERED THROUGH A NOVEL IMMUNOMETABOLIC AMPK-MTOR-NFKB PATHWAY

PREDICTING OUTCOMES & ANTI-VEGF RESPONSE IN DIABETIC EYES BY ADAPTIVE OPTICS SLO

CELL BIOLOGY OF DIABETIC MICROVASCULAR COMPLICATIONS

MODULATING THE ADAPTIVE IMMUNE RESPONSE TO ENHANCE THE EFFICIENCY OF IN VIVO GENE EDITING IN MUSCLE - PROJECT SUMMARY DUCHENNE MUSCULAR DYSTROPHY (DMD) IS A DEBILITATING AND INCURABLE MUSCLE WASTING DISEASE CAUSED BY INHERITED MUTATIONS IN THE DMD GENE, WHICH ENCODES THE MUSCLE STRUCTURAL PROTEIN DYSTROPHIN. THERAPEUTIC APPROACHES THAT SEEK TO CURE DMD MUST INCLUDE A STRATEGY TO REPAIR OR REPLACE THE MUTANT DYSTROPHIN PROTEIN. A RECENT SERIES OF PAPERS FROM MY LAB AND OTHERS DEMONSTRATED THE POTENTIAL FEASIBILITY OF USING ADENO- ASSOCIATED VIRUS (AAV) MEDIATED DELIVERY OF CRISPR-CAS9 NUCLEASES TO MAKE SITE SPECIFIC DNA CLEAVAGES IN THE DMD GENE WITHIN SKELETAL MUSCLE FIBERS, CARDIOMYOCTYES AND MUSCLE STEM CELLS OF LIVING ANIMALS. WE HAVE SHOWN THAT THIS IN VIVO GENE EDITING STRATEGY IS SUFFICIENT TO RESTORE DMD READING FRAME AND ENABLE PRODUCTION OF FUNCTIONAL DYSTROPHIN PROTEIN THAT CAN INCREASE MUSCLE SPECIFIC FORCE AND PROTECT DMD MUSCLE FROM CONTRACTION-MEDIATED DAMAGE. THIS STRATEGY PRESENTS SOME ADVANTAGES WITH RESPECT TO OTHER “EXON SKIPPING” APPROACHES, IN THAT IT ALLOWS EFFICIENT RECOVERY OF DYSTROPHIN PROTEIN IN BOTH THE SKELETAL MUSCLE AND HEART, AND, BECAUSE IT EFFECTIVELY TARGETS MUSCLE STEM CELLS AND IRREVERSIBLY MODIFIES THE DMD LOCUS, IT OFFERS POTENTIALLY PERMANENT RESTORATION OF DYSTROPHIN EXPRESSION IN DYSTROPHIC TISSUES. YET, DESPITE ITS PROMISE, THERE REMAIN SIGNIFICANT POTENTIAL CHALLENGES FOR CLINICAL APPLICATION OF IN VIVO GENE EDITING. IN PARTICULAR, THE BACTERIALLY-DERIVED CAS9 NUCLEASE REPRESENTS A POTENTIAL ANTIGENIC TARGET FOR THE HOST IMMUNE RESPONSE. IN FACT, OUR PRELIMINARY DATA INDICATE THAT CAS9 IS RAPIDLY AND ROBUSTLY RECOGNIZED BY THE IMMUNE SYSTEM WHEN ECTOPICALLY EXPRESSED IN THE MUSCLE OF IMMUNE COMPETENT MICE. IN ADDITION, ANALYSES OF HUMAN SERA INDICATE THAT MORE THAN HALF OF POTENTIAL HUMAN RECIPIENTS OF CAS9 THERAPIES HARBOR PRE-EXISTING ANTIBODY AND T CELL RESPONSES TO CAS9 ORTHOLOGS DUE TO PRIOR INFECTION WITH THE BACTERIA FROM WHICH THESE PROTEINS DERIVE. THUS, INDUCED OR PRE-EXISTING ANTI-CAS9 IMMUNE RESPONSES MAY LOWER THE EFFICIENCY OF THERAPEUTIC GENE EDITING IN MUSCLE BY CAUSING THE IMMUNE-MEDIATED ELIMINATION OF CAS9 TRANSDUCED, GENE- EDITED CELLS. MOREOVER, EMERGING DATA INDICATING THAT THE IMMUNE SYSTEM AND ITS PRODUCTS CAN MODULATE THE EXPRESSION OF AAV-ENCODED TRANSGENES AND OF COMPONENTS OF CELLULAR DNA DAMAGE RESPONSE PATHWAYS RAISE THE POSSIBILITY THAT ANTI-CAS9 IMMUNITY COULD ALTER BOTH THE DEGREE OF ON-TARGET DMD EDITING AS WELL AS THE FREQUENCY AND TYPES OF OFF-TARGET MODIFICATIONS INDUCED. IN THIS PROJECT, WE WILL TEST THE HYPOTHESES THAT ROBUST ANTI-CAS9 RESPONSES REDUCE THE PRODUCTION AND DURABILITY OF GENE-EDITED CELLS AND ALTER THE FREQUENCY OF POTENTIALLY GENOTOXIC EDITS, WHEREAS OPPOSING CAS9 IMMUNITY THROUGH GENETIC OR PHARMACOLOGIC MANIPULATION WILL INCREASE THE EFFICIENCY, SAFETY AND DURABILITY OF DYSTROPHIN RECOVERY BY GENE EDITING. THIS WORK WILL ESTABLISH WHETHER THE ANTI-CAS9 IMMUNE RESPONSE HELPS OR HINDERS IN VIVO GENE EDITING AND WILL BE CRITICALLY IMPORTANT FOR THE DESIGN OF FUTURE CLINICAL EFFORTS USING THIS APPROACH.

IDENTIFYING BIOMARKERS OF ALZHEIMERS IN INSULIN RESISTANT PATIENTS USING MRI

MEDIATORS AND MODIFIERS OF NF-KB IN INSULIN RESISTANCE

MATERNAL EXERCISE DURING LACTATION TO IGNITE INFANT METABOLISM - PROJECT SUMMARY/ABSTRACT BREASTFEEDING REDUCES RISK OF CHILDHOOD OBESITY AND DIABETES, BUT THE MECHANISMS REMAIN UNCLEAR. WE RECENTLY IDENTIFIED THAT 12,13-DIHOME, A SIGNALING LIPID CRUCIAL FOR ACTIVATING BROWN ADIPOCYTE THERMOGENESIS, IS PRESENT IN HUMAN MILK. WE REPORTED THAT HIGHER CONCENTRATION OF 12,13-DIHOME IN MILK AT 1-MO POSTPARTUM WAS ASSOCIATED WITH LOWER INFANT FAT MASS. FURTHERMORE, MILK LEVELS OF 12,13-DIHOME, AND OTHER METABOLITES INVOLVED IN ADIPOCYTE THERMOGENESIS, WERE SIGNIFICANTLY INCREASED AFTER MATERNAL EXERCISE. THUS, WE HYPOTHESIZE THAT MATERNAL EXERCISE DURING LACTATION PROTECTS AGAINST CHILDHOOD OBESITY BY UPREGULATING THERMOGENIC METABOLITES AND LIPIDS IN MILK. WE PROPOSE TO SYSTEMATICALLY COMPARE THE ACUTE VERSUS CHRONIC EFFECTS OF MATERNAL EXERCISE ON HUMAN MILK COMPOSITION, AND TO TEST HOW MATERNAL EXERCISE-INDUCED CHANGES IN MILK INFLUENCE METABOLIC ACTIVITY AND BODY COMPOSITION IN INFANTS. IN AIM 1, WE WILL MEASURE CHANGES IN THERMOGENIC METABOLITES AND LIPIDS IN HUMAN MILK IN RESPONSE TO A SINGLE EXERCISE SESSION AND TO AN 8-WEEK PHYSICAL ACTIVITY INTERVENTION. WE WILL RANDOMIZE 100 EXCLUSIVELY BREASTFEEDING MOTHER-INFANT PAIRS TO EITHER AN “ACTIVE” GROUP (TARGETING >8,000 STEPS/DAY) OR A CONTROL” GROUP (STANDARD OF CARE), WITH DAILY ACTIVITY MONITORING FROM 1 TO 3 MONTHS POSTPARTUM. ALL PARTICIPANTS WILL COMPLETE A SUPERVISED ACUTE EXERCISE BOUT AT 1 AND 3 MONTHS POSTPARTUM. MILK SAMPLES COLLECTED BEFORE AND AFTER EXERCISE WILL BE USED FOR METABOLOMIC AND LIPIDOMIC ANALYSES. IN AIM 1A, WE WILL DETERMINE HOW MILK COMPOSITION IS AFFECTED BY THE ACTIVITY INTERVENTION BY COMPARING THE “ACTIVE” VS. “CONTROL” GROUPS. THE GOAL OF AIM 1B IS TO MEASURE ACUTE CHANGES IN MILK COMPOSITION IN RESPONSE TO A SINGLE EXERCISE BOUT. WE WILL ALSO DETERMINE WHETHER THE 8-WEEK ACTIVITY PROGRAM ALTERS MILK COMPOSITION RESPONSES TO THE SINGLE EXERCISE SESSION. IN AIM 2, WE WILL TEST WHETHER CHRONIC AND ACUTE MATERNAL EXERCISE ALTERATIONS OF MILK-DERIVED MAMMARY EPITHELIAL CELLS (MECS) MEDIATE ADAPTIVE CHANGES IN HUMAN MILK COMPOSITION. BUILDING ON OUR PRELIMINARY DATA SHOWING THAT MRNA SIGNATURES IN MECS ARE ALTERED BY MATERNAL OBESITY AND EXERCISE, WE WILL ANALYZE MRNA SIGNATURES AND PROTEIN CONTENT FOR KEY ENZYMES AND SIGNAL TRANSDUCTION PATHWAYS FOR LIPID AND NUCLEOTIDE SUGAR METABOLITES, USING SAMPLES FROM THE ACUTE AND CHRONIC ACTIVITY EXPERIMENTS. IN AIM 3, WE WILL LINK MATERNAL EXERCISE-INDUCED CHANGES IN HUMAN MILK TO METABOLISM AND BODY COMPOSITION IN INFANTS OF PARTICIPANTS RECRUITED IN AIM 1. THE GOAL OF AIM 3A IS TO DEFINE THE IMMEDIATE METABOLIC IMPACT ON INFANTS FED WITH “AFTER-EXERCISE MILK” RELATIVE TO A FEEDING WITH “BEFORE-EXERCISE MILK”. THIS WILL INVOLVE MEASURING THERMOGENIC ACTIVATION VIA INFRARED THERMOGRAPHY, ENERGY EXPENDITURE WITH INDIRECT CALORIMETRY, AND ANALYZING THE INFANT PLASMA METABOLOME AND LIPIDOME AT 1 AND 3 MONTHS. IN AIM 3B, WE WILL COMPARE ADIPOSITY IN INFANTS OF MOTHERS IN THE ACTIVE VS. CONTROL GROUPS IN THE FIRST YEAR OF LIFE; OUTCOMES WILL INCLUDE BODY COMPOSITION (PEA POD AND DXA), BEIGE/BROWN FAT ANATOMY (MRI), AND INFANT GROWTH (Z-BMI). THESE STUDIES WILL PROVIDE KEY DATA TO GUIDE A LARGER CLINICAL TRIAL OF EXERCISE DURING LACTATION.

EVALUATING CLINICAL AND PATHOLOGICAL MARKERS OF COGNITIVE DECLINE AND ALZHEIMER'S DEMENTIA IN LONG-DURATION TYPE 1 DIABETES. - ABSTRACT THE BURDEN OF DIABETES-ASSOCIATED COGNITIVE DYSFUNCTION IS ON THE RISE AS BOTH TYPE 1 (T1D) AND TYPE 2 (T2D) DIABETES INCREASE WORLDWIDE, AND PEOPLE AFFECTED ARE LIVING LONGER. HOWEVER, WHILE COGNITIVE DYSFUNCTION, ALZHEIMER’S’ DISEASE AND RELATED DISORDERS (AD/ADRD) HAVE BEEN EXTENSIVELY STUDIED IN T2D, WITHIN T1D THIS REMAINS UNDERSTUDIED AND RESTRICTED TO YOUNGER POPULATIONS. DETAILED CLINICAL CHARACTERIZATION OF COGNITIVE DYSFUNCTION IN A LARGE COHORT WITH LONG-DURATION T1D HAS BEEN LIMITED GIVEN THAT LIVING LONGER THAN 55 YEARS IN THIS POPULATION IS ONLY A RECENT PHENOMENON. THIS PROPOSAL FOCUSES ON THE NECESSARY NEED TO UNDERSTAND SPECIFIC PATHOLOGY AND MECHANISMS UNDERLYING COGNITIVE DECLINE IN THOSE WITH LONG-DURATION T1D TO UNCOVER NOVEL PATHWAYS FOR INTERVENTION AND PREVENTION IN THIS POPULATION. THE JOSLIN 50-YEAR MEDALIST STUDY (“MEDALISTS”), A WELL-CHARACTERIZED COHORT OF INDIVIDUALS WITH ≥50 YEARS OF T1D, OFFERS A UNIQUE OPPORTUNITY TO STUDY POTENTIAL MARKERS OF COGNITIVE DECLINE AND AD/ADRD IN LONG-DURATION T1D. PRELIMINARY STUDIES SHOW THAT MEDALISTS HAD WORSE COGNITIVE FUNCTION AND LOWER BRAIN VOLUMES COMPARED TO NON-DIABETIC CONTROLS. SIGNIFICANT ASSOCIATIONS BETWEEN COGNITIVE IMPAIRMENT AND RETINAL NEURAL AND VASCULATURE CHANGES (ASSESSED BY OPTICAL COHERENCE TOMOGRAPHY [OCT] AND OCT-ANGIOGRAPHY [OCTA]) WERE ALSO OBSERVED. ADDITIONALLY, POST-MORTEM BRAIN HISTOPATHOLOGY REVEALED ONLY MILD-MODERATE AD-RELATED OR VASCULAR PATHOLOGY, UNLIKE THOSE WITH TYPE 2 DIABETES (T2D), SUPPORTING THE URGENT NEED TO BETTER CHARACTERIZE THE UNDERLYING PATHOPHYSIOLOGY OF COGNITIVE DECLINE IN T1D. THE OVERARCHING GOAL OF THIS R01 PROPOSAL IS TO ELUCIDATE THE DISTINCT PATHOPHYSIOLOGICAL MECHANISMS DRIVING COGNITIVE DECLINE IN PEOPLE WITH LONG-DURATION T1D, AND IDENTIFY PREDICTIVE MARKERS AND PATHWAYS THAT CAN INFORM IMPROVED SCREENING AND MANAGEMENT STRATEGIES IN THE FUTURE. SPECIFIC AIM 1. TO EVALUATE FUNCTIONAL AND PATHOLOGICAL MARKERS OF ALZHEIMER’S DISEASE AND COGNITIVE DECLINE IN LONG-DURATION T1D VIA COMPREHENSIVE CROSS-SECTIONAL AND LONGITUDINAL CLINICAL AND COGNITIVE ASSESSMENTS, BRAIN IMAGING, AND POST-MORTEM BRAIN GROSS AND HISTOPATHOLOGICAL EXAMINATIONS. WE AIM TO RECRUIT AN ADDITIONAL 100 MEDALISTS EACH FOR THE CROSS-SECTIONAL (TOTAL N=322), AND LONGITUDINAL (TOTAL N=148) STUDIES, AND TO PROCURE 25 ADDITIONAL POST-MORTEM BRAINS FOR HISTOPATHOLOGY (TOTAL N=51). A SUBSET OF MEDALISTS (N=20) WILL UNDERGO AMYLOID PET SCANS. PLASMA P-TAU217 WILL ALSO BE ASSAYED. THE ALZHEIMER’S DISEASE NEUROIMAGING INITIATIVE (ADNI) AND RUSH ALZHEIMER'S DISEASE CENTER (RADC) DATABASES WILL BE LEVERAGED FOR COMPARISONS WITH NON-DIABETIC INDIVIDUALS AND THOSE WITH T2D. SPECIFIC AIM 2. TO EVALUATE THE RELATIONSHIPS BETWEEN RETINAL NEUROVASCULATURE AND COGNITIVE DECLINE AND ALZHEIMER’S DISEASE IN LONG-DURATION T1D. THE 100 PARTICIPANTS WILL ALSO UNDERGO RETINAL IMAGING (OCT/OCTA) (TOTAL N=222).

XANTHINE OXIDASE INHIBITION RAISES INTRACELLULAR PURINES TO ACTIVATE AMPK, IMPROVE GLUCOSE CONTROL AND DECREASE FATTY LIVER AND ATHEROSCLEROSIS IN TY

EXERCISE REGULATION OF HUMAN ADIPOSE TISSUE

REGULATION OF C. ELEGANS SKN-1/NRF ACTIVITY BY THE UNFOLDED PROTEIN RESPONSE

ZEISS LASER SCAN MICROSCOPE (LSM) 510-DUO

CARDIAC AUTOIMMUNITY AS A MEDIATOR OF CARDIOVASCULAR OUTCOMES IN TYPE 1 DIABETES

RIPE3B1, AN IMPORTANT INSULIN GENE TRANSCRIPTION FACTOR

CARDIAC AUTOANTIBODIES AS BIOMARKERS FOR HEART DISEASE IN TYPE 1 DIABETES

TARGETING NEUTROPHILS AND LEUKOTRIENES TO TREAT TYPE 2 DIABETES

A PILOT STUDY OF FENOFIBRATE TO PREVENT KIDNEY FUNCTION LOSS IN TYPE 1 DIABETES - SUMMARY DESPITE IMPROVEMENTS IN THE PAST 20 YEARS IN GLYCEMIC AND BLOOD PRESSURE CONTROL, AND THE INTRODUCTION OF “RENO-PROTECTIVE” DRUGS SUCH AS RENIN-ANGIOTENSIN SYSTEM BLOCKERS (RASB), THE OVERALL INCIDENCE OF END- STAGE KIDNEY DISEASE (ESKD) IN TYPE 1 DIABETES (T1D) REMAINS HIGH. TO SEEK NEW TREATMENTS TO PREVENT DIABETIC KIDNEY DISEASE (DKD) AND/OR SLOW ITS PROGRESSION TO ESKD IN T1D, WE HAVE ESTABLISHED A UNIQUE CONSORTIUM OF HIGH-QUALITY ACADEMIC CENTERS, WHICH WE HAVE NAMED PERL (PREVENTING EARLY RENAL FUNCTION LOSS IN DIABETES) TO EMPHASIZE THE FOCUS ON INTERVENING RELATIVELY EARLY IN THE COURSE OF DKD IN T1D, WHEN RENAL DAMAGE CAN MORE LIKELY BE SLOWED OR STOPPED. FINDINGS FROM THE FIELD AND ACCORD TRIALS SUGGEST A RENO-PROTECTIVE EFFECT OF THE PPAR-ALPHA AGONIST FENOFIBRATE, RAISING THE EXCITING POSSIBILITY OF USING THIS INEXPENSIVE GENERIC DRUG TO PREVENT GFR DECLINE IN PERSONS WITH T1D. THESE DATA, HOWEVER, WERE OBTAINED THROUGH POST-HOC ANALYSES OF T2D POPULATIONS WITH CLINICAL CHARACTERISTICS OPTIMIZED FOR CVD STUDIES. THUS, A CLINICAL TRIAL SPECIFICALLY DESIGNED TO EVALUATE EFFECTS ON GFR DECLINE IS REQUIRED TO FIRMLY ESTABLISH A DKD INDICATION FOR FENOFIBRATE IN T1D. AS A FIRST STEP, AND IN RESPONSE TO FOA PAS-20-160 “SMALL R01S FOR CLINICAL TRIALS TARGETING DISEASES WITHIN THE MISSION OF NIDDK”, WE HAVE DESIGNED A PILOT STUDY INCLUDING 40 PARTICIPANTS WITH T1D AND EARLY-TO-MODERATE DKD, AT HIGH RISK OF ESKD, WHO WILL BE ENROLLED AT TWO OF THE PERL SITES AND RANDOMIZED IN A 1:1 RATIO TO TREATMENT WITH FENOFIBRATE OR PLACEBO FOR 18 MONTHS. THROUGH THIS PILOT STUDY WE WILL: 1. DEFINE THE NATURE OF THE ACUTE EFFECT OF FENOFIBRATE ON KIDNEY FUNCTION. IT REMAINS UNCLEAR WHETHER THE EGFR REDUCTION OBSERVED AT THE BEGINNING OF FENOFIBRATE TREATMENT IS AN ARTIFACT OF FENOFIBRATE-INDUCED CHANGES IN CREATININE PRODUCTION AND/OR RENAL TUBULAR HANDLING, OR CORRESPONDS TO AN ACTUAL REDUCTION IN GFR. WE WILL RESOLVE THIS CONTROVERSY, WHICH HAS CRUCIAL IMPLICATIONS FOR THE PIVOTAL TRIAL DESIGN, BY DIRECTLY MEASURING GFR BY PLASMA IOHEXOL DISAPPEARANCE – A METHODOLOGY IN WHICH PERL SITES ARE EXPERIENCED. 2. GENERATE FURTHER DATA ON THE LONG-TERM EFFECTS OF FENOFIBRATE ON GFR DECLINE IN PERSONS WITH T1D AND DKD WHO ARE AT HIGH RISK OF RAPID GFR DECLINE AND ESKD. THE POSITIVE EFFECTS OF FENOFIBRATE IN FIELD AND ACCORD WERE OBSERVED IN INDIVIDUALS WHO WERE NOT SELECTED FOR HAVING DKD AND WHO, IF UNTREATED, HAD A MEAN GFR DECLINE BARELY ABOVE THE PHYSIOLOGICAL DECLINE DUE TO AGING. TO MAKE A COMPELLING CASE FOR A PIVOTAL TRIAL FOR KIDNEY OUTCOMES, IT IS CRUCIAL TO GENERATE PRELIMINARY DATA ON THE EFFECTIVENESS AND SAFETY OF THIS DRUG IN PERSONS SELECTED FOR HAVING DKD AND BEING RAPIDLY PROGRESSING TOWARDS ESKD. 3. DETERMINE THE EFFECTS OF FENOFIBRATE ON BIOMARKERS OF INCREASED RISK OF FAST GFR DECLINE. A SALUTARY EFFECT OF FENOFIBRATE ON ONE OR MORE OF THESE BIOMARKERS WILL CORROBORATE ANY TREND OF A FENOFIBRATE BENEFIT IDENTIFIED IN AIM 2. WITH THE RESULTS OF THIS PILOT, WE WILL BE OPTIMALLY PREPARED TO APPLY TO NIDDK FOR SUPPORT OF A PIVOTAL TRIAL TO ESTABLISH A KIDNEY INDICATION FOR FENOFIBRATE IN T1D.

NEURAL PATHWAYS OF HYPOGLYCEMIA UNAWARENESS USING FMRI

IMAGING STRATEGIES TO MEASURE BROWN FAT AND ITS ACTIVITY

MECHANISMS OF EXERCISE EFFECTS IN OBESE HUMANS: SEX-SPECIFIC REGULATION OF NOVEL ADIPOSE TISSUE FUNCTION

IDENTIFYING TARGETS FOR PREVENTING NEUROCOGNITIVE COMPLICATIONS IN YOUTH WITH T1D - PROJECT SUMMARY/ABSTRACT EARLY CHILDHOOD IS A CRITICAL TIME IN BRAIN DEVELOPMENT FOR LATER HEALTH AND MATURATION. DURING THIS PERIOD OF RAPID BRAIN DEVELOPMENT, THE BRAIN MAY BE ESPECIALLY VULNERABLE TO DYSGLYCEMIA AND OTHER METABOLIC CONDITIONS ASSOCIATED WITH TYPE 1 DIABETES (T1D). THE CURRENT PROJECT SEEKS TO EVALUATE NEUROCOGNITIVE COMPLICATIONS IN PRE-PUBERTAL ONSET T1D AND IDENTIFY POTENTIAL RISK AND PROTECTIVE FACTORS OF BRAIN HEALTH. INVESTIGATORS AT JOSLIN DIABETES CENTER (JDC) AND BOSTON CHILDREN’S HOSPITAL (BCH) SEEK TO BECOME A JOINT CLINICAL CENTER IN THE PROPOSED CONSORTIUM TO INVESTIGATE NEUROCOGNITIVE COMPLICATIONS OF PEDIATRIC T1D. IN ADDITION, WE WILL COLLABORATE WITH BOSTON MEDICAL CENTER, AN INNER CITY SAFETY NET HOSPITAL, TO ENSURE INCLUSION OF UNDER-REPRESENTED AND UNDER- RESOURCED PARTICIPANTS. WE HAVE ASSEMBLED A MULTIDISCIPLINARY TEAM WITH EXPERTISE IN CLINICAL RESEARCH AND ADVANCED DIABETES TECHNOLOGY USE IN PEDIATRIC T1D, HEALTH EQUITY, PSYCHOSOCIAL AND COGNITIVE IMPLICATIONS OF T1D, PEDIATRIC NEUROCOGNITIVE TESTING, PEDIATRIC NEUROIMAGING, AND BRAIN DEVELOPMENT. IN ADDITION, WE HAVE ENGAGED A DIVERSE GROUP OF STAKEHOLDERS WHO WILL ACTIVELY PARTICIPATE IN THE DEVELOPMENT, DESIGN, AND EXECUTION OF THE STUDY AS WELL AS DISSEMINATION OF RESULTS. IN THIS CONSORTIUM, WE PROPOSE TO RECRUIT A PROSPECTIVE COHORT OF 600 (60 AT THE COMBINED JDC AND BCH SITE) PRE-PUBERTAL CHILDREN, AGED 4-8 YEARS WITH NEWLY DIAGNOSED T1D, REPRESENTATIVE OF THE RACIAL, ETHNIC, AND SOCIOECONOMIC DIVERSITY OF THE US, AND A NORMATIVE CONTROL GROUP OF 300 (30 AT JDC/BCH) HEALTHY CHILDREN MATCHED FOR AGE, SEX, RACE, AND ETHNICITY. CHILDREN WILL UNDERGO STRUCTURAL, DIFFUSION, AND RESTING STATE FUNCTIONAL MRI, NEUROCOGNITIVE AND PSYCHOLOGICAL ASSESSMENTS, AND CLINICAL EVALUATIONS SHORTLY AFTER DIAGNOSIS AND LONGITUDINALLY FOR 2 YEARS. STRUCTURAL AND FUNCTIONAL MRI MEASURES AND NEUROCOGNITIVE AND PSYCHOLOGICAL OUTCOMES OF CHILDREN WITH T1D WILL BE COMPARED WITH MATCHED NEUROTYPICALLY DEVELOPING CHILDREN FROM OUR NORMATIVE SAMPLE, SUPPLEMENTED WITH PUBLICLY AVAILABLE DATA IN LARGE PEDIATRIC MRI REPOSITORIES. TO FACILITATE COMPARISONS, IMAGING PROTOCOLS WILL BE ADAPTED FROM PUBLISHED STANDARD PROTOCOLS; INSTRUMENTS FOR ASSESSING COGNITIVE FUNCTION, PSYCHOSOCIAL VARIABLES, AND SOCIAL DETERMINANTS OF HEALTH AND COGNITION HAVE BEEN CURATED FROM STANDARDIZED COLLECTIONS COMPLEMENTED WITH SPECIFIC VALIDATED INSTRUMENTS. CLINICAL AND DIABETES MANAGEMENT DATA WILL BE ASSESSED VIA RECORD REVIEW AND CLOUD-BASED DATA CAPTURE INTEGRATING DEVICE DATA FROM CONTINUOUS GLUCOSE MONITORS (CGM), INSULIN PUMPS AND PENS, AND ACTIVITY MONITORS. AS A MEMBER OF THIS CONSORTIUM: WE AIM: (1) TO DESCRIBE THE IMPACT OF A PRE-PUBERTAL DIAGNOSIS OF T1D ON BRAIN DEVELOPMENT; (2) TO LINK FUNCTIONAL AND STRUCTURAL BRAIN CHANGES TO CLINICAL AND PERFORMANCE OUTCOMES; AND (3) TO ASSESS DETERMINANTS OF BRAIN HEALTH AND IDENTIFY TARGETS FOR PREVENTION OR EARLY INTERVENTION. IN THIS MPI APPLICATION, WE HOPE TO ADVANCE UNDERSTANDING OF CHILDHOOD BRAIN DEVELOPMENT, WITH POTENTIAL FOR DIRECT IMPLICATIONS FOR THE CLINICAL MANAGEMENT OF PEDIATRIC T1D TO PRESERVE NEUROCOGNITION.

TOLL RECEPTOR ACTIVATION OF NF-KB IN INSULIN RESISTANCE AND T2DM

TARGETING RENALASE FOR EARLY STAGE T1D TREATMENT - PROJECT SUMMARY/ABSTRACT TYPE 1 DIABETES (T1D) IS AN AUTOIMMUNE DISEASE THAT DESTROYS INSULIN-PRODUCING BETA CELLS IN THE PANCREAS, REQUIRING INSULIN INJECTIONS AND BLOOD GLUCOSE MONITORING, WHICH DO NOT REPLICATE THE PRECISE GLYCEMIC CONTROL OF FUNCTIONAL BETA CELLS NOR PREVENT DISEASE PROGRESSION AND COMPLICATIONS. IN LONG-STANDING T1D, MOST BETA CELLS ARE DESTROYED, WHICH REQUIRES THE REPLENISHMENT OF BETA CELL MASS TO RESTORE INSULIN PRODUCTION. THIS CAN BE ACHIEVED BY REGENERATING ENDOGENOUS BETA CELLS OR DIFFERENTIATING HUMAN EMBRYONIC STEM CELLS (ESCS) OR INDUCED PLURIPOTENT STEM CELLS (IPSCS) INTO BETA CELLS FOR TRANSPLANTATION, COMBINED WITH THERAPIES TO PREVENT AUTOIMMUNE DESTRUCTION OF THE NEWLY FORMED BETA CELLS. DIFFERENT THERAPEUTIC WINDOWS EXIST FOR INTERVENTION OR A POTENTIAL CURE, PARTICULARLY IN EARLY-ONSET OR NEWLY DIAGNOSED T1D WHERE SIGNIFICANT BETA CELLS REMAIN, PRESENTING AN OPPORTUNITY TO INTERVENE AND PRESERVE THESE CELLS, POTENTIALLY DELAYING OR PREVENTING FURTHER BETA CELL LOSS. RECENT FINDINGS SUGGEST THAT BETA CELL INJURY OR STRESS MAY SIGNIFICANTLY CONTRIBUTE TO IMMUNE- MEDIATED BETA CELL LOSS IN T1D, THUS THERAPIES AIMED AT REDUCING BETA CELL STRESS AND INJURY MAY AVERT IMMUNE TARGETING DURING THE PROGRESSION TO OVERT T1D. COMBINING BETA CELL THERAPIES WITH IMMUNE MODULATION MAY YIELD BETTER OUTCOMES, YET THE SPECIFIC TARGETS AND PATHWAYS FOR EFFECTIVE TREATMENT REMAIN UNCLEAR. RENALASE (RNLS) HAS EMERGED AS A PROMISING THERAPEUTIC TARGET IN T1D, BEING ASSOCIATED WITH T1D IN GENOME-WIDE ASSOCIATION STUDIES (GWAS) AND LINKED TO BETA CELL PROTECTION. LOSS OF RNLS FUNCTION IN BETA CELLS REDUCES ENDOPLASMIC RETICULUM (ER) AND OXIDATIVE STRESS, IMMUNE CELL INFILTRATION, AND NATURAL KILLER (NK) CELL ACTIVATION, PREVENTING AUTOIMMUNE DESTRUCTION. DESIGNING A BETTER STRATEGY FOR TARGETING RNLS ENZYMATIC ACTIVITY MAY OFFER A POTENTIAL THERAPEUTIC STRATEGY FOR T1D BY PROVIDING BETA CELL PROTECTION AGAINST STRESS AND AUTOIMMUNITY IN HUMANS. RNLS, KNOWN AS AN OXIDASE SIMILAR TO MONOAMINE OXIDASES (MAO), CAN BE BOUND BY SOME MAO INHIBITORS. WE FOUND THAT ONE OF THE FDA APPROVED MAO INHIBITORS, PARGYLINE, IS ABLE TO BIND TO RNLS AND PROTECT PANCREATIC BETA CELLS FROM STRESS AND AUTOIMMUNE DESTRUCTION. APPARENTLY, PARGYLINE MAY NOT BE SPECIFIC OR POTENT ENOUGH FOR RNLS. THEREFORE, DEVELOPING A MORE SPECIFIC AND POTENT RNLS INHIBITOR IS CRUCIAL. UTILIZING STRUCTURE-BASED DRUG DESIGN, THE GOAL IS TO CREATE A NEW CLASS OF COMPOUNDS FOR EARLY OR PREVENTIVE TREATMENT OF T1D. THIS RESEARCH INCLUDES UNCOVERING RNLS'S ROLE IN BETA CELL METABOLISM AND IMMUNE INTERACTIONS, CHARACTERIZING RNLS STRUCTURE AND ENZYMATIC FUNCTION FOR ROBUST ASSAY DEVELOPMENT, AND EVALUATING RNLS INHIBITORS BY BIOCHEMICAL, CELL, AND ANIMAL MODEL-BASED ASSAYS. THE EXPECTED OUTCOME IS THE DEVELOPMENT OF POTENT AND SELECTIVE RNLS INHIBITORS THAT ENHANCE BETA CELL SURVIVAL AND FUNCTION, REDUCE STRESS AND AUTOIMMUNE DESTRUCTION, AND DEMONSTRATE SAFETY AND EFFICACY IN HUMANIZED MOUSE MODELS, PROVIDING PROMISING THERAPEUTIC OPTIONS FOR T1D BY PROTECTING BETA CELLS FROM STRESS AND IMMUNE ATTACKS AT AN EARLY STAGE.

JOSLIN-BIDMC POSTBAC PROGRAM IN DIABETES AND METABOLISM - ABSTRACT THE UNITED STATES STRUGGLES TO CULTIVATE A BIOMEDICAL RESEARCH WORKFORCE THAT REFLECTS THE DIVERSITY OF ITS POPULATION. THIS CHALLENGE IS PARTICULARLY STRIKING IN THE FIELD OF DIABETES AND METABOLIC DISORDERS, WHICH DISPROPORTIONATELY IMPACTS RACIAL AND ETHNIC MINORITIES AND PEOPLE FROM DISADVANTAGED BACKGROUNDS. WORKFORCE DIVERSITY IS AN IMPORTANT GOAL FOR SOCIAL JUSTICE AND CONTRIBUTES TO ORGANIZATIONAL EFFECTIVENESS THROUGH INCREASED CREATIVITY, BROADER FOUNDATIONS FOR DECISION-MAKING, AND A SENSE OF INCLUSION AND BELONGING IN THE WORKPLACE. IN THE FIELD OF DIABETES AND METABOLISM, A DIVERSE WORKFORCE IS ALSO CRITICAL FOR BROADENING PARTICIPATION IN CLINICAL STUDIES. TO ADDRESS THIS PROBLEM, WE PROPOSE A NEW JOSLIN-BETH ISRAEL DEACONESS MEDICAL CENTER (BIDMC) POST-BAC PROGRAM IN DIABETES AND METABOLISM TO ATTRACT TALENTED INDIVIDUALS FROM DIVERSE BACKGROUNDS TO PURSUE FURTHER TRAINING AND ULTIMATELY, CAREERS IN BIOMEDICAL RESEARCH IN METABOLIC DISEASES. THIS PROGRAM WILL LEVERAGE JOSLIN AND BIDMC’S BROAD AND DEEP EXPERTISE IN DIABETES AND METABOLISM AND A HIGHLY EXPERIENCED CADRE OF MENTORS WHO WILL PROVIDE PARTICIPANTS WITH DEDICATED RESEARCH EXPERIENCES, MENTORING, COURSEWORK, AND PROFESSIONAL DEVELOPMENT ACTIVITIES. THE OVERALL GOAL OF OUR PROGRAM IS TO PROVIDE PARTICIPANTS WITH THE SCIENTIFIC, TECHNICAL, AND PROFESSIONAL SKILLS THEY WILL NEED TO PURSUE DOCTORAL DEGREES AND THE INSPIRATION TO DEDICATE THEIR TALENTS TO PREVENTION, TREATMENT, AND CURE OF DIABETES AND RELATED METABOLIC DISORDERS. TO EFFECTIVELY IDENTIFY PARTICIPANTS FROM DIVERSE BACKGROUNDS, OUR PROGRAM WILL PARTNER WITH THE BIOMEDICAL SCIENCES CAREER PROGRAM, A BOSTON-BASED NON-PROFIT ORGANIZATION DEDICATED TO PROVIDING MENTORSHIP AND CAREER DEVELOPMENT PROGRAMS TO INDIVIDUALS FROM UNDERREPRESENTED GROUPS AND DISADVANTAGED BACKGROUNDS TO PROMOTE THEIR SUCCESS IN BIOMEDICAL SCIENCE. WE WILL ALSO REACH POTENTIAL APPLICANTS THROUGH THE ONLINE PLATFORM, HANDSHAKE AND BY WORKING WITH THE UNIVERSITY OF MASSACHUSETTS AMHERST COLLEGE OF NATURAL SCIENCES CAREER & PROFESSIONAL DEVELOPMENT CENTER AND HARVARD COLLEGE LIFE SCIENCE RESEARCH ADVISORS.

AUTOIMMUNE MECHANISMS IN THE PROGRESSION OF CVD IN T1D

HUMAN BROWN ADIPOSE TISSUE AND ITS IMPACT ON METABOLISM

PATHWAYS OF MOTHER TO CHILD TRANSMISSION OF METABOLIC RISK

EFFECTS OF IKKB AND NFKB IN INSULIN RESISTANCE

ADIPOCYTE INSULIN SIGNALING IN METABOLISM AND AGING

ROLE OF BRADYKININ RECEPTOR 2 IN TPA STROKE THERAPY

THE ROLE OF PKCEPSILON IN DIABETIC RETINOPATHY

SEARCH OF DIABETES NEPHROPATHY GENES IN TYPE 1 DIABETES

SMALL ANIMAL HIGH RESOLUTION METABOLIC ANALYSIS SYSTEM

PRECISION MONITORING: UNDERSTANDING MOMENTARY AFFECT, GLUCOSE, AND SELF-CARE BEHAVIORS IN ADOLESCENTS AND YOUNG ADULTS WITH TYPE 1 DIABETES - PROJECT SUMMARY/ABSTRACT ADOLESCENTS AND YOUNG ADULTS (AYA) WITH TYPE 1 DIABETES (T1D) HAVE THE HIGHEST HEMOGLOBIN A1C VALUES OF ALL ACROSS THE LIFESPAN. CONTINUOUS GLUCOSE MONITORING (CGM) IS A MODERN APPROACH TO GLUCOSE ASSESSMENT THAT PROVIDES IN-THE-MOMENT INFORMATION ABOUT GLUCOSE LEVELS TO HELP OPTIMIZE TIME-IN-RANGE ([TIR] 70-180 MG/DL). RETROSPECTIVE REVIEW OF AMBULATORY GLUCOSE PATTERNS IS AN ESSENTIAL COMPONENT FOR IMPROVING DIABETES SELF-CARE. HOWEVER, MANY CGM USERS BECOME OVERWHELMED BY THE VOLUMINOUS CGM DATA AND RARELY REVIEW OR DOWNLOAD THEIR CGM. FURTHERMORE, INTERPRETATION OF CGM DATA SHOULD NOT OCCUR IN A VACUUM, RATHER ONE SHOULD CONSIDER CONTEXTUAL FACTORS THAT IMPACT SELF-CARE IN ORDER TO IMPROVE GLYCEMIC REGULATION. THUS, TO MAXIMIZE THE CLINICAL VALUE OF CGM IN AYA WITH T1D, A NOVEL, ECOLOGICALLY GROUNDED APPROACH IS NEEDED TO CONTEXTUALIZE CGM DATA WITH INFORMATION ON THE INTRA- AND INTER-PERSONAL CONTEXT IN WHICH GLUCOSE REGULATION OCCURS. IT IS TIMELY TO PAIR ECOLOGICAL MOMENTARY ASSESSMENT (EMA) WITH CGM TO DEVELOP AN INTERVENTION PROGRAM TO IMPROVE SELF-CARE, GLYCEMIA, AND PSYCHOSOCIAL OUTCOMES. WE WILL USE RIGOROUS METHODS TO CAPTURE DYNAMIC RELATIONSHIPS BETWEEN IN-THE-MOMENT RATINGS OF EMOTIONAL AND SOCIAL EXPERIENCES (USING EMA) AND CGM-RECORDED GLUCOSE VALUES, WHICH WILL BE COLLECTIVELY DISPLAYED ON A DASHBOARD, ENCAPSULATING THE EMOTIONAL AND SOCIAL CONTEXT OF AMBULATORY GLUCOSE (PSYCHOSOCIAL AMBULATORY GLUCOSE [PAG]). TRACKING AND REVIEWING OF PAG PATTERNS, ALONG WITH ONGOING STRUCTURED SUPPORT, WITHIN A MULTICOMPONENT INTERVENTION, WILL HELP AYA WITH T1D TO BETTER INTERPRET AND BEHAVIORALLY RESPOND TO OUT-OF-RANGE GLUCOSE VALUES, AND MAY SERVE AS A CATALYST TO IMPROVE A1C, TIR, AND SELF-CARE, AS WELL AS REDUCE DIABETES DISTRESS. AMIT SHAPIRA, PHD, PROPOSES A SERIES OF STUDIES WITH AN OVERARCHING GOAL OF DEVELOPING, ITERATIVELY REFINING, AND TESTING A PAG INTERVENTION IN AYA WITH T1D. THE PROPOSED 3 SPECIFIC AIMS ARE: 1) TO IDENTIFY SALIENT COMPONENTS OF A PAG DASHBOARD USING EMA THAT WILL COORDINATE THE DISPLAY OF EMOTIONAL AND SOCIAL CONTEXT COUPLED WITH CGM RECORDED DATA TO ENHANCE SELF-CARE OF AYA WITH T1D ABOVE TARGET A1C VALUES USING MIXED METHODS; 2) TO ADAPT AND BUILD UPON THE DASHBOARD FOR USE IN A BEHAVIORAL INTERVENTION THROUGH EXPERT FEEDBACK AND GUIDANCE FROM PERTINENT STAKEHOLDERS; AND 3) TO EXAMINE IF A PILOT RCT OF A REFINED PAG BEHAVIORAL INTERVENTION WILL BE ACCEPTABLE, FEASIBLE, AND SATISFACTORY TO PARTICIPANTS AND HAVE POTENTIAL TO IMPROVE DIABETES AND PSYCHOSOCIAL OUTCOMES COMPARED WITH PAG-ALONE. RESULTS FROM THESE STUDIES WILL INFORM A FULLY POWERED RCT FOR A FUTURE R01 APPLICATION. DR. SHAPIRA’S CAREER OBJECTIVE IS TO BECOME AN INDEPENDENT DIABETES BEHAVIORAL RESEARCHER FOCUSED ON OPTIMIZING SELF-CARE, INCLUDING USE OF CGM AND OTHER TECHNOLOGIES, TO IMPROVE OUTCOMES FOR PEOPLE WITH T1D. THE 5-YEAR, MENTORED CAREER DEVELOPMENT PROGRAM WILL PROVIDE HER WITH NECESSARY SKILLS IN DIABETES CARE (E.G., DIABETES TECHNOLOGIES), ADVANCED STATISTICAL METHODS, AND USABILITY AND QUALITATIVE RESEARCH TO LAUNCH HER INDEPENDENCE IN DIABETES BEHAVIORAL INVESTIGATION.

DEVELOPMENT OF AN INTERVENTION TO TARGET DIABETES DEVICE USE THROUGH AN IDENTITY FRAMEWORK: ACCPTECH (ADAPT AND COMMIT TO CGM AND PUMP TECHNOLOGY) - PROJECT SUMMARY/ABSTRACT DIABETES TECHNOLOGIES TO TREAT TYPE 1 DIABETES (T1D) ARE RAPIDLY ADVANCING: CONTINUOUS GLUCOSE MONITORS (CGMS) PROVIDE DATA ON GLUCOSE LEVELS AND TRENDS THROUGHOUT THE DAY WITHOUT THE BURDEN OF FINGERSTICKS; INSULIN PUMPS INCREASE EASE OF TAKING INSULIN WITHOUT INJECTIONS; AND HYBRID CLOSED-LOOP SYSTEMS AUTOMATE INSULIN DELIVERY BY SYNCING AN INSULIN PUMP AND CGM THROUGH A DOSING ALGORITHM TO ALLOW FOR REDUCED USER RESPONSIBILITIES. HOWEVER, DESPITE THESE ADVANCES, USE OF DIABETES TECHNOLOGIES REMAINS PARTICULARLY LOW IN YOUNG ADULTS (YAS) WITH T1D, AND MANY YAS EXPERIENCE SUBOPTIMAL GLUCOSE LEVELS, WHICH PLACES SUBSTANTIAL RISKS UPON THEIR LONG-TERM HEALTH. YAS REPORT THE MOST COMMON REASONS FOR THEIR AVOIDANCE OR DISCONTINUATION OF DIABETES DEVICE USE INCLUDE NOT LIKING TO WEAR ANYTHING ON THEIR BODY, DISLIKE OF HOW THE DEVICES LOOKS ON THEIR BODY, AND WORRIES ABOUT WHAT OTHERS MAY THINK. THESE CONCERNS AROUND PERSONAL AND SOCIAL PERCEPTIONS INTERSECT A MAJOR DEVELOPMENTAL TASK OF YOUNG ADULTHOOD: IDENTITY DEVELOPMENT. THIS K23 APPLICATION, LED BY DR. COMMISSARIAT, PROPOSES TO ADDRESS THE POTENTIAL CHALLENGES OF TECHNOLOGY USE BY APPLYING A FRAMEWORK OF HOW YAS “INCORPORATE” T1D INTO IDENTITY (E.G., THROUGH STIGMA MANAGEMENT, ADJUSTMENT TO PERCEIVED INTERFERENCE, AND BENEFIT-FINDING) IN ORDER TO DESIGN AND EVALUATE A NOVEL INTERVENTION TO INCREASE UPTAKE AND SUSTAINED USE OF DIABETES TECHNOLOGIES. THE 3 AIMS OF THE PROPOSED PROJECT ARE TO: 1) EXPLORE TRENDS OF TECHNOLOGY USE OVER TIME IN YAS USING ELECTRONIC HEALTH RECORD DATA; 2) IDENTIFY CORE INTERVENTION TARGETS THROUGH QUALITATIVE ANALYSIS OF THE INTERSECTION BETWEEN TECHNOLOGY AND IDENTITY; AND 3) CREATE, PILOT, AND EVALUATE THE ADAPT AND COMMIT TO CGM AND PUMP TECHNOLOGY (ACCPTECH) INTERVENTION TO ENHANCE DIABETES DEVICE USE IN YAS WITH T1D. THE PRIMARY OUTCOMES OF THE ACCPTECH PILOT RCT ARE FEASIBILITY AND ACCEPTABILITY, AND PRELIMINARY DATA WILL CONTRIBUTE TO OUR UNDERSTANDING OF HOW IDENTITY RELATES TO DEVICE USE, AND WHETHER ADDRESSING IDENTITY CHALLENGES CAN ENHANCE DEVICE USE IN YAS. THESE RESULTS WILL INFORM A FUTURE R01 APPLICATION OF A FULLY-POWERED RANDOMIZED CONTROLLED TRIAL TO ENHANCE DEVICE UPTAKE, USE, AND SUBSEQUENT GLYCEMIC AND PSYCHOSOCIAL OUTCOMES IN YAS WITH T1D. THE MENTORSHIP, EDUCATION, TRAINING, AND RESOURCES OUTLINED IN THIS PROPOSAL WILL SUPPORT DR. COMMISSARIAT’S CAREER DEVELOPMENT AND HER GOAL TO BECOME A SUCCESSFUL, INDEPENDENT BEHAVIORAL RESEARCHER FOCUSED ON UTILIZING DIABETES IDENTITY AS A MEANS TO IMPROVE DIABETES-RELATED MANAGEMENT AND OUTCOMES IN CHILDREN, ADOLESCENTS, AND YOUNG ADULTS WITH T1D.

MAPPING GENES FOR END-STAGE RENAL DISEASE IN TYPE 1 DIABETES

IMPROVING MANAGEMENT STRATEGIES FOR BLOOD PRESSURE AND LIPIDS IN PEDIATRIC T1D

COPAS BIOSORT FLOW CYTOMETER

IDENTIFYING METABOLIC MECHANISMS THAT REGULATE APPETITE AND FOODINTAKE - PROJECT SUMMARY UNDERSTANDING HOW APPETITE AND FOOD CONSUMPTION ARE REGULATED IS CRITICAL TO THE AGING FIELD. FROM AN INNOVATIVE HIGH-THROUGHPUT C. ELEGANS SCREEN OF TRANSCRIPTION FACTORS, IN WHICH WE IDENTIFIED REGULATORS OF FOOD CONSUMPTION (HERE TERMED FEEDING), WE DETERMINED THAT FEEDING IS DRAMATICALLY REDUCED BY KNOCKDOWN OF CRH-1 (CREB), WHICH IS INHIBITED BY AMP-ACTIVATED PROTEIN KINASE (AMPK). AMPK IS A KEY SENSOR OF LOW ENERGY STATES THAT INHIBITS GROWTH SIGNALS AND PROMOTES CATABOLIC PROCESSES AND IS OF GREAT INTEREST IN THE AGING FIELD. OUR EXCITING PRELIMINARY FINDINGS INDICATE THAT AMPK AND CERTAIN OTHER METABOLIC SIGNALS REGULATE FEEDING IN UNEXPECTED WAYS AND HAVE REVEALED A NEW BEHAVIOR PATTERN THAT IS A MAJOR REGULATOR OF FEEDING. THEY SUGGEST THAT: (1) IN C. ELEGANS AMPK SIGNALS “HUNGER” BY ACTING IN MULTIPLE TISSUES AND IN PART BY INHIBITING CRH-1. AMPK THEREBY INDUCES THE ANIMAL TO DWELL ON FOOD (“EATING”) BUT ALSO TO PERCEIVE THAT THIS FOOD IS INADEQUATE, SO THAT WHEN POSSIBLE IT WILL LEAVE IN SEARCH OF BETTER FOOD, PARADOXICALLY REDUCING FOOD CONSUMPTION. (2) THIS FOOD-LEAVING BEHAVIOR, WHICH WE TERM METABOLIC FOOD AVERSION, IS ALSO TRIGGERED BY OTHER STATES OF PERCEIVED NUTRITIONAL INADEQUACY, INCLUDING LACK OF THE MONO-UNSATURATED FATTY ACID (MUFA) OLEIC ACID (OA) OR OTHER SPECIFIC FAS. (3) IN MOST BUT NOT ALL CASES METABOLIC FOOD AVERSION CAN BE AVERTED BY OA SUPPLEMENTATION, SUGGESTING THAT AN OA-DERIVED LIPID SIGNAL INDICATES SATIETY OR CORRECTS CERTAIN METABOLIC IMBALANCES. (4) LIKE FOOD DWELLING, METABOLIC FOOD AVERSION DEPENDS UPON SEROTONIN SIGNALING, BUT IS RELATED TO A BEHAVIOR WHEREBY C. ELEGANS AVOIDS FOOD THAT IT PERCEIVES TO BE PATHOGENIC. IN THIS EXPLORATORY PROJECT WE WILL TEST AND EXTEND THESE INTRIGUING MODELS THROUGH TWO AIMS. IN AIM 1 WE WILL IDENTIFY SIGNALS THAT MEDIATE AMPK/CRH-1-REGULATED HUNGER BEHAVIORS. WE WILL EXPLORE HOW AMPK AND CRH-1 EXPRESSION IN DIFFERENT TISSUES INFLUENCES FEEDING AND INVESTIGATE THE INVOLVEMENT OF WELL- CHARACTERIZED SEROTONIN-MEDIATED, FOOD-INDUCED, AND OTHER SIGNALS IN THEIR REGULATION OF FOOD DWELLING AND AVERSION. IN AIM 2, WE WILL LEVERAGE TARGETED SCREENING AND FOLLOW-UP ANALYSES TO ELUCIDATE METABOLIC MECHANISMS THAT REGULATE FOOD AVERSION AND DWELLING. WE WILL COMPLETE A MEDIUM-SCALE GENE KNOCKDOWN SCREEN TO IDENTIFY METABOLIC PERTURBATIONS THAT INDUCE AVERSION THAT IS OR IS NOT SUPPRESSIBLE BY OA. WE WILL INVESTIGATE WHETHER AVERSION IS GENERALLY PAIRED WITH INCREASED FOOD DWELLING AND IS DEPENDENT UPON SIGNALING MECHANISMS IDENTIFIED IN AIM 1, AS IS TRUE FOR THE AMPK/CRH-1 PATHWAY. FINALLY, WE WILL ASK WHETHER SOME AVERSION EVENTS SIGNAL THROUGH AMPK AND BEGIN TO IDENTIFY TISSUES FROM WHICH AVERSION SIGNALS ORIGINATE. THIS PROJECT WILL IDENTIFY MECHANISMS THAT LINK METABOLIC DEFICITS TO SPECIFIC FEEDING BEHAVIORS: FOOD DWELLING AND SEEKING OF HIGHER QUALITY FOOD, PROVIDING FUNDAMENTAL BIOLOGICAL INSIGHTS AT A LEVEL OF CLARITY THAT WOULD BE POSSIBLE ONLY IN C. ELEGANS.

IDENTIFICATION OF PATHWAYS REQUIRED FOR HIGH GLUCOSE-INDUCED OXIDATIVE STRESS ASSOCIATED WITH DIABETIC EMBRYOPATHY BY CRISPR-GENERATED MUTATION SCREEN

IMPACT OF METABOLIC HEALTH IN TYPE 2 DIABETES ON SPERM EPIGENETIC MARKS IN HUMANS: A NOVEL INTERGENERATIONAL MECHANISM FOR METABOLIC DISEASE RISK TRANSMISSION

REGULATION OF SKN-1/NRF FUNCTIONS BY GERMLINE STEM CELLS

IMMUNE REGULATORY FUNCTION OF CD5

A PILOT CLINICAL TRIAL TO ASSESS FEASIBILITY, FACILITATORS AND BARRIERS OF CONTINUOUS GLUCOSE MONITORING IN ASIAN AMERICANS WITH TYPE 2 DIABETES - PROJECT SUMMARY/ABSTRACT RATES OF TYPE 2 DIABETES (T2D) ARE INCREASING BOTH NATIONALLY AND GLOBALLY. IN ADDITION TO KNOWN T2D COMPLICATIONS SUCH AS RETINOPATHY, NEPHROPATHY, NEUROPATHY, AND CARDIOVASCULAR DISEASE, T2D IS KNOWN TO AFFECT COGNITIVE IMPAIRMENT AND EVEN SEVERITY OF COVID-19 INFECTION. A FEW STUDIES HAVE SHOWN THE BENEFIT OF CONTINUOUS GLUCOSE MONITORING (CGM) DEVICES FOR BETTER GLYCEMIC MANAGEMENT IN T2D POPULATIONS. WHILE PREVENTION AND MANAGEMENT PROTOCOLS FOR T2D ARE UBIQUITOUS, THERE CONTINUE TO EXIST LARGE RACIAL/ETHNIC DISPARITIES AMONGST THE GENERAL US POPULATION. ASIAN AMERICANS (AA), SPECIFICALLY CHINESE-AMERICANS, PRESENT WITH MUCH HIGHER T2D PREVALENCE AND FACE DISPARITIES IN T2D CARE FOR THE FOLLOWING REASONS: 1) STIGMA ARISING FROM THE “MODEL MINORITY MYTH”, EXACERBATED BY THE RISE IN ANTI-ASIAN SENTIMENTS DURING THE COVID-19 PANDEMIC; 2) HIGHER T2D UNAWARENESS RATE; 3) CULTURAL AND LANGUAGE BARRIERS INCLUDING LIMITED DIGITAL LITERACY AND ENGLISH PROFICIENCY; AND 4) HISTORICAL EXCLUSION FROM T2D STUDIES, INCLUDING THOSE ON CGM DEVICES AND T2D, IN WHICH CULTURALLY-RELEVANT FACILITATORS AND BARRIERS TO CGM USE HAVE YET TO BE EVALUATED IN AAS. THIS STUDY WILL SPECIFICALLY EXAMINE HOW T2D COULD BE BETTER MANAGED IN CHINESE AMERICANS THROUGH A CGM INTERVENTION, AS COMPARED TO STANDARD FINGERSTICK GLUCOSE MONITORING (FSGM). WE WILL BE CONDUCTING A 6- MONTH, SINGLE-SITE, OPEN-LABELED RANDOMIZED CONTROLLED TRIAL EXAMINING CGM VERSUS NO CGM (FSGM) USE IN 1ST-GENERATION CHINESE AMERICANS. OUR SPECIFIC AIMS ARE: SP. AIM 1: IN A PILOT 6-MONTH RANDOMIZED CLINICAL TRIAL, WE WILL EXAMINE THE IMPACT OF CGM USE VS. NO CGM AMONG 1ST GENERATION CHINESE AMERICANS WITH T2D. SUB-AIM 1.1. EVALUATE FEASIBILITY (ADHERENCE AND CONSISTENCY) AND QUALITY OF LIFE MEASURES DURING CGM USE IN THIS POPULATION. SUB-AIM 1.2. GENERATE PRECISION ESTIMATES OF THE DISTRIBUTION OF THE SECONDARY OUTCOMES (6- MONTH GLYCEMIC CONTROL AND LIPID MARKERS) IN BOTH ARMS TO INFORM A FUTURE RANDOMIZED CLINICAL TRIAL (RCT). SP. AIM 2: IDENTIFY MULTI-LEVEL BARRIERS AND FACILITATORS OF CGM USE FOR CHINESE AMERICANS WITH T2D, USING A SOCIOECOLOGICAL FRAMEWORK (PATIENT-LEVEL, PROVIDER-LEVEL, AND COMMUNITY/ENVIRONMENT LEVEL). WE WILL EVALUATE THE IMPLEMENTATION PROCESS (FACILITATORS AND IMPEDIMENTS), RESOURCE REQUIREMENTS, AND INTERMEDIATE PATIENT ADHERENCE OUTCOMES FOR THE PROGRAM USING MIXED-METHODS APPROACHES. THESE WILL INFORM DESIGN OF CULTURALLY- TAILORED INTERVENTION FOR A LARGER RANDOMIZED CONTROLLED TRIAL.

NOVEL MECHANISMS FOR EXERCISE TRAINING EFFECTS ON GLUCOSE HOMEOSTASIS

DETERMINATION OF THE ROLE OF ADIPOCYTE INFLAMMATION IN OBESITY-INDUCED INSULIN RE

CHARACTERIZATION OF THE INDUCIBLE-BETA-CELL-SPECIFIC P53 KNOCKOUT MICE

MECHANISM-BASED BIOMARKERS FOR GLUCOSE-LOWERING IN TINSAL-T2D

ENDOTHELIAL CELL INSULIN AND INCREASED INTESTINAL TUMOR FORMATION IN OBESITY

TRANSITIONS IN CARE: EXAMINING THE PROCESS FOR YOUTH WITH TYPE 1 DIABETES

DIABETES DETECTION, PREVENTION, WA AT THE JOSLIN DIABETES CENTER

SPECIAL BD LSRFORTESSA

DIABETES DETECTION AND PREVENTION PROGRAM

DIABETES DETECTION AND PREVENTION PROGRAM

INITIATION OF AUTOIMMUNE DIABETES

BIOMARKERS FOR PROTECTION AGAINST COMPLICATIOS AND BETA-CELL DESTRUCTION

ROLE OF IGF-1 AND INSULIN RECEPTORS BETA-CELL SURVIVAL

DURABILITY AND MECHANISMS OF BARIATRIC SURGERY VS MEDICAL THERAPY IN OBESE T2D

GENOTYPE-DEPENDENT CARDIOVASCULAR AND ANTI-INFLAMMATORY EFFECTS OF FENOFIBRATE

ROLE OF LEPTIN IN ISLET GROWTH

ELUCIDATING ADIPOSITY IN A MOUSE MODEL OF PRENATAL UNDERNUTRITION

MULTI-OMICS APPROACH TO UNDERSTANDING THE BENEFICIAL EFFECTS OF EXERCISE IN DIABETES

GENETIC MODIFIERS OF MHC CLASS II PRESENTATION IN AUTOIMMUNE DIABETES

NAIVE T CELL HOMEOSTASIS: TREG SELECTION AND SURVIVAL

A MULTICENTER CLINICAL TRIAL OF ALLOPURINOL TO PREVENT GFR LOSS IN T1D

PANCREATIC BETA-CELL GLUCOSE TOXICITY AND MRNA TRANSLATION - ABSTRACT PANCREATIC Β-CELLS ARE SPECIALIZED TO CARRY OUT HIGH CAPACITY MRNA TRANSLATION THAT IS TIGHTLY REGULATED BY THE NUTRIENT ENVIRONMENT. HOWEVER, THESE FEATURES EXPOSE Β-CELLS TO SEVERAL VULNERABILITIES, INCLUDING DELETERIOUS ENDOPLASMIC RETICULUM STRESS AND TRANSLATION ERRORS THAT CAN CAN RESULT IN PRODUCTION OF DEFECTIVE RIBOSOME PRODUCTS (DRIPS), WHICH HAVE BEEN IMPLICATED IN AUTOIMMUNITY IN TYPE 1 DIABETES (T1D). WHEREAS, ACUTE GLUCOSE EXPOSURE ROBUSTLY INCREASES TRANSLATION OF INSULIN AND OTHER SECRETORY GRANULE PROTEINS, CHRONIC HIGH GLUCOSE DECREASES INSULIN TRANSLATION AND SECRETION EVEN PRIOR TO IMPACT ON GLOBAL TRANSLATION OR UPREGULATION OF ENDOPLASMIC RETICULUM STRESS. TO DETERMINE THE EXTENT OF TRANSLATIONAL CHANGES AT THIS EARLY STAGE OF Β-CELL DYSFUNCTION, I USED RIBOSOME PROFILING AND NASCENT PROTEOMICS IN MIN6 INSULINOMA CELLS TO ELUCIDATE THE GENOME- WIDE IMPACT OF SUSTAINED HIGH GLUCOSE ON Β-CELL MRNA TRANSLATION. SUSTAINED HIGH GLUCOSE CONDITIONS THAT SUPPRESSED INSULIN SECRETION DOWNREGULATED TRANSLATION OF INSULIN AND PROTEINS, SUCH AS SCGN, IDH2, VPS41, SLC2A2, IGF2, SLC30A8 AND PFKFB3, WHICH ARE INVOLVED IN INSULIN SECRETORY GRANULE FORMATION, EXOCYTOSIS, AND METABOLISM-COUPLED INSULIN SECRETION. TRANSLATION OF THESE MRNAS WAS ALSO DOWNREGULATED IN PRIMARY RAT AND HUMAN ISLETS FOLLOWING EX-VIVO INCUBATION WITH CHRONIC HIGH GLUCOSE, AND IN AN IN VIVO PARTIAL PANCREATECTOMY MODEL OF CHRONIC HYPERGLYCEMIA. TRANSLATIONAL DOWNREGULATION DECREASED CELLULAR ABUNDANCE OF THESE PROTEINS. SUBSEQUENT ANALYSIS OF ACTIVELY TRANSLATING RIBOSOMES AND PATHWAYS THAT REGULATE MRNA QUALITY DURING TRANSLATION SHOWED THAT SUSTAINED HIGH GLUCOSE CHANGES THE PROTEIN COMPOSITION OF TRANSLATING RIBOSOMES AND SUPPRESSES NONSENSE MEDIATED RNA DECAY (NMD). ALTERED RIBOSOME COMPOSITION AND ACTIVITY OF RNA DECAY PATHWAYS COULD RESULT IN CHANGES IN THE ABUNDANCE AND FIDELITY OF THE PROTEINS PRODUCED. I HYPOTHESIZE THAT HYPERGLYCEMIA-INDUCED REMODELING OF RIBOSOMES AND SUPPRESSION OF NMD ALTER EXPRESSION OF KEY Β-CELL GENES AND INCREASE NEO-ANTIGEN PRODUCTION. IN THIS PROPOSAL I WILL INVESTIGATE THE EXTENT TO WHICH SUSTAINED HIGH GLUCOSE-REMODELED RIBOSOMES CONTRIBUTE TO DYSREGULATION OF MRNA TRANSLATION IN Β-CELLS AND DETERMINE THE IMPACT OF REDUCED RNA SURVEILLANCE AND ALTERED TRANSLATION ON NEOANTIGEN PRODUCTION AND AUTOIMMUNITY IN T1D. THESE STUDIES COULD UNCOVER NOVEL THERAPEUTIC TARGETS FOR PREVENTION OF PROGRESSIVE Β-CELL FAILURE IN T1D AND FOR OPTIMIZING FUNCTIONALITY OF EX VIVO GENERATED Β-CELLS FOR CELL REPLACEMENT THERAPY.

INTERROGATING THE ROLE OF M6A MRNA METHYLATION IN THE AGING OF THE ?-CELL AND DIABETES - PROJECT SUMMARY ALTHOUGH THE ETIOPATHOGENESIS OF TYPE 1 (T1D) AND TYPE 2 DIABETES (T2D) IS DIFFERENT, DECREASED FUNCTIONAL SS-CELL MASS IS A CENTRAL FEATURE IN BOTH DISEASES. CONCERTED RESEARCH EFFORTS AIM AT DEVELOPING STRATEGIES TO IMPROVE SS-CELL SURVIVAL AND RESTORE SS-CELL FUNCTION IN PATIENTS WITH DIABETES. CELLULAR SENESCENCE IS A MAJOR HALLMARK OF AGING. RECENT WORK HAS DEMONSTRATED THAT SS-CELL SENESCENCE IS A COMMON CONTRIBUTOR TO TYPE 1 DIABETES (T1D) AND TYPE 2 DIABETES (T2D). RNA MODIFICATIONS ARE EMERGING AS IMPORTANT MODULATORS OF MANY HALLMARKS OF AGING. WE HAVE DISCOVERED THAT M6A MRNA METHYLATION IS ESSENTIAL FOR SS-CELL FUNCTION AND SURVIVAL, AND RECENTLY CHARACTERIZED THE M6A LANDSCAPE OF HUMAN T1D AND T2D ISLETS. OUR PRELIMINARY DATA SHOWS THAT THE M6A ERASER ALKBH5 IS UPREGULATED IN AGED HUMAN SS-CELLS AND DIFFERENTIALLY M6A METHYLATED GENES IN T1D AND T2D ARE ENRICHED FOR DNA DAMAGE RESPONSE AND SENESCENCE PATHWAYS COMPARED TO CONTROLS. THE GOAL OF THIS APPLICATION IS TO USE STATE-OF-THE ART METHODS TO CHARACTERIZE THE M6A METHYLOME OF THE AGING SS- CELL AND IDENTIFY M6A-REGULATED PATHWAYS THAT UNDERLIE THE ACCELERATED SS-CELL SENESCENCE IN DIABETES. I WILL TEST THE OVERARCHING HYPOTHESIS THAT AGING SS-CELLS EXHIBIT TRANSCRIPT-SPECIFIC M6A HYPOMETHYLATION AND CONSEQUENT UPREGULATION OF MRNAS INVOLVED IN DRIVING CELL SENESCENCE, AND THAT M6A IS THEREFORE A VALUABLE THERAPEUTIC TARGET FOR DIABETES. IN SPECIFIC AIM 1, I WILL CHARACTERIZE THE TEMPORAL M6A LANDSCAPE OF THE AGING SS-CELL. COMPLETION OF AIM 1, WHICH WILL TAKE PLACE DURING K99 PHASE, WILL PROVIDE ME WITH TRAINING IN ASPECTS OF AGING BIOLOGY AND FURTHER EXPERIENCE WITH BIOINFORMATICS ANALYSIS OF M6A DATA NECESSARY TO INDEPENDENTLY COMPLETE THE R00 PHASE AND FUTURE R01 SUBMISSIONS. IN SPECIFIC AIM 2, WHICH WILL SPAN THE K99 AND R00 PHASES, I WILL IDENTIFY THE MOLECULAR TARGETS OF ALKBH5 IN THE HUMAN SS-CELL TRANSITION TO SENESCENCE. COMPLETION OF THE SUB- AIM OF AIM 2 PROPOSED DURING THE K99 PHASE WILL PROVIDE ME WITH THE TRAINING IN PHOTOACTIVATABLE RIBONUCLEOSIDE- ENHANCED CROSSLINKING AND IMMUNOPRECIPITATION (PAR-CLIP) SEQUENCING AND DATA ANALYSIS NECESSARY TO COMPLEMENT AIM 1, AND INDEPENDENTLY COMPLETE AIM 2 DURING THE R00 PHASE. FOR SPECIFIC AIM 3, I WILL TARGET M6A LEVELS TO REDUCE SS-CELL SENESCENCE AND IMPROVE DIABETES IN VIVO. COMPLETION OF THIS AIM WILL ALLOW ME TO VALIDATE CANDIDATE GENES IDENTIFIED IN AIM’S 1 AND 2 AND TO TEST IN VIVO THE ROLE OF ALKBH5 IN DRIVING THE ACCELERATED SENESCENCE SEEN IN SS-CELLS IN DIABETES. THESE EXPERIMENTS ARE NOVEL AS THEY COMBINE OMICS-BASED APPROACHES, MOUSE GENETIC MODELS, PHYSIOLOGY, AND MOLECULAR BIOLOGY TO EXAMINE THE MECHANISTIC ROLE OF M6A, AND PARTICULARLY ALKBH5, IN SS-CELL SENESCENCE TRANSITION IN DIABETES.

ROLE OF THE TRANSCRIPTIONAL COREGULATOR TRIP BR2 IN ADIPOCYTE METABOLISM

ROLE OF CA2+/CALMODULIN KINASES IN SKELETAL MUSCLE GLUCOSE TRANSPORT AND GROWTH.

FOXOS IN THE REGULATION OF MUSCLE MICROPHAGY

DRIVERS OF HUMAN INSULIN RESISTANCE AND SEX-SPECIFIC ALTERATIONS - PROJECT SUMMARY/ABSTRACT INSULIN RESISTANCE IS A MAJOR RISK FACTOR IN THE DEVELOPMENT OF TYPE 2 DIABETES (T2D) AND IS ALSO PRESENT IN ONE- QUARTER OF THE GENERAL POPULATION IN A SEX-DEPENDENT MANNER, WHICH PREDISPOSES THESE INDIVIDUALS TO THE DEVELOPMENT OF T2D. THE MOLECULAR DETERMINANTS UNDERLYING INSULIN RESISTANCE AND SEX-SPECIFIC IMPACT REMAIN ELUSIVE. MS-BASED GLOBAL PHOSPHOPROTEOMICS USING MYOBLASTS DERIVED FROM INDUCED PLURIPOTENT STEM CELLS (IMYOS) TAKEN FROM INSULIN SENSITIVE (I-SEN) AND INSULIN RESISTANT (I-RES) INDIVIDUALS SHOWED A LARGE NETWORK OF DYSREGULATED PHOSPHORYLATIONS LINKED TO DIFFERENCES IN INSULIN SENSITIVITY. IN ADDITION, TRANSCRIPTOMIC AND PROTEOMIC ANALYSES OF THESE CELLS REVEALED MANY GENES DIFFERENTIALLY REGULATED BY INSULIN RESISTANCE. WHILE MANY ALTERATIONS WERE OBSERVED IN CELLS OF BOTH SEXES, I ALSO IDENTIFIED MANY CHANGES AT THE PHOSPHOPROTEOME, PROTEOME, AND GENE EXPRESSION LEVEL THAT WERE MODIFIED BY THE SEX OF THE PATIENT. PRELIMINARY ANALYSIS SUGGESTS THAT DNA METHYLATION MAY CONTRIBUTE TO THE SEX-SPECIFIC GENE EXPRESSION DIFFERENCES, BUT ITS ROLE IN INSULIN RESISTANCE CHANGES REMAINS UNCLEAR. BASED ON THESE FINDINGS AND THE IMPORTANCE OF INSULIN RESISTANCE IN THE DEVELOPMENT OF T2D, THE OVERALL GOAL OF THE GRANT IS TO IDENTIFY THE PRIMARY DRIVERS OF ALTERED INSULIN SIGNALING AND IMPAIRED GLUCOSE UPTAKE ABILITY IN INSULIN RESISTANCE. TO ACHIEVE THIS GOAL, THE RESEARCH AIMS ARE TO IDENTIFY THE UPSTREAM REGULATORS OF INSULIN RESISTANCE INCLUDING IDENTIFYING THE KINASES DRIVING THE ALTERED PHOSPHORYLATION IN INSULIN RESISTANCE AND IDENTIFYING THE DRIVERS OF DEFECTIVE GLUCOSE UPTAKE USING AN UNBIASED CRISPR SCREENING APPROACH. PHOSPHOPROTEOME BASED KINOME ANALYSIS WILL BE PERFORMED TO DETERMINE HOW THE KINASE BASED SIGNALING ALTERATIONS ARE RELATED TO CHANGES IN INSULIN RESISTANCE AND CELLULAR METABOLISM. A LOSS-OF-FUNCTION GENOME-WIDE UNBIASED CRISPR SCREEN WILL BE PERFORMED TO IDENTIFY WHICH GENES/PROTEINS AND/OR POTENTIAL KINASES CONTRIBUTE TO IMPAIRED GLUCOSE UPTAKE CHARACTERISTIC OF INSULIN RESISTANCE. THIS RESEARCH PROPOSAL WILL IDENTIFY PRIMARY DRIVERS OF INSULIN RESISTANCE AND PROVIDE MECHANISTIC INSIGHTS INTO DIFFERENT RISKS OF DEVELOPING METABOLIC DISEASES BETWEEN MEN AND WOMEN. IN ADDITION TO RESEARCH WORK, TRAINING AND CAREER DEVELOPMENT OBJECTIVES WILL ALSO BE ACCOMPLISHED DURING THE K01 CAREER DEVELOPMENT AWARD, INCLUDING EXPANDING KNOWLEDGE IN METABOLISM AND DIABETES RESEARCH, DEVELOPING PROFICIENCY IN COMPUTATIONAL ANALYSIS OF SEQUENCING DATA, AND DEVELOPING LEADERSHIP AND PROFESSIONAL SKILLS. THE TRAINING ENVIRONMENT AT JOSLIN DIABETES CENTER IS PHENOMENAL, GIVEN THAT JOSLIN HAS BEEN A PREEMINENT CENTER FOR DIABETES RESEARCH FOR OVER 110 YEARS AND THE MENTOR, DR. C. RONALD KAHN, IS AN INTERNATIONALLY RECOGNIZED INVESTIGATOR IN THE FIELD OF DIABETES RESEARCH. I HAVE AN INTEGRATIVE AND MULTIDISCIPLINARY TRAINING OPPORTUNITY TO RECEIVE INTELLECTUAL AND CAREER DEVELOPMENT ADVICE.

THE FUNCTION AND REGULATION OF TBC1D1 IN SKELETAL MUSCLE METABOLISM.

STARS-SRF PATHWAY- A NOVEL REGULATOR OF MUSCLE METABOLISM AND INSULIN RESISTANCE

BMP7 AND THE REGULATION OF CENTRAL AND PERIPHERAL ENERGY BALANCE

DESIGN OF A MULTICENTER CLINICAL TRIAL OF ALLOPURINOL TO PREVENT GFR LOSS IN T1D

NOVEL ROLES OF ADIPOSE TISSUE IN EXERCISE TRAINING EFFECTS ON GLUCOSE HOMEOSTASIS

REPLICATION ANALYSIS TO IDENTIFY ACUTE MOLECULAR RESPONSES TO EXERCISE THAT LEAD TO CHRONIC TRAINING ADAPTATIONS - PROJECT SUMMARY/ABSTRACT THE MOLECULAR TRANSDUCERS OF PHYSICAL EXERCISE CONSORTIUM (MOTRPAC) IS TASKED WITH DEFINING THE MOLECULAR FOOTPRINT THAT UNDERLIES THE IMMENSELY BENEFICIAL EFFECTS OF EXERCISE ON HEALTH. THE ANIMAL COMPONENT OF MOTRPAC CONSISTS OF TWO MAJOR STUDIES: ONE INVESTIGATING EXERCISE TRAINING AND THE OTHER THE EFFECTS OF A SINGLE BOUT OF ACUTE EXERCISE. THESE STUDIES USE FISCHER 344 RATS AT 6 AND 18 MONTHS OF AGE TO DETERMINE THE MOLECULAR EFFECTS OF EXERCISE TRAINING AND ACUTE EXERCISE ACROSS 19 DIFFERENT TISSUES. SEVEN DIFFERENT OMICS PLATFORMS ARE BEING USED TO DETERMINE EXERCISE EFFECTS ON AN ENORMOUS NUMBER OF FACTORS, INCLUDING BUT NOT LIMITED TO GENES, PROTEINS, PHOSPHOPROTEINS, AND METABOLITES. FOR THIS R03 PROJECT, WE WILL USE THE ACUTE EXERCISE STUDY DATA IN CONJUNCTION WITH THE TRAINING STUDY DATA TO IDENTIFY THE KEY MOLECULAR CHANGES THAT OCCUR IN RESPONSE TO A SINGLE BOUT OF ACUTE EXERCISE THAT RESULTS IN CHRONIC TRAINING-INDUCED ADAPTATIONS. THIS IS A HIGHLY SIGNIFICANT QUESTION BECAUSE THE IDENTIFICATION AND VALIDATION OF THESE ACUTE MOLECULAR RESPONSES COULD PROVIDE NOVEL TARGETS TO ACTIVATE THE HEALTH PROMOTING EFFECTS OF EXERCISE. WHILE THERE HAS BEEN A LONG-STANDING HYPOTHESIS IN THE EXERCISE SCIENCE FIELD THAT CHRONIC TRAINING ADAPTATIONS RESULT FROM THE ACCUMULATION OF ACUTE MOLECULAR RESPONSES THAT OCCUR WITH EACH INDIVIDUAL EXERCISE BOUT, MOST OF THESE STUDIES FOCUSED ON SINGLE GENES OR PROTEINS AND MOST HAVE ONLY BEEN DONE IN SKELETAL MUSCLE. HERE, USING THE VAST MOTRPAC DATA SETS, WE WILL BE ABLE TO ANALYZE THE GLOBAL ACUTE EXERCISE MOLECULAR RESPONSES IN CONJUNCTION WITH TRAINING ADAPTATIONS IN AN UNPRECEDENTED MANNER. TO ACHIEVE THIS GOAL, WE PROPOSE TO USE A NOVEL AND INNOVATIVE REPLICATION METHOD WE HAVE DEVELOPED TO IDENTIFY MOLECULAR CHANGES THAT OCCUR WITH BOTH ACUTE EXERCISE AND TRAINING. GIVEN THE ENORMITY OF MOTRPAC DATA AND THE ONE-YEAR LENGTH OF THE R03 PROJECT, DATA FROM TWO TISSUES WILL BE STUDIED, SUBCUTANEOUS WHITE ADIPOSE TISSUE (SCWAT) AND KIDNEY. THESE TISSUES WERE CHOSEN DUE TO THEIR ROLE IN METABOLIC HEALTH AND BECAUSE THEY UNDERGO SIGNIFICANT MOLECULAR CHANGES IN RESPONSE TO EXERCISE. THE SPECIFIC AIMS ARE: 1) TO IDENTIFY MOLECULAR ADAPTATIONS THAT REPLICATE BETWEEN ACUTE EXERCISE AND EXERCISE TRAINING IN SCWAT AND KIDNEY; AND 2) TO IDENTIFY MOLECULAR ADAPTATIONS WITHIN TISSUES THAT REPLICATE BETWEEN TRANSCRIPTOMIC/PHOSPHOPROTEOMIC RESPONSES TO ACUTE EXERCISE AND PROTEOME CHANGES IN TRAINING. WE ANTICIPATE GENERATING A LARGE DATABASE OF THESE POTENTIAL HEALTH-PROMOTING MOLECULES THAT WILL BE AN IMPORTANT RESOURCE FOR THE SCIENTIFIC COMMUNITY. FOR OUR LAB, AS PART OF THIS PROJECT, WE WILL USE PATHWAY ANALYSIS COMBINED WITH OUR PHYSIOLOGICAL KNOWLEDGE TO SELECT 5-10 CANDIDATES FROM EACH OF THE GENE, PROTEIN, PHOSPHOPROTEIN, AND METABOLITE OMES. IN FUTURE STUDIES, CANDIDATES WILL UNDERGO AN IN-DEPTH INVESTIGATION OF MECHANISMS OF ACTION AND UTILITY AS A THERAPEUTIC TARGET. THUS, THE DATA FROM THIS R03 PROJECT WILL PROVIDE A FOUNDATION FOR DEVELOPING THERAPEUTIC TARGETS FOR CHRONIC DISEASES AND UNDERSTANDING THE FUNDAMENTAL MOLECULAR MECHANISMS OF EXERCISE-INDUCED IMPROVEMENTS IN HEALTH.

INSULIN REGULATES AGING-ASSOCIATED GENES THROUGH CHROMATIN TOPOLOGY - PROJECT SUMMARY/ABSTRACT TITLE: INSULIN REGULATES AGING-ASSOCIATED GENES THROUGH CHROMATIN TOPOLOGY INSULIN SIGNALING PLAYS AN IMPORTANT ROLE IN CONTROLLING METABOLIC HOMEOSTASIS, WITH ITS DYSREGULATION LINKED TO VARIOUS METABOLIC DISEASES ASSOCIATED WITH AGING, INCLUDING TYPE 2 DIABETES (T2D) AND OBESITY. INHIBITING INSULIN SIGNALING HAS BEEN SHOWN TO PROLONG LIFESPAN AND POSTPONE AGING-ASSOCIATED DISEASES ACROSS VARIOUS ANIMAL SPECIES. HOWEVER, THE MECHANISM BY WHICH INSULIN ACTIVITY CONTROLS AGING AND RELATED METABOLIC SYNDROMES REMAIN UNCLEAR. IN MOUSE LIVER, A SIGNIFICANT PORTION OF INSULIN-REGULATED GENES ARE ASSOCIATED WITH AGING. AMONG THEM, THE EXPRESSION LEVEL OF TMTC2 (TRANSMEMBRANE AND TETRATRICOPEPTIDE REPEAT CONTAINING PROTEIN 2), A NOVEL CA2+ REGULATOR, SIGNIFICANTLY DECLINES WITH AGING, INSULIN TREATMENT AND INSULIN-ASSOCIATED METABOLIC CONDITIONS. THIS PROJECT HAS THREE OBJECTIVES. 1) AIM 1.1 (F99): THIS PROJECT DETERMINES HOW TMTC2 CONTROLS CELL RESPONSE TO STRESS AND SENESCENCE THROUGH CA2+ HOMEOSTASIS. TMTC2 WILL BE KNOCKED DOWN FOLLOWED BY THE IDENTIFICATION OF CELL RESPONSE TO METABOLIC STRESS. MECHANISTICALLY, CA2+ CHANNELS AND PUMPS INTERACTING WITH TMTC2 WILL BE DEFINED. 2) AIM 1.2 (F99): THIS PROJECT DEFINES HOW INSULIN REGULATES CHROMATIN TOPOLOGY THROUGH TRANSCRIPTION FACTOR (TF)-MEDIATED ALTERATIONS IN CTCF BINDING, SUBSEQUENTLY REGULATING THE EXPRESSION OF NOVEL AGING- ASSOCIATED GENES. CHROMATIN TOPOLOGY WILL BE DEFINED UPON INSULIN TREATMENT FOLLOWED BY THE IDENTIFICATION OF HOW TFS MEDIATES THIS PROCESS. 3) AIM 2 (K00): THIS PROJECT FURTHER IDENTIFIES HOW ALTERED DNA METHYLATION REMODELS THE GENOMIC TEMPLATE IN AGING CELLS TO ALTER CTCF BINDING AND CHROMATIN LOOPS, SHIFTING THE TRANSCRIPTIONAL IMPACT OF INSULIN AND PERMITTING AGING-ASSOCIATED METABOLIC DYSFUNCTION. DIFFERENTIAL DNA METHYLATION LANDSCAPE IN HEPATOCYTES FROM YOUNG AND OLD MICE WILL BE DEFINED, AND ITS FUNCTIONAL IMPLICATIONS IN THE ALTERATIONS OF CTCF OCCUPANCY AND CHROMATIN TOPOLOGY IN AGING WILL BE IDENTIFIED. THIS PROPOSAL HAS THE POTENTIAL TO PROVIDE NEW GENES OR DNA ELEMENTS AS TARGETS TO ACHIEVE LIVER REJUVENATION AND THE REVERSAL OF INSULIN RESISTANCE, WHICH IS ASSOCIATED WITH AGING. THIS PROJECT ALSO HAS THE POTENTIAL TO SHED LIGHT ON THE FUNDAMENTAL MECHANISM BY WHICH AGING SHIFTS CELL RESPONSE TO INSULIN.

TELOMERASE AS A MARKER OF BROWN AND WHITE ADIPOSE TISSUE STEM CELLS

DO A RANGE OF IMMUNE CELLS PARTICIPATE IN INSULIN RESISTANCE

STRUCTURAL BASIS FOR INSULIN RESISTANCE MEDIATED BY IRS-1 SERINE PHOSPHORYLATION

REGULATION AND PREVENTION OF OXIDATIVE STRESS BY SKN-1