Lankenau Institute For Medical Research

REGENERATIVE WOUND HEALING VIA INFLAMMATION-MODULATING BIOMATERIALS

NCI COMMUNITY ONCOLOGY RESEARCH PROGRAM (NCORP) COMMUNITY SITES

HUMAN MONOCLONAL ANTIBODIES THAT BIND BOTULINUM TOXINS

MAIN LINE HEALTH CCOP

DEVELOPMENT OF A WHOLE HEART MODEL OF THE J WAVE SYNDROMES AND NOVEL APPROACHES TO PHARMACOLOGIC MANAGEMENT OF ASSOCIATED LIFE-THREATENING ARRHYTHMIAS

IDO2 TARGETING IN PANCREATIC CANCER

MECHANISMS OF TUMOR INITIATION UPON DISRUPTION OF THE RB/E2F INTERACTION - ABSTRACT E2F (E2F1-8) TRANSCRIPTION FACTORS ARE CRITICAL REGULATORS OF CELL CYCLE AND THEIR ACTIVITY IS PHYSICALLY REGULATED BY RB FAMILY PROTEINS (RB, P107 AND P130). DISRUPTION OF THE RB/E2F INTERACTION IS A HALLMARK OF CANCER, AS DEFINED BY HANAHAN&WEINBERG. THE ACQUIRED RESISTANCE TO ANTIGROWTH SIGNALS RESULTING FROM THIS DISRUPTION IS THOUGHT TO BE NECESSARY FOR TUMOR INITIATION. HOWEVER, HOW DOES UNRESTRICTED E2F ACTIVITY INITIATES TUMORIGENESIS IN VIVO IS POORLY UNDERSTOOD AND REMAINS A FUNDAMENTAL GAP IN OUR UNDERSTANDING OF CANCER BIOLOGY. IN PARTICULAR, WHETHER THE DIFFERENTIATION STATUS AFFECT THE CAPACITY OF A CELL TO TRANSFORM UPON DISRUPTION OF RB/E2F INTERACTION IS UNKNOWN. IN ADDITION, WHETHER UNRESTRICTED E2F ACTIVATES OTHER ONCOGENIC FEATURES BESIDES ABERRANT PROLIFERATION IS STILL OBSCURE. HEPATOCELLULAR CARCINOMA (HCC) IS THE SECOND CANCER IN TERMS OF DEATH WORLDWIDE. LIMITED UNDERSTANDING OF THE FUNCTIONAL CONSEQUENCES FOR FREQUENT GENETIC EVENTS HAMPERS THE DEVELOPMENT OF EFFICIENT THERAPEUTICS. THE RB/E2F INTERACTION IS DISRUPTED IN THE VAST MAJORITY OF HCC, AS A CONSEQUENCE OF SEVERAL EVENTS THAT TARGET UPSTREAM COMPONENTS OF THE RB/E2F PATHWAY. THEREFORE, HCC IS A RELEVANT MODEL TO INVESTIGATE THE CONSEQUENCES OF UNRESTRICTED E2F ACTIVITY FOR CANCER INITIATION. ACCORDINGLY, PAN-LIVER INACTIVATION OF RB FAMILY GENES (TRIPLE KNOCK OUT, TKO) IS SUFFICIENT TO INITIATE HCC (TKO HCC) THAT RECAPITULATE MULTIPLE FEATURES OF THE HUMAN DISEASE. STUDIES OF CARCINOGEN-INDUCED MODELS OF HCC HAVE LED TO THE CONCLUSION THAT HEPATOCYTES ARE THE SOLE SOURCE OF HCC. SPECIFIC INACTIVATION OF RB FAMILY IN HEPATOCYTES TRIGGERS A SHORT PROLIFERATIVE BURST BUT FAILS TO INITIATE HCC. THIS RESULT CHALLENGES THE CURRENT HEPATO-CENTRIC MODEL IN THE FIELD AND SUGGESTS THAT OTHER LINEAGES CAN ALSO SERVE AS A CELL OF ORIGIN FOR HCC. ACCORDINGLY, INACTIVATION OF RB FAMILY IN MULTIPLE LIVER LINEAGES REVEALS THAT TKO HCC ARISES FROM A PERIDUCTAL PROGENITOR. IN PARTICULAR, OUR PRELIMINARY DATA INDICATES THAT UNRESTRICTED E2F ACTIVITY COUPLES ABERRANT PROLIFERATION WITH CELL FATE ALTERATION IN THIS POPULATION TO INITIATE HCC. BASED ON THESE DATA, WE PROPOSE TO:1) DETERMINE THE MOLECULAR MECHANISMS THAT ALTER THE CELL FATE OF A PERIDUCTAL PROGENITOR TO SERVE AS A CELL OF ORIGIN FOR TKO HCC. 2) DETERMINE THE INDIVIDUAL AND COMPOUND ROLE OF E2F FACTORS IN TKO HCC INITIATION AND DEVELOPMENT. 3) DETERMINE THERAPEUTIC VULNERABILITIES IN TKO HCC THAT COULD SERVE AS NOVEL TREATMENT FOR PATIENTS. WE BELIEVE THAT OUR PROPOSAL WILL ADDRESS FUNDAMENTAL QUESTIONS REGARDING THE ROLE OF E2F IN CANCER INITIATION, AS IDENTIFIED ABOVE. IN ADDITION, WE EXPECT THAT OUR RESULTS WILL ESTABLISH THAT DIFFERENT CELL LINEAGES CAN SERVE AS A CELL OF ORIGIN FOR HCC, WHICH WILL HAVE IMPORTANT CLINICAL IMPLICATIONS, IN PARTICULAR REGARDING THE CLASSIFICATION OF PATIENTS AND THE DEVELOPMENT OF THERAPIES TAILORED FOR DIFFERENT CLASSES OF HCC.

POLYAMINE-STIMULATED STEM CELL RECRUITMENT IN ARSENIC-INDUCED SKIN CANCER

IDO PATHWAYS IN INFLAMMATORY PATHOGENESIS AND TREATMENT OF RHEUMATOID ARTHRITIS

PROBING AN UNEXPLORED INTRACELLULAR PATHWAY IN DIABETES PATHOGENESIS - ABSTRACT DIABETIC NEPHROPATHY AND OTHER DIABETES COMPLICATIONS IMPOSE ENORMOUS BURDENS ON PATIENTS AND HEALTHCARE SYSTEMS, MAKING IT IMPERATIVE TO DEFINE ACTIONABLE ETIOLOGIC FACTORS AND DEVELOP EFFECTIVE, LOW-COST THERAPEUTIC INTERVENTIONS. NONENZYMATIC PROTEIN GLYCATION AND THE FORMATION OF ADVANCED GLYCATION END PRODUCTS (AGES) ARE STRONGLY IMPLICATED IN PATHOGENESIS. THE DRIVER OF AGE FORMATION IS 3-DEOXYGLUCOSONE (3DG), A HIGHLY REACTIVE DICARBONYL SPECIES THAT ALSO CAUSES ACUTE CELLULAR TOXICITIES BY DAMAGING ENZYMES AND DNA AND INFLAMING THE VASCULATURE. ACCORDINGLY, THE ABILITY TO ACCURATELY MEASURE 3DG LEVELS AND UNDERSTAND ITS ETIOLOGY ARE PARAMOUNT TO ELUCIDATING PATHOGENESIS, LIMITING ITS PATHOGENIC EFFECTS, AND IMPROVING CLINICAL MANAGEMENT OF DIABETIC COMPLICATIONS. ENDOGENOUS 3DG WAS DEEMED TO ARISE NONENZYMATICALLY FROM THE SLOW DISINTEGRATION OF GLYCATED PROTEINS IN THE BODY OR ABSORBED FROM INGESTED HEAT-PROCESSED FOODS. WE DEVELOPED NEW METHODS TO STUDY THE ENZYMATIC ACTIVITY OF FRUCTOSAMINE-3-KINASE (FN3K), AN ENZYME THOUGHT TO REPAIR GLYCATED PROTEINS AND PREVENT AGE, BUT AN END-PRODUCT OF FN3K ACTIVITY IS 3DG. WE DISCOVERED THAT 3DG LEVELS IN KIDNEY ARE HIGHER THAN PREVIOUSLY ANTICIPATED. OUR CORE HYPOTHESIS IS THAT FN3K-MEDIATED 3DG FORMATION IN CELLS IS A KEY PATHOGENIC DRIVER IN DIABETIC COMPLICATIONS. SPECIFIC AIM 1: WE WILL MEASURE 3DG ARISING IN TISSUES IN RELATIONSHIP TO PATHOGENESIS IN THE KK.CG-AY/J MURINE MODEL OF TYPE-2 DIABETES. SPECIFIC AIM 2: THE IMPACT OF A HIGH GLYCATION DIET ON 3DG LEVELS WILL BE MEASURED IN TISSUES SENSITIVE TO DIABETIC COMPLICATIONS, INCLUDING IN THE KIDNEY, HEART, AND LIVER OF THE DIABETIC MICE. SPECIFIC AIM 3: WE WILL DEFINE THE PHARMACODYNAMIC PROPERTIES AND MODES OF ACTION FOR MEGLUMINE, AN AGENT, ALREADY PROVEN SAFE, THAT WE DISCOVERED HAS UNRECOGNIZED MEDICINAL EFFECTS, HAVING PROVIDED NEPHROPROTECTION AND PREVENTED TRIGLYCERIDE ACCUMULATION IN DIABETIC MICE. SPECIFIC AIM 4: A SERIES OF FN3K ANTAGONISTS THAT WE DISCOVERED WILL BE CHARACTERIZED TO IDENTIFY A PRECLINICAL DRUG DEVELOPMENT CANDIDATE. THIS PROPOSAL OFFERS SEVERAL MAJOR INNOVATIVE ELEMENTS OF HIGH SIGNIFICANCE AND IMPACT IN DIABETES TRANSLATIONAL RESEARCH. AIM 1 WILL PROVIDE NEW DATA DEVELOPED WITH METHODOLOGY WE REFINED TO MEASURE FN3K ACTIVITY AND 3DG FORMATION MORE ACCURATELY, ADDRESSING KEY GAPS IN KNOWLEDGE. AIM 2 WILL EXPLORE THE LINKAGE BETWEEN INTRACELLULAR 3DG ELEVATION AND THE CONSUMPTION OF ‘WESTERN’ DIETS RICH IN FRUCTOSAMINES—THE SUBSTRATE FOR FN3K. DRUG SAFETY IS PARAMOUNT FOR ANY NEW DIABETES DRUG. THE DATA FROM AIM 3 WILL ACCELERATE THE DEVELOPMENT OF MEGLUMINE AS AN INNOVATIVE TREATMENT MODALITY—A COMPOUND PROVEN EXTREMELY SAFE FOR CHRONIC ADMINISTRATION—TO AMELIORATE DIABETIC NEPHROPATHY, FATTY LIVER, AND POTENTIALLY OTHER DIABETIC COMPLICATIONS. AIM 4 OFFERS OPPORTUNITY TO DELIVER FIRST-IN-CLASS ENZYME INHIBITORS AS POTENTIAL DRUG LEAD CANDIDATES. IN SUMMARY, THIS RESEARCH PROGRAM WILL ILLUMINATE AN UNEXPLORED INTRACELLULAR PATHWAY IN DIABETES PATHOGENESIS AND DELIVER UNPRECEDENTED TOOLS FOR BROADER RESEARCH INTO THE ROLE OF 3DG IN DIABETIC NEPHROPATHY AND OTHER DIABETES COMPLICATIONS.

DRUG-INDUCED REGENERATION AND RE-INNERVATION IN A MOUSE DIGIT AMPUTATION MODEL

IDO INHIBITORS FOR COMBINATORIAL CANCER THERAPY

(PQA2) MAMMALIAN REGENERATION, HIGH FAT DIETS, AND BREAST CANCER: A COMMON LINK

TARGETED NANOPARTICLE DNA THERAPY FOR OVARIAN CANCER

DEFINING THE CELLULAR AND MOLECULAR MECHANISM OF IDO2 FUNCTION DRIVING AUTOIMMUNE VS.PROTECTIVE IMMUNE RESPONSES - PROJECT SUMMARY AUTOIMMUNE DISEASES ARE CHRONIC INFLAMMATORY DISEASES THAT AFFECT MORE THAN 50 MILLION AMERICANS, LEADING TO PAIN, DISABILITY, AND INCREASED MORTALITY. WHILE DRUGS TO TREAT AUTOIMMUNE DISEASES EXIST, ALLEVIATION OF INFLAMMATORY SYMPTOMS IS OFTEN ASSOCIATED WITH SEVERE SIDE-EFFECTS, INCLUDING INCREASED SUSCEPTIBILITY TO INFECTIONS AND CANCER. OUR PROPOSAL ADDRESSES THE URGENT NEED TO IDENTIFY NEW THERAPEUTIC TARGETS TO ADDRESS THE UNDERLYING CAUSES AND ASSOCIATED SYMPTOMS OF THESE DEBILITATING DISEASES. IN THIS PROPOSAL, WE WILL INVESTIGATE INDOLEAMINE-2,3-DIOXYGENASE (IDO2) AS A NOVEL IMMUNOMODULATORY TARGET FOR AUTOIMMUNE DISEASES. IDO2 IS ONE OF TWO CLOSELY RELATED TRYPTOPHAN CATABOLIZING ENZYMES EXPRESSED PRIMARILY IN IMMUNE CELLS. USING A COMBINATION OF GENETIC AND PHARMACOLOGICAL STUDIES IN THE KRN MODEL OF INFLAMMATORY ARTHRITIS, WE IDENTIFIED IDO2 AS AN ESSENTIAL MEDIATOR OF AUTOANTIBODY-MEDIATED INFLAMMATORY RESPONSES AND DISCOVERED A NOVEL NON-ENZYMATIC FUNCTION OF IDO2 THAT DRIVES AUTOIMMUNE RESPONSES LEADING TO DISEASE. MECHANISTIC STUDIES IMPLICATE THE TRANSCRIPTION FACTOR RUNX1 AS A POTENTIAL COMPONENT OF THIS PREVIOUSLY UNKNOWN IDO2 SIGNALING PATHWAY. IDO2 APPEARS TO BE ESSENTIAL FOR ONLY SOME B CELL FUNCTIONS, AS IDO2 DEFICIENT MICE DEVELOP NORMAL IMMUNE CELL COMPARTMENTS AND MOUNT PRODUCTIVE IMMUNE RESPONSES TO CHALLENGE WITH SOME, BUT NOT ALL, MODEL ANTIGENS IN VITRO AND IN VIVO. A BETTER UNDERSTANDING OF IDO2 BIOLOGY, IN PARTICULAR THE ROLES THAT ENZYMATIC AND NON-ENZYMATIC ACTIVITY HAVE IN REGULATING IMMUNE FUNCTION IS NECESSARY TO DEVELOP IDO2 AS A POTENTIAL THERAPEUTIC TARGET FOR AUTOIMMUNE DISEASE. OUR WORKING HYPOTHESIS IS THAT IDO2 MODULATION OF IMMUNE RESPONSES INVOLVES BOTH ENZYMATIC AND NON-ENZYMATIC PATHWAYS WITH AUTOREACTIVE B CELL ACTIVATION/AUTOANTIBODY PRODUCTION DEPENDENT ON IDO2’S NON-ENZYMATIC FUNCTION. WE PROPOSE THAT THIS DIFFERENTIAL USE OF NON- ENZYMATIC VS. ENZYMATIC IDO2 PATHWAYS CAN BE SPECIFICALLY TARGETED AS A THERAPEUTIC STRATEGY TO INHIBIT IDO2 FUNCTION IN AUTOIMMUNE RESPONSES WITHOUT AFFECTING ITS ROLE IN PROTECTIVE IMMUNITY. IN THIS PROPOSAL, WE WILL USE PRECLINICAL MODELS OF AUTOIMMUNE ARTHRITIS, IDO2 AND RUNX1 CONDITIONAL KNOCKOUT MICE AND OVEREXPRESSING CELL LINES, AND WHOLE TRANSCRIPTOME RNA-SEQ ANALYSIS TO DEFINE THE CELLULAR AND MOLECULAR MECHANISM MEDIATING IDO2 NON-ENZYMATIC FUNCTION (AIM 1). WE WILL THEN USE CATALYTICALLY INACTIVE IDO2 KNOCK- IN MICE AND A NOVEL IDO2-TARGETING ANTIBODY, TOGETHER WITH IDO2-DEPENDENT MODELS OF AUTOIMMUNITY AND PROTECTIVE IMMUNITY, TO DETERMINE IF DIFFERENTIAL USE OF NON-ENZYMATIC VS. ENZYMATIC PATHWAYS DISTINGUISHES IDO2 FUNCTION IN AUTOIMMUNE AND PROTECTIVE IMMUNE RESPONSES (AIM 2). THE POTENTIAL LONG-TERM IMPACT OF THIS PROJECT WOULD MOVE TARGETING NON-ENZYMATIC IDO2 FUNCTION INTO DEVELOPMENT AS A NOVEL STRATEGY TO TREAT AUTOANTIBODY-MEDIATED AUTOIMMUNE DISEASES.

TARGETING NANOPARTICLE DNA DELIVERY TO PROSTATE TUMORS

POLYAMINES AND EPITHELIAL TUMORIGENESIS

DIRECTING THE FATE OF CELLS TO MYOGENIC LINEAGES

HISTONE MODIFICATIONS REGULATING PHENOTYPE PLASTICITY: ROLE OF NON-CANONICAL WNT SIGNALING

TARGETING INCREASED POLYAMINE TRANSPORT OF RESISTANT MELANOMAS

REGULATION OF MECHANICAL COUPLING IN AGING MYOCARDIUM - PROJECT SUMMARY AGE MODIFIES HEART STRUCTURE AND FUNCTION, EVEN IN THE ABSENCE OF OVERT CARDIOVASCULAR DISEASE. WITH AGE-RELATED CARDIAC REMODELING, THE HEART UNDERGOES EXTENSIVE CELLULAR AND MOLECULAR CHANGES THAT RESULT IN A STIFFER LESS COMPLIANT HEART THAT EXPERIENCES AN OVERALL DECLINE IN FUNCTION. YET, THE HUMAN HEART IS CAPABLE OF FUNCTIONING FOR DECADES DESPITE MINIMAL CELL TURNOVER OR REGENERATION, SUGGESTING THAT SOME MECHANISMS OF AGE-RELATED CARDIAC REMODELING MAY BE COMPENSATORY AND BENEFICIAL, HELPING SUSTAIN HEART FUNCTION OVER OUR LIFE SPAN. RECENT EVIDENCE INDICATES THE UPREGULATION OF CYTOSKELETAL PROTEINS MAY REPRESENT A CARDIOPROTECTIVE MECHANISM IN THE AGING HEART. IN THIS EXPLORATORY R21 PROPOSAL, WE HYPOTHESIZE THAT ENHANCING THE CYTOSKELETAL CONNECTION BETWEEN N-CADHERIN AND THE CORTICAL ACTIN NETWORK REPRESENTS AN ESSENTIAL COMPENSATORY MECHANISM TO MAINTAIN FORCE TRANSMISSION IN THE AGING MAMMALIAN HEART. TO TEST THIS HYPOTHESIS, THE REGULATION OF VINCULIN, A FORCE-SENSITIVE CYTOSKELETAL PROTEIN, WILL BE INVESTIGATED DURING CARDIAC AGING IN MICE. THE FOLLOWING INTERRELATED AIMS ARE PROPOSED: (1) TO TEST THE HYPOTHESIS THAT ABL KINASE IS REQUIRED TO MAINTAIN MECHANO- ELECTRICAL INTEGRITY IN THE AGING HEART. (2) TO TEST THE HYPOTHESIS THAT INHIBITION OF TYROSINE PHOSPHORYLATION OF VINCULIN WILL PERTURB N-CADHERIN-CYTOSKELETAL CONNECTION LEADING TO ACCELERATED CARDIAC AGING. THE PROPOSED STUDIES WILL ALLOW MECHANISTIC INSIGHT INTO HOW THE MAJOR PREDISPOSING FACTOR IN HEART DISEASE, AGE, AFFECTS THE REMODELING OF N- CADHERIN JUNCTIONS AND MECHANICAL COUPLING IN THE AGING HEART.

NEW DRUG DISCOVERY PARADIGMS FOR SYNUCLEINOPATHIES

COMPREHENSIVE FUNCTIONAL ASSESMENT OF THE HUMAN ANTIBODY RESPONSE TO ENTEROVIRUS 71

IMMUNOTOXINS BASED ON THE IGG RESPONSE TO AUTOLOGOUS ANTIGENS IN CANCER PATIENTS

NOVEL APPROACHES TO PHARMACOLOGIC MANAGEMENT OF LIFE-THREATENING ARRHYTHMIAS ASSOCIATED WITH THE J WAVE SYNDROMES

POTENTIAL ROLE OF FETAL STEM CELLS IN LUNG TUMOR DEVELOPMENT

IDO2 TARGETING FOR PANCREATIC CANCER TREATMENT

INVESTIGATING THE ROLE OF INDOLEAMINE 2,3-DIOXYGENASE (IDO) IN BREAST CANCER METASTASIS

OPPC TARGETING TO IMPROVE PANCREATIC CANCER TREATMENT

ROLE OF LONG NONCODING RNAS IN PROSTATE CANCER