Beckman Research Institute

CANCER CENTER SUPPORT GRANT

CITY OF HOPE LYMPHOMA SPORE

THE HUMAN ISLET DISTRIBUTION COORDINATING CENTER (UC4)

INNOVATIVE INFRASTRUCTURE TO ENHANCE THE CALIFORNIA TEACHERS STUDY

PHASE I MOLECULAR AND CLINICAL PHARMACODYNAMIC TRIALS ET-CTN

INTEGRATED ISLET DISTRIBUTION PROGRAM (U24) - 2021 - PROJECT SUMMARY / ABSTRACT HUMAN PANCREATIC ISLETS ARE AN ESSENTIAL RESEARCH RESOURCE FOR RESEARCH ON THE PREVENTION, TREATMENT, AND PATHOPHYSIOLOGY OF DIABETES MELLITUS. RECENT DATA HAVE HIGHLIGHTED IMPORTANT DIFFERENCES BETWEEN MURINE AND HUMAN ISLETS, SUBSTANTIATING THE CONTINUED NEED FOR ACCESS TO HUMAN ISLETS, AS THE GOLD STANDARD IN DIABETES RESEARCH. CITY OF HOPE (COH) IS APPLYING FOR THIS U24 RENEWAL TO REMAIN AS THE INTEGRATED ISLET DISTRIBUTION PROGRAM COORDINATING CENTER (IIDP CC) FOR THE NEXT 5 YEARS, TO CONTINUE TO PROVIDE DISTRIBUTION OF HUMAN CADAVERIC ISLETS AND ANCILLARY TISSUE FOR BIOMEDICAL RESEARCH TO RESEARCHERS WORLDWIDE. OUR PROPOSAL LEVERAGES THE SIGNIFICANT INVESTMENT MADE BY NIH OVER THE LAST 19 YEARS THAT HAS ESTABLISHED AND SUCCESSFULLY MAINTAINED THE IIDP AT COH. FROM QUALIFICATION AND AUDITING OF HIGH-QUALITY ISLET ISOLATION CENTERS (IICS), TO FORECASTING, TRACKING, AND MEETING THE NEEDS OF INVESTIGATORS, SINCE 2002 OUR EXPERIENCED TEAM HAS WORKED WITH 20 DIFFERENT ISLET ISOLATION LABORATORIES TO COORDINATE THE DISTRIBUTION OF OVER 330 MILLION ISLET EQUIVALENTS TO MORE THAN 400 INVESTIGATORS ACROSS 16 COUNTRIES SINCE 2002, SUPPORTING 767 PEER REVIEWED PUBLICATIONS. THROUGH THIS RENEWAL WE WILL CONTINUE TO SUBCONTRACT WITH OUR 5 HIGHLY QUALIFIED IICS TO ISOLATE AND DISTRIBUTE HUMAN ISLETS AND ANCILLARY TISSUE VIA OUR ADVANCED ELECTRONIC ISLET ALLOCATION SYSTEM (IAS). WE WILL CONTINUE TO MANAGE THE REVIEW PROCESS FOR ISLET RECEIPT, PILOT STUDIES, AND OPPORTUNITY POOL FUNDING. WE WILL FURTHER ENHANCE OUR IAS TO BROADCAST OFFERS ONLINE AND NOTIFY APPROVED WAITING RESEARCHERS OF ISLET AVAILABILITY, IN A FAIR, EQUITABLE AND TIME SENSITIVE MANNER. IIDP WILL CONTINUE TO MAINTAIN THE EXISTING COST RECOVERY SYSTEM THROUGH SUBSCRIPTION FEES COLLECTED FROM ISLET RESEARCHERS, WHICH HAS GARNERED A TOTAL OF $9,303,950 SINCE THE IMPLEMENTATION OF SUBSCRIPTION FEES TO OFFSET THE EXPENSES OF PANCREATIC PROCESSING FOR THE IICS. WE WILL CONTINUE TO CLOSELY MONITOR AND HELP TO IMPROVE THE QUALITY OF ISLETS DISTRIBUTED, THROUGH THE CONTINUATION OF THE HUMAN ISLET PHENOTYPING PROGRAM (HIPP) THAT CONDUCTS ASSAYS ON A SAMPLE FROM EACH ISLET ISOLATION. IIDP HAS JUST ADDED A HUMAN ISLET GENOTYPING INITIATIVE (HIGI) TO GENOTYPE EACH ISOLATION AS WELL. PHENOTYPING AND GENOTYPING DATA, AS WELL AS UNOS DATA, EXTENSIVE DONOR AND ISLET ISOLATION DATA, WILL BE MADE AVAILABLE TO APPROVED INVESTIGATORS THROUGH ONLINE ACCESS TO THE IIDP RESEARCH DATA REPOSITORY, WITH IIDP AND NIDDK APPROVAL OF APPLYING SCIENTISTS. INVESTIGATORS CAN EASILY SEARCH THE REQUIRED DATA, SELECT FILTER CRITERIA, SAVE THEIR SEARCHES, AND DOWNLOAD THE INTEGRATED IIDP DATA FOR EXPLORATORY ANALYSES. THROUGH OUR PROVEN STATE-OF-THE-ART ADMINISTRATIVE, BUSINESS, TECHNICAL, STATISTICAL, QUALITY ASSURANCE, AND INFORMATICS PROCESSES AND TOOLS, THE ACCESSIBILITY OF HUMAN ISLETS FOR INVESTIGATORS CONDUCTING ESSENTIAL DIABETES MELLITUS RESEARCH WILL BE SECURED. WE WILL CONTINUE TO PROVIDE AN INDISPENSABLE RESEARCH RESOURCE FOR THE DIABETES RESEARCH COMMUNITY BY ENSURING THAT THE IIDP REMAINS STABLE, TECHNOLOGICALLY ADVANCED, CONTINUALLY ENHANCED, AND FULLY RESPONSIVE TO THE ISLET NEEDS OF THE RESEARCH COMMUNITY, PROMOTING THE NEXT GENERATION OF SCIENTIFIC EXPERIMENTATION TOWARD THE PREVENTION AND TREATMENT OF DIABETES.

CALIFORNIA TEACHERS STUDY

ANTIBODY TARGETED RADIATION AND IMMUNOTHERAPY FOR THE TREATMENT OF SOLID TUMORS

HUMAN ISLET RESEARCH ENHANCEMENT CENTER FOR THE HUMAN ISLET RESEARCH NETWORK

ONCOLOGY RESEARCH CAREER DEVELOPMENT PROGRAM

PALLIATIVE CARE FOR QUALITY OF LIFE AND SYMPTOM CONCERNS IN LUNG CANCER

COMBATING SUBCLONAL EVOLUTION OF RESISTANT CANCER PHENOTYPES

NEW BIOSPECIMENS TO ENHANCE RESEARCH IN THE CALIFORNIA TEACHERS STUDY COHORT

HEMATOPOIETIC CELL TRANSPLANTATION FOR HEMATOLOGIC MALIGNANCIES

PRECISION APPROACHES TO REFINING TP53-ASSOCIATED CANCER RISK

NONCODING RNA BIOMARKERS FOR NONINVASIVE AND EARLY DETECTION OF PANCREATIC CANCER

RNA DIRECTED SILENCING AND EXCISION OF HIV-1 AND CCR5

HUMAN NATURAL KILLER CELLS: ADVANCING BIOLOGY AND CLINICAL APPLICATIONS

DIFFERENTIAL MECHANISMS FOR RORGAMMAT-REGULATED THYMOCYTE DEVELOPMENT AND TH17 DI

INTEGRATION OF PALLIATIVE CARE FOR CANCER PATIENTS ON PHASE I TRIALS

TRANSCRIPTIONAL REGULATION BY ANGIOTENSIN II IN VASCULAR SMOOTH MUSCLE CELLS

CIRCULATING EXTRACELLULAR VESICLES AS A MARKER OF HUMAN TYPE 1 DIABETES PATHOGENESIS - PROJECT SUMMARY/ABSTRACT (30 LINES OR LESS) THE PREMISE FOR THE PROPOSED RESEARCH STEMS FROM PRECEDENCE IN OTHER DISEASES, SUCH AS CANCER, CARDIOVASCULAR, NEURODEGENERATIVE, AND MOST RELEVANT TO THE CURRENT PROPOSAL, AUTOIMMUNE DISEASES, IN WHICH CIRCULATING EXTRACELLULAR VESICLES (CEVS) PLAY A ROLE IN PATHOPHYSIOLOGY AND ARE IMPORTANT BIOMARKERS FOR EARLY DETECTION. HOWEVER, LITTLE IS KNOWN CONCERNING THE ROLE OF CEVS IN HUMAN TYPE 1 DIABETES (T1D). OUR OVERALL HYPOTHESIS IS THAT CEVS HAVE THE POTENTIAL TO BE USED AS BIOMARKERS FOR EARLY PRE-DISEASE DETECTION OF T1D, BASED ON THEIR DISTINCT MOLECULAR AND FUNCTIONAL PHENOTYPE IN T1D AND PRE-DISEASE STAGES, COMPARED TO HEALTHY OR LOW RISK INDIVIDUALS. OUR SPECIFIC AIMS ARE: 1) TO IDENTIFY THE DISTINCT PROTEIN AND RNA CARGO UNIQUE TO CEVS AT DIFFERENT STAGES OF T1D DISEASE DEVELOPMENT; AND 2) TO INVESTIGATE THE EFFECT OF CEVS FROM SUBJECTS AT DIFFERENT STAGES OF DISEASE PROGRESSION ON IMMUNE AND BETA CELL PHENOTYPES AND ELUCIDATE THE FUNCTIONAL RELEVANCE OF THEIR DISTINCT MOLECULAR CARGO. TO ADDRESS THESE AIMS, WE HAVE ASSEMBLED A TEAM OF INVESTIGATORS WITH HIGHLY RELEVANT EXPERTISE AND TECHNIQUES. WE PROPOSE TO USE LONGITUDINAL SAMPLES FROM THE ENVIRONMENTAL DETERMINANTS OF DIABETES IN THE YOUNG (TEDDY) STUDY OF CHILDREN WITH T1D ASSOCIATED GENETIC RISK TO IDENTIFY CRITICAL TIMEPOINTS AND UNDERLYING MECHANISMS MEDIATING, A) THE EARLIEST STAGES OF PATHOGENESIS PRECEDING AAB APPEARANCE OF THE FIRST ISLET AUTOANTIBODY, AND B) THE PERIOD OF SEROCONVERSION FROM SINGLE TO MULTIPE AAB+ OR REMAINING SINGLE AAB+ AND C) THE PERIOD AFTER MULTIPLE AAB APPEARANCE WITH A HIGHLY VARIABLE RATE OF PROGRESSION TO HYPERGLYCEMIA. WE HAVE THE EXPERTISE AND TECHNICAL ABILITY TO ISOLATE CEVS FROM PLASMA, PERFORM PROTEOMIC AND RNASEQ ANALYSIS ON EVS, AND PERFORM IMMUNE AND BETA CELL RELATED FUNCTIONAL ASSAYS. OUR RESEARCH PLAN IS TO GENERATE CEVS FROM DONORS AT DIFFERENT STAGES OF T1D DISEASE PROGRESSION, TO IDENTIFY THE UNIQUELY PACKAGED PROTEIN AND RNA CARGO FROM THESE CEVS, TO EVALUATE THE EFFECTS OF THE CEVS ON IMMUNE CELL FUNCTIONAL PHENOTYPE AND ISLET HEALTH AND ELUCIDATE THE FUNCTIONAL RELEVANCE OF THE DISTINCT MOLECULAR CARGO TARGETS. THESE STUDIES WILL YIELD NOVEL MECHANISTIC INSIGHTS INTO EARLY DISEASE PATHOGENESIS AND IDENTIFY POTENTIAL NOVEL BIOMARKERS FOR T1D INITIATION AND PROGRESSION.

INTESTINAL STEM CELL CONSORTIUM COORDINATING CENTER

TRANSFORMING GROWTH FACTOR BETA1, MICRORNAS AND DIABETIC NEPHROPATHY

FUNCTIONAL ANALYSIS OF FEN-1 NUCLEASE IN GENOME STABILITY AND CANCERS

INFLAMMATORY GENE REGULATION IN DIABETIC CONDITIONS

REGULATION OF IONIZING RADIATION INDUCED DNA DAMAGE RESPONSE

ROLE OF NUCLEASES IN RNA PRIMER REMOVAL AND MUTAGENESIS

ROLE OF AUTOREACTIVITY IN PATHOGENESIS OF CHRONIC GVHD

DIGITAL HEALTH INTERVENTION FOR SELF-MANAGEMENT AND TELEMONITORING IN CHIMERIC ANTIGEN RECEPTOR-T CELL THERAPY

COMBINATORIAL USE OF ANTI-HIV RNA-BASED THERAPEUTICS

A HUMAN IPSC-BASED CELL THERAPY FOR CANAVAN DISEASE - PROJECT SUMMARY CANAVAN DISEASE (CD) IS A RARE, AUTOSOMAL RECESSIVE NEURODEVELOPMENTAL DISORDER THAT AFFECTS CHILDREN FROM INFANCY. MOST CHILDREN WITH INFANTILE-ONSET CD, THE MOST PREVALENT FORM OF THE DISEASE, WILL DIE WITHIN THE FIRST DECADE OF LIFE. THERE IS NEITHER A CURE NOR A STANDARD TREATMENT FOR THIS DISEASE. CD IS CAUSED BY GENETIC MUTATIONS IN THE ASPARTOACYLASE (ASPA) GENE, WHICH ENCODES A METABOLIC ENZYME SYNTHESIZED BY OLIGODENDROCYTES IN THE BRAIN. ASPA BREAKS DOWN N-ACETYL-ASPARTATE (NAA), AN AMINO ACID DERIVATIVE IN THE BRAIN. THE CYCLE OF PRODUCTION AND BREAKDOWN OF NAA APPEARS TO BE CRITICAL FOR MAINTAINING THE WHITE MATTER OF THE BRAIN, WHICH CONSISTS OF NERVE FIBERS COVERED BY MYELIN. INDICATIONS OF CD INCLUDE LACK OF ASPA ACTIVITY, ACCUMULATION OF NAA, MYELINATION DEFECT, AND SPONGY DEGENERATION (VACUOLATION) IN THE BRAIN. THE CLINICAL SYMPTOMS INCLUDE IMPAIRED MOTOR FUNCTION AND MENTAL RETARDATION. THERE IS CURRENTLY NO APPROVED THERAPY FOR THIS CONDITION. THEREFORE, THERE IS A CLEAR, UNMET MEDICAL NEED FOR AN EFFECTIVE THERAPY FOR CD. THE DEVELOPMENT OF HUMAN INDUCED PLURIPOTENT STEM CELL (HIPSC) TECHNOLOGY HAS OPENED EXCITING AVENUES FOR CELL THERAPY. IN OUR PRELIMINARY STUDIES, WE HAVE USED HIPSC TECHNOLOGY TO GENERATE CD PATIENT IPSCS AND DIFFERENTIATED THESE IPSCS INTO NEURAL PROGENITOR CELLS (CD INPCS). WE THEN INTRODUCED A FUNCTIONAL ASPA GENE INTO CD INPCS THROUGH LENTIVIRAL TRANSDUCTION TO GENERATE GENETICALLY ENGINEERED FUNCTIONAL ASPA-CONTAINING INPCS, TERMED ASPA INPCS. IN ORDER TO MOVE THE ASPA INPC CELL PRODUCT TO THE CLINIC, WE DEVELOPED CURRENT GOOD MANUFACTURING PRACTICE (CGMP)-COMPATIBLE PROCESS TO MANUFACTURE THE ASPA INPCS. THE RESULTANT ASPA INPCS GENERATED FROM THREE DIFFERENT CD PATIENTS WERE TESTED IN A CD MOUSE MODEL FOR EFFICACY AND SAFETY. AFTER BEING TRANSPLANTED INTO BRAINS OF CD MICE, THE ASPA INPCS PROVIDED SUSTAINED ASPA ACTIVITY, LED TO SIGNIFICANTLY LOWER NAA LEVEL, CONSIDERABLE RESCUE OF SPONGY DEGENERATION AND MYELINATION DEFECT IN THE BRAIN, AND SUBSTANTIALLY IMPROVED MOTOR FUNCTION IN THE TRANSPLANTED CD MICE. IMPORTANTLY, NO TUMORIGENESIS OR OTHER ADVERSE EFFECT WAS OBSERVED IN THE TRANSPLANTED MICE. THESE ROBUST PRECLINICAL DATA PROVIDE STRONG RATIONALE FOR THE PROPOSED STUDY. THE OBJECT OF THE PROPOSED RESEARCH IS TO ESTABLISH A HIPSC-BASED CELL THERAPY FOR CD. THE CELL PRODUCTS HAVE PROVEN IN PRECLINICAL STUDIES TO BE LONG LASTING AND EFFICACIOUS WITH A FAVORABLE SAFETY PROFILE. WE PROPOSE THE FOLLOWING SPECIFIC AIMS: AIM 1: TO CONDUCT IND-ENABLING QUALIFICATION RUNS AND PERFORM FINAL MANUFACTURING OF THE ASPA INPC CELL PRODUCTS. AIM 2: TO PERFORM DEFINITIVE PRECLINICAL EFFICACY AND SAFETY/TUMORIGENICITY STUDIES OF THE ASPA INPC CELL PRODUCTS. AIM 3: TO OBTAIN IND APPROVAL. AIM 4: TO CONDUCT A PHASE I CLINICAL TRIAL TO ESTABLISH THE SAFETY AND FEASIBILITY OF ADMINISTERING THE ASPA INPC CELL PRODUCTS TO CD PATIENTS. THIS STUDY COULD LEAD TO THE DEVELOPMENT OF A NOVEL CELL THERAPY FOR CD AND DEMONSTRATE THE FEASIBILITY OF USING HIPSC-BASED CELL PRODUCTS FOR THE TREATMENT OF SIMILAR DISEASES.

REGULATION OF GLUCOSE HOMEOSTASIS BY MUNC18 PROTEINS

HUMAN ISLET RESEARCH NETWORK (HIRN) COORDINATING CENTER

THE ROLE OF INTESTINAL MICROBIOTA IN GRAFT-VERSUS-HOST DISEASE

SUMO MODIFICATION AND DNA DAMAGE RESPONSE IN CANCER THERAPY

MUCUS-DEGRADING INTESTINAL BACTERIA AND TOXICITIES OF HEMATOPOIETIC CELL TRANSPLANTATION - OVERALL PROGRAM SUMMARY: MUCUS-DEGRADING INTESTINAL BACTERIA AND TOXICITIES OF HEMATOPOIETIC CELL TRANSPLANTATION THE MICROBIAL COMMUNITY (MICROBIOTA) IN THE HUMAN GUT PLAYS A CENTRAL ROLE IN DIGESTION OF DIETARY FIBER, PROVIDING NUTRIENTS TO THE GUT MICROBIOTA AS WELL AS TO THE HOST THROUGH FERMENTATION PRODUCTS LIKE SHORT-CHAIN FATTY ACIDS. A MINORITY OF GUT BACTERIA CAN ALSO UTILIZE NUTRIENTS DERIVED FROM MUCUS, A COMPLEX MIXTURE OF SECRETED, MUCIN GLYCOPROTEINS THAT FORM A LAYER OVER THE EPITHELIUM TO PROTECT IT FROM MICROBIAL ENCROACHMENT. THE MOST ABUNDANT MUCIN UTILIZERS ARE MEMBERS OF THE GENERA AKKERMANSIA AND BACTEROIDES. ALLOGENEIC HEMATOPOIETIC CELL TRANSPLANTATION (ALLO HCT) IS AN IMPORTANT TREATMENT MODALITY PERFORMED FOR A VARIETY OF BENIGN AND MALIGNANT HEMATOLOGICAL CONDITIONS. PATIENTS RECEIVE SYSTEMIC CYTOTOXIC CONDITIONING, FOLLOWED BY INFUSION OF ALLOGENEIC HEMATOPOIETIC CELLS. IN TWO RECENT MANUSCRIPTS CO-AUTHORED BY THE PROGRAM PROJECT GRANT (PPG) PROJECT AND CORE LEADERS, MUCIN-UTILIZING INTESTINAL BACTERIA WERE FOUND TO CONTRIBUTE TO INTESTINAL GRAFT-VERSUS-HOST DISEASE (GVHD) THAT IS AGGRAVATED BY MEROPENEM ANTIBIOTIC TREATMENT, AND NEUTROPENIC FEVER (NF). THIS IS A REVISED PPG PROPOSAL WITH 3 INTEGRATED PROJECTS ADDRESSING HOW MUCIN-UTILIZING INTESTINAL BACTERIA CONTRIBUTE TO INTESTINAL GVHD. RECENT PRELIMINARY DATA FROM EXPERIMENTS PERFORMED IN MICE IDENTIFIED THAT AKKERMANSIA AND BACTEROIDES COMBINE TO COMPROMISE THE COLONIC MUCUS LAYER, PRODUCE HYPOTHERMIA IN MODELS OF HCT CONDITIONING, AND AGGRAVATE INTESTINAL GVHD. THE PROJECTS OF THIS PPG WILL EXTEND THESE FINDINGS, IMPROVING OUR UNDERSTANDING OF HOW DIET AND METABOLISM MODULATE EXPRESSION OF MUCIN-DEGRADING ENZYMES IN AKKERMANSIA AND BACTEROIDES. PROJECT 1 BUILDS UPON EXPERTISE IN CLINICAL ALLO HCT MICROBIOME PROFILING AND PRECLINICAL MODELING TO DEVELOP TRANSLATIONAL APPROACHES. PROJECT 2 INCORPORATES AN UNDERSTANDING OF THE NATURAL LANDSCAPE OF AKKERMANSIA GENETIC HETEROGENEITY, COUPLED WITH AN EXTENSIVE LIBRARY OF FUNCTIONAL AKKERMANSIA MUTANTS TO DEEPLY CHARACTERIZE THE ROLE OF AKKERMANSIA IN INTESTINAL GVHD. FINALLY, PROJECT 3 OFFERS A COMPREHENSIVE ANALYSIS OF THE EFFECTS OF SPECIFIC DIETARY GLYCANS ON MUCIN-DEGRADING BACTEROIDES AND GENETIC UNDERPINNINGS REGULATING EXPRESSION OF MUCIN-DEGRADING ENZYMES. SUPPORTING THESE ARE 2 UNIQUE CORES. CORE A WILL PROVIDE ADMINISTRATIVE SERVICES, MICROBIOME- SPECIALIZED BIOSTATISTICAL SUPPORT, CLINICAL SPECIMEN AND DATA COLLECTION FROM PATIENTS UNDERGOING ALLO HCT AT TWO CENTERS, AND BIONUTRITIONAL SUPPORT FOR COLLECTION OF DIETARY DATA SUPPLEMENTED BY METAGENOMIC SEQUENCING OF FECAL CHLOROPLAST DNA. CORE B WILL PERFORM RANDOM AND TARGETED BACTERIAL MUTAGENESIS TO SUPPORT HIGH THROUGHPUT MUTANT SCREENS AS WELL AS TESTING SPECIFIC HYPOTHESES, GENERATE FLUORESCENTLY LABELLED GLYCANS TO VISUALIZE GLYCAN UPTAKE AND DEGRADATION AT THE SINGLE CELL LEVEL, AND BUILD A DATABASE OF MUCIN-DEGRADING BACTERIAL ENZYMES THAT WILL SERVE AS A RESOURCE FOR THE INTESTINAL MICROBIOME COMMUNITY.

CONFORMAL TOTAL BODY AND MARROW IRRADIATION FOR LEUKEMIA

MENOPAUSAL TRANSITION - A WINDOW OF SUSCEPTIBILITY FOR THE PROMOTION OF BREAST CANCER BY ENVIRONMENTAL EXPOSURES

TARGETING MICRORNAS TO ERADICATE LEUKEMIA STEM CELLS

UNMASKING THE ROLES OF VIRAL GLYCOPROTEINS IN ORAL TRANSMISSION OF KSHV - PROJECT SUMMARY MORE THAN 44,000 NEW CASES OF KAPOSI SARCOMA (KS) ARE REPORTED GLOBALLY EACH YEAR, 84% OF WHICH OCCUR IN AFRICA. THIS AND OTHER KAPOSI SARCOMA-ASSOCIATED HERPESVIRUS (KSHV)-INDUCED MALIGNANCIES PREDOMINATE IN PEOPLE WITH ACQUIRED OR IATROGENIC IMMUNODEFICIENCIES. ALTHOUGH KSHV CAN BE DETECTED IN OTHER HUMAN BODY FLUIDS, ITS FREQUENT DETECTION IN SALIVA IN GROUPS BOTH WITH AND WITHOUT RISK OF SEXUALLY TRANSMITTED INFECTIONS (E.G., CHILDREN) SUGGESTS THAT THE ORAL CAVITY IS THE SITE OF PRIMARY ACQUISITION. HOWEVER, THE MECHANISM OF KSHV ORAL TRANSMISSION IN VIVO, PARTICULARLY THE CRITICAL VIRAL ENVELOPE GLYCOPROTEINS (GPS) REQUIRED FOR VIRAL ENTRY, REMAINS UNRESOLVED. SEVERAL KSHV–HOST INTERACTIONS HAVE BEEN IDENTIFIED, BUT ALL PRIOR EXPERIMENTS WERE PERFORMED IN VITRO AND HAVE NOT BEEN VALIDATED IN VIVO DUE TO PRIOR LACK OF AN APPROPRIATE ANIMAL MODEL. THROUGH COLLABORATION WITH THE WISCONSIN NATIONAL PRIMATE RESEARCH CENTER, OUR LABORATORY HAS ACCESS TO THE COMMON MARMOSET (CALLITHRIX JACCHUS, CJ), A RECENTLY DEVELOPED KSHV NON-HUMAN PRIMATE MODEL THAT IS SUSCEPTIBLE TO KSHV ORAL INFECTION, AND UNDER IMMUNOSUPPRESSION ACQUIRES KS-LIKE SKIN LESIONS. THE OBJECTIVE OF THIS APPLICATION IS TO ELUCIDATE THE MINIMUM GPS REQUIRED TO INITIATE PRIMARY ORAL INFECTION IN VIVO, AS A PREREQUISITE TO SELECTING KEY GPS FOR DEVELOPING AN EFFECTIVE PROPHYLACTIC VACCINE CANDIDATE. THIS APPLICATION BUILDS ON DR. OGEMBO’S RECENTLY COMPLETED NCI K01 CA184388-05 RESEARCH ON KSHV ENTRY MECHANISMS AND VACCINE DEVELOPMENT. RECENTLY, WE SHOWED THAT IN VITRO, THE KSHV GLYCOPROTEIN GH/GL IS ESSENTIAL FOR VIRAL INFECTION OF EPITHELIAL, ENDOTHELIAL, AND FIBROBLASTS CELLS, BUT NOT B CELLS. NOTABLY, WE AND OTHERS HAVE ALSO SHOWN THAT BOTH MONOCLONAL AND POLYCLONAL ABS TO KSHV GLYCOPROTEINS GB, GH/GL, AND GPK8.1, CAN NEUTRALIZE KSHV INFECTION OF DIVERSE PERMISSIVE HUMAN CELLS IN VITRO. BUILDING ON THIS SUCCESS, WE GENERATED KSHV DELETION MUTANTS LACKING THE FOUR GLYCOPROTEINS THOUGHT TO BE CRITICAL FOR VIRAL ENTRY (GB, GH/GL, GPK8.1) AND VARIOUS MONOCLONAL ANTIBODIES SPECIFIC TO THESE GPS. IN THIS PROJECT, WE WILL USE HUMAN EX VIVO SAMPLES AND THE CJ KSHV MODEL TO TEST THE HYPOTHESIS THAT GB AND GH/GL ARE CRITICAL FOR KSHV IN VIVO ORAL TRANSMISSION. THE PREMISE OF OUR PROPOSAL IS BUILT ON STRONG EVIDENCE THAT 1) KSHV CAN INFECT CJ, WHICH DEVELOP KS-LIKE SKIN LESIONS, AND 2) ABS AGAINST THE KSHV GLYCOPROTEINS GB AND GH/GL CAN NEUTRALIZE KSHV INFECTION IN VITRO AND EX VIVO. FURTHERMORE, THE PERMISSIVENESS TO KSHV INFECTION OF HUMAN CELLS EX VIVO AND CJ MAKES THESE PLATFORMS IDEAL TO TEST THE KSHV GP REQUIREMENTS FOR INFECTION. SUCCESSFUL COMPLETION OF THE PROPOSED STUDY WILL ELUCIDATE THE MINIMUM KSHV GPS REQUIRED FOR PRIMARY INFECTION IN EX VIVO AND IN VIVO MODELS, ADVANCING OUR LONG-TERM GOAL OF DEFINING THE INITIAL STEPS IN KSHV INFECTION OF HUMANS AND THE ROLE OF ANTIBODIES IN PROTECTING AGAINST THE EARLY STEPS OF KSHV TRANSMISSION. THIS WILL ULTIMATELY INFORM DESIGN AND DEVELOPMENT OF PROPHYLACTIC VACCINES THAT CAN PREVENT KSHV INFECTION AND ITS ASSOCIATED CANCERS.

NEURAL STEM CELL MEDIATED CE-CPT11 THERAPY FOR NEUROBLASTOMA

DEVELOPMENT OF SMALL MOLECULE INHIBITORS AND BIOLOGIC AGENTS FOR TREATMENT OF GLIOBLASTOMA USING INTRACEREBRAL MICRODIALYSIS AND SIGNATURES OF VULNERABILITY - PROJECT SUMMARY - OVERALL THE OVERALL GOAL OF THIS GLIOBLASTOMA (GBM) THERAPEUTICS NETWORK (GTN) U19 APPLICATION FROM CITY OF HOPE, TRANSLATIONAL GENOMICS RESEARCH INSTITUTE, AND UNIVERSITY OF ALABAMA AT BIRMINGHAM IS TO DEVELOP SUPERIOR TREATMENTS FOR PATIENTS WITH GBM, THE MOST COMMON AND AGGRESSIVE PRIMARY BRAIN TUMORS IN ADULTS. EFFECTIVE TREATMENTS REMAIN ELUSIVE AND PATIENTS ARE RARELY CURED WITH STANDARD THERAPIES. THIS GTN U19 APPLICATION EMBODIES A UNIQUE COMBINATION OF APPROACHES DESIGNED TO SIGNIFICANTLY ADVANCE THE TREATMENT OF PATIENTS WITH GBM BY ADDRESSING TUMOR HETEROGENEITY, BLOOD-BRAIN BARRIER PENETRATION, AND THE IMMUNOSUPPRESSIVE GBM TUMOR MICROENVIRONMENT. THE THREE PROPOSED RESEARCH PROJECTS WILL TRANSLATE THERAPEUTIC AGENTS FROM PRECLINICAL DEVELOPMENT, THROUGH IND-ENABLING STUDIES, AND INTO PHASE I CLINICAL STUDIES IN ADULT PATIENTS WITH GBM. EACH PROJECT IS BASED ON NOVEL MOLECULAR PRECLINICAL STUDIES WITH SMALL-MOLECULE INHIBITORS AND IMMUNOMODULATORY AGENTS THAT USE SIGNATURE-GUIDED ASSESSMENT AND TREATMENTS. SPECIFIC GOALS OF THE PROJECTS ARE: PROJECT 1. DEVELOP AND CLINICALLY TEST AN ENGINEERED ONCOLYTIC HERPES VIRUS EXPRESSING A FULL- LENGTH ANTI-CD47 MONOCLONAL ANTIBODY FOR TREATMENT OF GBM. PROJECT 2. DEVELOP AND CLINICALLY TEST TASQUINIMOD AS AN ADJUNCT TO ENHANCE THE EFFICACY OF ANTI-GBM IMMUNOTHERAPIES ADMINISTERED PERI-OPERATIVELY. PROJECT 3. DEVELOP AND CLINICALLY TEST A MOLECULAR “SIGNATURES OF VULNERABILITY” GUIDED TREATMENT OF GBM WITH NEDDYLATION INHIBITOR PEVONEDISTAT. IN ADDITION, THIS U19 APPLICATION PROPOSES STRATEGIES THAT WILL ADDRESS MAJOR BARRIERS IN DRUG DEVELOPMENT BY INCORPORATING TWO INNOVATIVE RESEARCH TOOLS: 1) INTRACEREBRAL MICRODIALYSIS TO RATIONALLY SELECT APPROPRIATE SYSTEMICALLY ADMINISTERED THERAPIES FOR TESTING IN GBM PATIENTS AND 2) NEXT GENERATION EXOME AND TRANSCRIPTOME SEQUENCING TO IDENTIFY MOLECULAR “SIGNATURES OF VULNERABILITY” THAT CAN GUIDE APPROPRIATE PATIENT SELECTION FOR CLINICAL TRIAL ENROLLMENT. THESE ANALYTICAL CAPABILITIES WILL ENABLE US TO QUANTIFY CNS DRUG PENETRATION AND DISSECT GENOMIC HETEROGENEITY IN TUMOR AND STROMAL CELLS IN THE PROPOSED CLINICAL TRIALS. ALSO, TWO OF THE PROPOSED PROJECTS LEVERAGE CITY OF HOPE’S GMP FACILITIES TO MANUFACTURE BIOLOGICAL AGENTS AND SMALL MOLECULES THAT WILL BE TESTED IN ADULT GBM PATIENTS FOR THE FIRST TIME. IN SUMMARY, THE INNOVATIVE PROJECTS AND SHARED RESOURCES CORES IN THIS APPLICATION COMBINE OUR STRENGTHS IN BASIC, TRANSLATIONAL, AND CLINICAL RESEARCH IN A HIGHLY COLLABORATIVE SETTING THAT PROMOTES THE SHARING OF IDEAS, RESULTS, RESOURCES, AND CLINICAL POPULATIONS TO DEVELOP EFFECTIVE TREATMENTS FOR GBM. IF SUCCESSFUL, DATA GENERATED BY THESE STUDIES HAVE THE POTENTIAL TO TRANSFORM THE TREATMENT OF ADULT GBM PATIENTS BY INTRODUCING NEW AGENTS THAT CIRCUMVENT TUMOR HETEROGENEITY AND IMMUNOSUPPRESSION.

VALIDATION OF EPIGENOMIC BIOMARKERS FOR THYROID CANCER DIAGNOSTICS - PROJECT SUMMARY/ABSTRACT EACH YEAR, UP TO 50,000 PATIENTS IN THE UNITED STATES RECEIVE UNNECESSARY THYROIDECTOMIES. THESE UNNECESSARY THYROIDECTOMIES ARE DUE TO DIFFICULTIES IN PREOPERATIVELY DISTINGUISHING BENIGN THYROID NODULES FROM THYROID CANCERS. IN PRELIMINARY DATA, WE DEVELOPED AN EPIGENETIC TEST THAT SHOWS PROMISE TO DISTINGUISH BENIGN VERSUS MALIGNANT THYROID NODULES. HERE, WE WILL RIGOROUSLY TEST AND VALIDATE THE ABILITY OF OUR EPIGENETIC BIOMARKERS TO EVALUATE THE BIOLOGIC AGGRESSIVENESS OF THYROID NODULES AND DETERMINE WHETHER THE NEW EPIGENETIC TESTING WILL IMPROVE THYROID NODULE MANAGEMENT TOWARDS THE ERADICATION OF UNNECESSARY THYROIDECTOMIES. CURRENT MOLECULAR DIAGNOSTICS FOR INDETERMINATE THYROID NODULES, WHILE PROVIDING SOME IMPROVEMENT, HAVE NOT ELIMINATED THE UNNECESSARY THYROIDECTOMIES. CURRENT MOLECULAR DIAGNOSTICS ARE BASED ON MOLECULAR DIFFERENCES BETWEEN NORMAL THYROID TISSUE AND THYROID CANCER. HOWEVER, BENIGN THYROID NODULES CAN CONTAIN MANY MOLECULAR ALTERATIONS INCLUDING GENE FUSIONS AND MUTATIONS. AS A RESULT, OVER HALF OF THYROID NODULES WITH A SIGNIFICANT CANCER RISK ACCORDING TO THE CURRENT MOLECULAR CLASSIFIERS ARE FOUND TO BE BENIGN AFTER THYROIDECTOMY. IN OUR PUBLISHED PRELIMINARY DATA, WE PERFORMED A GENOME-WIDE DNA METHYLATION ANALYSIS OF 109 SURGICALLY EXCISED THYROID NODULES AND ADJACENT BENIGN TISSUE. WE FOUND THAT THE DNA METHYLATION PATTERN IN BENIGN NODULES IS DIFFERENT FROM THYROID CANCER AND NORMAL THYROID. BASED ON THE DNA METHYLATION PATTERN SPECIFIC TO BENIGN NODULES AND THE DNA METHYLATION PATTERN SPECIFIC TO THYROID CANCER, WE DEVELOPED THE DIAGNOSTIC DNA METHYLATION SIGNATURE (DDMS) APPROACH TO DISTINGUISH BETWEEN BENIGN VERSUS MALIGNANT NODULES. IN A RETROSPECTIVE PILOT STUDY PERFORMED UNDER 1R21CA223367, WE DEVELOPED DDMS FURTHER (DDMS-2). WE TESTED THE ABILITY OF THE DDMS-2 ASSAY TO DISTINGUISH BENIGN FROM MALIGNANT SURGICALLY EXCISED THYROID NODULES (N=121). IN THIS PILOT STUDY, DDMS- 2 HAD AN ESTIMATED POSITIVE PREDICTIVE VALUE (PPV) OF 96% AND A NEGATIVE PREDICTIVE VALUE (NPV) OF 98%. GUIDED BY OUR PRELIMINARY DATA, WE HYPOTHESIZE THAT DDMS (I) CAN BE SUCCESSFULLY USED FOR MOLECULAR THYROID CANCER DIAGNOSTICS OF PRE-OPERATIVE THYROID NODULE ASPIRATIONS; (II) WILL HAVE SUPERIOR PERFORMANCE IN COMPARISON TO CURRENT THYROID CANCER MOLECULAR TESTING AND (III) CAN AFFECT PHYSICIAN DECISION-MAKING TOWARDS ELIMINATION OF UNNECESSARY THYROIDECTOMIES. WE WILL ACCOMPLISH OUR OVERALL OBJECTIVE BY PURSUING THE FOLLOWING SPECIFIC AIMS: AIM 1: TO PERFORM ANALYTICAL VALIDATION OF THE DDMS-2 ASSAY. AIM 2: TO DETERMINE THE DDMS-2 ACCURACY IN A PROSPECTIVE COHORT OBTAINED FROM 7 MEDICAL CENTERS AND CONTAINING 1450 THYROID NODULE ASPIRATIONS INCLUDING 800 ASPIRATIONS WITH INDETERMINATE CYTOPATHOLOGY. AIM 3: TO COMPARE THE DIAGNOSTIC ACCURACY BETWEEN DDMS- 2 AND TWO CURRENT THYROID CANCER MOLECULAR DIAGNOSTIC APPROACHES AND TO EVALUATE HOW THE KNOWLEDGE OF THE DDMS-2 RESULTS IMPACTS CLINICAL MANAGEMENT OF THYROID NODULES. THE DEVELOPMENT OF A MORE ACCURATE ASSAY TO DISTINGUISH BENIGN AND MALIGNANT THYROID NODULES WILL ADDRESS CURRENT CLINICAL LIMITATIONS AND REDUCE THE NUMBER OF NEEDLESS THYROIDECTOMIES AND ASSOCIATED MORBIDITIES.

GERIATRIC ONCOLOGY RESEARCH INFRASTRUCTURE TO IMPROVE CLINICAL CARE

ROLE OF THE XBP1S/GFAT1 AXIS IN PATHOLOGICAL CARDIAC REMODELLING

KSHV SUBUNIT VACCINE CANDIDATES TO ELICIT POTENT HUMORAL IMMUNE REPONSES AGAINST KSHV INFECTION - PROJECT SUMMARY KAPOSI SARCOMA-ASSOCIATED HERPESVIRUS (KSHV) HAS BEEN CLASSIFIED AS A DIRECT CARCINOGEN BY THE INTERNATIONAL AGENCY FOR RESEARCH ON CANCER BECAUSE OF ITS ABILITY TO CAUSE KAPOSI SARCOMA (KS) AND TWO RARE TYPES OF B-CELL LYMPHOMA. KS FREQUENTLY OCCURS AMONG IATROGENIC OR HIV/AIDS-INDUCED IMMUNOSUPPRESSED INDIVIDUALS. TO DATE, THERE IS NO LICENSED KSHV VACCINE THAT CAN PREVENT PRIMARY INFECTION AND SUBSEQUENT MALIGNANCIES. OUR OBJECTIVE IN THIS APPLICATION IS TO OPTIMIZE THE INCLUSION OF KEY KSHV GLYCOPROTEINS (GPS), WHICH ARE INVOLVED IN EPITHELIAL, ENDOTHELIAL, FIBROBLAST AND B-CELL ENTRY, INTO MULTIVALENT SUBUNIT VACCINE CANDIDATES THAT CAN STIMULATE NEUTRALIZING ANTIBODY (NAB) IMMUNE RESPONSES TO PREVENT OR LIMIT KSHV INFECTION AND KSHV+ CANCERS IN VIVO. THIS APPLICATION BUILDS ON MY RECENTLY COMPLETED NCI K01 CA184388-05 RESEARCH ON KSHV ENTRY MECHANISMS AND VACCINE DEVELOPMENT. RECENTLY, WE SHOWED THAT IN VITRO, THE KSHV GLYCOPROTEIN GH IS ESSENTIAL FOR VIRAL INFECTION OF EPITHELIAL, ENDOTHELIAL, AND FIBROBLASTS CELLS, BUT NOT B CELLS. NOTABLY, WE AND OTHER HAVE ALSO SHOWN THAT BOTH MONOCLONAL AND POLYCLONAL ABS TO KSHV GLYCOPROTEINS K8.1, GB, AND GH/GL CAN NEUTRALIZE KSHV INFECTION OF DIVERSE PERMISSIVE HUMAN CELLS IN VITRO. BUILDING ON THIS SUCCESS, WE HAVE GENERATED QUADRIVALENT VIRUS-LIKE PARTICLES (KSHV-LPS) INCORPORATING THE FOUR GLYCOPROTEINS CRITICAL FOR VIRAL ENTRY (K8.1, GB AND GH/GL). IN THIS APPLICATION WE WILL USE WILD-TYPE AND HUMANIZED MICE AND COMMON MARMOSET (CALLITHRIX JACCHUS) MODELS TO TEST THE HYPOTHESIS THAT PURIFIED KSHV-LPS DELIVERED DIRECTLY OR THROUGH IMMUNIZATION WITH A MODIFIED VACCINIA ANKARA VECTOR (MVA-KSHV-LPS) WILL ELICIT ROBUST PROTECTIVE NAB RESPONSES TO KSHV INFECTION AND ITS ASSOCIATED MALIGNANCIES. THE PREMISE OF OUR PROPOSAL IS BUILT ON STRONG EVIDENCE THAT 1) INFECTION WITH KSHV DOES NOT OCCUR DURING EARLY CHILDHOOD, AS IS TYPICAL FOR OTHER HERPESVIRUSES, OPENING A WINDOW OF OPPORTUNITY FOR VACCINATION AND 2) ABS AGAINST THE KSHV GLYCOPROTEINS K8.1, GB, AND GH/GL CAN NEUTRALIZE KSHV INFECTION. FURTHERMORE, THE PERMISSIVENESS OF HUMANIZED MICE AND MARMOSETS TO KSHV INFECTION OFFERS AN IDEAL PLATFORM TO TEST CANDIDATE VACCINES. THUS, A POLYVALENT VACCINE THAT INDUCES PROPHYLACTIC NEUTRALIZING ABS RESPONSES WILL NOT ONLY BE AN INVALUABLE CANDIDATE VACCINE IN PREVENTING KSHV INFECTION, BUT ALSO OF UTMOST IMPORTANCE IN PREVENTING KSHV-ASSOCIATED DISEASES. WE WILL PROVIDE EVIDENCE FOR THE SAFETY OF OUR CANDIDATE KSHV VACCINE BASED ON THREE PRE-CLINICAL ANIMAL MODELS AS PREREQUISITE DATA FOR AN IND APPLICATION FOR A PHASE I CLINICAL TRIAL. IN THE LONG TERM, THE SUCCESS OF OUR APPROACH WILL INTRODUCE A NEW VACCINE TO THE MARKET WITH A POTENTIAL FOR REDUCING GLOBAL INCIDENCE OF KSHV+ MALIGNANCIES (>44,000 CASES/YEAR), AND THE POSSIBILITY OF LIMITING KSHV INFECTION AND ASSOCIATED MALIGNANCIES IN DEVELOPING COUNTRIES OR ERADICATING THEM FROM DEVELOPED COUNTRIES WHERE KSHV SEROPREVALENCE IS <10%.

PHASE I MOLECULAR AND CLINCAL PHARMACODYNAMIC TRIALS

GERMLINE AND TUMOR GENOMIC ANALYSES OF BREAST CANCER IN LATINAS

TECHNOLOGY-ENABLED ACTIVATION OF SKIN CANCER SCREENING FOR HEMATOPOIETIC CELL TRANSPLANTATION SURVIVORS AND THEIR PRIMARY CARE PROVIDERS

SELF-MANAGEMENT TO OPTIMIZE SURVIVORSHIP CARE AND OUTCOMES IN LUNG AND COLORECTAL CANCER

1/2 DRUG DEVELOPMENT AND CAPACITY BUILDING: A UCR/COH-CCC PARTNERSHIP - THE UNITED STATES (U.S.) HAS LED THE WORLD IN DRUG DISCOVERY FOR OVER 50 YEARS. WHILE THIS IS A SIGNIFICANT ACCOMPLISHMENT, U.S. DRUGS HAVE BEEN ALMOST EXCLUSIVELY OPTIMIZED AND TESTED IN NON-HISPANIC EUROPEAN- AMERICANS AND BEEN MINIMALLY EVALUATED IN MEN- OR WOMEN-OF-COLOR. AS A RESULT, DRUGS THAT WORK WELL IN NON- HISPANIC EUROPEAN-AMERICANS (ANGLOS), MAY HAVE UNEXPECTED TOXICITY OR DECREASED EFFICACY IN MOST OF THE WORLD’S POPULATION. DISPARITIES IN U.S. DRUG DEVELOPMENT OCCUR THROUGHOUT THE ENTIRE DRUG DISCOVERY PIPELINE. ONLY A SMALL NUMBER OF BASIC SCIENTISTS ARE LATINO/HISPANIC- OR AFRICAN-AMERICAN. INITIAL DRUG DEVELOPMENT AND OPTIMIZATION TAKES PLACE IN CELL LINES DERIVED FROM ANGLOS. LESS THAN 2% OF PHYSICIANS CONDUCTING CLINICAL TRIALS ARE LATINO/HISPANIC- OR AFRICAN-AMERICAN6. MOST CLINICAL TRIALS PARTICIPANTS ARE ANGLOS. YET, NEW DRUGS ARE FDA APPROVED FOR LATINOS/HISPANICS- AND AFRICAN-AMERICANS/AFRICANS WITHOUT SUFFICIENT TESTING. IT IS UNACCEPTABLE THAT DRUGS ARE DEVELOPED BY, AND OPTIMIZED FOR, ONLY A FRACTION OF OUR CITIZENS. IN THIS U54 PARTNERSHIP WE AIM TO DEVELOP THE RESOURCES, INFRASTRUCTURE, AND TRAINING TO MENTOR THE NEXT GENERATION OF RESEARCHERS THAT REFLECT THE DIVERSITY OF OUR CATCHMENT AREA. BUILDING ON OUR SUCCESSFUL P20 GRANT, HERE IN THIS U54 PARTNERSHIP, UCR AND COHCCC AIM TO DEVELOP THE COLLABORATIONS, RESOURCES, AND TRAINING PROGRAMS TO REDUCE DISPARITIES IN DRUG DEVELOPMENT THROUGHOUT THE ENTIRE DRUG DEVELOPMENT PIPELINE. OUR GOAL IS FOR THIS PROGRAM TO BECOME A FOCAL POINT FOR UCR AND COHCCC TO MENTOR AND TRAIN A DIVERSE FORCE OF CANCER BIOLOGISTS AND ADDRESS THE DISPARITIES IN CANCER THERAPEUTICS AND DRUG DEVELOPMENT. ALREADY, OUR P20 HAS FOSTERED JOINT R01 GRANTS, K01 GRANTS, AND PRE-/POST-DOCTORAL FELLOWSHIPS. BOTH INSTITUTIONS ARE HIGHLY COMMITTED - COHCCC CONTRIBUTED OVER $800K TO OUR P20 GRANT AND WILL CONTRIBUTE $250K/YEAR TO ENSURE THE SUCCESS OF THIS U54 PARTNERSHIP. AIM 1 WILL STRENGTHEN UCR’S CANCER RESEARCH CAPACITY AND DEVELOP THE RESOURCES TO INCREASE UCR/COHCCC’S ABILITY TO JOINTLY DEVELOP THERAPEUTIC AGENTS OPTIMIZED FOR THE DIVERSE POPULATIONS IN OUR CATCHMENT AREA. AIM 2 WILL INCREASE THE CAPACITY OF UCR AND COHCCC TO JOINTLY DEVELOP DRUGS THAT TARGET DISPARITIES IN SURVIVAL AFFECTING THE DIVERSE INDIVIDUALS LIVING IN OUR SOUTHERN CALIFORNIA COMMUNITIES. AIM 3 WILL PROVIDE THE TRAINING, OPPORTUNITY, AND MENTORSHIP TO ENSURE THAT THE NEXT GENERATION OF THERAPEUTIC SCIENTISTS AND CLINICAL TRIALISTS REFLECT THE DIVERSITY OF SOUTHERN CALIFORNIA.

CANCER GENETICS PROFESSIONAL EDUCATION IN A GLOBAL COMMUNITY OF PRACTICE

LEVERAGING DIGITAL HEALTH SOLUTIONS TO REDUCE LEARNING AND FUNCTIONAL DISPARITIES IN CHILDREN WITH CANCER - PROJECT SUMMARY/ABSTRACT AS THE POPULATION OF CHILDHOOD CANCER SURVIVORS GROWS, THE IMPACT OF LONG-TERM TREATMENT-RELATED SIDE EFFECTS ALSO GROWS. CANCER TREATMENTS FOR CHILDREN WITH LEUKEMIA AND LYMPHOBLASTIC LYMPHOMA (LL) HAVE LONG-LASTING TOXIC EFFECTS ON THE DEVELOPING BRAIN, KNOWN AS NEUROCOGNITIVE LATE EFFECTS, WHICH CAN IMPACT THE CHILDREN’S LEARNING, FUNCTION, AND ABILITY TO ACHIEVE INDEPENDENCE AS ADULTS. HOWEVER, THESE PROBLEMS ARE OFTEN UNTREATED IN LEUKEMIA AND LL SURVIVORS, CREATING A MEDICALLY UNDERSERVED POPULATION. RECOGNIZING THE IMPACT OF FAMILIAL FACTORS ON SURVIVOR OUTCOMES, WE DESIGNED A PARENT-DIRECTED TRAINING INTERVENTION (THE HIGH-INTENSITY INTERVENTION PROGRAM, HIP), WHICH TEACHES PARENTS ABOUT BRAIN DEVELOPMENT AND NEUROCOGNITIVE LATE EFFECTS; TRAINS THEM ON TOOLS TO IMPROVE THEIR CHILD’S BEHAVIOR AND COGNITIVE FUNCTIONING; PROVIDES TIPS FOR ESTABLISHING BETTER LEARNING ENVIRONMENTS AND EFFECTIVE STUDY HABITS; AND HELPS THEM TO MANAGE STRESS FOR THEMSELVES AND THEIR CHILDREN. PILOT TRIALS OF THE HIP IN ENGLISH- AND SPANISH-SPEAKING FAMILIES DEMONSTRATED EFFICACY BUT ALSO REVEALED CRITICAL BARRIERS TO SUCCESS, INCLUDING TRAVEL TIME AND SCHEDULING CHALLENGES. DIGITAL HEALTH TECHNOLOGIES OFFER TRANSFORMATIVE SOLUTIONS TO IMPROVE THE EFFICIENCY, QUALITY, AND CONVENIENCE OF HEALTHCARE DELIVERY. HOWEVER, “EHEALTH” HAS NOT BEEN APPLIED TO A PARENT-DIRECTED INTERVENTION TO IMPROVE EDUCATIONAL OUTCOMES FOR CHILDREN. THEREFORE, WE PROPOSE TO TEST A NEW EHEALTH VERSION OF OUR ENGLISH/SPANISH INTERVENTION (HIP-EHEALTH) IN A RANDOMIZED CLINICAL TRIAL OF 180 PARENT/CHILD DYADS FROM 4 SITES IN CALIFORNIA. HIP-EHEALTH ADDRESSES THE REMAINING BARRIERS TO HIP ACCESS AND INCORPORATES IMPROVEMENTS RECOMMENDED BY PARENTS IN OUR EARLIER TRIALS. WE WILL DELIVER HIP-EHEALTH FROM A SINGLE CENTRAL SITE (CITY OF HOPE) THROUGH A HIPAA-COMPLIANT STUDY WEBSITE THAT HOSTS ZOOM VIDEOCONFERENCING FOR HIP SESSIONS; DIGITIZED STUDY CONTENT; SUPPLEMENTAL MULTIMEDIA CONTENT; LINKS TO THE AWARD-WINNING IXL LEARNING ENVIRONMENT; GAMIFICATION FEATURES AND AUTOMATED REMINDERS TO INCREASE ENGAGEMENT; AND ROBUST USER ANALYTICS. OTHER ADVANCES INCLUDE A STREAMLINED 4-SESSION PROGRAM, AN ENHANCED BOOSTER PHASE, AND GREATER INCLUSION OF THE CHILDREN. WE HYPOTHESIZE THAT HIP-EHEALTH WILL PRODUCE GREATER IMPROVEMENTS IN CHILD AND PARENT OUTCOMES THAN A LOWER-INTENSITY PROGRAM (LIP) THAT MIMICS THE USUAL CARE PROVIDED TO SURVIVORS OF PEDIATRIC BRAIN TUMORS (I.E., A SINGLE MEETING TO DISCUSS THE CHILD’S NEUROPSYCHOLOGICAL TESTING RESULTS AND PROVIDE RECOMMENDATIONS FOR OPTIMAL LEARNING). OUR STUDY AIMS TO: (1) EVALUATE THE EFFECTIVENESS OF HIP-EHEALTH ON THE LEARNING AND SCHOOL PERFORMANCE OF PEDIATRIC CANCER SURVIVORS; (2) EVALUATE THE EFFECTIVENESS OF HIP-EHEALTH ON THE “PRO-LEARNING” EFFICACY OF THEIR PARENTS; (3) INVESTIGATE THE EXTENT TO WHICH THE PARENTS’ EFFICACY AND/OR CHILDREN’S USE OF ONLINE LEARNING ACTIVITIES IS ASSOCIATED WITH CHANGES IN THE CHILDREN’S SCHOOL PERFORMANCE; AND (4) ASSESS FACTORS THAT IMPACT THE PARENTS’ ABILITY TO COMPLETE THE INTERVENTION. WE ANTICIPATE THIS STUDY WILL HELP SHAPE A SCALABLE AND EFFECTIVE THERAPY THAT IS EASILY INTEGRATED INTO STANDARD CARE.

THE BIOLOGY AND DIAGNOSIS OF HNPCC

A MULTIMEDIA SELF-MANAGEMENT INTERVENTION TO PREPARE FAMILY CAREGIVERS AND PATIENTS FOR LUNG CANCER SURGERY

ROLE OF RECEPTOR FOR ADVANCED GLYCATION END PRODUCT (RAGE) PATHWAY IN BRAIN TUMOR

STRUCTURAL BASIS OF G-PROTEIN SELECTIVITY IN GPCRS USING MULTISCALE DYNAMICS

INVESTIGATION OF THE ROLES OF NUCLEAR RECEPTOR FXR IN HEPATOCELLULAR

COMBINED BREAST MRI/BIOMARKER STRATEGIES TO IDENTIFY AGGRESSIVE BIOLOGY

EPIGENETIC DAMAGE IN WOMEN LIVING IN LA FOOD-DESERT ZIP CODES

CLINICAL EVALUATION OF CHLOROTOXIN-REDIRECTED CHIMERIC ANTIGEN RECEPTOR (CAR) T CELLS FOR TREATMENT OF GLIOBLASTOMA

MECHANISM OF ESTROGEN INDEPENDENT PROLIFERATION IN ER+ BREAST CANCER CELLS - ABSTRACT COMBINING CYCLIN-DEPENDENT KINASE (CDK) INHIBITORS WITH ENDOCRINE THERAPY IMPROVES OUTCOMES FOR METASTATIC ESTROGEN RECEPTOR POSITIVE (ER+), HER2 NEGATIVE, BREAST CANCER PATIENTS. HOWEVER, THE VALUE OF THIS COMBINATION IN POTENTIALLY CURABLE EARLIER STAGE PATIENTS IS VARIABLE. OUR PRELIMINARY RESULTS EXAMINED THE EVOLUTIONARY TRAJECTORIES OF EARLY STAGE BREAST CANCER TUMORS USING SINGLE CELL TRANSCRIPTOMIC PROFILING OF SERIAL TUMOR BIOPSIES FROM A CLINICAL TRIAL OF PREOPERATIVE ENDOCRINE THERAPY ALONE (LETROZOLE) OR IN COMBINATION WITH THE CELL CYCLE INHIBITOR RIBOCICLIB. RESISTANT TUMORS WITH ACCELERATED LOSS OF ESTROGEN SIGNALING SHOW UP-REGULATION OF THE JNK PATHWAY, WHILE THOSE THAT MAINTAIN ESTROGEN SIGNALING DURING THERAPY SHOW POTENTIATION OF CDK4/6 ACTIVATION CONSISTENT WITH ERBB4 AND ERK SIGNALING UP-REGULATION. CELL CYCLE RECONSTRUCTION IDENTIFIED THAT TUMORS CELLS CAN REACTIVATE DURING COMBINATION TREATMENT, INDICATING STRONGER SELECTION FOR A PROLIFERATIVE STATE. WE HYPOTHESIZE THAT RESISTANCE TO CDK4/6 INHIBITION IN EARLIER STAGE BREAST CANCER IS DRIVEN BY JNK MAPK PATHWAY STIMULATION AND REACTIVATION OF THE CELL CYCLE THROUGH PROMOTION OF CDK6 EXPRESSION OR DECREASED CELL CYCLE INHIBITOR FUNCTION. IN AIM 1, WE WILL USE A NEW MECHANISTIC MODEL OF CDK4/6 REGULATION BY CELL CYCLE INHIBITORS AND PROMOTERS (CIP) THAT COUPLES ESTROGEN AND JNK SIGNALING WITH CELL CYCLE PROGRESSION TO MEASURE THE MECHANISMS DRIVING CELL CYCLE ACTIVATION IN A SERIES OF ISOGENIC CELL LINES SENSITIVE AND RESISTANT TO CDK4/6 AND ENDOCRINE INHIBITORS AND IN PATIENT TUMOR CELLS. THIS ANALYSIS WILL REVEAL HOW DISTINCT SIGNALING PATHWAYS CONTRIBUTE TO CELL CYCLE REACTIVATION DURING ESTROGEN, CDK4/6 AND JNK INHIBITION TREATMENTS AND PROVIDE SIGNATURES OF EACH RESISTANT MECHANISM ACROSS CELL TYPES, OVER TIME AND BETWEEN SYSTEMS. AIM 2 LEVERAGES OUR COLLECTION OF PATIENT TUMORS FROM THE FELINE CLINICAL TRIAL TO DISCOVER THE INTRACELLULAR AND INTRATUMORAL RESISTANCE MECHANISMS DRIVING PROLIFERATION. FUNDAMENTAL RESISTANCE MECHANISMS WILL BE MEASURED IN OVER ~300,000 PATIENT CELLS FROM 360 TUMOR SAMPLES USING SINGLE CELL RNA SEQUENCING DATA ALREADY IN HAND TO IDENTIFY CORE INTRACELLULAR SIGNALING STATES THAT ACT ALONE OR IN CONCERT TO DRIVE PROLIFERATION. NEXT, THE POPULATION OF CELLS WITHIN EACH TUMOR WILL BE ANALYZED TO QUANTIFY INTRATUMORAL HETEROGENEITY AND HOW RESISTANT POPULATIONS DIFFER IN GROWING OR SHRINKING TUMORS DURING DRUG TREATMENT. APPLYING CIP TO PROJECT PROLIFERATION ACROSS PATIENT TUMOR CELLS WILL ALLOW PREDICTION OF INHIBITOR STRATEGIES THAT MOST EFFECTIVELY BLOCK INTRACELLULAR AND INTRATUMORAL PROLIFERATION. LASTLY, AIM 3 WILL APPLY A SERIES OF JNK PATHWAY DRUGS WITH CLINICAL POTENTIAL TO DESIGN AND TEST TREATMENT STRATEGIES THAT MAINTAIN DURABLE INHIBITION OF PROLIFERATION IN ER+ CANCER CELLS. ITERATIVE FEEDBACK BETWEEN MATHEMATICAL MODELS AND PATIENT/EXPERIMENTAL DATA SERVES TO PROVIDE A DEEP UNDERSTANDING OF CELL CYCLE REGULATION AND MECHANISMS OF DYSREGULATION LEADING TO RESISTANCE. TOGETHER, THESE EXPERIMENTS WILL REVEAL THE BALANCE BETWEEN ESTROGEN AND ALTERNATIVE MEDIATED JNK SIGNALING, AND THEIR ROLES IN RESISTANCE AND PROVIDE A GUIDE FOR THERAPEUTIC REGIMES WITH MORE DURABLE CONTROL OF CANCER CELL PROLIFERATION.

INFORMATION FLOW AND STATE TRANSITIONS AT THE SYSTEM AND MULTI-DIMENSIONAL SCALES IN LEUKEMIA PROGRESSION

RISK FACTORS FOR MOLECULAR SUBTYPES OF NHL A PROSPECTIVE EVALUATION

CMVPEPVAX TO PROTECT HCT RECIPIENTS FROM CYTOMEGALOVIRUS INFECTION

DIAGNOSTIC ASSAY SYSTEMS FOR BOTULINUM NEUROTOXINS

PITT-CAL ETCTN PK RESOURCE LABORATORY

ENHANCING THE INTRACELLULAR FUNCTIONING OF ANTI-HIV RNAS

CARDIOVASCULAR RESERVE CAPACITY IN SURVIVORS OF HEMATOPOIETIC CELL TRANSPLANTATION

EXERCISE AS INTERCEPTION THERAPY IN PRIMARY HIGH RISK CANCER

A PHASE 1 STUDY TO EVALUATE CHIMERIC ANTIGEN RECEPTOR (CAR) T CELLS TARGETING TAG72 IN PATIENTS WITH RECURRENT EPITHELIAL OVARIAN CANCER - PROJECT SUMMARY PATIENTS WITH RECURRENT EPITHELIAL OVARIAN CANCER (EOC) HAVE A POOR PROGNOSIS WITH A POST-RELAPSE MEDIAN SURVIVAL OF APPROXIMATELY 30 MONTHS AND LIMITED THERAPEUTIC OPTIONS, THUS PRESENTING A FUNDAMENTAL UNMET MEDICAL NEED. PROGRESS IN IMMUNOTHERAPY ACROSS A BROAD RANGE OF TUMOR TYPES PROVIDES HOPE THAT IMMUNOLOGICAL APPROACHES MAY IMPROVE OUTCOMES FOR PATIENTS WITH EOC. PARTICULARLY, A TYPE OF IMMUNOTHERAPY CALLED CHIMERIC ANTIGEN RECEPTOR (CAR) T CELL THERAPY RETRAINS THE IMMUNE SYSTEM TO TARGET CANCERS BY RECOGNIZING SPECIFIC CANCER MARKERS. EOC PRESENTS SEVERAL CHALLENGES TO EFFECTIVE CAR T CELL IMMUNOTHERAPY, INCLUDING POOR TUMOR SITE INFILTRATION, ACTIVATION, INADEQUATE FUNCTION AND PERSISTENCE OF THESE T CELLS WITHIN THE HARSH PERITONEAL TUMOR MICROENVIRONMENT. ADDITIONALLY, THERE ARE A LACK OF EFFECTIVE CAR T CELL TARGETS ON THE SURFACE OF ADVANCED EOC TUMOR CELLS. OUR GOAL IS TO DEVELOP EFFECTIVE THERAPIES AGAINST METASTATIC EOC, WITH A SPECIFIC FOCUS ON REGIONAL DELIVERY OF CAR T CELL THERAPIES TO TREAT PERITONEAL METASTASIS. TAG72 IS HIGHLY OVER- EXPRESSED IN EOC AND OTHER SOLID TUMORS WITH LITTLE OR NO EXPRESSION IN NORMAL TISSUES, MAKING IT AN IDEAL TARGET FOR CAR T CELL THERAPY. OUR TEAM AT CITY OF HOPE HAS DEVELOPED AND COMPLETED LABORATORY TESTING OF A TAG72- TARGETING CAR T CELL THERAPY. OUR PRECLINICAL DATA ALSO SUPPORTS SUPERIOR ANTI-TUMOR ACTIVITY WHEN TAG72 CAR T CELLS ARE ADMINISTERED REGIONALLY BY INTRAPERITONEAL DELIVERY VERSUS SYSTEMICALLY BY INTRAVENOUS DELIVERY, LIKELY DUE TO DIRECT AND IMMEDIATE ANTIGEN CAR T CELL ACCESS TO TUMOR CELLS. THE HYPOTHESIS IS THAT REGIONALLY- ADMINISTERED TAG72-CAR T CELLS WILL BE SAFE AND MEDIATE ANTI-TUMOR EFFECTS, WHICH WILL BE ASSESSED IN THE FOLLOWING SPECIFIC AIMS: 1) EVALUATE SAFETY AND FEASIBILITY OF REGIONAL INTRAPERITONEAL DELIVERY OF TAG72-CAR T CELLS IN PATIENTS WITH ADVANCED EOC IN A PHASE 1 CLINICAL TRIAL; 2) ASSESS CAR T CELL-MEDIATED IMMUNE LANDSCAPE CHANGES THAT MAY INDICATE THERAPEUTIC RESPONSE OR RESISTANCE; AND 3) INVESTIGATE PATHWAYS OF TUMOR RESISTANCE AND CAR T CELL-INDUCED TUMOR EVOLUTION. OUR PROGRAM HAS INCORPORATED AN INNOVATIVE USE OF PRE-CONDITIONING REGIMENS TO OUR SOLID TUMOR CAR T CELL THERAPIES, REGIONAL ROUTES OF CAR T CELL ADMINISTRATION, AND A FULLY- OPTIMIZED TAG72-CAR CONSTRUCT. THESE FEATURES AIM TO IMPROVE THE POTENCY AND SELECTIVITY OF TARGETING TAG72+ TUMORS WHILE POTENTIALLY MINIMIZING IMMUNE RESPONSES THAT LIMIT PERSISTENCE AND/OR FUNCTION OF TAG72-CAR T CELLS. THIS APPROACH IS SIGNIFICANT IN THAT IT WILL EXPAND OUR THERAPEUTIC PORTFOLIO FOR EOC AND OTHER SOLID TUMORS.

MOLECULAR MECHANISMS OF BRCA1-DEPENDENT DNA DAMAGE RESPONSE AND TUMORIGENESIS

DNA REPAIR GENE MUTATIONS AND PROSTATE CANCER

TARGETING P38 GAMMA SIGNALING TO ADVANCE CUTANEOUS T CELL LYMPHOMA THERAPY

EXAMINING THE IMPACT OF STRUCTURAL RACISM ON AFRICAN AMERICAN NON-SMALL CELL LUNG CANCER MUTATIONAL SIGNATURES AND OUTCOMES - ABSTRACT NON-HISPANIC BLACK/AFRICAN AMERICAN (AA) INDIVIDUALS DEVELOP NON-SMALL CELL LUNG CANCER (NSCLC) 5 YEARS EARLIER THAN THEIR NON-HISPANIC WHITE (NHW) COUNTERPARTS. AAS ALSO HAVE A HIGHER LUNG CANCER INCIDENCE AND MORTALITY RATES. THE ROLE OF STRUCTURAL RACISM IN THE DEVELOPMENT AND MAINTENANCE OF THESE DISPARITIES HAS BEEN UNDERSTUDIED. OUR LONG-TERM GOAL IS TO ACHIEVE HEALTH EQUITY WITHIN NSCLC THROUGH INVESTIGATIONS THAT BENEFIT THE AA COMMUNITY. THE OBJECTIVE OF THIS PROPOSAL IS TO DETERMINE THE IMPACT OF RACISM-RELATED SOCIO- ENVIRONMENTAL FACTORS, INCLUDING AIR POLLUTION AND RESIDENTIAL SEGREGATION ON THE MUTAGENIC PROCESS IN NSCLC TUMORS OF AA PATIENTS. THE CENTRAL HYPOTHESIS IS THAT MEASURABLE STRUCTURAL RACISM STRESSORS INDUCE CHANGES IN MUTATIONAL PROCESSES THAT NEGATIVELY IMPACT DISEASE PROGRESSION IN AA PATIENTS WITH EARLY-STAGE NSCLC. OUR STUDY RATIONALE IS THAT GENOMIC DATA FROM AA NSCLC SPECIMENS IS LACKING, WHICH HINDERS UNDERSTANDING OF THE DISEASE ETIOLOGY AND PROGRESSION IN THIS VULNERABLE POPULATION. FOR EXAMPLE, ONLY 82 OF 1,053 PATIENTS IN THE CANCER GENOME ATLAS REPORTED IDENTIFYING AS AFRICAN AMERICAN. IN ADDITION, IT IS IMPERATIVE TO DETERMINE THE EXTENT TO WHICH RACISM-RELATED SOCIO-ENVIRONMENTAL FACTORS AFFECT THE DISTRIBUTION OF MUTATIONAL SIGNATURES IN NSCLC TUMORS IN AA PATIENTS IN ORDER TO IDENTIFY SPECIFIC PATHWAYS LINKING STRUCTURAL RACISM STRESSORS TO WORSE OUTCOMES AND YIELD TARGETS FOR INTERVENTION AND BIOMARKERS FOR RISK STRATIFICATION. WE PROPOSE TO SURVEY 200 AA PATIENTS WITH STAGE I–II NSCLC IDENTIFIED FROM CALIFORNIA AND GEORGIA CANCER REGISTRIES AND 100 AA FROM DETROIT TO DETERMINE THEIR EXPOSURE TO STRUCTURAL RACISM STRESSORS, WHICH WE WILL CORRELATE WITH DATA FROM THE WHOLE-GENOME SEQUENCING (WGS) OF THEIR TUMOR TISSUE. OUR AIMS ARE TO: (1) DETERMINE THE EXTENT TO WHICH EXPOSURE TO STRUCTURAL RACISM OVER TIME IS LINKED WITH DIFFERENCES IN NSCLC TUMOR EVOLUTION BY CHARACTERIZING THE TYPES OF MUTATIONS, THE ORDER OF THEIR ACQUISITION, AND THE ACTIVITY OF MUTATIONAL PROCESSES; (2) DEFINE THE EFFECT OF STRUCTURAL RACISM STRESSORS ON EARLY RECURRENCE (WITHIN 2 YEARS OF SURGERY) IN STAGE I-II AA NSCLC PATIENTS AND (3) DETERMINE THE DISTRIBUTION OF THE GENOMICS OF NEVER-SMOKING AAS WITH NSCLC COMPARED TO THE SHERLOCK-LUNG STUDY AND IF THIS DISTRIBUTION IS AFFECTED BY THE STRESSORS OF INTEREST. THIS INNOVATIVE STUDY WILL BE ONE OF THE FIRST TO USE WGS, TUMOR EVOLUTION, AND GEOSPATIAL ANALYSIS TECHNIQUES TO IDENTIFY CAUSAL PATHWAYS LINKING STRUCTURAL RACISM STRESSORS TO THE MUTAGENIC PROCESSES OF NSCLC, PROVIDING A MUCH-NEEDED INTEGRATION OF SOCIAL SCIENCE WITH MOLECULAR OUTCOMES. WE EXPECT THAT THE STRUCTURAL RACISM STRESSORS WILL INDUCE DIFFERENCES IN TIMING OF DRIVER MUTATION DEVELOPMENT THAT PROMOTE EARLY NSCLC RECURRENCE. THIS STUDY WILL ELUCIDATE HOW RACISM-RELATED SOCIO-ENVIRONMENTAL FACTORS AFFECT AA NSCLC BIOLOGY, WHICH IS EXPECTED TO REVEAL THE MECHANISMS UNDERLYING DISPARATE RACE-BASED NSCLC OUTCOMES, ULTIMATELY LEADING TO IDENTIFICATION OF NEW TARGETS AND DEVELOPMENT OF NOVEL INTERVENTIONS TO REDUCE HEALTH DISPARITIES.

UNDERSTANDING THE INCREASED RISK OF CHILDHOOD ACUTE LYMPHOBLASTIC LEUKEMIA IN LATINOS - ABSTRACT THIS APPLICATION IS BEING SUBMITTED IN RESPONSE TO THE NOTICE OF SPECIAL INTEREST (NOSI) IDENTIFIED AS NOT-CA-22-056. ACUTE LYMPHOBLASTIC LEUKEMIA (ALL) IS THE MOST COMMON PEDIATRIC CANCER AND, DESPITE ADVANCES IN TREATMENT, IT IS STILL ONE OF THE LEADING CAUSES OF CHILDHOOD DEATH IN THE UNITED STATES (US), AND SURVIVORS FACE SIGNIFICANT LIFELONG TREATMENT-RELATED MORBIDITIES. CHILDREN OF LATINO ETHNICITY HAVE THE HIGHEST AND FASTEST-INCREASING RISK OF ALL IN THE US, AND HAVE LOWER SURVIVAL THAN NON-LATINO WHITES; HOWEVER, THIS ETHNIC DISPARITY IN INCIDENCE AND OUTCOME IS NOT FULLY UNDERSTOOD. ELUCIDATING THE INCREASED RISK OF ALL IN LATINO CHILDREN MAY REVEAL NOVEL INSIGHTS IN THE ETIOLOGY OF ALL IN BOTH LATINO AND NON-LATINO POPULATIONS, AND MAY HIGHLIGHT POTENTIAL AVENUES FOR DISEASE PREVENTION. WE HYPOTHESIZE THAT GERMLINE GENETIC VARIATION PLAYS AN ESSENTIAL ROLE IN THE INCREASED ALL RISK IN LATINOS, THAT THIS RISK IS IMPARTED VIA NATIVE AMERICAN ANCESTRY, AND THAT ALL RISK ALLELES WERE SELECTED IN NATIVE AMERICANS DURING EUROPEAN COLONIZATION OF THE AMERICAS DUE TO THEIR BENEFICIAL EFFECTS ON IMMUNE RESPONSE TO NEW INFECTIONS. WE HAVE ASSEMBLED THE LARGEST EVER CASE- CONTROL SET OF CHILDHOOD ALL IN LATINOS, INCLUDING OVER 5,400 CASES AND 27,000 CONTROLS FROM THREE INDEPENDENT STUDIES IN CALIFORNIA, PLUS STUDIES IN TEXAS, CHILDREN’S ONCOLOGY GROUP/ST. JUDE CHILDREN’S RESEARCH HOSPITAL, AND GUATEMALA. IN OUR FIRST AIM, WE WILL PERFORM THREE COMPLEMENTARY APPROACHES TO DISCOVER NOVEL COMMON RISK LOCI ASSOCIATED WITH CHILDHOOD ALL: I) A GENOME-WIDE ASSOCIATION STUDY (GWAS) META-ANALYSIS, II) ADMIXTURE MAPPING TO CAPITALIZE ON THE RECENTLY ADMIXED NATURE OF LATINO GENOMES, AND III) A TRANSCRIPTOME- WIDE ASSOCIATION STUDY TO IDENTIFY NOVEL LOCI AND TO PINPOINT CAUSAL GENES AT KNOWN AND NOVEL RISK REGIONS. IN OUR SECOND AIM, WE WILL CHARACTERIZE THE GENETIC VARIANTS IN TERMS OF THEIR ASSOCIATION WITH LOCAL NATIVE AMERICAN ANCESTRY AND WHETHER THEY EXHIBIT EVIDENCE OF DIRECTIONAL NATURAL SELECTION ON THE NATIVE AMERICAN BRANCH OF THE HUMAN POPULATION TREE. WE WILL ALSO CHARACTERIZE THE AGGREGATE EFFECTS OF COMMON VARIANTS ON CHILDHOOD ALL RISK AND HOW THIS VARIES BY ETHNICITY, VIA COMPREHENSIVE MODELING OF POLYGENIC RISK SCORES (PRS) FOR ALL IN LATINOS AND IN NON-LATINO WHITES. FINALLY, WE WILL INCORPORATE OUR GENETIC FINDINGS INTO EPIDEMIOLOGIC ANALYSES, BY ACCOUNTING FOR COMMON (PRS) GENETIC VARIANTS IN ALL RISK MODELS FOR IMMUNE- RELATED RISK FACTORS IN LATINOS AND NON-LATINO WHITES, INCLUDING CESAREAN DELIVERY AND IN UTERO CYTOMEGALOVIRUS INFECTION, BOTH OF WHICH HAVE SHOWN STRONGER EFFECTS ON ALL RISK IN LATINOS AND ARE POTENTIALLY MODIFIABLE RISK FACTORS. THE RESULTS OF THIS STUDY WILL SHED LIGHT ON THE ETIOLOGY OF CHILDHOOD ALL IN GENERAL AND OF THE INCREASED RISK OF ALL IN LATINO CHILDREN, WHICH WILL HELP TO ALLEVIATE THIS ETHNIC DISPARITY AND MAY INFORM FUTURE APPROACHES FOR CHILDHOOD LEUKEMIA PREVENTION.

PATHOGENESIS, PREVENTION AND TREATMENT OF CORTICOSTEROID-RESISTANT GUT GVHD - PROJECT SUMMARY: ALLOGENEIC HEMATOPOIETIC CELL TRANSPLANTATION (ALLO-HCT) CAN CURE HEMATOLOGICAL MALIGNANCIES, BUT THIS TREATMENT CAUSES GRAFT-VERSUS-HOST DISEASE (GVHD), AN IMMUNE RESPONSE OF DONOR CELLS AGAINST THE RECIPIENT. ANTI-INFLAMMATORY CORTICOSTEROID MEDICATIONS CAN OFTEN CONTROL GVHD, BUT IN SOME CASES, GVHD IS RESISTANT TO THIS TREATMENT, ESPECIALLY WHEN IT AFFECTS THE INTESTINAL TRACT. THE GOAL OF THIS PROJECT IS TO UNDERSTAND THE MECHANISMS THAT CAUSE STEROID-RESISTANT (SR) GUT GVHD AND TO DEVELOP EFFECTIVE APPROACHES TO PREVENT OR TREAT SR-GUT-GVHD. RESULTS WITH A NEWLY ESTABLISHED MURINE MODEL HAVE DEMONSTRATED THAT SR-GUT- GVHD IS INDUCED BY PROLONGED ADMINISTRATION OF CORTICOSTEROIDS. CORTICOSTEROID TREATMENT INDUCES EXPANSION OF IL-22-PRODUCING TH/TC22-LIKE CELLS, IL-22-DEPENDENT ABNORMALITIES IN GUT MICROORGANISMS, AND REDUCED NUMBERS OF ANTI-INFLAMMATORY CX3CR1HI MONONUCLEAR PHAGOCYTES (MNP) THAT REGULATE T CELL EXPANSION AND CONTROL BACTERIAL TRANSLOCATION. INFORMATION FROM OUR PRELIMINARY STUDIES AND OTHER INVESTIGATORS SUGGESTS THAT CORTICOSTEROID TREATMENT INHIBITS THE POLYAMINE-HYPUSINE PATHWAY IN T AND MNP CELLS AND REGULATES THE FIDELITY OF T CELL LINEAGE DIFFERENTIATION AND THE EXPANSION OF PRO- AND ANTI-INFLAMMATORY MNP CELLS. OUR STUDIES ALSO SUGGEST THAT STEROID TREATMENT INCREASES CEACAM-1 EXPRESSION BY INTESTINAL EPITHELIAL CELLS, THEREBY INCREASING BACTERIAL TRANSCYTOSIS AND UNFAVORABLY ALTERING THE BALANCE BETWEEN PROINFLAMMATORY CX3CR1LO MNP AND ANTI- INFLAMMATORY CX3CR1HI MNP CELLS. THESE CHANGES TRIGGER A FEEDFORWARD PATHOGENIC LOOP CONSISTING OF EXPANDING IL-22-PRODUCING TH/TC22-LIKE CELLS, IL-22-DEPENDENT DYSBIOSIS, AND INCREASED NUMBERS OF PROINFLAMMATORY CX3CR1LO MNP WITH DECREASED NUMBERS OF REGULATORY CX3CR1HI MNP, LEADING TO FULL-BLOWN SR-GUT-GVHD. TO TEST THIS HYPOTHESIS, THIS APPLICATION PROPOSES 3 AIMS. AIM 1 WILL DETERMINE WHETHER CORTICOSTEROID INHIBITION OF THE POLYAMINE-HYPUSINE PATHWAY IN T CELLS LEADS TO EXPANSION OF TH/TC22-LIKE CELLS WITH LINEAGE INFIDELITY. AIM 2 WILL DETERMINE WHETHER CORTICOSTEROID INHIBITION OF POLYAMINE-HYPUSINE PATHWAY IN CX3CR1+ MNP CELLS AUGMENT EXPANSION OF PROINFLAMMATORY CX3CR1LO MNP WITH REDUCED NUMBERS OF ANTI- INFLAMMATORY CX3CR1HI MNP IN RESPONSE TO CHALLENGE BY DYSBIOSIS. AIM 3 WILL DETERMINE WHETHER OBSERVATIONS IN MURINE MODELS REFLECT THE PATHOGENESIS OF SR-GUT-GVHD IN HUMANS AND WHETHER REVERSAL OF ABNORMALITIES IN THE POLYAMINE-HYPUSINE PATHWAY AND WHETHER BLOCKING BACTERIAL INTERACTION WITH CEACAM-1 ON INTESTINAL EPITHELIAL CELLS BY ANTI-CEACAM-1 MAB PREVENTS AND REVERSES SR-GUT-GVHD. RELEVANCE: BY ELUCIDATING IN-DEPTH MECHANISTIC UNDERSTANDING OF THE PATHOGENESIS LEADING TO SR-GUT-GVHD IN A WELL CHARACTERIZED MURINE MODEL AND BY ASSESSING THE RELEVANCE OF THE EXPERIMENTAL RESULTS IN COLON TISSUE FROM PATIENTS WITH SR-GUT-GVHD, RESULTS OF THIS PROJECT COULD IDENTIFY POTENTIALLY EFFECTIVE APPROACHES FOR TRANSLATIONAL TESTING IN HUMANS.

(PQ11) IMMUNE MODULATORY EFFECTS OF WHITE BUTTON MUSHROOM IN PROSTATE CANCER - A TRANSLATIONAL RESEARCH

FUNCTIONAL ANALYSIS OF STRUCTURE SPECIFIC NUCLEASES IN GENOME STABILITY AND CANCERS

ENGINEERED ANTI-PSCA ANTIBODIES FOR IMMUNOPET AND TARGETED THERAPY OF PANCREATIC CANCER - PANCREATIC CANCER REMAINS AMONG THE MOST LETHAL OF SOLID TUMORS, DUE TO LATE DIAGNOSIS AND A HIGH PROBABILITY OF METASTATIC SPREAD. EFFECTIVE NEW SYSTEMIC TREATMENTS ARE NEEDED IN ORDER TO IMPROVE OUTCOMES IN PATIENTS. TARGETED RADIONUCLIDE THERAPY HAS DEMONSTRATED EFFECTIVENESS CANCER THERAPY, NOTABLY WITH THE SUCCESS OF 177LU- DOTATATE IN NEUROENDOCRINE TUMORS INCLUDING THOSE OF THE PANCREAS. PROSTATE STEM CELL ANTIGEN (PSCA) IS UPREGULATED IN 60-80% OF PANCREATIC ADENOCARCINOMAS, MAKING IT A PROMISING TARGET FOR ANTIBODY-DIRECTED THERAPY. AN ENGINEERED ANTIBODY FRAGMENT, THE A2 SCFV-FC2 DM HAS BEEN SPECIFICALLY DESIGNED FOR OPTIMIZED DELIVERY OF THERAPEUTIC RADIONUCIDES IN PANCREATIC CANCER. IT IS BASED ON A HUMANIZED, HIGH-AFFINITY ANTI-PSCA ANTIBODY, AND CONTAINS FC MUTATIONS ENGINEERED TO FOSTER RAPID BLOOD CLEARANCE VIA THE HEPATOBILIARY ROUTE. AS A RESULT, RADIATION DOSE TO KEY ORGANS/TISSUES (BONE MARROW AND KIDNEY) IS MINIMIZED, ENABLING EFFECTIVE DELIVERY OF AN ALPHA- OR BETA-EMITTING RADIONUCLIDE TO TUMORS. IN AIM 1, BIODISTRIBUTION STUDIES WILL BE UNDERTAKEN IN MOUSE MODELS, IN ORDER TO CONFIRM THE EXPECTED TUMOR TARGETING AND HEPATIC CLEARANCE. FORMAL DOSE ESTIMATIONS WILL BE MADE FOR THE SCFV-FC2 DM RADIOLABELED WITH EITHER 177LU OR 225AC FOR THERAPY. AIM 2 WILL EXPLORE THE POTENTIAL EFFICACY OF THE ANTI-PSCA SCFV-FC2 DM IN MOUSE MODELS OF PANCREATIC CANCER, INCLUDING SUBCUTANEOUS XENOGRAFTS OF HUMAN PANCREATIC TUMOR CELLS, A SYNGENEIC MODEL OF KPC-PSCA TUMORS IN HUPSCA KNOCK-IN MICE, AND PATIENT-DERIVED PANCREATIC TUMOR XENOGRAFT MODELS. THE RELATIVE EFFICACIES AND TOXICITIES OF THE ALPHA-EMITTER 225AC AND BETA-EMITTER 177LU WILL BE ANALYZED IN ORDER TO PREPARE FOR FUTURE CLINICAL THERAPY STUDIES. IN AIM 3, CLINICAL PRODUCTION AND CONJUGATION OF THE ANTI-PSCA SCFV-FC2 DM WILL BE PERFORMED, TESTING CONDUCTED, AND AN IND FILED. FINALLY, IN AIM 4 WE WILL CONDUCT A FIRST-IN-HUMAN IMAGING STUDY USING 64CU-DOTA-ANTI-PSCA SCFV- FC2 DM IN PATIENTS WITH ADVANCED PANCREATIC CANCER, TO EVALUATE THE TARGETING, AND CLEARANCE PROPERTIES AND POTENTIAL RADIATION DOSE DELIVERY OF THIS NOVEL ENGINEERED ANTIBODY FRAGMENT. RESULTS FROM THIS CLINICAL IMMUNOPET STUDY WILL BE CENTRAL TO GUIDING FUTURE DEVELOPMENT OF A RADIOIMMUNOTHERAPY AGENT THAT CAN BE IMPLEMENTED IN A THERANOSTIC APPROACH TO PANCREATIC CANCER.

CLINICAL EVALUATION OF IL13RA2-TARGETED CAR T CELL THERAPY IN COMBINATION WITH NIVOLUMAB IN PATIENTS WITH RECURRENT MALIGNANT GLIOMA

ENHANCING THE INTRACELLULAR FUNCTIONING OF ANTI-HV RNAS

MULTIFUNCTIONAL IMMUNOPET TRACERS FOR PANCREATIC AND PROSTATE CANCER

EXPRESSION OF ANTI-HIV SIRNA IN BLOOD CELLS

TARGETING PAK1 TO IMPROVE FUNCTIONAL BETA-CELL MASS AND INSULIN SENSITIVITY

EXTREME WEATHER, AIR POLLUTION, AND STROKE AMONG AN AGING FEMALE POPULATION - ABSTRACT THIS APPLICATION IS IN RESPONSE TO THE PAR-19-250: “ENVIRONMENTALINFLUENCES ON AGING: EFFECTS OF EXTREME WEATHERAND DISASTER EVENTS ON AGING POPULATIONS .”AIR POLLUTANTS ARE ESPECIALLY DETRIMENTAL FOR AGING POPULATIONS; EXPOSURE TO AIR POLLUTION AS MEASURED BY AMBIENT PARTICULATE MATTER (E.G., PM2.5) HAS BEEN LINKED TO DISEASES THAT INCREASE WITH AGE, INCLUDING CARDIOVASCULAR DISEASES (CVD) AND STROKE. OLDER WOMEN (WHO OUTNUMBER MEN 3:2) ARE PARTICULARLY SUSCEPTIBLE TO CVD ENDPOINTS AND TO STROKE RISK IN PARTICULAR; STROKE RISK IN WOMEN DOUBLES IMMEDIATELY FOLLOWING MENOPAUSE. EXTREME WEATHER EVENTS SUCH AS WILDFIRES AND PROLONGED DROUGHT IN THE LAST DECADE ALONE HAVE ADVERSELY AFFECTED AIR POLLUTION EXPOSURE IN STATES SUCH AS CALIFORNIA. A NUMBER OF METROPOLITAN COUNTIES IN THE STATE HAVE PARTICULATE POLLUTION LEVELS ABOVE FEDERAL AND STATE AMBIENT STANDARDS, AND DURING EXTREME WEATHER EVENTS, THESE LEVELS RIVAL THAT OF THE WORST CITIES IN THE WORLD. OUR OVERALL STUDY OBJECTIVE IS TO EVALUATE THE INTERSECTION OF EXTREME WEATHER EVENTS AND AIR POLLUTION IN AN AGING FEMALE POPULATION WHO ARE AT INCREASED RISK FOR CVDS, AND IN PARTICULAR, AT PEAK SUSCEPTIBILITY FOR STROKE. IN AIM1, WE WILL EVALUATE THE ACUTE EFFECTS FROM WILDFIRE EVENTS BY ASCERTAINING STROKE EVENTS WITHIN GEOGRAPHICALLY AFFECTED AREAS BASED ON SATELLITE IMAGERY. ELEVATED PM2.5 EXPOSURE ESTIMATES RESULTING FROM A WILDFIRE EVENT IN THE AFFECTED AREAS WILL BE ASSOCIATED WITH HOSPITALIZATIONS (INCLUDING EMERGENCY ROOM VISITS) FROM STROKE AND CVD. IN AIM 2, WE WILL DETERMINE THE ROLE OF SPECIFIC PM2.5 COMPONENTS IN STROKE RISK AND MORTALITY. EMPLOYING COMPLEMENTARY SATELLITE- AND SOURCE-BASED APPROACHES, WE WILL IDENTIFY KEY SOURCES OF PM2.5 AND ITS CHEMICAL CONSTITUENTS THAT ARE ATTRIBUTABLE TO STROKE. LEVERAGING 25-YEARS OF FOLLOW-UP FROM THE STUDY POPULATION, THIS AIM WILL PERMIT US TO DELINEATE COMPONENTS OF AIR POLLUTION FROM DROUGHT AND WILDFIRE EVENTS VERSUS OTHER (E.G., TRANSPORTATION, INDUSTRIAL) EXPOSURES. FOR BOTH AIMS, SELECT CVD ENDPOINTS (E.G., MYOCARDIAL INFARCTION) WILL ALSO BE EXPLORED. IN AIM 3, WE WILL EVALUATE THE ASSOCIATION BETWEEN PM2.5 EXPOSURE WITH SERUM IMMUNE MARKERS AMONG 2000 PARTICIPANTS WHOSE EXPOSURES REFLECT A CROSS-SECTION OF EXPOSURE DURING THE 2015 DROUGHT AND WILDFIRE SEASON. THIS AIM WILL INFORM THE PURPORTED BIOLOGIC UNDERPINNING FOR STROKE AND CVD RISK, BY KEY PM2.5 SOURCES AND CONSTITUENTS. TO SUCCESSFULLY ACCOMPLISH THESE AIMS, WE WILL LEVERAGE LONGITUDINAL DATA FROM THE CALIFORNIA TEACHERS STUDY, A GEOGRAPHICALLY-DEFINED POPULATION-BASED COHORT STUDY OF 133,479 WOMEN WHOSE 25 YEAR FOLLOW-UP SPANS: (A) KEY PERIODS OF STROKE AND CVD SUSCEPTIBILITY IN WOMEN, AND (B) ELEVATED LEVELS OF AIR POLLUTION EXPOSURE IN CALIFORNIA WHERE THERE HAVE BEEN PROLONGED PERIODS OF DROUGHT AND NUMEROUS WILDFIRES.

REDUCING RISK OF ANTHRACYCLINE-RELATED HEART FAILURE AFTER CHILDHOOD CANCER

ENVIRONMENTAL CHEMICAL BODY BURDEN AND PROSPECTIVE BREAST CANCER RISK IN THE CANCER PREVENTION STUDY-3 COHORT - TODAY THERE ARE MORE THAN 85,000 EPA-REGISTERED SYNTHETIC CHEMICALS, BUT ONLY 10% HAVE BEEN TESTED FOR CARCINOGENICITY IN ANIMAL STUDIES. OF THOSE TESTED, ~200 HAVE SHOWN EVIDENCE OF CAUSING CANCER AND/OR AN INCREASE IN MAMMARY TUMORS. HOWEVER, FEW OF THE CHEMICALS HAVE BEEN EVALUATED IN HUMAN STUDIES, AND RESULTS IN HUMANS HAVE BEEN INCONCLUSIVE. OUR STUDY WILL FOCUS ON CHEMICAL BODY BURDEN OF PER- AND POLY-FLUOROALKYL SUBSTANCES (PFAS), AND CONCOMITANT RISK OF DEVELOPING INVASIVE BREAST CANCER DURING THE MENOPAUSAL TRANSITION. THESE “FOREVER CHEMICALS” ARE PERVASIVE, ENDURING, AND OF HIGH PUBLIC INTEREST, YET STUDIES OF THEIR POSSIBLE RELATIONSHIP TO BREAST CANCER HAVE BEEN LIMITED. WE HYPOTHESIZE THAT ELEVATED BODY-BURDEN LEVELS OF THESE ENDOCRINE-DISRUPTING CHEMICALS (EDCS) WILL INCREASE THE RISK OF DEVELOPING INVASIVE BREAST CANCER, AND THAT ALTERATIONS IN DNA METHYLATION (DNAM) AND THE BREAST MICROENVIRONMENT ARE MECHANISMS THAT LINK THESE CHEMICAL EXPOSURES AND BREAST CANCER. WE WILL TEST THIS HYPOTHESIS USING A THREE-PRONGED APPROACH. IN SPECIFIC AIM 1, WE WILL CONDUCT A PROSPECTIVE STUDY OF WOMEN IN THE AMERICAN CANCER SOCIETY CANCER PREVENTION STUDY-3 (CPS-3) TO ASSESS THE ASSOCIATION BETWEEN BODY BURDEN OF PFAS DURING THE MENOPAUSAL TRANSITION AND SUBSEQUENT DEVELOPMENT OF INVASIVE BREAST CANCER. WE WILL MEASURE PFAS LEVELS IN PLASMA SAMPLES COLLECTED 1 TO 7 YEARS BEFORE INVASIVE BREAST CANCER DIAGNOSIS IN 1000 CPS-3 PARTICIPANTS AND 1000 MATCHED, CANCER-FREE CPS-3 PARTICIPANTS, ALL BETWEEN 40 AND 57 YEARS OF AGE AT BLOOD DRAW. IN SPECIFIC AIM 2, STUDYING THE SAME PARTICIPANTS, WE WILL MEASURE DNAM USING INFINIUM METHYLATIONEPIC BEADCHIPS AND CONDUCT AN EPIGENOME- WIDE ASSOCIATION STUDY (EWAS) TO IDENTIFY DNAM CHANGES ASSOCIATED WITH LEVELS OF PFAS IN THE CANCER-FREE PARTICIPANTS. WE THEN WILL DETERMINE THE ASSOCIATION BETWEEN THE PFAS-ASSOCIATED DNAM CHANGES AND RISK OF DEVELOPING BREAST CANCER. THIS VALUABLE DNAM DATA ALSO CAN BE USED LATER FOR OTHER OUTCOMES, INCLUDING EXPOSURE TO OTHER EDCS. IN SPECIFIC AIM 3, WE WILL DETERMINE THE DIRECT EFFECTS OF PFAS ON TISSUE- AND MOLECULAR- LEVEL STATES ASSOCIATED WITH SUSCEPTIBILITY TO CANCER INITIATION IN GENOMICALLY-WELL-CHARACTERIZED PRIMARY HUMAN BREAST MAMMARY EPITHELIAL CELLS (HMECS). IDENTIFYING THESE MECHANISMS IN PRIMARY BREAST CELLS IS CRITICALLY IMPORTANT, AS PFAS MECHANISMS OF ACTION GENERALLY DIFFER BY TISSUE. WE WILL USE 2-D CULTURES TO DETERMINE THE EFFECTS OF SHORT-TERM EXPOSURES AND 3-D CULTURES TO DEFINE THE EFFECTS OF PROTRACTED CHEMICAL EXPOSURES ON CHANGES IN EPITHELIAL LINEAGE CONSISTENT WITH ACCELERATED AGING AND AGE-RELATED MOLECULAR CHANGES IN GENOME METHYLATION, LINEAGE-SPECIFIC TRANSCRIPTION, AND CYTOKERATIN PROTEINS. RESULTS FROM THIS MULTI-DISCIPLINARY APPROACH WILL ADVANCE OUR UNDERSTANDING OF THE EFFECTS OF PFAS EXPOSURES ON RISK OF DEVELOPING BREAST CANCER DURING AN IMPORTANT WINDOW OF SUSCEPTIBILITY. ULTIMATELY, WE HOPE RESULTS FROM OUR PROPOSED PROJECT WILL IDENTIFY AN INTEGRATED BIOLOGICAL SIGNATURE OF ENVIRONMENTAL EXPOSURE, DELIVER MECHANISTIC INSIGHTS INTO BREAST CANCER DEVELOPMENT FROM EDCS, AND INFORM FUTURE STUDIES FOR PREVENTION STRATEGIES TO REDUCE OR MITIGATE EXPOSURES.

INTEGRATED IMMUNOTHERAPY FOR BREAST CANCER

THE ROLE OF THE INTESTINAL MICROBIOME IN CANCER IMMUNOTHERAPY - ABSTRACT THE GUT MICROBIOTA CONSISTS OF A COMMUNITY OF DIVERSE MICROBES AND HAS MANY EFFECTS ON HUMAN (PATHO)PHYSIOLOGY. MICROBIOME COMPOSITION HAS BEEN ASSOCIATED WITH MANY DISEASES, BUT CAUSAL INFERENCE IS OFTEN LACKING. PRECLINICAL AND CLINICAL STUDIES HAVE DEMONSTRATED THAT THE INTESTINAL MICROBIOTA CAN REGULATE INNATE AND ADAPTIVE IMMUNITY, INCLUDING T CELL AND ANTITUMOR IMMUNITY AFTER ALLOGENEIC HEMATOPOIETIC CELL TRANSPLANTATION (ALLO-HCT) AND CHECKPOINT BLOCKADE. MY LAB HAS FOCUSED ON THE ROLE OF GUT MICROBIOTA IN OUTCOMES OF ALLO-HCT AND IMMUNOTHERAPY. FOR EXAMPLE, WE SHOWED THAT MICROBIOTA COMPOSITION UNDERGOES SIGNIFICANT AND FREQUENT CHANGES DURING ALLO-HCT AND THAT LOWER INTESTINAL MICROBIOTA DIVERSITY IS ASSOCIATED WITH INCREASED MORTALITY. WE ALSO FOUND THAT DOMINANCE BY CERTAIN SPECIES, MOST FREQUENTLY ENTEROCOCCUS, IS ASSOCIATED WITH LETHAL GRAFT-VERSUS-HOST DISEASE (GVHD); THAT EXPOSURE TO CERTAIN ANTIBIOTICS IS ASSOCIATED WITH WORSE OUTCOMES FOLLOWING ALLO-HCT AND CHIMERIC ANTIGEN RECEPTOR T CELL (CART) THERAPY; AND THAT HEMATOPOIETIC RECONSTITUTION IS ASSOCIATED WITH THE PRESENCE OF BENEFICIAL FLORA. THESE STUDIES HAVE BEEN TRANSLATED INTO CLINICAL TRIALS USING AUTOLOGOUS FECAL MICROBIOTA TRANSPLANT, ADMINISTRATION OF DEFINED BACTERIAL CONSORTIA, AND ANTIBIOTIC STEWARDSHIP TO SPARE AND/OR RESTORE THE COMMENSAL FLORA. THE OVERARCHING HYPOTHESIS OF THIS PROPOSAL IS THAT THE INTESTINAL MICROBIOME IS AN IMPORTANT MODULATOR OF INNATE AND ADAPTIVE IMMUNITY IN THE SETTING OF CANCER IMMUNOTHERAPY. WHILE IMMUNOTHERAPIES ARE CURATIVE IN SOME RECIPIENTS, IMPROVING THEIR EFFICACY AND ABATING TOXICITIES ARE UNMET NEEDS IN ONCOLOGY. THE MAJOR GOALS ARE TO IMPROVE CANCER IMMUNOTHERAPY BY TARGETING THE INTESTINAL MICROBIOME BASED ON PRECLINICAL AND CLINICAL STUDIES. EXAMPLES OF OUR ONGOING AND PLANNED STUDIES INCLUDE: A) DEVELOPMENT OF A NEW PIPELINE FOR MICROBIOME ANALYSIS, B) PRECLINICAL AND CLINICAL PROJECTS REGARDING INTESTINAL MICROBIOME AND CART, C) NEW TECHNIQUES TO ANALYZE THE EFFECTS OF DIET AND DRUGS ON THE INTESTINAL MICROBIOME, AND D) PRECLINICAL AND CLINICAL STUDIES REGARDING IMMUNE MODULATION BY BILE ACIDS, AS AN EXAMPLE HOW WE STUDY THE MECHANISMS BY WHICH THE INTESTINAL MICROBIOME CAN MODULATE IMMUNITY AND CANCER IMMUNOTHERAPY. WE HAVE ORGANIZED A MULTICENTER GLOBAL CONSORTIUM TO COLLECT FECAL SAMPLES (FUNDED SEPARATELY FROM THIS APPLICATION) ALONG WITH A NOVEL MULTI-OMIC APPROACH TO INTEGRATE PATIENT, MICROBIOME, AND TUMOR PROFILING MODALITIES USING A COMPUTATIONAL PLATFORM (MSK-MIND) FOR DATA HARMONIZATION AND MACHINE LEARNING. THESE INVESTIGATIONS WILL BE PERFORMED VIA PERPETUAL DIALOGUE BETWEEN WORK WITH MICE AND HUMANS: HUMAN STUDIES ENABLE US TO OBSERVE CORRELATIONS, DEVELOP HYPOTHESES, AND TEST THERAPEUTIC STRATEGIES; ANIMAL STUDIES ENABLE US TO ESTABLISH OR REFUTE CAUSAL RELATIONSHIPS BETWEEN MICROBIOTA AND HOST IMMUNOLOGY AND TO OBTAIN MECHANISTIC INSIGHTS. THESE DATA WILL INFORM THE FUTURE DEVELOPMENT OF CLINICAL TRIALS TO TEST THERAPEUTIC STRATEGIES TO ENHANCE EFFICACY AND DECREASE TOXICITY IN PATIENTS RECEIVING CANCER IMMUNOTHERAPY, SUCH AS CART AND ALLO-HCT.

NATIONAL ISLET CELL CONSORTIUM COORDINATING CENTER

MOLECULAR EPIDEMIOLOGY OF NON-HODGKIN LYMPHOMA PROGNOSIS AND PREVENTION

APOE2 IN ALZHEIMER'S DISEASE: THE NEUROPROTECTIVE EFFECTS AND MECHANISMS

SCALABLE TOOLS TO EFFECTIVELY TRANSLATE GENOMIC DISCOVERIES INTO THE CLINIC

EXTRACELLULAR VESICLE-MEDIATED ISLET IMMUNE CROSS TALK IN TYPE 1 DIABETES PATHOGENESIS - PROJECT SUMMARY/ABSTRACT (30 LINES OR LESS) THE PREMISE FOR THE PROPOSED RESEARCH STEMS FROM PRECEDENCE IN OTHER DISEASES, SUCH AS CANCER, CARDIOVASCULAR, NEURODEGENERATIVE, AND MOST RELEVANT TO THE CURRENT PROPOSAL, AUTOIMMUNE DISEASES, IN WHICH EXTRACELLULAR VESICLES (EVS) PLAY A ROLE IN THE PATHOPHYSIOLOGY AND ARE IMPORTANT BIOMARKERS FOR EARLY DETECTION. HOWEVER, IN HUMAN TYPE 1 DIABETES (T1D), LITTLE IS KNOWN CONCERNING EVS IN CELLULAR COMMUNICATION. OUR OVERALL HYPOTHESIS IS THAT AUTOCRINE-PARACRINE INTERACTIONS, MEDIATED THROUGH EVS, BETWEEN THE ISLETS AND ISLET-INFILTRATING IMMUNE CELLS IN THE PANCREAS CONTRIBUTE TO THE DEVELOPMENT AND PROGRESSION OF T1D. OUR SPECIFIC AIMS ARE: 1) TO ASSESS THE FUNCTIONAL IMPACT OF ISLET-INFILTRATING T-CELL DERIVED EVS (T-EV) FROM T1D AND AUTOANTIBODY+ (AAB+) DONORS ON ISLET HEALTH AND ON DISTINCT IMMUNE CELL POPULATIONS; 2) TO INVESTIGATE THE CONTRIBUTION OF STRESSED OR T1D ISLET-DERIVED EVS (I-EV) ON IMMUNE CELL PHENOTYPE AND ISLET HEALTH; AND 3) TO DECIPHER THE DIFFERENTIAL PROTEIN CARGO IN T-EV AND I-EV FROM T1D AND AAB+ DONORS. TO ADDRESS THESE AIMS, WE HAVE ASSEMBLED A TEAM OF INVESTIGATORS WITH HIGHLY RELEVANT EXPERTISE, TECHNIQUES AND UNIQUE RESOURCES OF CELL LINES AND TISSUE SAMPLES. FROM 36 HUMAN TISSUE DONORS WITH T1D OR AAB POSITIVITY, WE HAVE >600 T CELL LINES GROWN DIRECTLY FROM THE INDIVIDUAL ISLETS OF PANCREATA. WE HAVE THE EXPERTISE AND TECHNICAL ABILITY TO ISOLATE EV FROM ISLETS (I-EV) AND FROM ISLET-INFILTRATING T CELL LINES (T-EV) FROM T1D DONORS. OUR PRELIMINARY DATA INDICATES I-EV AND T-EV HAVE BOTH PARACRINE AND AUTOCRINE EFFECTS ON ISLET HEALTH AND IMMUNE CELL PHENOTYPE. OUR RESEARCH PLAN IS TO GENERATE T-EV AND I-EV FROM DONORS WITH T1D OF DISTINCT DURATIONS OR POSITIVITY FOR ISLET AUTOANTIBODIES, TO EVALUATE THEIR EFFECTS ON ISLET HEALTH AND IMMUNE CELL FUNCTION, AND TO DETERMINE THE UNIQUELY PACKAGED PROTEIN CARGO FROM THESE EVS WHOSE MOLECULAR COMPOSITION REFLECTS THE PATHOPHYSIOLOGIC STATE OF THE DISSEMINATING CELL. THESE STUDIES WILL YIELD IMPORTANT INFORMATION CONCERNING THE COMMUNICATION BETWEEN IMMUNE CELL POPULATIONS AND ISLETS VIA EVS IN THE PATHOGENESIS OF T1D, AND POTENTIAL BIOMARKERS OR THERAPEUTIC TARGETS FOR T1D.

THE EPIDEMIOLOGY OF IMMUNE RESPONSES IN COLORECTAL CANCER

OPTIMIZING GVHD PREVENTION WITH SYSTEMS PHARMACOLOGY MODELS

PEPTIDE VACCINE TO PREVENT CMV DISEASE AFTER HSCT

EXPERIMENTAL-COMPUTATIONAL SYNTHESIS OF ALTERED IMMUNE SIGNALING IN BREAST CANCER

TREATMENT OF BCR/ABL CAUSED LEUKEMIAS WITH FTIS

RANDOMIZED TRIAL OF EXERCISE THERAPY ON MARKERS OF PROGRESSION IN LOCALIZED PROSTATE CANCER: - ABSTRACT WIDESPREAD USE OF SERUM PROSTATE-SPECIFIC ANTIGEN (PSA) SCREENING RESULTS IN 50% OF NEW CASES OF PROSTATE CANCER BEING DIAGNOSED WITH LOW-GRADE LOCALIZED DISEASE. STANDARD OF CARE FOR THESE CASES IS TO DEFER IMMEDIATE TREATMENT IN FAVOR OF ACTIVE SURVEILLANCE (AS), A LOW-TOXICITY MANAGEMENT WHICH ENTAILS CLOSE MONITORING WITH PSA TESTS, REPEAT PROSTATE BIOPSIES AND MULTI-PARAMETRIC MRI (MPMRI). AROUND 30% OF MEN ON AS PROGRESS OR ELECTIVELY UNDERGO DEFINITIVE TREATMENT WITHIN TWO YEARS OF DIAGNOSIS. EFFICACIOUS LOW-COST, LOW-TOXICITY STRATEGIES TO LIMIT EXIT FROM AS ARE NEEDED TO REDUCE OVER-TREATMENT AND IMPROVE HEALTH-RELATED QUALITY OF LIFE (QOL). ACCORDINGLY, IN THIS PROPOSAL WE WILL TEST WHETHER AND HOW EXERCISE ALTERS PROSTATE CANCER MOLECULAR LANDSCAPE. USING A TWO-ARM RCT DESIGN, 122 MEN INITIATING AS (E.G., INACTIVE, DIAGNOSTIC BIOPSY <6 MONTHS) WILL BE RANDOMLY ALLOCATED (1:1 RATIO) TO EXERCISE THERAPY INTERVENTION (N=61) OR USUAL CARE (PLUS GENERAL PHYSICAL ACTIVITY ADVICE; N=61) FOR 12 MONTHS. EXERCISE THERAPY WILL CONSIST OF HOME-BASED, TREADMILL WALKING FIVE DAYS EACH WEEK AT 65-75% OF THEIR PERSONAL BASELINE EXERCISE CAPACITY. ALL SESSIONS WILL BE IMPLEMENTED USING TELEMED-X, A NOVEL TELEMEDICINE PLATFORM PIONEERED IN OUR PROGRAM. OUR CENTRAL HYPOTHESIS IS THAT EXERCISE THERAPY WILL REDUCE THE NUMBER OF NIMBOSUS FEATURES TO COLLECTIVELY INHIBIT CLINICAL PROGRESSION. THIS CENTRAL HYPOTHESIS IS TESTED IN THREE DISTINCT, BUT COMPLEMENTARY AIMS: AIM 1. DETERMINE EFFECTS OF EXERCISE THERAPY ON MOLECULAR NIMBOSUS HALLMARKS; AIM 2. DETERMINE EFFECTS ON THE RADIOLOGIC AND PATHOLOGIC NIMBOSUS HALLMARKS; AND AIM 3. DELINEATE THE PRECISION OF ESTIMATING RATES OF CLINICAL PROGRESSION AT 18 MONTHS.

TRANSCRIPTIONAL CONTROL, TARGETED MODIFICATION, AND EXCISION OF HIV IN THE BRAIN

PHAMACOLOGICAL MODULATION OF EPIGENETIC CHANGES IN AML

MYELOID CELL-SELECTIVE, OLIGONUCLEOTIDE-BASED STAT3 INHIBITION COMBINED WITH TOTAL MARROW AND LYMPHOID IRRADIATION FOR IMMUNOTHERAPY OF ACUTE MYELOID LEUKEMIA - PROJECT SUMMARY THERE HAS BEEN LITTLE OR NO LONG-TERM IMPROVEMENT IN OUTCOMES FOR PATIENTS WITH TREATMENT-REFRACTORY ACUTE MYELOID LEUKEMIA (AML). THE RECENTLY DEVELOPED IMAGE-GUIDED RADIATION STRATEGY, TOTAL MARROW AND LYMPHOID IRRADIATION (TMLI) DELIVERS HIGH RADIATION DOSES TO MAJOR LEUKEMIA RESERVOIRS WHILE SPARING NORMAL TISSUES. ALTHOUGH TMLI PRIOR TO ALLOGENEIC HEMATOPOIETIC CELL TRANSPLANTATION (HCT) IMPROVED PATIENTS’ 2-YEAR OVERALL SURVIVAL (OS) RATE FROM <10% TO 48%, RELAPSES REMAINED COMMON. SUCH RADIATION RESISTANCE IS A CONSEQUENCE OF BOTH INTRINSIC CANCER CELL PROPERTIES AND THE EXTRINSIC INFLUENCE OF THE TUMOR MICROENVIRONMENT. IT WAS PREVIOUSLY DEMONSTRATED THAT RADIATION-INDUCED CELL DEATH CAUSES THE RELEASE OF DANGER SIGNALS RECRUITING TOLL-LIKE RECEPTOR- 9 (TLR9)-POSITIVE MYELOID CELLS, WHICH JUMP-START TUMOR VASCULARIZATION AND REGROWTH. THESE CANCER-PROMOTING, RATHER THAN IMMUNOSTIMULATORY, EFFECTS ARE MEDIATED BY TLR9-MEDIATED SECRETION OF CYTOKINES SUCH AS IL-6, THEREBY LEADING TO ACTIVATION OF SIGNAL TRANSDUCER AND ACTIVATOR OF TRANSCRIPTION 3 (STAT3). STAT3 IS A MULTIFACETED ONCOGENE AND A CENTRAL IMMUNE CHECKPOINT REGULATOR OPERATING IN AML CELLS AS WELL AS IN TUMOR- ASSOCIATED MYELOID CELLS IN PATIENTS. HOWEVER, IT REMAINS AN ELUSIVE TARGET, WITH NO FDA-APPROVED DIRECT SMALL MOLECULE STAT3 INHIBITORS. TO OVERCOME THIS CHALLENGE, WE PREVIOUSLY DEVELOPED A STRATEGY TO DELIVER OLIGONUCLEOTIDE STAT3 INHIBITORS, SUCH AS SIRNA OR DECOY DNA, SPECIFICALLY INTO COMMONLY TLR9-POSITIVE AML AND NORMAL MYELOID CELLS. THE NUCLEASE-RESISTANT, SECOND-GENERATION CPG-STAT3 DECOY INHIBITOR (CSI-2) INJECTED INTRAVENOUSLY SHOWED EFFICACY IN TARGETING STAT3, SUPPRESSING LEUKEMIA CELL SURVIVAL AND/OR INDUCING IMMUNE RESPONSES AGAINST MODERATE BURDEN OF HUMAN AND MOUSE AML IN VIVO. THE HYPOTHESIS IS THAT COMBINING THE IMMUNOSTIMULATORY CSI-2 STRATEGY WITH CONDITIONING TMLI TREATMENTS WILL IMPROVE TREATMENT EFFICACY AGAINST AML EVEN AT HIGH BURDEN (>50% LEUKEMIC BLASTS IN THE BONE MARROW) BY PROVIDING TIME FOR THE GENERATION OF ADAPTIVE T-CELL DRIVEN IMMUNE RESPONSES. THE PRELIMINARY RESULTS DEMONSTRATED THAT THE TMLI REGIMEN CAN IMPROVE UPTAKE OF CSI-2 BY AML, THEREBY REDUCING LEUKEMIA-INITIATING POTENTIAL, AUGMENTING AML IMMUNOGENICITY, AND THEREBY INDUCING POTENT CD8+ T CELL-MEDIATED ANTILEUKEMIC IMMUNE RESPONSES. WE PROPOSE TO: 1. ELUCIDATE THE MOLECULAR MECHANISMS OF TMLI/CSI-2 EFFECT ON AML CELL DIFFERENTIATION; 2. OPTIMIZE TMLI TO MAXIMIZE THE EFFECT ON LEUKEMIC BONE MARROW VASCULAR STRUCTURE, CSI-2 DELIVERY, LEUKEMOGENIC POTENTIAL, AND IMMUNOGENICITY; 3. ASSESS THE EFFICACY AND CELLULAR MODE-OF-ACTION OF THE TMLI/CSI-2 COMBINATION TREATMENT COMPARED TO EITHER TREATMENT ALONE IN HUMAN OR MOUSE AML MODELS IN HUMANIZED OR SYNGENEIC MICE, RESPECTIVELY. THE OVERARCHING GOAL OF THIS INTERDISCIPLINARY PROPOSAL IS TO PRODUCE A CLINICALLY RELEVANT, EFFECTIVE, AND SAFE COMBINATORIAL RADIATION-IMMUNOTHERAPY FOR PATIENTS WITH RELAPSED/RECURRENT AML, REPRESENTING THE HIGHEST UNMET NEED IN CANCER THERAPY.

THE ROLE AND THERAPEUTIC POTENTIAL OF IGF2BP2 IN MLL-REARRANGED LEUKEMIA - PROJECT SUMMARY (ABSTRACT): TITLE: THE ROLE AND THERAPEUTIC POTENTIAL OF IGF2BP2 IN MLL-REARRANGED LEUKEMIA. BACKGROUND: N6-METHYLADENOSINE (M6A) MODIFICATION IS THE MOST ABUNDANT INTERNAL MODIFICATION IN EUKARYOTIC MESSENGER RNAS (MRNAS) AND PLAYS ROLES IN MANY NORMAL BIOPROCESSES. EVIDENCE IS EMERGING THAT THE ABERRATION IN M6A MODIFICATION AND THE ASSOCIATED MACHINERY ALSO PLAYS IMPORTANT ROLES IN VARIOUS TYPES OF CANCERS. ACUTE MYELOID LEUKEMIA (AML) IS ONE OF THE MOST COMMON FORMS OF HEMATOPOIETIC MALIGNANCIES WITH VARIOUS CYTOGENETIC AND MOLECULAR ABNORMALITIES. THE MIXED-LINEAGE LEUKEMIA (MLL)-REARRANGED (MLLR) AML IS A COMMON AND FATAL SUBTYPE OF AML, WHICH ACCOUNTS FOR 5%-10% OF “DE NOVO” AML CASES AND 10%-15% OF THERAPY-RELATED LEUKEMIA (T-AML) CASES. MLLR AML PATIENTS ARE ASSOCIATED WITH POOR OUTCOMES, WITH A 5-YEAR OVERALL SURVIVAL (OS) RATE OF ~30%. THEREFORE, THERE IS A CRITICAL UNMET MEDICAL NEED TO DEVELOP IMPROVED THERAPEUTICS FOR MLLR AML TREATMENT. THE LEUKEMIA STEM/INITIATING CELLS (LSCS/LICS) ARE CONSIDERED TO BE THE ROOT CAUSE FOR THE TREATMENT FAILURE AND RELAPSE OF AML. COLLECTIVELY, IT IS CRITICAL TO BETTER UNDERSTAND THE MOLECULAR MECHANISMS UNDERLYING MLLR AML PATHOGENESIS AND LSC/LIC SELF-RENEWAL, WHICH MAY LEAD TO THE DEVELOPMENT OF IMPROVED NOVEL THERAPEUTIC STRATEGIES TO TREAT MLLR AML. OUR PRELIMINARY DATA SHOWED THAT IGF2BP2, WHICH ENCODES AN M6A READER, IS SPECIALLY OVEREXPRESSED IN MLLR AML AND ITS INCREASED EXPRESSION IS ASSOCIATED WITH A POOR PROGNOSIS IN AML PATIENTS. OUR PRELIMINARY FUNCTIONAL STUDIES SUGGEST THAT IGF2BP2 LIKELY PLAYS A CRITICAL ONCOGENIC ROLE AS AN M6A READER IN PROMOTING MLLR AML PATHOGENESIS. IGF2BP2 IS ALSO EXPRESSED AT A SIGNIFICANTLY HIGHER LEVEL IN MLLR LSCS/LICS COMPARED TO HEALTHY HEMATOPOIETIC STEM/PROGENITOR CELLS (HSPCS) AND BULK MLLR AML CELLS, IMPLYING A ROLE OF IGF2BP2 IN MLLR LSC/LIC SELF-RENEWAL. IN ADDITION, WE HAVE DEVELOPED A POTENT INHIBITOR (NAMELY CWI1-2) THAT TARGETS IGF2BP2 DIRECTLY AND EXHIBITS HIGH ANTI-LEUKEMIA EFFICACY AS SHOWN IN OUR PRELIMINARY STUDIES. HYPOTHESIS: IGF2BP2 PLAYS AN ESSENTIAL ROLE IN MLLR AML PATHOGENESIS AND LSC/LIC SELF-RENEWAL, AND THAT PHARMACOLOGICAL INHIBITION OF IGF2BP2 CAN LEAD TO EFFECTIVE TREATMENT OF MLLR AML. SPECIFIC AIMS: 1) DETERMINE THE ROLE OF IGF2BP2 IN MLLR AML PATHOGENESIS AND LSC/LIC SELF-RENEWAL; 2) DECIPHER THE MOLECULAR MECHANISM UNDERLYING THE ROLE OF IGF2BP2 IN MLLR AML; AND 3) ASSESS THE THERAPEUTIC POTENTIAL OF PHARMACOLOGICALLY TARGETING IGF2BP2 IN TREATING MLLR AML. POTENTIAL IMPACT: OUR PROPOSED STUDIES ARE OF HIGH NOVELTY AND HIGH SIGNIFICANCE IN BOTH BASIC RESEARCH AND TRANSLATIONAL MEDICINE, WHICH WILL SUBSTANTIALLY ADVANCE OUR UNDERSTANDING OF THE BIOLOGY OF MLLR LEUKEMIA, AND MAY ALSO RESULT IN THE DEVELOPMENT OF EFFECTIVE NOVEL THERAPEUTICS FOR THE TREATMENT OF MLLR LEUKEMIA.

CUTANEOUS T CELL LYMPHOMA: A PARADIGM FOR DISSECTING SUSCEPTIBILITY AND RESISTANCE TO CHECKPOINT INHIBITION THERAPY

HCMV VACCINE PRODUCED FROM BAC-MVA THAT BLOCKS EPITHELIAL AND FIBROBLAST ENTRY

TARGETING FTO TO TREAT ACUTE MYELOID LEUKEMIA

MOLECULAR PROGRAMMING OF SALIVARY GLAND GENE EXPRESSION

COMBINATIONAL TARGETING THE FEED FORWARD EPIGENETIC CIRCUITRY IN MIXED LINEAGE LEUKEMIA

INTEGRATION OF EPIDEMIOLOGY, PATHOLOGY, IMMUNOLOGY AND OUTCOMES IN COLORECTAL CANCER - ABSTRACT MACHINE LEARNING HAS THE POTENTIAL TO TRANSFORM PATHOLOGIC DIAGNOSIS AND TO ADDRESS VERY LIMITED ACCESSIBILITY OF EXPERT PATHOLOGY IN LOW-INCOME COUNTRIES. ROUTINE HISTOLOGY IMAGES OF SOLID TUMORS CONTAIN AN IMMENSE NUMBER OF VISUAL FEATURES THAT CAN BE EXTRACTED AND PROCESSED BY ARTIFICIAL INTELLIGENCE TOOLS LIKE MACHINE LEARNING, WHICH EXCELS AT BASIC IMAGE ANALYSIS TASKS SUCH AS TUMOR DETECTION. IN ADDITION, MACHINE LEARNING CAN ALSO PREDICT CLINICALLY RELEVANT FEATURES DIRECTLY FROM HISTOLOGY IMAGES INCLUDING MICROSATELLITE INSTABILITY AND IMMUNE FEATURES THAT INDEPENDENTLY PREDICT PROGNOSIS RESPONSE TO THERAPY. THIS LARGE, MULTICULTURAL, RACIALLY AND ETHNICALLY DIVERSE STUDY USES IMAGES OF WHOLE SLIDES FROM ROUTINELY COLLECTED CLINICAL SPECIMENS AND APPLIES COMPUTATIONAL PATHOLOGY METHODS AND DIGITAL SPATIAL EXPRESSION PROFILING TO QUANTIFIABLY IMPROVE CRC DIAGNOSIS, PROGNOSIS AND PREDICTIVE MODELS TOGETHER WITH CLINICAL, EPIDEMIOLOGIC AND GENETIC DATA. THE STUDY GOALS WILL BE ACCOMPLISHED THROUGH THREE SPECIFIC AIMS. IN AIM 1, WE WILL APPLY NOVEL MACHINE LEARNING ALGORITHMS FROM WHOLE SLIDE IMAGES TO REPRODUCIBLY IDENTIFY MSI, HISTOPATHOLOGIC AND IMMUNE FEATURES OF COLORECTAL CANCER IN RACIALLY/ETHNICALLY DIVERSE POPULATIONS. WE WILL STUDY H&E SLIDES FROM 6,751 CRC CASES, DIGITIZING EXISTING SLIDES FROM 5,551 CRC CASES AND 1,200 NEW CASES OF CRC WITH CONTEMPORANEOUS CLINICAL AND EPIDEMIOLOGIC DATA. THEN, WE WILL APPLY DEEP LEARNING METHODS TO ACCURATELY IDENTIFY HISTOPATHOLOGIC FEATURES AND IMMUNE CHARACTERISTICS OF CRC. WE WILL USE A ROBUST TRAINING VALIDATION, AND TESTING DESIGN (70%/15%/15%) TO ENSURE THE RIGOR AND REPRODUCIBILITY OF OUR FINDINGS. IN AIM 2, WE WILL TEST WHETHER MACHINE LEARNING ALGORITHMS THAT PREDICT MSI AND IMMUNE FEATURES RELATED TO CRC PROGNOSIS IMPROVE WITH THE ADDITION OF CLINICAL, EPIDEMIOLOGIC, AND GERMLINE GENETIC DATA. WE WILL USE MACHINE LEARNING STATISTICAL METHODS TO TEST WHETHER ALGORITHMS DEVELOPED IN AIM 1 IMPROVE PREDICTION OF OVERALL SURVIVAL AND RESPONSE TO THERAPY WITH THE ADDITION OF SUPPLEMENTAL INFORMATION BEYOND WHOLE SLIDE DIGITAL IMAGES. FINALLY, IN AIM 3, WE WILL COMPARE THE INFORMATION DERIVED FROM DIGITAL SPATIAL PROFILING OF EXPRESSED PROTEINS IN COLORECTAL TUMORS WITH THE INFORMATION DERIVED FROM IMMUNOSCORE QUANTIFICATION OF LYMPHOCYTE POPULATIONS AT THE TUMOR CENTER (CT) AND THE INVASIVE MARGIN (IM), AND EXPLORE WHETHER THESE MEASURES IMPROVE THE MODELS DEVELOPED IN AIMS 1 AND 2 IN A SUBSET OF SAMPLES. WE WILL PERFORM GEOMX DIGITAL SPATIAL PROFILING OF 56 PROTEINS EXPRESSED IN 150 STAGE I-III TNM COLORECTAL CANCERS TO COMPARE THE PERFORMANCE OF DIGITAL SPATIAL PROFILING TO IMMUNOSCORE, A SCORING SYSTEM RELYING EXCLUSIVELY ON EXPRESSION PATTERNS OF CD3+ AND CD8+ T CELLS. THIS STUDY TAKES ADVANTAGE OF PATHOLOGIC, EPIDEMIOLOGIC, CLINICAL, IMMUNOLOGIC AND GERMLINE GENETIC DATA FROM RACIALLY/ETHNICALLY DIVERSE CRC PATIENTS FROM CALIFORNIA, DETROIT, NEW YORK, FLORIDA, PUERTO RICO, ISRAEL AND SPAIN. OUR OVERARCHING GOAL IS TO IMPROVE THE EFFICIENT DIAGNOSIS OF COLORECTAL CANCER WITH CLINICALLY IMPACTFUL IMMUNE PROFILES.

REPURPOSING LEFLUNOMIDE TO DELAY PROGRESSION OF SMOLDERING MULTIPLE MYELOMA IN AFRICAN AMERICANS - PROJECT SUMMARY SMOLDERING MULTIPLE MYELOMA (SMM) IS AN ASYMPTOMATIC PRECURSOR OF ACTIVE MULTIPLE MYELOMA (MM), AND 50 PERCENT OF PATIENTS MEETING CRITERIA FOR HIGH-RISK DISEASE WILL DEVELOP ACTIVE MM WITHIN 2 YEARS. DESPITE RECENT ADVANCES IN TREATMENT FOR MM, AFRICAN AMERICANS HAVE EXPERIENCED RACIAL DISPARITIES IN DISEASE OUTCOME AND BEAR SIGNIFICANTLY GREATER DISEASE BURDEN. THE REASONS FOR THIS RACIAL DISPARITY ARE UNCLEAR AND MAY BE DUE TO BIOLOGICAL DIFFERENCES, DIAGNOSTIC AND TREATMENT DELAYS, AND/OR UNEQUAL ACCESS TO HEALTH CARE. ALTHOUGH SOME DRUGS USED TO TREAT MM HAVE SHOWN PROMISE IN IMPROVING PROGRESSION FREE SURVIVAL COMPARED TO OBSERVATION, THIS ADVANTAGE HAS NOT BEEN ESTABLISHED FOR AFRICAN AMERICAN PARTICIPANTS. IN ADDITION, BECAUSE THESE DRUGS CARRY A PRICE IN TERMS OF TOXICITY AND ECONOMIC BURDEN, THEIR ROLE IN TREATING PRE-MYELOMA CONDITIONS IS CONTROVERSIAL. THIS HIGHLIGHTS THE URGENT NEED FOR NEW APPROACHES WITH NOVEL MECHANISMS OF ACTION THAT CAN BE USED SUCCESSFULLY LONG-TERM TO PREVENT DISEASE PROGRESSION. TO MEET THIS NEED, WE PROPOSE TO EVALUATE LEFLUNOMIDE, A COMMERCIALLY AVAILABLE ORAL IMMUNOSUPPRESSIVE AGENT THAT HAS BEEN FDA-APPROVED SINCE 1998 FOR THE TREATMENT OF RHEUMATOID ARTHRITIS. OUR PRECLINICAL DATA INDICATE THAT CLINICALLY ACHIEVABLE CONCENTRATIONS OF LEFLUNOMIDE: A) INDUCE FAVORABLE IMMUNOLOGICAL CHANGES ABLE TO DELAY MM PROGRESSION IN IMMUNOCOMPETENT MM MICE AND B) DOWNREGULATE EXPRESSION OF THE MASTER REGULATORY MM ONCOGENE C-MYC AT THE MRNA AND PROTEIN LEVELS IN MM CELLS. MOREOVER, WE SAW ENCOURAGING RESULTS IN OUR RECENTLY COMPLETED PHASE I CLINICAL TRIAL OF SINGLE AGENT LEFLUNOMIDE IN RELAPSED/REFRACTORY MM PATIENTS IN WHICH SAFETY AND DISEASE STABILIZATION WERE SEEN IN NINE OF ELEVEN PATIENTS, INCLUDING TWO AFRICAN AMERICAN PATIENTS WHO HAD STABLE DISEASE LASTING FOR OVER A YEAR. THEREFORE, WE HYPOTHESIZE THAT LEFLUNOMIDE, AS A SINGLE AGENT, WILL BENEFIT PATIENTS WITH HIGH-RISK SMM BY PREVENTING OR DELAYING PROGRESSION TO SYMPTOMATIC MM. WE PROPOSE TO 1) DETERMINE THE ANTI-MYELOMA ACTIVITY OF SINGLE AGENT LEFLUNOMIDE IN A PHASE 2 CLINICAL TRIAL IN AFRICAN AMERICAN AND EUROPEAN AMERICAN PATIENTS WITH HIGH-RISK SMM; 2) CHARACTERIZE THE TEMPORAL RELATIONSHIP BETWEEN SERUM CONCENTRATION OF TERIFLUNOMIDE, THE ACTIVE METABOLITE OF LEFLUNOMIDE, AND DISEASE STATUS AND THE IMPACT OF GENETIC POLYMORPHISMS ON TERIFLUNOMIDE CONCENTRATION; AND 3) DETERMINE THE RELATIONSHIP BETWEEN LEFLUNOMIDE, IMMUNOLOGICAL CHANGES, AND DISEASE STATUS, AND CHANGES IN C-MYC SIGNATURE. SUCCESSFUL COMPLETION OF THESE STUDIES WOULD PROVIDE THE FIRST INSIGHT INTO THE UNDERLYING MECHANISM OF HOW LEFLUNOMIDE MODULATES THE IMMUNE SYSTEMS OF AFRICAN AMERICAN AND EUROPEAN AMERICAN PATIENTS WITH HIGH-RISK SMM AND HOW THESE CHANGES AFFECT RESPONSE TO TREATMENT AND DISEASE PROGRESSION. FURTHERMORE, SHOWING LEFLUNOMIDE TO BE ACTIVE IN DELAYING OR PREVENTING PROGRESSION OF SMM TO ACTIVE DISEASE WOULD PROVIDE A WELL-TOLERATED ALTERNATIVE FOR PATIENTS WITH HIGH-RISK SMM, AND RESULTS COULD BE EXTRAPOLATED TO OTHER PATIENT POPULATIONS.

ROLE OF 12-LIPOXYGENASE IN DIABETIC NEPHROPATHY

ASPIRIN AND CANCER PREVENTION IN LYNCH SYNDROME: FROM CELL TO POPULATION DATA

REOLYSIN-BASED COMBINATION THERAPY IN RELAPSED MULTIPLE MYELOMA

THE ROLE OF MIR-142 IN THE TRANSFORMATION OF CLONAL HEMATOPOIETIC DISORDERS INTO AML - CHRONIC CLONAL BLOOD DISORDERS SUCH AS MYELOPROLIFERATIVE NEOPLASMS (MPN) AND CHRONIC PHASE (CP) CHRONIC MYELOGENOUS LEUKEMIA (CML) MAY OVER TIME TRANSFORM, RESPECTIVELY, INTO SECONDARY (S) ACUTE MYELOID LEUKEMIA (AML) AND BLAST CRISIS (BC) CML, WHICH ARE POORLY RESPONSIVE TO CURRENTLY AVAILABLE THERAPIES, INCLUDING ALLOGENEIC STEM CELL TRANSPLANTATION. THUS, THE AVAILABILITY OF NOVEL AND MORE EFFECTIVE TREATMENTS IS A TRUE UNMET NEED FOR THESE PATIENTS. MICRORNAS (MIRNAS) ARE SMALL NON-CODING RNAS THAT TARGET MESSENGER RNAS AND REGULATE THE CORRESPONDING PROTEIN LEVELS. MIR142, ENCODING MIR-142, IS A HIGHLY CONSERVED “GENE”, EXPRESSED AT HIGH LEVELS IN HEMATOPOIETIC CELLS AND IS INVOLVED IN THE DEVELOPMENT AND FUNCTION OF MYELOID, LYMPHOID AND MEGAKARYOCYTE- ERYTHROID PROGENITORS. MIR142 HAS BEEN FOUND MUTATED AND/OR DOWNREGULATED BOTH IN LYMPHOMA AND AML. FURTHERMORE, MIR-142 KNOCK-OUT (KO) CAUSES IMPAIRED HEMATOPOIESIS IN ZEBRA FISH AND MICE, WITH EXPANSION OF HEMATOPOIETIC STEM AND PROGENITOR CELLS (HSPCS) AND DECREASED HEMATOPOIETIC OUTPUT. WE RECENTLY DEMONSTRATED THAT MIR-142 KO IN MOUSE MODELS WITH CLONAL MYELOPROLIFERATIVE DISORDERS (MPDS; I.E., FLT3-ITD+ MPN OR CP CML) PROMPTS TRANSFORMATION INTO AN AML-LIKE DISEASE AND CONFERS A SIGNIFICANTLY SHORTER SURVIVAL TO THESE ANIMALS. OUR DATA SUPPORT A ROLE OF MIR-142 DEFICIT IN DEREGULATION OF THE METABOLISM OF CLONAL HEMATOPOIETIC STEM CELLS (HSCS), WITH A SWITCH TO HIGHER LEVELS OF OXIDATIVE PHOSPHORYLATION (OXPHOS) VIA INCREASED FATTY ACID OXIDATION (FAO); THESE CHANGES LIKELY PLAY A KEY ROLE IN THE TRANSFORMATION OF CLONAL HSCS INTO LEUKEMIC STEM CELLS (LSCS). WE DEMONSTRATED THAT RESCUE OF MIR-142 DEFICIT WITH A NOVEL MIR-142 MIMIC COMPOUND (CPG-M-MIR-142) REDUCED OXPHOS LEVELS AND VIABILITY OF LSCS, DECREASED LSC BURDEN AND ACTIVITY AND PROLONGED SURVIVAL OF TREATED BC CML MICE. THUS, THE CENTRAL HYPOTHESIS OF THIS PROPOSAL IS THAT THE UNDERSTANDING OF THE CELLULAR AND MOLECULAR BASIS OF MIR-142 DOWNREGULATION AND ITS IMPACT ON THE TRANSFORMATION OF CLONAL MPD INTO AGGRESSIVE AML-LIKE DISEASE WILL ALLOW US TO DESIGN AND OPTIMIZE NOVEL TREATMENTS TO COMPENSATE FOR THE MIR-142 DEFICIT AND PREVENT AND CURE MPD TRANSFORMATION. WE PROPOSE THE FOLLOWING SPECIFIC AIMS (SAS): SA#1: TO DEFINE THE ROLE OF MIR-142 DEFICIT IN THE SAML/BC CML TRANSFORMATION. SA#2: TO DISSECT THE MOLECULAR MECHANISMS THROUGH WHICH MIR-142 DEFICIT CONTRIBUTES TO SAML/BC CML TRANSFORMATION. SA#3: TO INVESTIGATE THE PHARMACOKINETIC (PK), PHARMACODYNAMIC (PD) AND THERAPEUTIC IMPACT OF A SYNTHETIC CPG-M-MIR-142 THAT WILL RESCUE MIR-142 DEFICIT IN SAML/BC CML.

BAFF-R CAR T CELL THERAPY FOR ALL - PROJECT SUMMARY B-CELL ACUTE LYMPHOBLASTIC LYMPHOMA (B-ALL) IS A NEOPLASM OF B-CELL LYMPHOID PRECURSORS THAT TYPICALLY AFFECTS CHILDREN, BUT OCCURS IN ADULTS AS WELL. WHILE INTENSIVE MULTI-AGENT CHEMOTHERAPY IS HIGHLY EFFECTIVE IN THE PEDIATRIC POPULATION, OUTCOMES REMAIN POOR IN ADULTS AND HIGH-RISK PATIENTS. THE RECENT INTRODUCTION OF BLINATUMOMAB AND CD19-DIRECTED CAR T THERAPY HAS TRANSFORMED THE CARE OF PATIENTS WITH RELAPSED AND REFRACTORY (R/R) B-ALL. HOWEVER, AN INCREASING NUMBER OF REPORTS DESCRIBE A HIGH RATE OF POST-TREATMENT RELAPSE DUE TO ACQUIRED RESISTANCE TO THESE IMMUNOTHERAPIES, NOTABLY VIA DOWN-REGULATION OR LOSS OF CD19 SURFACE EXPRESSION. CAR T CELLS THAT WERE RECENTLY DEVELOPED AGAINST ANOTHER TARGET, CD22, SHOWED SIMILAR SHORTCOMINGS WITH RELAPSES ASSOCIATED WITH DIMINISHED ANTIGEN EXPRESSION. THE SEARCH FOR ALTERNATIVE TARGETS IS THEREFORE ESSENTIAL TO OVERCOMING ANTIGEN ESCAPE. TO ADDRESS THIS ISSUE, WE HAVE DEVELOPED CAR T CELLS AGAINST A NEW B-ALL TARGET, THE B-CELL ACTIVATING FACTOR RECEPTOR (BAFF-R). BAFF-R IS A MARKER OF B CELLS THAT IS ALSO FUNCTIONALLY EXPRESSED IN B-ALL, INCLUDING IN PATIENTS WITH CD19-NEGATIVE RELAPSE. ALTHOUGH ONE OF BAFF-R’S LIGANDS (BAFF) HAS BEEN SUCCESSFULLY TARGETED FOR THE TREATMENT OF AUTOIMMUNE DISEASES, WE ARE THE FIRST TO HAVE DEVELOPED BAFF-R CAR T CELLS THAT ARE EFFECTIVE AGAINST B-CELL MALIGNANCIES IN VIVO, INCLUDING IN CD19-NEGATIVE MOUSE MODELS. BECAUSE BAFF-R IS A CRITICAL REGULATOR OF B-CELL FUNCTION AND SURVIVAL, WE CAN EXPECT THAT THE TUMOR’S ABILITY TO ESCAPE THERAPY BY DOWN-REGULATION OF BAFF-R WILL BE LIMITED. WHILE WE ANTICIPATE CLINICAL EFFICACY OF BAFF-R CAR T CELL THERAPY, DUAL TARGETING OF CD19 AND BAFF-R CAN PREVENT THE EMERGENCE OF RESISTANCE AND IMPROVE CLINICAL OUTCOMES. THEREFORE, WE ARE ALSO DEVELOPING BISPECIFIC CD19- BAFF-R CAR T CELLS THAT WE AIM TO RAPIDLY TRANSLATE FROM THE BENCH TO THE BEDSIDE. IN SPECIFIC AIM 1, WE WILL EVALUATE BAFF-R CAR T CELL THERAPY IN A FIRST-IN-HUMAN CLINICAL TRIAL IN PATIENTS WITH R/R B-ALL WHO ARE INELIGIBLE FOR OR HAVE FAILED PRIOR CD19-DIRECTED THERAPY. THE TRIAL, CURRENTLY OPEN AT CITY OF HOPE, WILL USE OUR TN/MEM- DERIVED MANUFACTURING PLATFORM, WHICH YIELDS A NAÏVE/MEMORY T-CELL ENRICHED T CELL PRODUCT AND HAS SHOWN REMARKABLE EFFICACY AND TOLERABILITY IN B-ALL. WE WILL CONDUCT EXTENSIVE CORRELATIVE STUDIES USING CUTTING-EDGE TECHNOLOGIES, SUCH AS ASSESSING THE KINETICS OF THE CAR T CELLS AND THAT OF DIVERSE CYTOKINES, THERAPEUTIC EFFECT, AND POTENTIAL MECHANISMS OF RELAPSE AND ANTIGEN ESCAPE. IN SPECIFIC AIM 2, WE WILL ESTABLISH THE THERAPEUTIC EFFICACY OF BISPECIFIC CD19-BAFF-R CAR T CELLS AGAINST MIXED (CD19-NEGATIVE + BAFF-R- NEGATIVE) B-ALL TUMOR MODELS THAT MIMIC THE HETEROGENOUS TUMOR PHENOTYPE LEADING TO RESISTANCE. WE WILL ALSO PERFORM EXTENSIVE TESTING OF CGMP-GRADE BISPECIFIC CD19-BAFF-R CAR T CELLS THAT IS REQUIRED PRIOR TO SUBMISSION OF AN IND APPLICATION TO THE FDA. OUR PROPOSAL ADDRESSES THE URGENT NEED FOR NEW THERAPEUTIC OPTIONS IN PATIENTS WITH R/R B-ALL AND COULD ALSO BENEFIT PATIENTS WITH OTHER B-CELL MALIGNANCIES.

TARGETED INHIBITION OF HIV-1 LATENCY

ROLE OF AUTOREACTIVITY IN PATHOGENESIS OF CHRONIC GRAFT VERSUS HOST DISEASE - ALLOGENEIC HEMATOPOIETIC CELL TRANSPLANTATION (ALLO-HCT) IS A CURATIVE THERAPY FOR PATIENTS WITH HEMATOLOGICAL MALIGNANCIES (I.E. LEUKEMIA AND LYMPHOMA), THROUGH GRAFT VERSUS LEUKEMIA/LYMPHOMA (GVL) EFFECT MEDIATED BY ALLOREACTIVE DONOR T CELLS. HOWEVER, GRAFT-VERSUS-HOST DISEASE (GVHD) CAUSED BY THE SAME ALLOREACTIVE T CELLS REMAINS A MAJOR OBSTACLE. CHRONIC GVHD (CGVHD), A SYSTEMIC AUTOIMMUNE-LIKE SYNDROME, REMAINS A MAJOR CAUSE OF MORBIDITY AND MORTALITY IN LONG-TERM SURVIVORS OF ALLO-HCT. DURING THE PAST DECADE, WE HAVE USED MURINE AND HUMANIZED MURINE MODELS OF CGVHD AND BIOSPECIMENS FROM CGVHD PATIENTS TO DEMONSTRATE THAT CD4+ T CELLS AND B CELLS AND THEIR INTERACTIONS IN GVHD TARGET TISSUES SUCH AS THE LIVER, LUNG AND SKIN PLAY A CRITICAL ROLE IN THE PATHOGENESIS AND PERSISTENCE OF THE DISEASE. THE PREMISE OF THIS NEW PROPOSAL IS THAT DEEPER MECHANISTIC UNDERSTANDING OF THE INTERACTIONS BETWEEN NON-CIRCULATING TISSUE- RESIDENT CD4+ MEMORY T (TRM) AND TISSUE-RESIDENT B (BRM) CELLS IN GVHD TARGET TISSUES WILL IDENTIFY NEW THERAPEUTIC TARGETS FOR PREVENTION OR TREATMENT OF CGVHD. IN AIM 1, WE WILL DETERMINE WHETHER CD4+ TRM CELLS INITIATE FORMATION OF TERTIARY LYMPHOID STRUCTURES (TLS) IN GVHD TARGET TISSUES, IN WHICH LY108+ STEM CELL-LIKE PROGENITORS MAINTAIN THE POOL OF TRM AND BRM TO PERPETUATE CGVHD PATHOGENESIS. WE WILL ALSO DEVELOP NOVEL CELL-PENETRATING PS-CAS9-GRNA TO TARGET STAT3 IN DONOR T CELLS TO PREVENT DEVELOPMENT OF PATHOGENIC CD4+ TRM AND CGVHD WHILE PRESERVING GVL ACTIVITY, BECAUSE OUR RECENT STUDIES INDICATE THAT STAT3 IS REQUIRED FOR TRM DEVELOPMENT. IN AIM 2, WE WILL DETERMINE WHETHER B CELL HELPER PD-1HIPSGL1LOCD4+ TRH CELLS INTERACT WITH T-BET+ BRM CELLS TO PRODUCE AUTOANTIBODIES THAT AUGMENT SYSTEMIC TISSUE DAMAGE. IN AIM 3, WE WILL DETERMINE WHETHER NON-B CELL HELPER PSGL1HICD4+ TRM CELLS INTERACT WITH DENDRITIC CELLS, MACROPHAGES, AND FIBROBLASTS IN CGVHD TARGET TISSUES TO MEDIATE DAMAGE AND FIBROSIS IN AN IL-33/ST2 PATHWAY-DEPENDENT MANNER. THESE STUDIES HAVE HIGH BIOLOGICAL AND TRANSLATIONAL SIGNIFICANCE AND MAY LEAD TO A PARADIGM SHIFT IN OUR UNDERSTANDING OF CGVHD PATHOGENESIS AND TO DEVELOPMENT OF NOVEL APPROACHES FOR PREVENTING AND TREATING CGVHD.

BIOLOGIC AND THERAPEUTIC SIGNIFICANCE OF MIR-155 IN AML

PREPARING ONCOLOGY ADVANCED PRACTICE NURSES AS GENERALISTS IN PALLIATIVE CARE

TARGETING TUMOR-ASSOCIATED MACROPHAGES FOR TRIPLE-NEGATIVE BREAST CANCER TREATMENT - PROJECT SUMMARY/ABSTRACT TRIPLE-NEGATIVE BREAST CANCER (TNBC) IS CHARACTERIZED BY THE LACK OF ESTROGEN RECEPTOR, PROGESTERONE RECEPTOR AND HUMAN EPIDERMAL GROWTH FACTOR RECEPTOR 2, ALL OF WHICH ARE IMPORTANT THERAPEUTIC TARGETS. TNBC IS THE MOST DIFFICULT-TO-TREAT SUBGROUP OF BREAST CANCERS AND IS RESISTANT TO MANY CURRENT CANCER THERAPIES. THE PRESENT SITUATION OF POOR PROGNOSIS WITH LIMITED THERAPY OPTIONS IN TNBC EMPHASIZES AN URGENT NEED FOR MORE EFFECTIVE THERAPEUTICS. THE ABILITY TO ESCAPE FROM THE SURVEILLANCE BY THE IMMUNE SYSTEM IS REGARDED AS ONE OF THE ESSENTIAL HALLMARKS OF CANCER CELLS. RECENT EXCITING DISCOVERIES HAVE IDENTIFIED MANY IMPORTANT SIGNALS AND MECHANISMS MEDIATING CANCER CELL IMMUNE EVASION. IMMUNOTHERAPIES HAVE BEEN DEVELOPED TO TARGET THESE SIGNALS, REVOLUTIONIZING THE TREATMENT OF A VARIETY OF HUMAN CANCERS. TUMOR-ASSOCIATED MACROPHAGES (TAMS) REPRESENT THE MAJOR COMPONENTS OF THE TUMOR MICROENVIRONMENT IN TNBC. RECENT STUDIES DEMONSTRATE THAT THE BLOCKADE OF A “DON’T EAT ME” SIGNAL CD47 LEADS TO DIRECT PHAGOCYTOSIS OF LIVING CANCER CELLS BY MACROPHAGES, AND SIGNIFICANTLY INHIBITS THE ENGRAFTMENT OF VARIOUS MALIGNANT HEMATOPOIETIC AND SOLID TUMOR CELLS IN MICE THAT LACK T, B, AND NK CELLS, INDICATING A CRITICAL ROLE OF MACROPHAGES IN CANCER IMMUNOSURVEILLANCE. TARGETING TAMS IN THE TUMOR MICROENVIRONMENT REPRESENTS A NEW CLASS OF PROMISING CANCER IMMUNOTHERAPY. WHILE INDUCING ANTICANCER FUNCTIONS OF TAMS HOLDS CONSIDERABLE PROMISE FOR CANCER TREATMENT, THERE ARE SEVERAL BARRIERS THAT NEED TO BE OVERCOME TO ACHIEVE DESIRED EFFICACY FOR TREATING TNBC. IN PRELIMINARY STUDIES, WE FOUND THAT TAMS CAN BE REPROGRAMMED BY SMALL MOLECULE ANTINEOPLASTIC COMPOUNDS TO INDUCE THEIR PHAGOCYTIC ABILITY AGAINST TNBC CELLS. HOWEVER, THE UNDERLYING MOLECULAR MECHANISMS REGULATING THE REPROGRAMMING OF MACROPHAGES REMAINS UNCLEAR. THE OVERALL OBJECTIVE OF THE PROPOSED STUDY IS TO UNDERSTAND THE UNDERLYING MECHANISMS OF MACROPHAGE-MEDIATED IMMUNOSURVEILLANCE IN TNBC AND TO DEVELOP STRATEGIES TO EFFECTIVELY TREAT TNBC BY EXPLOITING TUMORICIDAL ROLES OF TAMS, WITH A COMBINATION OF IN VITRO AND IN VIVO PRECLINICAL TNBC MODELS. IN AIM1, WE WILL ASSESS THE EFFICACY OF REPROGRAMMING MACROPHAGES IN TNBC TREATMENT BY USING METASTATIC TNBC MODELS AND CHEMOTHERAPY-RESISTANT PATIENT-DERIVED XENOGRAFT MODELS. IN AIM2, WE WILL STUDY THE MOLECULAR MECHANISMS BY WHICH MACROPHAGES ARE REPROGRAMMED BY DISSECTING THE FUNCTIONS AND ROLES OF PATTERN RECOGNITION RECEPTOR SIGNALING PATHWAYS IN MACROPHAGE REPROGRAMMING AND CHARACTERIZING TAM SUBGROUPS IN TNBC TUMORS. IN AIM3, WE WILL DETERMINE THE EFFECTS OF TARGETING MACROPHAGE CELL SURFACE MOLECULAR MACHINERY ON ACTIVATING TAMS FOR TNBC TREATMENT. SUCCESSFUL COMPLETION OF THE PROPOSED STUDIES SHOULD SHED LIGHT ON THE BASIC PRINCIPLES OF CANCER CELL IMMUNE EVASION AND INSPIRE THE DEVELOPMENT OF NOVEL THERAPEUTICS FOR TNBC TREATMENT.

LONG NON-CODING RNA-MEDIATED CHROMATIN REMODELING IN ANGIOGENESIS

INTEGRATIVE ROLE OF RPS6KB1 IN PATHOLOGICAL CARDIAC REMODELING - PROJECT SUMMARY HEART FAILURE IS A LEADING CAUSE OF MORBIDITY AND MORTALITY WORLDWIDE. HYPERTENSION IS ONE OF THE MOST IMPORTANT RISK FACTORS OF HEART FAILURE. DESPITE PARAMOUNT INTERESTS AND URGENT CLINICAL NEEDS, OUR UNDERSTANDING OF THE MECHANISMS OF HEART FAILURE DEVELOPMENT REMAINS LIMITED. TO ACCOMMODATE THE ELEVATED DEMAND OF CARDIAC CONTRACTILITY UNDER HIGH BLOOD PRESSURE, THE HEART MOUNTS AN ACUTE REACTION THROUGH CARDIOMYOCYTE HYPERTROPHIC GROWTH. THIS ONCE ADAPTIVE RESPONSE MAY DECOMPENSATE AND PROGRESS INTO HEART FAILURE. THE LONG- TERM GOAL OF THIS PROJECT IS TO IDENTIFY NOVEL MECHANISMS OF THE PATHOLOGICAL TRANSITION FROM ADAPTIVE CARDIAC HYPERTROPHY TO HEART FAILURE AND EXPLORE THERAPEUTIC INTERVENTIONS. WITH AN UNBIASED PHOSPHOPROTEOMIC ANALYSIS, RPS6KB1 WAS IDENTIFIED AS ONE OF THE MOST PROMINENT KINASES IN CARDIOMYOCYTE HYPERTROPHIC GROWTH. RPS6KB1 IS A CLASSICAL DOWNSTREAM TARGET OF MTOR, HOWEVER, ITS ROLE IN HYPERTENSIVE HEART DISEASE REMAINS INCOMPLETELY UNDERSTOOD. MOREOVER, RPS6KB1, WITH ITS COMPLEX ACTIVATION MECHANISM, MAY INTEGRATE SIGNALS FROM MULTIPLE UPSTREAM PATHWAYS, ABOVE AND BEYOND THE MTOR SIGNALING. PRELIMINARY STUDIES SHOWED THAT RPS6KB1 DELETION IN THE HEART SUPPRESSES ADAPTIVE CARDIAC HYPERTROPHIC GROWTH AND CONSEQUENTLY, CARDIOMYOPATHY AND HEART FAILURE ARE ACCELERATED UNDER PRESSURE OVERLOAD. BASED ON THESE FINDINGS, A CENTRAL HYPOTHESIS IS FORMULATED THAT RPS6KB1 IS AN ESSENTIAL PLAYER IN ADAPTIVE HYPERTROPHIC GROWTH BY INTEGRATING SIGNALS FROM MULTIPLE UPSTREAM PATHWAYS. ADDITIONAL PRELIMINARY DATA SHOWED THAT RPS6KB1 IS DIRECTLY PHOSPHORYLATED BY ERK, INDEPENDENT OF THE MTOR SIGNALING. A NOVEL THREONINE SITE OF PHOSPHORYLATION BY ERK WAS LATER IDENTIFIED BY MASS SPECTROMETRY. MORE IMPORTANTLY, ERK-MEDIATED PHOSPHORYLATION OF RPS6KB1 IS REQUIRED FOR FULL RPS6KB1 ACTIVATION. FURTHER PILOT TESTS DEMONSTRATED THAT RPS6KB1 IS SUBJECTED TO ANOTHER POST-TRANSLATIONAL MODIFICATION, K63 POLYUBIQUITINATION. LYSINE 85 WAS IDENTIFIED AS THE CRITICAL SITE FOR THIS MODIFICATION THROUGH MULTIPLE MUTAGENESIS ASSAYS, AND LYS85ALA MUTATION STRONGLY DIMINISHED BOTH UBIQUITINATION AND ACTIVATION OF RPS6KB1. IN THIS GRANT APPLICATION, THE ROLE OF ERK-MEDIATED PHOSPHORYLATION OF RPS6KB1 WILL BE FURTHER CHARACTERIZED, AND ITS IMPLICATIONS IN ADAPTIVE HYPERTROPHIC GROWTH AND HEART FAILURE WILL BE ILLUSTRATED USING BOTH GAIN- AND LOSS-OF-FUNCTION MOUSE MODELS. IN ADDITION, THE RELEVANCE AND SIGNIFICANCE OF LYS85 UBIQUITINATION IN HYPERTENSIVE HEART DISEASE WILL BE DELINEATED BY IDENTIFICATION OF THE RESPONSIBLE E3 LIGASE AND EVALUATION OF THE NOVEL LYS85ALA KNOCK-IN MOUSE MODEL. MOREOVER, THE INTERPLAY BETWEEN THIS NOVEL PHOSPHORYLATION AND THIS NEW UBIQUITINATION IN CARDIAC HYPERTROPHY WILL BE INTERROGATED. PRIMARY CARDIOMYOCYTE CULTURE WILL BE EMPLOYED TO COMPLEMENT IN VIVO ANIMAL MODELS AT THE MECHANISTIC LEVEL. ELUCIDATION OF THE ROLE OF RPS6KB1 AS AN INTEGRATIVE PLAYER IN PATHOLOGICAL CARDIAC REMODELING WILL ADVANCE OUR UNDERSTANDING OF HYPERTENSIVE HEART DISEASE AND HEART FAILURE AND PAVE A WAY FOR NOVEL, MORE EFFECTIVE THERAPEUTIC DESIGN.

EXOSOMAL BIOMARKERS FOR THE NONINVASIVE DETECTION OF COLORECTAL CANCER

VASCULAR REMODELING IN THE BONE MARROW LEUKEMIC NICHE: A THERAPEUTIC TARGET?

INTRACEREBROVENTRICULAR (ICV) ADMINISTRATION OF CD19-TARGETING CHIMERIC ANTIGEN RECEPTOR (CAR) T CELLS FOR TREATMENT OF PRIMARY CENTRAL NERVOUS SYSTEM LYMPHOMA - PROJECT SUMMARY PRIMARY CENTRAL NERVOUS SYSTEM LYMPHOMA (PCNSL) IS A RARE HEMATOLOGIC MALIGANCY IN WHICH NON-HODGKIN LYMPHOMA (NHL) INITIALLY PRESENTS IN THE CENTRAL NERVOUS SYSTEM (CNS). THERAPEUTIC OPTIONS FOR PCNSL ARE LIMITED; STANDARD OF CARE HIGH-DOSE METHOTREXATE-CONTAINING REGIMENS HAVE BEEN UNCHANGED FOR OVER 40 YEARS, AND ARE NOT CURATIVE IN MOST PATIENTS. CHIMERIC ANTIGEN RECEPTOR (CAR) T CELL THERAPY TARGETING CD19 (CD19- CAR T CELLS) IS A POWERFUL FORM OF IMMUNOTHERAPY THAT HAS AN ESTABLISHED SAFETY PROFILE WHEN DELIVERED INTRAVENOUSLY (IV) TO TREAT PATIENTS WITH NHL. OUR CLINICAL PLATFORM FOR MANUFACTURING CD19-CAR T CELLS AT CITY OF HOPE (COH) HAS BEEN EVALUATED IN A SERIES OF PHASE 1 CLINICAL TRIALS FOR B CELL ACUTE LYMPHOBLASTIC LEUKEMIA (ALL) AND FOR NHL. TO DATE, ALL PREVIOUS AND ONGOING CD19-CAR T CELL TRIALS HAVE INFUSED THE CAR T CELL PRODUCT IV. WE HAVE PRELIMINARY EVIDENCE THAT IV-ADMINISTERED CD19-CAR T CELLS CAN BE DETECTED IN THE CNS AND HAVE ANTI-TUMOR ACTIVITY IN TREATING PATIENTS WITH PCNSL. HOWEVER, THE EFFICACY OF IV CAR T CELL THERAPIES FOR PATIENTS WITH PCNSL IS LIMITED, POSSIBLY DUE TO POOR TRAFFICKING OF CAR T CELLS FROM BLOOD TO CNS THAT MAY RESULT IN REDUCED ACTIVITY AGAINST PCNSL COMPARED TO SYSTEMIC NHL. IN PHASE 1 TRIALS AT COH, LOCOREGIONAL DELIVERY OF CAR T CELLS TO TREAT CNS MALIGNANCIES SUCH AS GLIOBLASTOMA HAS LED TO IMPROVED OUTCOMES. STUDIES IN OUR ANIMAL MODELS SHOW IMPROVED DISEASE RESPONSE, DURABILITY AND RESISTENCE TO TUMOR RECHALLENGE USING INTRACEREBROVENTRICULAR (ICV)- VS IV- DELIVERED CD19-CAR T CELLS IN XENOGRAFT MOUSE MODELS OF CNS LYMPHOMA. THUS, TO OPTIMIZE THE EFFICACY OF CD19-CAR T CELLS AND IMPROVE THE OUTCOMES OF PATIENTS WITH PCNSL, WE PROPOSE TO ADMINISTER CD19-CAR T CELLS VIA ICV DELIVERY. WE HYPOTHESIZE THAT ICV-DELIVERED CD19-CAR T CELLS WILL BE SAFE AND DEMONSTRATE HIGH ANTI-PCNSL ACTIVITY. IN SPECIFIC AIM 1, WE WILL CONDUCT A PHASE 1 CLINICAL TRIAL TO ASSESS THE SAFETY AND ACTIVITY, AND DETERMINE THE RECOMMENDED PHASE 2 DOSE (RP2D) OF ICV DELIVERED CD19-CAR T CELLS IN PARTICIPANTS WITH PCNSL. IN SPECIFIC AIM 2, WE WILL CONDUCT A SERIES OF CORRELATIVE STUDIES TO ASSESS MECHANISMS OF TOXICITY, CAR T CELL PERSISTANCE, TRAFFICKING TO THE PERIPHERAL BLOOD, IMMUNE CELL PHENOTYPE, AND EFFECT ON TUMOR. WE PLAN TO EXAMINE THE EFFECTS OF ICV-DELIVERED CAR T CELLS ON NORMAL CD19+ B CELLS IN THE PERIPHERAL BLOOD TO DETERMINE WHETHER CAR T CELLS TRAFFIC FROM THE CNS TO THE BLOOD, AS WE EXPECT BASED ON OUR PRECLINICAL ANIMAL MODELS OF PCNSL. ACTIVITY AGAINST NORMAL B CELLS IN THE BLOOD WOULD INDICATE THAT ICV DELIVERY COULD BE A VIABLE TREATMENT OPTION FOR PATIENTS WITH SECONDARY CNS LYMPHOMA, WHO HAVE BOTH CNS AND SYSTEMIC DISEASE. SUCCESSFUL COMPLETION OF THE PROPOSED CLINICAL TRIAL AND CORRELATIVE STUDIES WILL EXPAND THERAPEUTIC OPTIONS FOR PATIENTS WITH PCNSL AND COULD INFORM THE DESIGN OF A POTENTIAL SUBSEQUENT CLINICAL TRIAL TO EVALUATE THE SAFETY AND EFFICACY OF TREATING PATIENTS WITH SECONDARY CNS LYMPHOMA.

HUMAN ISLET RESEARCH NETWORK BIOINFORMATICS CENTER

ENHANCING THE BREADTH AND EFFICACY OF THERAPEUTIC VACCINES FOR BREAST CANCER

OPTIMIZING BUSULFAN: EFFICACY, TOXICITY, AND PHARMACOMETABOLOMICS

CLINICAL AND BIOLOGICAL PREDICTORS OF CHEMOTHERAPY TOXICITY IN OLDER ADULTS

LIMITED COMPETITION FOR THE CONTINUATION OF THE INTESTINAL STEM CELL CONSORTIUM COORDINATING CENTER

TRANSLATION OF A LUNG CANCER PALLIATIVE CARE INTERVENTION FOR CLINICAL PRACTICE

YES 2 SUCCESS

TARGETING FLT3 FOR THE TREATMENT OF RELAPSED OR REFRACTORY ACUTE MYELOID LEUKEMIA - PROJECT SUMMARY CHIMERIC ANTIGEN RECEPTOR (CAR)-ENGINEERED T CELL THERAPY HAS REVOLUTIONIZED TREATMENT FOR CERTAIN B CELL MALIGNANCIES, BUT SIMILAR SUCCESSES FOR ACUTE MYELOID LEUKEMIA (AML) HAVE NOT YET BEEN SHOWN, ALTHOUGH INTERIM RESULTS OF EARLY PHASE CLINICAL TRIALS, INCLUDING OUR OWN, ARE PROMISING. WE HAVE CONSTRUCTED A CAR THAT TARGETS THE TYPE III RECEPTOR TYROSINE KINASE (RTK) CALLED FLT3. UTILIZING A PATIENT-DERIVED XENOGRAFT (PDX) MODEL OF FLT3(+) HUMAN AML, WE HAVE SHOWN THAT INFUSION OF HUMAN FLT3-CAR T CELLS PROLONGED SURVIVAL OF AML- BEARING MICE IN THE ABSENCE OF OTHER THERAPIES. FURTHER, FLT3-CAR T CELLS DID NOT AFFECT ENGRAFTMENT OR SURVIVAL OF HEMATOPOIETIC STEM CELLS (HSCS) IN MICE BEARING AML. THE EFFECTIVENESS OF FLT3-CAR T CELL THERAPY IN AML WILL DEPEND ON (1) SURFACE ANTIGEN DENSITY OF FLT3 ON AML BLASTS, INCLUDING LEUKEMIC STEM CELLS (LSC) RELATIVE TO NORMAL HSCS, AND (2) THE ABILITY OF OTHER IMMUNE EFFECTOR CELLS TO CONTRIBUTE TO THE ERADICATION OF AML IN VIVO. TO THIS END, WE HAVE DISCOVERED THAT TREATMENT OF AML BLASTS WITH AN RTK INHIBITOR (TKI) UPREGULATES THE EXPRESSION OF FLT3 ON THE AML BLAST AND THE LSC IN VIVO, RELATIVE TO FLT3 EXPRESSION ON NORMAL HSCS. TO ADVANCE A SECOND CELLULAR THERAPEUTIC INTERVENTION FOR AML, WE HAVE EXPRESSED THE FLT3-CAR IN HUMAN NATURAL KILLER (NK) CELLS TO GENERATE FLT3-CAR NK CELLS, WHICH DEMONSTRATED POTENT ANTI-LEUKEMIC ACTIVITY AGAINST FLT3(+) AML. COLLECTIVELY, THESE DISCOVERIES HAVE LED US TO CONCLUDE THAT A PROGRAM DIRECTED AGAINST FLT3(+) AML HAS STRONG RATIONALE, IS INNOVATIVE AND COULD RESULT IN A SIGNIFICANT DECLINE IN MORTALITY FOR A SUBSET OF AML PATIENTS. THEREFORE, THE LONG-TERM OBJECTIVE OF THIS PROPOSAL IS TO PERFORM BOTH PRECLINICAL AND CLINICAL STUDIES THAT WILL BEST DEFINE AN OPTIMAL STRATEGY TO REDUCE MORTALITY FROM AML WITH FLT3-CAR CELLULAR THERAPY, EITHER ALONE OR IN COMBINATION WITH TKIS. OUR CENTRAL HYPOTHESIS IS THAT TARGETING RELAPSED/REFRACTORY FLT3(+) AML WITH FLT3-CAR T CELLS OR FLT3-CAR NK CELLS IN COMBINATION WITH TKIS WILL IMPROVE OUTCOMES IN AML. IN THIS PROPOSAL, WE WILL ASSESS THE FEASIBILITY, SAFETY AND TOXICITY OF PERFORMING A PHASE I STUDY OF HUMAN FLT3-CAR T CELL THERAPY DIRECTED AGAINST FLT3(+) AML (AIM 1), WE WILL DETERMINE THE MECHANISM BY WHICH TKIS UPREGULATE THE SURFACE DENSITY EXPRESSION OF FLT3 ON LSCS AND HSCS (AIM 2), AND WE WILL OPTIMIZE A FLT3-CAR NK CELL PLATFORM AND ASSESS ITS FUNCTIONALITY AGAINST AML ALONE OR COMBINED WITH TKI (AIM 3). TO ACCOMPLISH THESE OBJECTIVES, WE HAVE BEGUN CLINICAL MANUFACTURING OF FLT3-CAR T CELLS TO TREAT ELIGIBLE PATIENTS WITH REFRACTORY OR RELAPSED AML; WE WILL UTILIZE BOTH HUMAN AML CELL LINES AND PATIENTS' AML BLASTS IN VITRO AS WELL AS IN VIVO ALONG WITH OUR PDX MODEL FOR OUR CORRELATIVE AND PRECLINICAL STUDIES EVALUATING FLT3-CAR T AND FLT3-CAR NK CELLS IN COMBINATION WITH TKI. UPON CONCLUSION, WE WILL UNDERSTAND HOW BEST TO OPTIMIZE CELLULAR IMMUNE THERAPY TO CURE AML. FURTHER INSIGHT INTO THIS PROCESS, AS WILL RESULT FROM THE IMPLEMENTATION AND COMPLETION OF THIS PROPOSAL IS IMPACTFUL AS IT WILL ULTIMATELY LEAD TO A REDUCTION IN MORTALITY FOR SELECT PATIENTS SUFFERING FROM AML.

BCL11B ACTIVATION AS AN APPROACH FOR ENHANCING THE EFFICACY OF IMMUNOTHERAPY - OUR GOAL IS TO INVESTIGATE OVEREXPRESSION OF THE T-LINEAGE TRANSCRIPTION FACTOR (TF) BCL11B AS A NOVEL STRATEGY TO ENHANCE: 1) T-CELL RECONSTITUTION AFTER ALLOGENEIC HEMATOPOIETIC STEM CELL TRANSPLANTATION (HSCT), AND 2) THE EFFICACY OF ANTICANCER CHIMERIC ANTIGEN RECEPTOR (CAR) T-CELLS. HSCT IS A CURATIVE THERAPY FOR MANY LEUKEMIAS BY ITSELF OR AS A POST-CAR CONSOLIDATION THERAPY. HOWEVER, THE GENERATION OF T-CELLS FROM DONOR HEMATOPOIETIC STEM AND PROGENITOR CELLS (HSPC) TAKES MANY MONTHS MAKING LIFE THREATENING INFECTIONS AND LEUKEMIA RELAPSE MAJOR CHALLENGES IN HSCT. WHILE CAR T-CELLS INDUCE HIGH REMISSION RATES IN CD19+ LEUKEMIAS, POOR T-CELL FUNCTION AND PERSISTENCE AND T-CELL EXHAUSTION DUE TO INHIBITION BY THE TUMOR MICROENVIRONMENT REMAIN MAJOR OBSTACLES TO THE CURATIVE EFFICACY OF CAR T-CELLS IN LEUKEMIA AND SOLID TUMORS. SPECIES RELATED DIFFERENCES IN THE REGULATION OF T-CELL DIFFERENTIATION BY TF AND THE POOR UNDERSTANDING OF MECHANISMS IN HUMAN T-CELL DIFFERENTIATION HAVE BEEN HURDLES TO THE DEVELOPMENT OF APPROACHES TO ENHANCE T-CELL DIFFERENTIATION AND FUNCTION. THE TUMOR SUPPRESSOR TF BCL11B IS REQUIRED FOR THE REPRESSION OF ALTERNATIVE (NON-T) LINEAGE POTENTIALS BUT DOES NOT PLAY A ROLE IN THE INDUCTION OF T-LINEAGE GENE EXPRESSION DURING THE INITIAL STAGES OF T-CELL DIFFERENTIATION OF MURINE HSPC. IN CONTRAST, WE SHOWED THAT BCL11B IS CRITICAL FOR BOTH THE INDUCTION OF THE T- LINEAGE PROGRAM AND REPRESSION OF ALTERNATIVE LINEAGE PROGRAMS DURING THE INITIAL STAGES OF HUMAN T-CELL DIFFERENTIATION. WE NOW HAVE NOVEL PRELIMINARY IN VITRO DATA THAT LENTIVIRAL BCL11B OVEREXPRESSION: 1) EXPEDITES T-CELL DIFFERENTIATION FROM HUMAN HSPC INCLUDING THE GENERATION OF MATURE T-CELLS, AND 2) ENHANCES THE FUNCTION, PROMOTES DIFFERENTIATION INTO CELLS WITH A CENTRAL MEMORY PHENOTYPE, AND DELAYS EXHAUSTION OF HUMAN T-CELLS. INTEGRATED ANALYSIS OF FUNCTIONAL, CHIP-SEQ, AND SINGLE CELL RNA-SEQ DATA REVEALED NOTCH3 AND IRF8 AS SPECIES SPECIFIC CANDIDATE TARGETS OF BCL11B IN HUMANS. OF NOTE, BCL11B OVEREXPRESSION STUDIES HAVE NOT BEEN POSSIBLE IN MURINE HSPC DUE TO TOXICITY. BASED ON THESE DATA, WE HYPOTHESIZE THAT TRANSPLANTATION OF HSPC ENGINEERED TO OVEREXPRESS BCL11B WILL ENHANCE POST-HSCT T-CELL RECONSTITUTION. BCL11B OVEREXPRESSION WILL INCREASE THE EFFICACY OF CAR T-CELLS BY ENHANCING THEIR FUNCTION AND PERSISTENCE AND AMELIORATING EXHAUSTION. WE WILL TEST THE HYPOTHESIS THROUGH THE FOLLOWING AIMS: 1.1) DETERMINE THE EPIGENETIC EFFECTS OF BCL11B ON T- CELL GENES AND THE ROLE OF BCL11B MEDIATED REGULATION OF NOTCH3 (1.2) AND IRF8 (1.3) IN HUMAN T-CELL DIFFERENTIATION. 1.4) DEFINE THE EFFICACY OF BCL11B OVEREXPRESSING HUMAN HSPC FOR THE ENHANCEMENT OF POST- HSCT T-CELL RECONSTITUTION IN HUMANIZED MOUSE MODELS, AND 2) DEFINE THE EFFECTS OF BCL11B OVEREXPRESSION ON ANTI-CANCER EFFICACY, PERSISTENCE, AND EXHAUSTION OF HUMAN CAR T-CELLS IN LEUKEMIA AND NEUROBLASTOMA MODELS. THESE STUDIES COULD REVEAL NEW FUNCTIONS OF BCL11B AND LEAD TO BCL11B ENGINEERED CELL THERAPIES THAT IMPROVE OUTCOMES IN LEUKEMIA AND SOLID TUMORS. THIS PROPOSAL IS INNOVATIVE BECAUSE IT BUILDS ON OUR WORK DEFINING SPECIES SPECIFIC EFFECTS OF BCL11B IN HUMANS TO ADDRESS KEY BARRIERS IN HSCT AND CAR T-CELL THERAPY.

IN VIVO ANALYSIS OF RORGAMMA MEDIATED FUNCTIONS

UV DAMAGE, REPAIR, AND MUTAGENESIS

EXPLOITING CAVEOLAE-DEPENDENT ALBUMIN ENDOCYTOSIS TO OPTIMIZE THERAPY IN PANCREATIC CANCER

COMPUTATIONALLY GUIDED DESIGN OF THERMOSTABLE MUTANTS OF NEUROTENSIN RECEPTOR1

CELL-SELECTIVE CPG-STAT3 INHIBITORS FOR RADIOIMMUNOTHERAPY OF MALIGNANT GLIOMA

EARLY LIFE EXPOSURE TO ENDOCRINE DISRUPTING CHEMICALS: EPIGENOMIC AND TRANSCRIPTOMIC NETWORKS AND RAGE IN LIVER DISEASE - PROJECT SUMMARY BISPHENOL S (BPS), BISPHENOL F (BPF), AND DIISONONYLPHTHALATE (DINP) ARE INCREASINGLY REPLACING THE ENDOCRINE DISRUPTING CHEMICALS BISPHENOL A (BPA) AND DI-2-ETHYLHEXYLPHTHALATE (DEHP), RESPECTIVELY. HOWEVER, BPS, BPF, AND DINP HAVE NOT UNDERGONE SAFETY TESTING. PERINATAL EXPOSURES (IN UTERO AND DURING LACTATION) TO BPA AND DEHP ARE ASSOCIATED WITH THE DEVELOPMENT OF NON-ALCOHOLIC FATTY LIVER DISEASE (NAFLD). NAFLD, A COMMON DISEASE IN CHILDREN, IS BEING DIAGNOSED AT INCREASINGLY YOUNGER AGES AND ITS PREVALENCE IS INCREASING IN LATE ADOLESCENCE. WHILE BPA AND DEHP HAVE BEEN EXTENSIVELY STUDIED IN THIS CONTEXT, BPS/BPF AND DINP HAVE NOT. BPS EXPOSURE PROMOTED NAFLD PROGRESSION IN ZEBRAFISH, AND BPF SERUM LEVELS ARE HIGHER IN NAFLD PATIENTS THAN IN CONTROLS. BPS AND BPF LEVELS WERE ASSOCIATED WITH INCREASED PREVALENCE OF OBESITY IN CHILDREN, A RISK FACTOR FOR NAFLD. DINP CAUSED LIPIDOMIC DISRUPTION IN NEONATAL MICE, WHILE PHTHALATE EXPOSURE CAUSED HEPATIC STEATOSIS IN ADULT MICE. THUS, LIKE BPA AND DEHP, EARLY-LIFE BPS/BPF AND DINP EXPOSURE MAY PROMOTE NAFLD DEVELOPMENT/PROGRESSION. TO DATE, THE MECHANISMS BY WHICH BISPHENOLS AND PHTHALATES, PARTICULARLY AS A MIXTURE, TRIGGER THESE LONG-TERM METABOLIC CONSEQUENCES ARE ELUSIVE. HERE, WE WILL TEST THE HYPOTHESIS THAT EARLY LIFE EXPOSURE TO A BISPHENOL/PHTHALATE MIXTURE ACTIVATES RAGE SIGNALING PATHWAYS, TRIGGERS A TRAINED IMMUNE RESPONSE IN PERIPHERAL MONOCYTES, AND REPROGRAMS THE EPIGENOME/TRANSCRIPTOME IN LIVER AND IN PERIPHERAL MONOCYTES, THEREBY PRIMING OFFSPRING FOR EXAGGERATED METABOLIC DYSFUNCTION UPON EXPOSURE TO WESTERN DIET. THIS VICTER CONSORTIUM WILL ADDRESS THIS NOVEL HYPOTHESIS THROUGH INTEGRATED STUDIES IN MICE AND IN HUMAN SUBJECTS.

ROLE OF STAT3 IN TUMOR IMMUNE EVASION AND IMMUNE SUPPRESSION

NOVEL RNA-DIRECTED THERAPY FOR THE TREATMENT OF ACUTE MYELOID LEUKEMIA

AGE-RELATED SHIFTS IN EPITHELIAL LINEAGES AND TISSUE HOMEOSTASIS IN MAMMARY GLAND

THE ROLE AND MECHANISM OF METTL3/METTL14-MEDIATED RNA MODIFICATION IN THE PATHOGENESIS AND DRUG-RESISTANCE OF AML

TARGETING TET1 SIGNALING TO TREAT ACUTE MYELOID LEUKEMIA

CITY OF HOPE SOUTHWEST ONCOLOGY GROUP CLINICAL TRIALS

ROLE OF TREFOIL FACTOR FAMILY PROTEINS IN BETA CELL FUNCTION. - MODIFIED PROJECT SUMMARY/ABSTRACT SECTION IN THE UNITED STATES, NEARLY 10% OF THE POPULATION (MORE THAN 30 MILLION) HAS TYPE 2 DIABETES (T2D) AND THIS NUMBER IS GROWING NOT ONLY IN OLDER ADULTS BUT ALSO IN CHILDREN, TEENS, AND YOUNG ADULTS. THE ALARMING AND CONTINUED INCREASE OF T2D INCIDENCE, ESPECIALLY AMONG YOUNG AMERICANS, CREATES AN URGENT NEED FOR NEW THERAPEUTICS AND INTERVENTIONS. ONE OF THE UNDERLYING CONDITIONS LEADING TO T2D IS THE DYSFUNCTION OF BETA CELLS. WHILE DISCOVERING THE MOLECULAR CHANGES IN BETA CELLS LEADING TO THEIR DYSFUNCTION HAS BEEN A FOCUSED EFFORT IN THE FIELD, LESS IS KNOWN ABOUT HOW THE MICROENVIRONMENT, SUCH AS SECRETED FACTORS FROM EXOCRINE PANCREAS, MAY ALSO CONTRIBUTE TO PATHOGENESIS. TREFOIL FACTOR FAMILY PEPTIDES (TFF1, TFF2, AND TFF3) ARE UNIQUE SECRETED PROTEINS IN THAT THEY CONTAIN SIX CYSTEINE RESIDUES THAT FORM THREE INTRAMOLECULAR DISULFIDE BRIDGES, WHICH MAKES THEM MORE STABLE AND RESISTANT TO PROTEASES AND OTHER HARSH CONDITIONS. TREFOIL FACTORS HAVE BEEN SHOWN TO PLAY ESSENTIAL ROLES IN MAINTAINING THE INTEGRITY OF GUT AND LUNG EPITHELIUM, AND IN REPAIRING EPITHELIAL CELLS AFTER INJURY. TREFOIL FACTORS ARE EXPRESSED IN THE PANCREAS, BUT THEIR ROLES ARE NOT WELL UNDERSTOOD. WE HAVE PRELIMINARY DATA REVEALING THAT TFF2 IS EXPRESSED BY THE EXOCRINE PANCREAS AND THE KNOCKOUT OF THIS GENE IN THE PANCREAS LEADS TO BETA CELL DYSFUNCTION IN ADULT MICE. HOWEVER, THE MOLECULAR AND CELLULAR MECHANISMS BY WHICH TFF2 EXERTS ON BETA CELLS ARE UNKNOWN. OUR CENTRAL HYPOTHESIS IS THAT TFF2 PROTECTS AGAINST THE DAMAGING EFFECTS OF DIABETOGENIC STRESS. WE WILL USE OUR PANCREAS-SPECIFIC KNOCKOUT MOUSE MODEL OF TFF2 AND PRIMARY HUMAN TISSUES/CELLS FOR OUR STUDIES. IN AIM 1, WE WILL EXAMINE HOW THE LACK OF TFF2 IN THE MURINE PANCREAS AFFECTS BETA CELL DYSFUNCTION IN AGED MICE AND UNDER DIABETOGENIC CONDITIONS. IN AIM 2, WE WILL CLARIFY THE DISTRIBUTION PATTERNS OF TREFOIL FACTOR FAMILY PROTEINS AND THEIR RECEPTORS IN HUMAN PANCREATIC TISSUES AND STUDY THE CROSSTALK BETWEEN EXOCRINE AND ISLET CELLS VIA TFF2 BY USING IN VITRO MODEL SYSTEMS. WE WILL USE INNOVATIVE TOOLS TO TEST NOVEL HYPOTHESES ON THE ACTIONS ABOUT TREFOIL FACTORS WITHIN A TRANSLATION-FOCUSED INSTITUTIONAL ENVIRONMENT AT CITY OF HOPE. THIS WORK WILL POSITIVELY IMPACT DIABETES RESEARCH BY ADDRESSING A PROMISING CANDIDATE SECRETORY FACTOR FOR DIABETES PREVENTION/TREATMENT.

MODELING ALZHEIMER?S DISEASE USING DIRECTLY REPROGRAMMED ISOGENIC NEURAL CELLS

A PHASE I STUDY OF INTRACRANIALLY ADMINISTERED CARBOXYLESTERASE-EXPRESSING NEURAL STEM CELLS IN COMBINATION WITH INTRAVENOUS IRINOTECAN IN PATIENTS W

OVERCOMING IBRUTINIB RESISTANCE IN MANTLE CELL LYMPHOMA - PROJECT ABSTRACT IBRUTINIB, A COVALENT INHIBITOR OF BRUTON TYROSINE KINASE (BTK) AND SIMILAR INHIBITORS OF B-CELL RECEPTOR (BCR)- ASSOCIATED KINASES ARE SHIFTING THE THERAPEUTIC LANDSCAPE IN B-CELL NEOPLASIA. HOWEVER, THE DEVELOPMENT OF RESISTANCE IN PATIENTS TREATED WITH THESE AGENTS IS ASSOCIATED WITH RAPIDLY PROGRESSIVE DISEASE AND DEATH. MUTATIONS IN BTK AND ITS DOWNSTREAM TARGET PHOSPHOLIPASE C2 HAVE BEEN IMPLICATED IN RESISTANCE TO IBRUTINIB IN CHRONIC LYMPHOCYTIC LEUKEMIA (CLL). WHILE SUCH MUTATIONS ARE FOUND IN A SUBSET OF MANTLE CELL LYMPHOMA (MCL) PATIENTS, THEY DO NOT ACCOUNT FOR ALL PATTERNS OF IBRUTINIB RESISTANCE. RECENT STUDIES HAVE IMPLICATED NUCLEAR FACTOR-B (NFB) AS A POTENTIAL CRITICAL COMPONENT OF RESISTANCE IN MCL. THE PATHOGENIC CONTRIBUTION OF NFB IS SUPPORTED BY IDENTIFICATION OF A) ACTIVATION OF NFB SIGNALING PATHWAYS WITHIN THE LYMPH NODE MICROENVIRONMENT; B) CONSTITUTIVE NFB ACTIVITY RESULTING FROM RECURRENT MUTATIONS IN TRAF2, BIRC3 AND RELA; AND C) MUTATIONS IN GENES AFFECTING NFB SIGNALING IN PRIMARY IBRUTINIB-REFRACTORY MCL SAMPLES. THUS, TUMOR-INTRINSIC AND MICROENVIRONMENT-DRIVEN NFB ACTIVATION MAY UNDERLIE A SIGNIFICANT FRACTION OF IBRUTINIB FAILURES. HOWEVER, THE ROLE OF NFB HAS NOT BEEN STUDIED PROSPECTIVELY IN THIS SETTING. WE HYPOTHESIZE THAT NFB AND ITS TRANSCRIPTIONAL TARGETS ARE UPREGULATED IN PATIENTS WHO EXHIBIT RESISTANCE TO IBRUTINIB. UNDER AIM 1, WE WILL CONDUCT A PROSPECTIVE STUDY TO DETERMINE FUNCTIONAL SIGNIFICANCE OF NFB ACTIVATION IN IBRUTINIB RESISTANCE IN MCL. WE WILL COMPREHENSIVELY EVALUATE NFB SIGNALING IN PRIMARY MCL CELLS OBTAINED FROM BONE MARROW AND/OR LYMPH NODES PRIOR TO THE START OF IBRUTINIB, ON THERAPY AND AT RELAPSE. GIVEN THE EMERGING IMPORTANCE OF NFB IN MCL, AN OPTIMAL THERAPEUTIC APPROACH WOULD INCORPORATE TARGETING NFB ACTIVATION. IN EVALUATING THE NFB PATHWAY AS A THERAPEUTIC TARGET, WE DISCOVERED THAT IT CAN BE NEUTRALIZED VIA BLOCKADE OF UPSTREAM REGULATORS IN THE UBIQUITIN-PROTEASOME SYSTEM (UPS). OUR DATA INDICATE THAT NFB ACTIVITY MAY BE BLOCKED BY TARGETING NEDD8-ACTIVATING ENZYME (NAE; A UBIQUITIN-LIKE MODIFIER) IN NEOPLASTIC B-CELLS, RESULTING IN CELL APOPTOSIS. WE HYPOTHESIZE THAT THERAPEUTIC DISRUPTION OF NFB BY BLOCKING THE NAE WILL OVERCOME IBRUTINIB RESISTANCE IN MCL. THUS, AIM 2 WILL EVALUATE THERAPEUTIC TARGETING OF THE E1 LIGASE, NAE, IN IBRUTINIB- RESISTANT MCL. WE WILL ELUCIDATE FUNCTIONAL EFFECTS OF INHIBITING NAE ON NFB SIGNALING, CELL APOPTOSIS IN MCL CELL LINES AND PRIMARY IBRUTINIB-RESISTANT CELLS IN VITRO AND IN A MURINE MCL XENOGRAFT MODEL. FINALLY, AIM 3 WILL ASSESS THE NAE INHIBITION-MEDIATED DEREGULATION OF NFB IN NEOPLASTIC B CELLS AND T CELLS OBTAINED FROM PATIENTS WHO RECEIVE TREATMENT WITH PEVONEDISTAT AND IBRUTINIB ON A CLINICAL TRIAL. OUR STUDY WILL TEST NOVEL APPROACHES TO OVERCOME IBRUTINIB RESISTANCE THROUGH NFB INACTIVATION, AND THUS SIGNIFICANTLY IMPACT MCL THERAPEUTICS.

INTEGRATIVE ANALYSES OF SATURATION CRISPR PROTEIN SCAN

THE FUNCTION AND UNDERLYING MECHANISM OF TET1 IN MYELODYSPLASTIC SYNDROMES

EPIGENETIC MARKERS OF COMPLICATIONS AND METABOLIC MEMORY IN THE DCCT/EDIC COHORT.

THE ROLE OF MIR-142 IN REGULATORY T CELL DEVELOPMENT AND FUNCTION

IL-15 CHARACTERIZATION THROUGH EXPERIMENTAL IMMUNOLOGY

DEVELOP AGE-RELEVANT GLIAL CELLULAR MODELS USING HUMAN DIRECTLY REPROGRAMMED CELLS - PROJECT SUMMARY BRAIN AGING IS CHARACTERIZED BY REDUCED COGNITIVE CAPACITIES, LEARNING AND MEMORY. THE EXACT MECHANISMS OF BRAIN AGING REMAIN ELUSIVE. BECAUSE AGING IS THE GREATEST RISK FACTOR FOR MAJOR DEBILITATING NEURODEGENERATIVE DISORDERS, INCLUDING ALZHEIMER'S DISEASE (AD), IT IS IMPORTANT TO UNCOVER MECHANISMS UNDERLYING BRAIN AGING IN ORDER TO DEVELOP EFFECTIVE THERAPIES FOR AGE-RELATED NEURODEGENERATIVE DISEASES. AD IS THE MOST COMMON NEURODEGENERATIVE DISORDER AND A LEADING CAUSE OF DISABILITY AND DEATH. HOWEVER, THE PRECISE MECHANISMS UNDERLYING AD PATHOGENESIS REMAINS TO BE ELUCIDATED. ALTHOUGH MANY TRANSGENIC MOUSE MODELS HAVE BEEN GENERATED FOR AD RESEARCH AND THESE MODELS ARE IMPORTANT FOR OUR UNDERSTANDING OF THE PATHOLOGICAL BASIS OF THE DISEASE, IT IS INCREASINGLY RECOGNIZED THAT THERE ARE SIGNIFICANT SPECIES DIFFERENCES BETWEEN MOUSE AND HUMAN NEURAL CELLS. THEREFORE, THERE IS AN URGENT NEED TO ESTABLISH HUMAN DISEASE MODELING PLATFORMS TO COMPLEMENT STUDIES IN ANIMAL MODELS FOR AD RESEARCH. DIRECT REPROGRAMMING IS A CELLULAR REPROGRAMMING TECHNOLOGY, WHICH ALLOWS DIRECT CONVERSION OF ONE TYPE OF SOMATIC CELLS, SUCH AS FIBROBLASTS, INTO ANOTHER TYPE OF SOMATIC CELLS, SUCH AS NEURONS. IT HAS BEEN SHOWN THAT DIRECT REPROGRAMMING ENABLES GENERATION OF HUMAN NEURONS THAT POSSESS KEY ELEMENTS OF CELLULAR AGING. THEREFORE, DIRECTLY REPROGRAMMED CELLS COULD PROVIDE A HUMAN CELLULAR PLATFORM FOR US TO MODEL BRAIN AGING AND AGE-RELATED LATE-ONSET DISEASES, SUCH AS LATE-ONSET AD (LOAD). THE OBJECTIVE OF THIS PROPOSAL IS TO DEVELOP HUMAN AGE-RELEVANT GLIAL CELLULAR MODELS USING DIRECT REPROGRAMMING TECHNOLOGY, IN ORDER TO RECAPITULATE AGE-ASSOCIATED PHENOTYPES IN BRAIN AGING AND NEURODEGENERATION AND UNCOVER NOVEL UNDERLYING MECHANISMS. INCREASING EVIDENCE SUGGESTS THAT ASTROCYTES PLAY IMPORTANT ROLES IN BRAIN HEALTH AND PATHOGENESIS OF NEURODEGENERATIVE DISEASES. THEREFORE, WE PROPOSE TO ESTABLISH CELLULAR MODELS FOR BRAIN AGING AND AD USING ASTROCYTES DIRECTLY REPROGRAMMED FROM FIBROBLASTS OF AGED SUBJECTS AND LOAD PATIENTS AND CO-CULTURES OF ASTROCYTES WITH OTHER BRAIN CELL TYPES, INCLUDING MICROGLIA, OLIGODENDROCYTES, AND NEURONS. WE HYPOTHESIZE THAT CELLULAR AGING REGULATES ASTROCYTE FUNCTION AND CELL-CELL INTERACTIONS TO MODULATE BRAIN AGING PHENOTYPES AND LOAD PATHOLOGIES. ACCORDINGLY, WE PROPOSE THE FOLLOWING SPECIFIC AIMS:AIM 1: TO GENERATE AGE-ASSOCIATED ASTROCYTES THROUGH DIRECT REPROGRAMMING AND EVALUATE HOW CELLULAR AGING REGULATES ASTROCYTIC FUNCTIONS. AIM 2: TO DETERMINE WHETHER AND HOW ASTROCYTIC CELLULAR AGING MODULATES NEUROINFLAMMATION AND NEURONAL PHENOTYPES. AIM 3: TO DETERMINE WHETHER AND HOW ASTROCYTIC CELLULAR AGING REGULATES OPC PROPERTIES AND MYELINATION. THE PROPOSED STUDIES WILL LIKELY HELP TO DEFINE ROLES OF GLIAL CELLULAR AGING IN BRAIN FUNCTIONAL DETERIORATION DURING AGING AND UNCOVER THE UNDERLYING MECHANISMS, WHICH COULD LEAD TO THE DEVELOPMENT OF NOVEL STRATEGIES TO MAINTAIN BRAIN HEALTH AND REDUCE RISK FOR AD. THE KNOWLEDGE GAINED FROM THIS STUDY COULD HELP US TO DESIGN NOVEL THERAPEUTIC STRATEGIES FOR AD.

EXOSOMAL BIOMARKERS FOR THE EARLY DETECTION OF HEPATOCELLULAR CARCINOMA - PROJECT SUMMARY: LIVER CANCER IS ESTIMATED TO AFFLICT 42,230 INDIVIDUALS AND RESULT IN APPROXIMATELY 30,230 DEATHS IN THE US IN 2021. THE INCIDENCE RATES FOR LIVER CANCER HAVE MORE THAN TRIPLED SINCE 1980, MAKING IT THE 5TH LEADING CAUSE OF CANCER- RELATED DEATHS IN THE US. APPROXIMATELY THREE-FOURTHS OF LIVER CANCER CASES ARE HEPATOCELLULAR CARCINOMA (HCC). AS FOR ALL CANCERS, DETECTION OF HCC AT AN EARLIER STAGE IS CRITICAL TO ELICIT THE BEST CHANCE OF A CURE. EARLY DETECTION OF HCC HAS SIGNIFICANT POTENTIAL TO REDUCE MORTALITY RATES, DUE TO THE SIGNIFICANT EFFICACY OF LOCAL TREATMENTS FOR EARLY-STAGE DISEASE VS. SYSTEMIC THERAPY FOR ADVANCED-STAGE CANCERS. ALTHOUGH SURVEILLANCE OF PATIENTS AT HIGH- RISK FOR HCC (E.G., THOSE WITH CHRONIC HEPATITIS OR LIVER CIRRHOSIS [LC]) IS WIDELY PERFORMED, THE POPULATION OF PATIENTS WITH HCC WITHOUT VIRAL ETIOLOGIES IS INCREASING BECAUSE OF INSUFFICIENT SCREENING FOR HCC. AT PRESENT, ALPHA-FETOPROTEIN (AFP) IS THE MOST WIDELY USED BLOOD TUMOR MARKER; AND HEPATIC ULTRASOUND IS A LOW-COST IMAGING METHOD FOR SURVEILLANCE OF HCCS. HOWEVER, BOTH APPROACHES HAVE LIMITED SENSITIVITY AND SPECIFICITY FOR DETECTING EARLY-STAGE HCC, HIGHLIGHTING THE IMPERATIVE NEED TO DEVELOP ROBUST BIOMARKERS FOR THE EARLY DETECTION OF HCC. ACCUMULATING EVIDENCE INDICATES THAT DYSREGULATION OF MICRORNAS (MIRNAS) OCCURS IN ALL HUMAN CANCERS, INCLUDING HCC. AS BIOMARKERS, MIRNAS ARE MORE RESILIENT THAN MRNAS, AND ARE FREQUENTLY DEREGULATED EVEN IN THE EARLIEST STAGES OF NEOPLASIA. FURTHERMORE, THE RECENT DISCOVERY THAT CANCERS ACTIVELY EXCRETE SMALL EXTRACELLULAR VESICLES, CALLED EXOSOMES, HAS BROUGHT ADDITIONAL ENTHUSIASM TO THIS BURGEONING TRANSLATIONAL RESEARCH TOPIC. WHILE EXOSOMES ARE CONSIDERED TO REFLECT THEIR RESPECTIVE CELLS-OF-ORIGIN, THEIR USE IN BIOMARKER RESEARCH HAS BEEN HAMPERED DUE TO LACK OF STANDARDIZED PROTOCOLS FOR THEIR ISOLATION AND PURIFICATION, USE OF CELL LINE-DERIVED, BUT NOT PATIENT-DERIVED SPECIMENS FOR BIOMARKER DISCOVERY; AND LACK OF BIOMARKER DISCOVERY IN CANCER-DERIVED EXOSOMES FROM MATCHED TISSUES AND PLASMA SPECIMENS. FURTHERMORE, DESPITE THE PERCEPTION THAT EXOSOMAL-MIRNAS (EXO-MIRNAS) MAY BE SUPERIOR TO CIRCULATING CELL-FREE MIRNAS (CF-MIRNAS), NO STUDIES HAVE UNDERTAKEN AN EFFORT TO DIRECTLY COMPARE THESE TWO TYPES, TO SUPPORT OR NEGATE THEIR SUPERIORITY AS DISEASE BIOMARKERS. IN THIS PROPOSAL, WE WILL ADDRESS THESE CONCERNS BY UNDERTAKING THE FOLLOWING SPECIFIC AIMS. AIM 1: DISCOVER CANDIDATE CF-MIRNA AND EXO-MIRNA BIOMARKERS USING SMALL RNA-SEQ IN MATCHED TISSUE AND PLASMA FROM PATIENTS WITH EARLY-STAGE HCC VS. CONTROLS. AIM 2: DEVELOP A BIOMARKER PANEL COMPOSED OF CF-MIRNAS AND EXO-MIRNAS FOR THE IDENTIFICATION OF PATIENTS WITH HCC IN AN INDEPENDENT COHORT. AIM 3: CLINICALLY VALIDATE THE OPTIMIZED PANEL OF NON-INVASIVE PLASMA MIRNA BIOMARKERS IN A LARGE PROSPECTIVE COHORT OF PATIENTS WITH HCC. IF SUCCESSFUL, THIS PROPOSAL WILL PROVIDE MOLECULAR CHARACTERIZATION OF CELL-FREE AND EXOSOMAL MIRNAS AS LIQUID BIOPSY BIOMARKERS, WHICH MAY ALLOW EARLY-DETECTION OF HCC USING A NON-INVASIVE, AND INEXPENSIVE ASSAY.

OKAZAKI FRAGMENT MATURATION: MUTAGENESIS AND CELL SURVIVAL - ABSTRACT THE LONG-TERM GOAL OF THIS PROJECT IS TO DEFINE THE MOLECULAR MECHANISMS OF AN ERROR-PRONE, STRESS-INDUCED OKAZAKI FRAGMENT MATURATION (OFM) PATHWAY BY WHICH CANCER CELLS COUNTERACT REPLICATION STRESS AND SURVIVE. REPLICATION STRESS IS A HALLMARK OF CANCER CELLS AND HAS BEEN CONSIDERED THE ACHILLES' HEEL FOR CANCER TREATMENT SUCH AS RADIO- AND CHEMOTHERAPY. UNDER ELEVATED TEMPERATURE STRESS, YEAST CELLS MUTANT FOR FLAP ENDONUCLEASE 1 (FEN1 IN HUMANS OR RAD27 IN YEAST) ACTIVATE DNA DAMAGE RESPONSE PATHWAYS TO BLOCK CELL PROLIFERATION AND INDUCE CELL SENESCENCE AND DEATH; HOWEVER, A SUBPOPULATION OF CELLS CAN OVERCOME THESE BARRIERS AND ESCAPE OTHERWISE LETHAL CONDITIONS. GENOME-WIDE MUTATIONS AND REARRANGEMENTS HAVE BEEN SUGGESTED AS A MAJOR MOLECULAR MECHANISM THAT DRIVES THIS EVOLUTION. HOWEVER, HOW SUCH SPONTANEOUS MUTATIONS ARE ACQUIRED IN CELLS UNDER REPLICATION STRESS IS A LONG-STANDING QUESTION. RECENTLY, WE IDENTIFIED AN ERROR-PRONE, 3' FLAP OFM PATHWAY THAT IS ACTIVATED IN RESPONSE TO STRESS TO SUPPORT CELL SURVIVAL AND FUEL CELLULAR EVOLUTION; ITS INDUCTION LEADS TO GENOME-WIDE MUTAGENESIS AND SUPPRESSION OF RESTRICTIVE GROWTH TEMPERATURE-INDUCED LETHALITY, A PROCESS MIMICKING THAT OF CANCER CELLS ACQUIRING DRUG RESISTANCE. THIS LED US TO A MODEL IN WHICH OFM CAN GO IN TWO WAYS, WHICH MAY DICTATE THE FATE OF CELLS, INCLUDING HUMAN CANCER CELLS: A 5' FLAP-BASED, ERROR-FREE PROCESS OR AN ALTERNATIVE 3' FLAP-BASED, STRESS-INDUCED, AND ERROR-PRONE PROCESS. HOWEVER, KEY COMPONENTS THAT DRIVE SUCH FLAP DYNAMICS REMAIN UNDEFINED. THE OBJECTIVES OF THE PROPOSED PROJECT ARE TO DEFINE THE KEY ENZYMES THAT CATALYZE 3' FLAP FORMATION AND CLEAVAGE IN MAMMALIAN CELLS AND TO PROVIDE PROOF OF CONCEPT THAT SUPPRESSING ALTERNATIVE 3' FLAP OFM CAN PREVENT DRUG RESISTANCE IN HUMAN CANCER CELLS. FURTHER PRELIMINARY DATA GATHERED TO SUPPORT THIS GRANT APPLICATION SHOW THAT 3' FLAP OFM IS CONSERVED IN BOTH YEAST AND HUMAN CELLS. WE OBSERVED THAT ANTI-CANCER EGFR TYROSINE KINASE INHIBITORS ACTIVATED THE ATM/CHK2 DNA DAMAGE CHECKPOINTS IN HUMAN LUNG CANCER CELLS. USING YEAST GENETIC SCREENING, WE IDENTIFIED PIF1 (PIF1 IN HUMANS) AND SGS1 (BLM AND WRN IN HUMANS) AS HELICASES FOR 5' TO 3' FLAP TRANSFORMATION AND RAD1 (XPF IN HUMANS) AND MUS81 (MUS81 IN HUMANS) AS 3' NUCLEASES FOR 3' FLAP CLEAVAGE, IN ADDITION TO THE 3' NUCLEASE ACTIVITY OF POL . THEREFORE, OUR CENTRAL HYPOTHESIS IS THAT UNPROCESSED 5' FLAPS IN MAMMALIAN CELLS ACTIVATE ATM/ATR AND CHK1/2 SIGNALING TO RECRUIT AND STIMULATE PIF1, BLM, AND WRN AND/OR OTHER HELICASES FOR TRANSFORMING 5' FLAPS INTO 3' FLAPS FOR NUCLEOLYTIC DEGRADATION BY 3' NUCLEASES INCLUDING POL , XPF, AND/OR MUS81, AND THAT BLOCKING THE 3' FLAP OFM PATHWAY WILL SUPPRESS DNA MUTATIONS AND THUS PREVENT DRUG RESISTANCE. TO TEST THIS, WE WILL: I) DETERMINE THE ROLES OF HELICASES PIF1, BLM, AND WRN IN 3' FLAP FORMATION AND INDUCTION OF ALTERNATIVE OFM; II) DEFINE THE FUNCTIONAL DISTRIBUTION OF 3' NUCLEASES POL , XPF, AND MUS81 IN PROCESSING 3' FLAPS WITH OR WITHOUT SECONDARY STRUCTURES DURING 3' FLAP OFM; AND III) DEFINE THE EXTENT TO WHICH STRESS-ACTIVATED ATM/CHK2 SIGNALING INDUCES 3' FLAP OFM AND MUTATIONS TO SUPPORT CANCER CELL SURVIVAL AND PROMOTE DRUG RESISTANCE.

TARGETING PROTEIN ACETYLATION AS A THERAPEUTIC APPROACH FOR MDS

EPIGENETIC DYSREGULATION OF TRANSPOSONS IN OBESITY

THE ROLE OF ADIPOCYTE URIDINE BIOSYNTHESIS IN OBESITY AND DIABETES PROGRESSION

TARGETING PROTEIN ARGININE METHYLTRANSFERASES TO ERADICATE ACUTE MYELOID LEUKEMIA - PROJECT SUMMARY TREATMENT OUTCOMES FOR PATIENTS WITH ACUTE MYELOID LEUKEMIA (AML) HAVE CONTINUED TO LAG BEHIND OUTCOMES REPORTED FOR OTHER HEMATOLOGICAL MALIGNANCIES LIKE ACUTE LYMPHOBLASTIC LEUKEMIA (ALL), IN PART BECAUSE OF THE RELATIVELY SLOW DEVELOPMENT OF IMMUNOTHERAPY (INCLUDING IMMUNE CHECKPOINT INHIBITORS [ICI]) COMPARED WITH ALL. HOWEVER, TRANSLATION OF CURRENTLY AVAILABLE IMMUNOTHERAPIES TO AML TREATMENT HAS BEEN CHALLENGING, AND NOVEL AML-TARGETING DRUGS WITH IMMUNE-STIMULATING ACTIVITY ARE GREATLY NEEDED. THE CGAS-STING SIGNALING IS A MAJOR PATHWAY THAT PROMOTES AN EXTRINSIC TYPE I INTERFERON (IFN-I) RESPONSE, WHICH POTENTLY PRIMES T CELLS IN “IMMUNE- COLD” CANCERS, INCLUDING AML. MY LABORATORY HAS BEEN ACTIVE IN DEFINING THE FUNCTION OF PROTEIN ARGININE METHYLTRANSFERASES (PRMTS) IN LEUKEMIA. HEREIN, OUR PRELIMINARY STUDIES ON THE BASIS OF PRIMARY AML CELLS AS WELL AS MURINE AML MODELS REVEAL AN ANTI-AML IMMUNE ACTIVATION SEEN AFTER INHIBITION OF PRMT9, A MOST RECENTLY DEFINED PRMT. SPECIFICALLY, PRMT9 INHIBITION STIMULATED LEUKEMIA-INTRINSIC CGAS, AS EVIDENCED BY CGAMP (AN IMMUNOTRANSMITTER) PRODUCTION, AND INDUCED A LEUKEMIA-ELIMINATING IFN-I RESPONSE IN MURINE AML- MICROENVIRONMENT (ME). WHEN COMBINED WITH AN ICI (PD1 INHIBITOR), THE IN-HOUSE PRMT9 INHIBITOR LD2 ERADICATED AML IN ANIMAL MODELS. THUS, WE HYPOTHESIZE THAT PRMT9 ACTIVITY ALLOWS AML CELLS TO EVADE IMMUNE SURVEILLANCE BY REPRESSING CGAS-STING ACTIVITY IN THE ME, AND THAT LD2, ALONE OR COMBINED WITH AN ICI (PD1 INHIBITOR), ELICITS T CELL ACTIVITY TO ELIMINATE AML CELLS. EXTRACELLULAR CGAMP, WHICH IS HYDROLYZED BY ENPP1, AND STING ACTIVATION IN HOST IMMUNE CELLS, ARE REPORTEDLY ESSENTIAL FOR IMMUNITY. THUS, IN AIM 1, USING TWO AML TRANSPLANT MODELS, WE WILL DETERMINE WHETHER DOWNREGULATING CGAMP (VIA EITHER ENPP1-OVEREXPRESSION OR CGAS-KNOCKOUT (KO) IN AML CELLS) OR STING-KO IN THE IMMUNE CELL COMPARTMENT OF RECIPIENT MICE WOULD REVERSE PRMT9 INHIBITION-INDUCED AML REGRESSION. IN AIM 2, WE WILL DETERMINE THE MECHANISMS UNDERLYING CANCER- INTRINSIC CGAS ACTIVATION SEEN AFTER PRMT9 INHIBITION. SPECIFICALLY, WE WILL DEFINE PRMT9 TARGETS THAT WHEN UNMETHYLATED UNDERLIE CGAS ACTIVATION. IN AIM 3, WE WILL ASSESS ANTI-AML ACTIVITY OF LD2 ALONE OR COMBINED WITH A PD1 INHIBITOR, USING AML MOUSE MODELS. MOREOVER, TO DEFINE POTENTIAL SHIFTS IN IMMUNE CELL TYPES/STATES AFTER PRMT9 INHIBITION, WE WILL PERFORM SINGLE-CELL TRANSCRIPTOME ANALYSIS ON AML PDX CELLS FROM HUMANIZED MOUSE TREATED WITH LD2 OR THE COMBINATION. WE ARE THE FIRST TO IDENTIFY PRMT9 AS A DRUGGABLE IMMUNE ACTIVATION TARGET AGAINST CANCER. IF SUCCESSFUL, THIS WORK WOULD SUPPORT COMBINING A PRMT9 INHIBITOR WITH ICIS AS A THERAPY FOR AML, IN WHICH SINGLE ICI THERAPY HAS VERY LIMITED EFFECTS.

DEVELOPMENTAL THERAPEUTICS AND POTENTIAL TREATMENT TARGETING RALBP1 FOR CASTRATION-RESISTANT PROSTATECARCINOMA

REGULATION OF BETA-CELL HOMEOSTASIS BY DNA METHYLATION AND HYDROXYMETHYLATION.

INFLAMMATION AS DETERMINANT OF CLONAL SELECTION IN MPN PROGRESSION

THE ROLE AND MECHANISM OF FTO IN LEUKEMOGENESIS AND DRUG RESPONSE

ROLE OF PROTEIN ARGININE METHYLTRANSFERASE 9 IN ACUTE MYELOID LEUKEMIA MAINTENANCE

IDENTIFYING AND ENGAGING A UNIVERSAL ADJUVANT FOR BREAKING MACROPHAGE IMMUNE TOLERANCE IN CANCER - PROJECT SUMMARY/ABSTRACT THE ABILITY OF CANCER CELLS TO EVADE IMMUNOSURVEILLANCE IS ONE OF THE HALLMARKS OF CANCER. UNDERSTANDING THE MECHANISMS BY WHICH T, B AND NK CELLS DETECT AND ATTACK CANCER CELLS AND DEVELOPING APPROACHES TO BREAK TUMOR IMMUNE TOLERANCE HAVE REVOLUTIONIZED THE TREATMENT OF VARIOUS HUMAN CANCERS. MACROPHAGES ARE THE MAJOR COMPONENTS OF PRIMARY TUMORS AND METASTATIC SITES, AND EMERGING STUDIES HAVE DEMONSTRATED THAT THE ESCAPE FROM MACROPHAGE IMMUNOSURVEILLANCE IS A CRITICAL IMMUNE EVASION MECHANISM OF CANCER CELLS. RECENT PARADIGM-SHIFTING DISCOVERIES HAVE BEGUN TO REVEAL THAT INDUCING MACROPHAGE-MEDIATED CANCER CELL RECOGNITION AND PHAGOCYTOSIS, A PROCESS TERMED PROGRAMMED CELL REMOVAL (PRCR), IS CRUCIAL TO TUMOR SURVEILLANCE AND ELIMINATION. A “DON’T EAT ME” SIGNAL CD47 HAS BEEN IDENTIFIED TO BE UPREGULATED ON A VARIETY OF MALIGNANT HEMATOPOIETIC AND SOLID TUMOR CELLS, AND BLOCKADE OF CD47 ENABLES THE SUSCEPTIBILITY OF CANCER CELLS TO PRCR. RECENT PROGRESS IN CLINICAL TRIALS DEMONSTRATED SIGNIFICANT THERAPEUTIC POTENTIAL OF RESTORING PRCR AS A PROMISING CANCER IMMUNOTHERAPY. WHILE INDUCING PRCR HOLDS CONSIDERABLE PROMISE IN TREATING MULTIPLE TYPES OF CANCERS, ITS UNDERLYING MECHANISMS REMAIN UNCLEAR, WHICH HAVE HINDERED THE DEVELOPMENT OF PRCR-BASED THERAPY TO ACHIEVE ITS MAXIMAL EFFICACY. THE OVERALL OBJECTIVE OF THIS RESEARCH PROPOSAL IS TO DEVELOP STRATEGIES TO PROMOTE MACROPHAGE-MEDIATED CANCER SURVEILLANCE AND ELIMINATION BY BREAKING MACROPHAGE TOLERANCE, AND TO DETERMINE THE EFFICACY AND MECHANISMS OF SUCH STRATEGIES IN PRCR-BASED CANCER IMMUNOTHERAPY. THE PROPOSED STUDIES FOCUS ON IDENTIFYING AND ENGAGING ADJUVANTS TO AUGMENT THE ANTICANCER EFFICACY OF PRCR-BASED THERAPY, AND DISSECTING THE MOLECULAR MECHANISMS OF PRCR ACTIVATION AND MACROPHAGE-CANCER CELL INTERACTION. AIM1 IS TO EXAMINE THE ROLES OF PRCR ADJUVANTS IN ENHANCING THE EFFICACY OF CD47-BLOCKING ANTIBODIES, DETERMINE TO WHAT EXTENT THE ADJUVANTS BREAK MACROPHAGE IMMUNE TOLERANCE TO AUGMENT PRCR-BASED THERAPY, AND DECIPHER THE COMPOSITION AND PHENOTYPE OF TUMOR-ASSOCIATED MACROPHAGES UPON THE TREATMENT OF THE ADJUVANTS, WITH IN VITRO AND IN VIVO PRECLINICAL CANCER MODELS. AIM2 IS FOCUSED ON UNDERSTANDING THE MOLECULAR MECHANISMS OF THE ADJUVANTS IN AUGMENTING THE EFFICACY OF PRCR-BASED THERAPY, BY IDENTIFYING THE MOLECULAR MACHINERY INVOLVED IN CANCER CELL PHAGOCYTOSIS AND THE FUNCTION AND REGULATION OF PRCR IMMUNE CHECKPOINTS IN MEDIATING MACROPHAGE- CANCER CELL INTERACTION. A COMBINATION OF IN VITRO AND IN VIVO CELL BIOLOGICAL, BIOCHEMICAL, BIOINFORMATIC, AND IMMUNOLOGICAL METHODS WILL BE USED FOR THE PROPOSED STUDIES. THE INVESTIGATION OF THE UNDERLYING MECHANISM OF PRCR ACTIVATION AND REGULATION WILL ENABLE A BETTER UNDERSTANDING OF THE BASIC PRINCIPLES OF CANCER DEVELOPMENT AND INSPIRE THE INVENTION OF MORE EFFECTIVE CANCER IMMUNOTHERAPIES. SUCCESSFUL COMPLETION OF THIS STUDY WILL LEAD TO THE IDENTIFICATION OF NOVEL FUNCTIONAL COMPONENTS MEDIATING CANCER IMMUNE EVASION AND WILL IMMEDIATELY PROVIDE NOVEL THERAPEUTIC OPPORTUNITIES FOR A VARIETY OF CANCERS.

HYPERGLYCEMIA-INDUCED DNA DAMAGE AS A DRIVER OF GENOMIC INSTABILITY

CONTROL OF B CELL DEVELOPMENT AND EFFECTOR FUNCTIONS BY MICRORNA-142

MECHANISMS FOR SALIVARY TUMOR EPITHELIAL-MESENCHYMAL TRANSITIONS

EVALUATION OF THE GROUP DECISION-MAKING PROCESS OF CLINICAL GUIDELINE PANELS

ELUCIDATING THE ROLE OF DNAPKCS IN CHROMOSOMAL BREAK END JOINING AND CLASTOGEN RESISTANCE - SUMMARY. KINASE INHIBITORS TARGETING THE CATALYTIC SUBUNIT OF THE DNA-DEPENDENT PROTEIN KINASE (DNAPKCS) ARE BEING DEVELOPED TO SENSITIZE TUMORS TO CLASTOGENIC (CHROMOSOMAL-BREAKING) AGENTS, SUCH AS RADIOTHERAPY. THE DNAPKCS KINASE INHIBITOR M3814 (EMD SERONO) IS IN CLINICAL TRIALS, AND OTHER NEW POTENT INHIBITORS HAVE BEEN REPORTED. ELUCIDATING THE ROLE OF DNAPKCS IN THE REPAIR OF CHROMOSOMAL BREAKS IS SIGNIFICANT BECAUSE IT WILL ENABLE THE IDENTIFICATION OF THE CONTEXTS (E.G. TUMOR GENETIC BACKGROUND AND TREATMENT REGIMENS) IN WHICH DNAPKCS KINASE INHIBITORS ARE MOST EFFECTIVE, AND THEREBY DEVELOP BIOMARKERS TO PREDICT TUMOR RESPONSE. DNAPKCS HAS LONG BEEN RECOGNIZED AS IMPORTANT FOR PROMOTING RADIORESISTANCE, BUT ITS ROLE IN CHROMOSOMAL BREAK REPAIR HAS REMAINED ELUSIVE. NAMELY, DNAPKCS ASSOCIATES WITH FACTORS IN THE CANONICAL NON-HOMOLOGOUS END JOINING (C-NHEJ) PATHWAY FOR DNA DOUBLE-STRAND BREAK (DSB) REPAIR BUT IS NOT REQUIRED FOR ALL C-NHEJ EVENTS (E.G. IS LARGELY DISPENSABLE FOR V(D)J RECOMBINATION SIGNAL EJ). FURTHERMORE, UNTIL RECENTLY, SPECIFIC ASSAYS TO DETECT C-NHEJ OF CHROMOSOMAL DSBS, APART FROM V(D)J RECOMBINATION, HAVE NOT BEEN AVAILABLE, DUE TO PARTIAL REDUNDANCY WITH THE ALTERNATIVE EJ (ALT-EJ) PATHWAY. TO ADDRESS THIS GAP IN TECHNOLOGY, OUR LABORATORY RECENTLY IDENTIFIED A SPECIFIC HALLMARK OF C-NHEJ. NAMELY, WE REPORTED THAT SEVERAL C-NHEJ FACTORS (I.E. XLF, KU70, AND XRCC4) ARE REQUIRED FOR EJ BETWEEN BLUNT-ENDED DSBS (INDUCED BY THE CAS9 NUCLEASE) THAT ARE JOINED WITHOUT CAUSING INSERTION/DELETION MUTATIONS (INDELS), I.E. FOR ACCURATE/NO INDEL EJ. IN PRELIMINARY DATA, WE SHOW THAT DNAPKCS IS PARTIALLY REQUIRED FOR NO INDEL EJ, BUT ITS ROLE IS SUBSTANTIALLY MAGNIFIED IN CELLS WITH XLF REDUCED-FUNCTION MUTATIONS (HYPOMORPHS), INCLUDING MUTATIONS IN AN XLF MOTIF IMPORTANT FOR BINDING THE KU HETERODIMER AND DNA. BUILDING ON THESE FINDINGS, WE WILL DEFINE THE ROLE OF DNAPKCS IN CHROMOSOMAL EJ AND CLASTOGEN RESISTANCE. IN AIM 1, WE HYPOTHESIZE THAT XLF AND DNAPKCS FUNCTION SYNERGISTICALLY TO ENSURE NO INDEL EJ, AND THIS SYNERGY IS CRITICAL FOR CLASTOGEN RESISTANCE. WE WILL ALSO EXAMINE THE INTERPLAY BETWEEN THESE TWO FACTORS IN DNA REPLICATION, AND DEFINE THE KEY DOMAINS OF XLF AND DNAPKCS IMPORTANT FOR EACH OF THESE FUNCTIONS. IN AIM 2, WE INVESTIGATE THE INTERPLAY BETWEEN DNAPKCS AND A SET OF KU-BINDING FACTORS, AS WELL AS ALT-EJ, ON CLASTOGEN RESISTANCE AND GENOME STABILITY. WE TEST WHETHER DNAPKCS FUNCTIONS SYNERGISTICALLY NOT ONLY WITH XLF, BUT ALSO WITH EACH OF FOUR KU-BINDING FACTORS (CYREN/MRI, PAXX, APLF, AND WRN), FOR NO INDEL EJ, CLASTOGEN RESISTANCE, AND ROBUST RECRUITMENT OF KU TO DNA DAMAGE. THESE STUDIES WILL FILL A MAJOR GAP IN OUR UNDERSTANDING OF C-NHEJ MECHANISMS. FINALLY, WE POSIT THAT COMBINED DISRUPTION OF DNAPKCS AND ALT-EJ CAUSES ABERRANT DSB END PROCESSING, LEADING TO LARGE DELETIONS, AND/OR PERSISTENT, UNREPAIRED DSBS, THEREBY CAUSING REDUCED CLONOGENIC SURVIVAL AND/OR CLASTOGEN SENSITIZATION. IN SUMMARY, WE WILL TEST THE OVERALL HYPOTHESIS THAT DNAPKCS PROMOTES ACCURATE EJ (I.E. NO INDEL EJ), THAT ITS ROLE IN SUCH EJ IS MAGNIFIED BY DEFECTS IN A SET OF KU-BINDING FACTORS, AND THAT IT IS CRITICAL FOR GENOME STABILITY IN CELLS DEFICIENT IN ALT-EJ.

TARGETING TRANSCRIPTIONAL REGULATORS FOR IMMUNOTHERAPY OF ACUTE MYELOID LEUKEMIA

ELUCIDATE AND MODULATE CELL SIGNALING IN NK CELLS FOR GLIOMA TREATMENT

DNA DAMAGE RESPONSE AND ONCOGENIC SIGNALING

EPIGENOMIC MODIFICATIONS IN MAMMALIAN NEUROGENESIS

MICROENVIRONMENTAL CUES CONTROL PANCREAS CELL FATE AND BETA-CELL MATURATION

NUCLEAR MECHANISMS OF VSMC GENE REGULATION AND CELLULAR INTERACTIONS IN DIABETES

STEM CELL/NANOPARTICLE CONSTRUCTS FOR TARGETED OVARIAN CANCER THERAPY

BIOMARKERS, MECHANISMS AND MODULATION OF OXIDATIVE STRESS ASSOCIATED RISK FACTORS IN CARCINOGENESIS - ABSTRACT CANCER IS THE SECOND MOST COMMON CAUSE OF DEATH IN DEVELOPED NATIONS, AND INCIDENCE IS RISING AMONG DEVELOPING NATIONS. AN ESTIMATED 70% OF CANCERS ARE ATTRIBUTABLE TO “MODIFIABLE” RISK FACTORS, INCLUDING OBESITY, CHRONIC INFLAMMATORY DISEASES, AND POOR DIET, ALL OF WHICH HAVE BEEN ASSOCIATED WITH INCREASED OXIDATIVE STRESS. THESE ARE NOT THEMSELVES “CARCINOGENETIC”, BUT THEY ARE THOUGHT TO ACT AS “CANCER PROMOTERS”, INCREASING THE PROBABILITY OF DEVELOPING CANCER. WITH ADVANCES IN WHOLE GENOME SEQUENCING AND DEVELOPMENT OF COMPUTATIONAL TECHNIQUES TO EXAMINE THE CANCER GENOME, WE CAN NOW USE MATHEMATICAL PROFILING OF SOMATIC MUTATIONAL PROFILES (TERMED MUTATIONAL SIGNATURES) TO IDENTIFY POTENTIAL CAUSES UNDERLYING A GIVEN TUMOR (E.G., SMOKING VERSUS UV LIGHT). IT REMAINS UNCLEAR, HOWEVER, IF THERE IS A MUTATIONAL SIGNATURE THAT IS A BIOMARKER OF CUMULATIVE LIFETIME EXPOSURE TO REACTIVE OXYGEN SPECIES (ROS)-MEDIATED DNA DAMAGE AND IF THIS CORRELATES WITH CANCER-ASSOCIATED LIFESTYLE FACTORS. HERE, WE WILL UTILIZE CUTTING-EDGE MULTI-OMIC PROFILING AND MOLECULAR BIOLOGY AND COMPUTATIONAL TOOLS TO BETTER UNDERSTAND THE CONTRIBUTION/MECHANISM OF OXIDATIVE STRESS AS A CANCER PROMOTER. TO EXAMINE THE CORRELATION OF ROS MUTATIONAL SIGNATURE LEVELS AND INFLAMMATION-RELATED CANCER RISK FACTORS, WE WILL PERFORM WHOLE GENOME SEQUENCING AND MUTATIONAL SIGNATURE ANALYSIS OF A LARGE COHORT OF COLORECTAL TUMORS FROM PATIENTS WITH DETAILED, LONGITUDINALLY COLLECTED LIFESTYLE DATA (E.G., SMOKING, CALORIC INTAKE, RED MEAT INTAKE, EXERCISE LEVEL, ETC.) COLLECTED BY THE MOLECULAR EPIDEMIOLOGY OF COLORECTAL CANCER STUDY. WE WILL ALSO EVALUATE WHETHER ACCUMULATION OF ROS-GENERATED MUTATIONS IS BIASED TOWARD CTCF BINDING LOCI AND WHETHER CHROMOSOMAL ARCHITECTURE IS MODIFIED BY EXPOSURE TO CARCINOGENS OR CANCER- ASSOCIATED PROCESSES, THEREBY MEDIATING UNIQUE “EPIGENOMIC SIGNATURES”. THESE AIMS WILL ALSO PROVIDE DATA THAT CAN BE USED IN THE DEVELOPMENT OF TWO NOVEL COMPUTATIONAL TOOLS FOR THE ANALYSIS OF CANCER DRIVER MUTATIONAL SIGNATURES AND EPIGENOMIC SIGNATURES OF CARCINOGEN EXPOSURE. FINALLY, WE WILL TEST THE MOLECULAR AND CLINICAL BENEFIT OF INTERMITTENT FASTING DURING DAILY RADIATION THERAPY BASED ON THE HYPOTHESIS THAT LIFESTYLE FACTORS COULD MODULATE SUSCEPTIBILITY TO ROS MUTAGENESIS. PATIENT- REPORTED QUALITY OF LIFE AND CLINICIAN-REPORTED ADVERSE EVENTS, AS WELL AS MOLECULAR ASSAY FOR TISSUE-SPECIFIC LEVELS OF ROS-ASSOCIATED DNA DAMAGE, WILL ALLOW US TO ASSESS WHETHER INTERMITTENT FASTING CAN REDUCE NORMAL TISSUE TOXICITY. SUCCESSFUL COMPLETION OF THE PROPOSED RESEARCH WILL PROVIDE A COMPREHENSIVE EXAMINATION OF THE EPIDEMIOLOGY AND MECHANISM OF ROS-MEDIATED DNA DAMAGE IN HUMAN CANCERS AND DEMONSTRATE THE SAFETY AND POTENTIAL EFFICACY OF INTERMITTENT FASTING AS A CLINICALLY TRANSLATABLE AND EASILY ADAPTABLE APPROACH TO REDUCING BOTH ACUTE AND CHRONIC SIDE EFFECTS ASSOCIATED WITH RADIOTHERAPY.

COOPERATION OF SF3B1 MUTATIONS AND ATM DELETIONS IN THE PATHOGENESIS OF CHRONIC LYMPHOCYTIC LEUKEMIA

REGULATION OF N6-METHYLADENOSINE (M6A) DEPOSITION BY ARGININE METHYLATION

TARGETING THE MICROENVIRONMENT/ONCOGENE COOPERATION TO TREAT POOR PROGNOSIS T-ALL - BACKGROUND. DESPITE THE SIGNIFICANT SUCCESS OF RECENT THERAPIES, ACUTE LYMPHOBLASTIC LEUKEMIA (ALL) REMAINS THE SECOND LEADING CAUSE OF CHILDHOOD DEATH. THE DISCORDANCE BETWEEN THERAPEUTIC IMPROVEMENTS AND POOR OUTCOMES IS PARTIALLY CAUSED BY THE DIFFICULTY OF SALVAGING RELAPSED DISEASE. WHILE OUTCOMES HAVE IMPROVED IN DE NOVO TREATMENT, DISMAL RATES OF OVERALL SURVIVAL (LESS THAN 25%) WERE OBSERVED IN RELAPSED SUBTYPES OF ALL. THE CLINICAL ARMAMENTARIUM FOR TREATING RELAPSED OR REFRACTORY T-ALL MUST BE SUPPORTED WITH NEW OPTIONS. STRATEGY. WHILE MOST OF T-ALL CASES EXHIBIT GAIN-OF-FUNCTION MUTATIONS IN NOTCH SIGNALING, THERAPIES AGAINST NOTCH HAVE NOT FULFILLED THEIR CLINICAL PROMISE. TO IDENTIFY ALTERNATIVE TARGETS FOR NEW THERAPIES, WE PROPOSE TO DEFINE HOW CELL-INTRINSIC ONCOGENIC EVENTS INTEGRATE WITH EXTERNAL SIGNALS FROM THE MICROENVIRONMENT. RECENT STUDIES POINT TO THE FUNCTIONAL IMPACT OF “STROMAL” SIGNALS IN LEUKEMIA BIOLOGY. HOWEVER, ONE SIGNIFICANT GAP IN THIS KNOWLEDGEBASE IS HOW MICROENVIRONMENTAL FACTORS BECOME ESSENTIAL FOR LEUKEMOGENESIS AND MAINTENANCE. PRELIMINARY RESULTS. ACCORDING TO OUR RESULTS IN PRIMARY T-ALL CELLS, ACTIVATING MUTATIONS IN NOTCH FAILED TO SATURATE NOTCH SIGNALING: NOTCH SIGNAL STRENGTH INCREASED WHEN T-ALL CELLS ENCOUNTER NOTCH LIGANDS WITHIN THE MICROENVIRONMENT – E.G. INTERLEUKIN 7 (IL-7). THE INCREASED STRENGTH OF NOTCH SIGNALING CORRELATED WITH INCREASED SURFACE EXPRESSION OF IL-7RA BY DIRECT TRANSCRIPTIONAL ACTIVATION OF THE IL-7RA PROMOTER, RESULTING IN T-ALL HYPER- RESPONSIVENESS TO IL-7. IL-7 ALSO INDUCED THE CELL CYCLE REGULATOR SKP2, ACTIVATED STAT5, AND (SURPRISINGLY) STAT3. PRIMARY T-ALL CELLS SHOWED PERSISTENT STAT3 ACTIVATION AND OUR RESULTS SUGGEST STAT3 DELETION IMPAIRS T-ALL LEUKEMOGENESIS. HYPOTHESIS. THESE DATA SUPPORT SIGNIFICANT INTERPLAY BETWEEN ONCOGENIC FACTORS AND THE MICROENVIRONMENT. ACCORDING TO OUR HYPOTHESIS, INTERPLAY BETWEEN MICROENVIRONMENTAL SIGNALS (IL-7), NOTCH SIGNALING, SKP2, AND STAT3 FORM A RECIPROCAL POSITIVE FEEDBACK LOOP THAT IS ESSENTIAL FOR T-CELL LEUKEMOGENESIS; THIS AXIS ALSO COMPENSATES FOR THE ACTION OF STANDARD THERAPIES IN RELAPSED AND REFRACTORY DISEASE. APPROACH. TO TEST THIS, WE PROPOSE: 1) TO DETERMINE THE TEMPORAL REQUIREMENT FOR STAT3 DELETION IN INITIATION, PROGRESSION, AND RELAPSE IN T-ALL BY USING A MODEL OF INDUCIBLE GENETIC DELETION OF STAT3 IN COMBINATION WITH A MODEL OF NOTCH-INDUCED T-ALL. 2) TO MAP HOW T-ALL DEVELOPMENT IS AFFECTED BY NOTCH/IL-7/STAT3/SKP2 SIGNALING CIRCUITRY BY USING OVEREXPRESSION AND GENE SILENCING APPROACHES TO DEFINE THE RECIPROCAL REGULATION OF NOTCH, STAT3, AND SKP2. 3) TO IDENTIFY THE IMPACT OF INHIBITING NOTCH/STAT/SKP2 CIRCUITRY IN RELAPSED T-ALL BY TESTING BOTH PRE-CLINICAL AND CLINICAL INHIBITORS OF STAT SIGNALING AND SKP2 INHIBITORS IN PRE-CLINICAL PDX MODELS OF T-ALL. IMPACT. SUCCESSFUL COMPLETION OF THIS PROPOSED WORK WILL: 1) DEFINE HOW COOPERATION BETWEEN ONCOGENIC SIGNALING AND THE MICROENVIRONMENT AFFECTS THERAPY OF RELAPSED AND REFRACTORY T-ALL; 2) BUILD A FOUNDATION FOR VALIDATING NEW MOLECULAR TARGETS IN RELAPSED AND REFRACTORY T-ALL; 3) PROVIDE A PROOF-OF-PRINCIPLE FOR AN ALTERNATIVE STRATEGY IN WHICH ENTIRE MOLECULAR CIRCUITS ARE CONSIDERED DURING THE DEVELOPMENT OF THERAPIES.

CITY OF HOPE CORE CLINICAL CENTER FOR THE BLOOD AND MARROW TRANSPLANT CLINICAL TRIALS NETWORK

TET2-MEDIATED EPITRANSCRIPTOMIC REGULATION IN LEUKEMIA MICROENVIRONMENT - PROJECT SUMMARY: BACKGROUND: ACUTE MYELOID LEUKEMIA (AML) IS ONE OF THE MOST AGGRESSIVE TYPES OF HEMATOPOIETIC MALIGNANCIES WITH VARIOUS GENETIC ALTERATIONS. TEN-ELEVEN TRANSLOCATION 2 (TET2), AN ENZYME INVOLVED IN DNA DEMETHYLATION, IS DELETED OR MUTATED IN 15-20% OF AML PATIENTS. THOSE PATIENTS WITH TET2 DEFICIENCY ARE POORLY RESPONSIVE TO CURRENTLY AVAILABLE THERAPEUTIC REGIMENS, LEADING TO MORE ADVERSE OUTCOMES THAN PATIENTS WITH OTHER AML SUBTYPES. THUS, IT IS URGENT TO IDENTIFY NEW THERAPEUTIC TARGET(S) AND DEVELOP NOVEL EFFECTIVE APPROACHES TO TREAT TET2-DEFICIENT AMLS. TET2 LOSS IN MICE FACILITATES THE SELF-RENEWAL OF HEMATOPOIETIC STEM CELLS (HSCS) AND LEUKEMIC STEM/INITIATING CELLS (LSCS/LICS). THE LSCS/LICS RESIDE IN A SPECIALIZED MICROENVIRONMENT CALLED “NICHE” IN THE BONE MARROW (BM) TO SUPPORT THEIR SURVIVAL AND SELF-RENEWAL. THERE ARE SEVERAL CRITICAL GAPS IN OUR CURRENT KNOWLEDGE OF THE MOLECULAR MECHANISM UNDERLYING LSC/LIC HOMING AND OF THE ROLE OF TET2 DEFICIENCY IN THE BM MICROENVIRONMENT. MEANWHILE, EVIDENCE IS EMERGING TO SUPPORT A NOVEL FUNCTION FOR TET2-MEDIATED OXIDATION OF METHYL-5-CYTOSINE (M5C) IN RNAS, INCLUDING MESSENGER RNA (MRNA). HOWEVER, IT IS UNKNOWN WHETHER AND (IF SO) HOW TET2-MEDIATED RNA M5C DEMETHYLATION CONTRIBUTES TO LEUKEMOGENESIS. OUR PRELIMINARY STUDY SHOWED THAT TET2 DEFICIENCY STIMULATES THE TETRASPANIN 13 (TSPAN13)/C-X-C MOTIF CHEMOKINE RECEPTOR 4 (CXCR4) AXIS TO FACILITATE AML HOMING/MIGRATION INTO THE BM MICROENVIRONMENT, GIVING RISE TO INCREASED LSC/LIC SELF-RENEWAL AND FAST LEUKEMOGENESIS IN VIVO. TET2 DEFICIENCY-MEDIATED INCREASE OF MRNA M5C MODIFICATION IN TSPAN13 IS RECOGNIZED BY Y-BOX BINDING PROTEIN 1 (YBX1), WHICH IN TURN STABILIZES TSPAN13 TRANSCRIPT AND INCREASES ITS EXPRESSION, THEREBY ACTIVATING THE CXCR4 SIGNALING. PHARMACOLOGICAL INHIBITION OF CXCR4 SUPPRESSES LSC/LIC HOMING INTO THE BM MICROENVIRONMENT AND SHOWS A SYNERGISTIC EFFECT WITH HYPOMETHYLATING AGENTS IN KILLING TET2-DEFICIENT AMLS. THESE RESULTS LEAD TO OUR CENTRAL HYPOTHESIS THAT TET2-MEDIATED MRNA M5C DEMETHYLATION IS INVOLVED IN REPROGRAMMING BM MICROENVIRONMENT. GUIDED BY STRONG PRELIMINARY DATA, WE PROPOSE THREE SPECIFIC AIMS TO TEST OUR HYPOTHESIS: (1) DETERMINE THE DEFINITIVE ROLE OF TET2 IN THE HOMING OF LSCS/LICS INTO BM MICROENVIRONMENT; (2) CHARACTERIZE THE MRNA M5C-DEPENDENT AND FUNCTIONALLY ESSENTIAL TARGETS OF TET2 AND DECIPHER THE MOLECULAR MECHANISMS UNDERLYING THE ROLE OF TET2 IN LSC/LIC HOMING AND SELF-RENEWAL; AND (3) ASSESS THE THERAPEUTIC POTENTIAL OF TARGETING THE TET2/CXCR4 AXIS IN HIGH-RISK TET2-DEFICIENT AMLS. OVERALL, OUR PROPOSED STUDIES WILL SUBSTANTIALLY ADVANCE OUR UNDERSTANDING OF THE FUNDAMENTAL BIOLOGY OF TET2-MEDIATED EPITRANSCRIPTOMIC CHANGES IN BM MICROENVIRONMENT AND MAY RESULT IN THE DEVELOPMENT OF NOVEL EFFECTIVE APPROACHES TO TREAT AMLS WITH TET2 DEFICIENCY. THUS, OUR PROJECT IS OF HIGH NOVELTY AND SIGNIFICANCE IN BOTH BASIC RESEARCH AND TRANSLATIONAL MEDICINE.

CHARACTERIZING AND TARGETING THE NOVEL IL-15- AKT-XBP1S PATHWAY IN NK CELLS

EXPLORING THE PRECLINICAL RELEVANCE OF THERAPEUTIC RADIOLABELED DARATUMUMAB (ANTI-CD38) IN COMBINATION WITH ANTI-CS1 CAR T CELLS AS A NOVEL COMBINATORIAL TREATMENT FOR MULTIPLE MYELOMA

TARGETED RADIATION AND IMMUNOCYTOKINE THERAPY FOR CEA POSITIVE MALIGNANCIES - SUMMARY/ABSTRACT IMAGE GUIDED RADIOTHERAPY (IGRT) OR SYSTEMICALLY ADMINISTERED RADIONUCLIDE THERAPY ARE ATTRACTIVE APPROACHES THAT CAN PERTURB THE TUMOR MICROENVIRONMENT (TME) SO THAT TARGETED IMMUNOTHERAPY CAN CONVERT A LARGELY IMMUNORESISTANT INTO AN IMMUNOSUSCEPTIBLE TUMOR. ALTHOUGH THIS APPROACH HAS BEEN CLINICALLY EXPLORED USING HIGH OR LOW OR HIGH PLUS LOW DOSE IGRT PLUS UNTARGETED CHECKPOINT IMMUNOTHERAPY, FURTHER IMPROVEMENTS ARE WARRANTED AND REQUIRE TESTING IN APPROPRIATE CLINICAL STUDIES AND PRECLINICAL MODELS. IN THIS APPLICATION WE PROPOSE THREE AIMS TO TEST OUR HYPOTHESIS THAT IGRT AND OR TARGETED ALPHA THERAPY (TAT) FOLLOWED IMMEDIATELY BY IMMUNOCYTOKINE (ICK), A FORM OF TARGETED IMMUNOTHERAPY, WILL LEAD TO INCREASED TUMOR INFILTRATION OF IFN+ CD8S (TEFF) AND DECREASED FOXP3+ CD4S (TREG). THIS HYPOTHESIS IS SUPPORTED BY TWO STUDIES TARGETING CEA, A MAJOR MARKER OF SOLID TUMOR MALIGNANCIES (EG. COLON AND BREAST), IN CEA TRANSGENIC MICE THAT ARE IMMUNOCOMPETENT, EXPRESS CEA IN NORMAL TISSUES, AND ARE TOLERANT TO CEA. WE NOW PROPOSE TO TEST THIS HYPOTHESIS IN TWO CLINICAL TRIALS, AIMS 1 (IGRT + ICK) AND 2 (TAT), AND PERFORM IMMUNOCORRELATE STUDIES FOR TEFF AND TREG CELLS, AMONG OTHERS, ON PRE- AND POST-THERAPY BIOPSIES. AIM 2 IS A TAT ONLY STUDY AS A PRELUDE TO A THIRD CLINICAL TRIAL COMBINING TAT PLUS ICK, THAT WILL BE INITIATED AT THE END OF THE PROJECT PERIOD. IN AIM 3, WE WILL REFINE OUR ANIMAL STUDIES TO DETERMINE TME CHANGES IMMEDIATELY AFTER SEQUENTIAL THERAPY FOR BOTH IGRT PLUS ICK OR TAT PLUS ICK, BY A COMBINATION OF PET IMAGING FOR CEA, CD8S AND STROMAL CELLS BY FIBROBLAST ACTIVATED PROTEIN (FAPI), ALONG WITH FLOW AND IHC ANALYSIS OF TUMORS, LNS AND SPLEEN. WE EXPECT THE RESULTS OF AIMS 1 AND 2 TO FURTHER GUIDE AIM 3, AND THE RESULTS OF AIM 3 TO GUIDE FUTURE TRIALS THAT INCORPORATE IGRT AND/OR TAT PLUS ICK IN CEA POSITIVE MALIGNANCIES.

THE ROLE OF CEACAM1 IN ONCOGENIC B-CELL RECEPTOR SIGNALING AND IMMUNOTHERAPY IN MANTLE CELL LYMPHOMA - ABSTRACT MANTLE CELL LYMPHOMA (MCL) ACCOUNTS FOR 6-8% OF ALL NON-HODGKIN LYMPHOMAS (NHLS). WHILE SUBSTANTIAL THERAPEUTIC ADVANCES HAVE BEEN ACHIEVED FOR OTHER NHLS, MCL REMAINS AN INCURABLE LYMPHOMA, THE REASON OF WHICH IS NOT KNOWN. MCL PATIENTS HAVE A DISMAL PROGNOSIS WITH A MEDIAN OVERALL SURVIVAL OF 3-5 YEARS. STANDARD OF CARE INCLUDES IBRUTINIB, A SMALL MOLECULE INHIBITOR OF THE B-CELL RECEPTOR (BCR)-PROXIMAL TYROSINE KINASE BTK. HOWEVER, ONE-THIRD OF MCL PATIENTS DO NOT RESPOND TO THE DRUG. EVEN INITIALLY IBRUTINIB- SENSITIVE PATIENTS INVARIABLY DEVELOP RESISTANCE; HOWEVER, THE MECHANISMS OF IBRUTINIB-RESISTANCE ARE NOW CLEAR. SINCE MECHANISTIC INSIGHT INTO ONCOGENIC BCR SIGNALING IN DLBCL AND CLL ENABLED THE DEVELOPMENT OF HIGHLY EFFECTIVE TREATMENT APPROACHES, THIS PROPOSAL WILL ADDRESS THE MECHANISMS OF ONCOGENIC BCR- SIGNALING IN MCL. IN AN INTEGRATED FUNCTIONAL ANALYSIS COMBINING A GENOME-WIDE CRISPR-CAS9 LIBRARY, GENE EXPRESSION PROFILING AND BCR SIGNAL TRANSDUCTION STUDIES, WE HAVE UNCOVERED CEACAM1 AS A CENTRAL COMPONENT OF ONCOGENIC BCR SIGNALING THAT IS ESSENTIAL IN MCL BUT NOT IN NORMAL B CELLS OR OTHER B-CELL MALIGNANCIES. AS A TRANSMEMBRANE PROTEIN, CEACAM1 IS EXPRESSED ON THE SURFACE OF ACTIVATED LYMPHOCYTES AND CARRIES TWO IMMUNORECEPTOR TYROSINE-BASED INHIBITORY MOTIFS (ITIMS) ON ITS CYTOPLASMIC TAIL. OWING TO RECRUITMENT OF THE INHIBITORY PHOSPHATASE SHP1 TO THE ITIMS, CEACAM1 FUNCTIONS AS A REGULATOR OF T-CELL RECEPTOR (TCR) SIGNALING IN T CELLS, HOWEVER, ITS FUNCTION IN NORMAL B CELLS AND MCL IS NOT KNOWN. UNEXPECTEDLY, OUR PRELIMINARY DATA SHOWED THAT CEACAM1 FUNCTION INDUCED A NET INCREASE OF BCR SIGNALING, LEADING TO INCREASED SURVIVAL AND PROLIFERATION OF MCL CELLS IN VITRO AND IN VIVO. OUR MECHANISTIC STUDIES REVEALED THAT CEACAM1 RECRUITED THE ACTIN-BINDING PROTEIN FILAMIN A TO THE PLASMA MEMBRANE MICRODOMAINS AND ACTIVATED THE BCR-PROXIMAL KINASE LYN AFTER ANTIGEN ENGAGEMENT. FURTHERMORE, SUPER- RESOLUTION CONFOCAL MICROSCOPY REVEALED THAT CEACAM1 PROMOTED REORGANIZATION OF THE ACTIN CYTOSKELETAL NETWORK FOLLOWING BCR CROSS-LINKING. LEVERAGING THE CLINICAL GRADE CELL THERAPEUTICS FACILITY AT CITY OF HOPE, WE DESIGNED AND VALIDATED A NOVEL CEACAM1 CHIMERIC ANTIGEN RECEPTOR (CAR) ENGINEERED IN PRIMARY HUMAN T CELLS. THE CAR-T CELLS WERE HIGHLY ACTIVE IN ELIMINATING CEACAM1+ MCL BUT LACKED REACTIVITY AGAINST OTHER CELL TYPES. BASED ON OUR DISCOVERY OF CEACAM1 AS A CRITICAL BCR SIGNALING COMPONENT IN MCL AND THE SUCCESSFUL DEVELOPMENT OF CEACAM1 CAR-T CELLS, WE HYPOTHESIZE THAT CEACAM1 FUNCTIONS AS A CENTRAL DRIVER OF ONCOGENIC BCR ACTIVITY AND REPRESENTS A NOVEL THERAPEUTIC TARGET IN MCL. THE FOLLOWING SPECIFIC AIMS WILL TEST AND REFINE THE CONCEPT OF CEACAM1-BASED THERAPIES FOR MCL: AIM 1) DEFINE THE MECHANISTIC ROLE OF CEACAM1 IN ONCOGENIC BCR SIGNALING IN MCL; AIM 2) DISSECT THE ROLE OF CEACAM1 IN NEW GENETIC MOUSE MODELS FOR MCL; AIM 3) VALIDATION OF CAR T-CELL STRATEGIES TARGETING CEACAM1 IN REFRACTORY MCL. RESULTS FROM THE PROPOSED STUDIES ARE EXPECTED TO PROVIDE: 1) NEW INFORMATION ON THE CENTRAL ROLE OF CEACAM1 IN ONCOGENIC BCR SIGNALING AND MCL, AND 2) PRE-CLINICAL VALIDATION OF NOVEL IMMUNOTHERAPEUTIC STRATEGIES TARGETING CEACAM1 IN REFRACTORY MCL.

AGING AND THE UNSTABLE EPIGENOME

DEVELOPING BRAF MUTANT AND BRAF WILD-TYPE SELECTIVE STRATEGIES FOR RADIOSENSITIZATION IN ANAPLASTIC THYROID CANCER

FATTY ACIDS AND THEIR RECEPTORS-MEDIATED TUMOR METASTASIS AND PROGRESSION

PD-L1 INTERACTS WITH CD80 AND PD-1 TO REGULATE GVHD AND GVL ACTIVITY

THE ROLE OF ALKBH5-MEDIATED RNA DEMETHYLATION IN THE MAINTENANCE OF GENOMIC STABILITY IN HSPCS - ABSTRACT MYELODYSPLASTIC SYNDROMES (MDS) ARE A GROUP OF DIVERSE MALIGNANT HEMATOLOGICAL DISORDERS THAT ORIGINATE FROM HEMATOPOIETIC STEM CELLS (HSCS). INCREASED LEVELS OF REACTIVE OXYGEN SPECIES (ROS) AND DNA DAMAGE ARE COMMONLY DETECTED IN HEMATOPOIETIC CELLS FROM MDS PATIENTS. AN ELEVATED LEVEL OF ROS, GENERATED FROM EITHER ENDOGENOUS OR EXOGENOUS SOURCES INCLUDING ONCOGENE ACTIVATION, LEADS TO LOSS OF QUIESCENCE AND SELF-RENEWAL OF HSCS. ROS-INDUCED DNA DAMAGE SPEEDS UP THE AGING PROCESS OF STEM CELLS AND CONTRIBUTES TO THE MUTAGENESIS ASSOCIATED WITH CANCER DEVELOPMENT. M6A RNA METHYLATION PLAYS A SIGNIFICANT ROLE IN MULTIPLE BIOLOGICAL PROCESSES BY INTRODUCING ANOTHER LAYER OF POST-TRANSCRIPTIONAL REGULATION OF GENE EXPRESSION WITHIN CELLS. THE GOAL OF THIS PROJECT IS TO ELUCIDATE THE SIGNIFICANT ROLE OF ALKBH5-MEDIATED EPIGENETIC REGULATION IN THE MAINTENANCE OF GENOMIC STABILITY IN HEMATOPOIETIC STEM/PROGENITOR CELL (HSPCS) DURING OXIDATIVE STRESS, AND HOW DEREGULATION OF ALKBH5 CONTRIBUTES TO PROMOTION OF LEUKEMIC TRANSFORMATION OF HSPCS IN THE INITIATION AND DEVELOPMENT OF MDS. WE FOUND THAT ROS SIGNIFICANTLY INCREASED GLOBAL M6A RNA METHYLATION IN HUMAN CELL LINES, AND THAT THE ELEVATION OF M6A MRNA METHYLATION IS REQUIRED FOR RAPIDLY REPAIRING ROS-INDUCED DNA LESIONS AND PREVENTING CELL DEATH. INTERESTINGLY, WE FOUND THAT ALKBH5, THE M6A RNA DEMETHYLASE, IS RESPONSIBLE FOR ROS-INDUCED ELEVATION OF M6A MRNA METHYLATION. ROS INDUCED POST- TRANSLATIONAL MODIFICATION OF ALKBH5, AND INHIBITED THE DEMETHYLASE ACTIVITY OF ALKBH5. WE SHOWED THAT FORCED EXPRESSION OF ALKBH5 INHIBITED ROS-INDUCED M6A MRNA METHYLATION AND SIGNIFICANTLY DELAYED REPAIR OF ROS-INDUCED DNA DAMAGE. THUS, WE HYPOTHESIZE THAT ABERRANT EXPRESSION OF ALKBH5 DISRUPTS HSPC FUNCTIONS BY NEGATIVELY INFLUENCING GENOME INTEGRITY AND SURVIVAL OF HSPCS, THEREBY CONTRIBUTING TO LEUKEMIC TRANSFORMATION OF HSPCS DURING THE INITIATION AND DEVELOPMENT OF MDS. IN THIS PROPOSAL, WE WILL DETERMINE 1) THE ROLE AND UNDERLYING MECHANISM OF ALKBH5 IN THE MAINTENANCE OF GENOMIC STABILITY IN HSPCS IN RESPONSE TO OXIDATIVE STRESS; 2) THE EFFECTS OF ALKBH5/ALKBH5 OVEREXPRESSION ON THE MAINTENANCE OF MOUSE AND HUMAN PRIMARY HSPCS DURING ROS STRESS IN VIVO; AND 3) WHETHER ALKBH5/ALKBH5 IS REQUIRED FOR THE MAINTENANCE OF PRE-LEUKEMIC STEM CELLS (PRE-LSCS) IN MDS. OUR STUDY WILL PROVIDE NEW INSIGHTS INTO NOVEL MECHANISMS OF MDS DEVELOPMENT AND EPITRANSCRIPTIONAL REGULATION OF GENE EXPRESSION IN HSPCS IN RESPONSE TO OXIDATIVE STRESS. ADDITIONALLY, OUR STUDY WILL PROVIDE THE FIRST SET OF EVIDENCE TO SUPPORT A SIGNIFICANT ROLE OF ALKBH5- MEDIATED M6A MRNA DEMETHYLATION IN THE MAINTENANCE OF NORMAL HSPCS AND PRE-LEUKEMIC STEM CELL (PRE- LSCS).

BILE ACIDS AND METABOLIC SURGERY

THE ROLE OF O-GLCNACYLATION IN DNA DAMAGE REPAIR AND CANCER THERAPY