WU250381S
SCOPE OF WORK PROJECT TITLE: DRUGGING NRF2 TO IMPROVE RADIATION THERAPY IN HEAD AND NECK SQUAMOUS CELL CARCINOMA START/END DATES: 12/19/2024-11/30/2029 MPI NAME: DR. BERNARD WEISSMAN PROJECT DESCRIPTION: THE NRF2 TRANSCRIPTION FACTOR IS ACTIVE IN MORE THAN 30% OF HEAD & NECK SQUAMOUS CELL CARCINOMAS (HNSCC), OWING TO PATHWAY ACTIVATING MUTATIONS AND MUTATION-INDEPENDENT MECHANISMS. NRF2 DRIVES A GENE EXPRESSION PROGRAM THAT MITIGATES OXIDATIVE AND ELECTROPHILIC STRESS, REPROGRAMS AND ENABLES CANCER CELL METABOLISM, AND SUPPRESSES IMMUNE CELL INFILTRATION. IN HNSCC PATIENTS, NRF2 ACTIVITY PORTENDS A POOR PROGNOSIS; ITS MUTATIONAL ACTIVATION IS PREDICTIVE FOR RESISTANCE TO RADIATION AND LOCAL REGIONAL FAILURE. THOUGH FREQUENTLY ACTIVE IN HUMAN CANCERS, MOUSE MODELS SUGGEST THAT CONSTITUTIVE NRF2 ACTIVITY IS NOT SUFFICIENT FOR ONCOGENESIS, BUT RATHER NRF2 SYNERGIZES WITH ONCOGENES AND TUMOR SUPPRESSORS TO DRIVE TUMOR PROGRESSION. DESPITE GREAT PROMISE, PROVEN AND EFFICACIOUS NRF2 THERAPEUTIC INHIBITORS DO NOT YET EXIST IN THE CLINIC. WE HYPOTHESIZE THAT CHEMICAL SUPPRESSION OF NRF2 ACTIVITY WILL SENSITIZE HNSCC TO RT. OUR PREVIOUS FUNCTIONAL GENOMIC, PROTEOMIC, AND SMALL MOLECULE SCREENS REVEALED THERAPEUTIC TARGETS AND PHARMACOLOGICAL AGENTS THAT SUPPRESS NRF2 ACTIVATION IN HNSCC. HERE WE FOCUS ON TWO IN VIVO EFFICACIOUS, POTENT AND MECHANISTICALLY DISTINCT NRF2 CHEMICAL INHIBITORS. FIRST, WE PREVIOUSLY REPORTED THE ANCHOR CLASS OF KEAP1 MUTANTS WHICH MAINTAIN ENZYMATIC ACTIVITY FOR NRF2 UBIQUITYLATION BUT FAIL TO DEGRADE THE NRF2 PROTEIN. WORKING WITH VIVIDION THERAPEUTICS, WE SOUGHT CYSTEINE-REACTIVE MOLECULES TO REVERSE THE ANCHOR PHENOTYPE. THIS RESULTED IN VVD065, A HIGHLY SELECTIVE AND POTENT SMALL MOLECULE THAT INDUCES NRF2 UBIQUITYLATION, DEGRADATION AND RADIOSENSITIVITY. SECOND, OUR DRUG SCREENS DISCOVERED PYRIMETHAMINE (PYR), A FDA APPROVED MEDICINE, AS AN EFFICACIOUS INHIBITOR OF NRF2 SIGNALING. WHEN ADMINISTERED TO OUR GENETICALLY ENGINEERED MOUSE MODEL (GEMM) CARRYING AN INDUCIBLE ACTIVATING NRF2E79Q MUTATION, PYR REVERSED ESOPHAGEAL AND ORAL CAVITY HYPERPLASIA WITH NO OBSERVED TOXICITY. SUBSEQUENT STRUCTURE-ACTIVITY-RELATIONSHIP OPTIMIZATION RESULTED IN 30-FOLD MORE POTENT SMALL MOLECULE WE CALL WCDD115. BECAUSE NRF2 ACTIVITY DRIVES RESISTANCE TO RADIATION THERAPY (RT), OUR PRIMARY GOAL IS TO LEVERAGE THESE TWO NOVEL NRF2 INHIBITORS AS SYNTHETIC COMBINATION AGENTS FOR RT IN HUMAN AND MOUSE MODELS OF HNSCC. IMPORTANTLY FOR THIS WORK, WE RECENTLY REPORTED THE FIRST NRF2-DRIVEN GEMM OF SPONTANEOUS ORAL SCC. OUR EXPERIMENTS USE HUMAN HNSCC XENOGRAFTS, MOUSE SYNGENEIC TUMOR GRAFTS, OUR NRF2-DRIVEN ORAL CAVITY SCC GEMM, AND HUMAN AND MOUSE 3-D SPHEROID CULTURES TO TEST IF VVD065 OR PYR/WCDD115 SYNTHETICALLY COMBINE WITH RT TO SUPPRESS TUMOR GROWTH WHILE MINIMIZING TOXICITY TO NORMAL TISSUES. WE WILL ALSO IDENTIFY PATIENT STRATIFYING MOLECULAR SIGNATURES PREDICTIVE OF SUSCEPTIBILITY TO NRF2 CHEMICAL INHIBITION. TOGETHER, THESE STUDIES WILL PRODUCE FOUNDATIONAL DATA FOR THE DOSING, SCHEDULING, EFFICACY AND TOXICITY OF NRF2 INHIBITORS AS SYNTHETIC COMBINATION AGENTS WITH RT IN PRE-CLINICAL HNSCC MODELS. THE IMPACT OF THIS RESEARCH WILL BE AN IMPROVED RESPONSE OF HNSCC TO RADIATION THERAPY. SPECIFICALLY, DR. WEISSMAN HAS STUDIED THE ROLE OF TUMOR SUPPRESSOR GENES IN HUMAN CANCER DEVELOPMENT FOR THE MAJORITY OF HIS CAREER. HE BRINGS OVER 41 YEARS OF EXPERIENCE IN MOUSE MODELS INCLUDING XENOGRAFTS AND GENETICALLY ENGINEERED MOUSE MODELS (GEMMS), CANCER CELL BIOLOGY, CELL TRANSFORMATION AND CHARACTERIZATION OF CHANGES IN TUMOR PROGRESSION AND METASTASIS. HE WILL CO-DIRECT THE DESIGN, PERFORMANCE, AND ANALYSIS OF THE PROPOSED STUDIES, IN BOTH SPECIFIC AIMS, INCLUDING GENERATION AND CHARACTERIZATION OF OSCC GEMMS, TISSUE COLLECTION, HISTOLOGY, IHC, AND PATHOLOGY. IN ADDITION, DR. BEN MAJOR (CONTACT PI) AND DR. BERNARD WEISSMAN (MPI) HAVE BEEN COLLABORATING FOR >15 YEARS. THEY PREVIOUSLY SERVED AS MULTI-PRINCIPAL INVESTIGATORS (MPIS) ON A NRF2 AND KINASE CENTRIC RO1 AND A RESEARCH PROGRAM IN LUNG CANCER AND HNSCC. THEIR WORK HAS DISCOVERED NEW MECHANICS OF NRF2 SIGNALING, THERAPEUTIC SMALL MOLECULE INHIBITORS AND SURPRISING NEW BIOLOGY IN VARIOUS NRF2 MOUSE MODELS OF SCC. THEIR COLLABORATIVE SYNERGY IS SUPPORTED BY JOINT PUBLICATIONS (N=13), PRELIMINARY DATA, NEW DRUGS (PYR, VVD065), AND THE SUCCESS OF THEIR STUDENTS AND POSTDOCS (TWO HAVE BEEN CO-MENTORED). THIS NEW R01 GRANT WILL CONTINUE THIS WORK LEVERAGING TWO NOVEL NRF2 INHIBITORS TO SYNTHETICALLY COMBINE WITH RADIATION THERAPY TO IMPROVE HNSCC PATIENT OUTCOME. AIM 1: TEST IF VVD065 INHIBITS NRF2-DRIVEN RESISTANCE TO RT. VVD065 IS A POTENT, HIGHLY SPECIFIC AND MECHANISTICALLY EXCITING KEAP1 AGONIST, WHICH RESULTS IN RAPID NRF2 LOSS. WE SHOW THAT VVD065 INHIBITS NRF2 ACTIVE CANCER GROWTH ACROSS NUMEROUS IN VITRO AND IN VIVO HNSCC MODELS. SINCE THE A0, WE DISCOVERED THAT VVD065 ENHANCES RT-INDUCED SUPPRESSION OF TUMOR GROWTH IN SYNGENEIC HNSCC MODELS. HERE WE WILL FURTHER EVALUATE VVD065 AS A SYNTHETIC COMBINATION AGENT TO RT. WE WILL OPTIMIZE RT DOSE AND SCHEDULING IN HUMAN AND MOUSE HNSCC MODELS, INCLUDING A NOVEL NRF2-DRIVEN AND NRF2-DEPENDENT HNSCC GEMM. SPECIFICALLY, WE RECENTLY PUBLISHED THAT CDKN2AFL/FL;TP53FL/FL;NRF2E79Q (CPNE79Q) MICE, BUT NOT TP53FL/FL;CDKN2AFL/FL;NRF2+/+ (CP) MICE, DEVELOP ORAL CAVITY SCC (OSCC). MOLECULAR CHARACTERIZATIONS WILL INCLUDE W.BLOT, OIS-PRM TARGETED MS, RNASEQ/QPCR, ROS QUANTIFICATION, DNA DAMAGE, FLOW CYTOMETRY AND IHC. FINALLY, WE WILL CONNECT CANCER MUTATION GENOTYPE IN KEAP1 AND CUL3 WITH NRF2 ACTIVATION, VVD065 SENSITIVITY AND RT:VVD065 SYNERGY. UNC TASKS FOR AIM 1: 1) GENERATE CDKN2AFL/FL;TP53FL/FL;NRF2E79Q (CPNE79Q) MICE, CDKN2AFL/FL;TP53FL/FL;NRF2E79Q (CPND29H) MICE AND TP53FL/FL;CDKN2AFL/FL;NRF2+/+ (CP) MICE. 2) CHARACTERIZE THE EFFECTS OF NRF2 INHIBITORS OR OTHER DRUGS, +/- RADIATION, ON THE ORAL MUCOSA OF CPNE79Q, CPND29H, AND CP MICE 3) CHARACTERIZE CPNE79Q, CPND29H, AND CP MICE FOR TUMOR DEVELOPMENT AND SURVIVAL 4) TREAT CPNE79Q, CPND29H, AND CP MICE WITH RADIATION, +/- VVD065 AND OTHER NRF2 INHIBITORS OR DRUGS/REAGENTS, CHOSEN BY THE MPIS. UNC DELIVERABLES FOR AIM 1: 1) EFFICACY STUDIES OF RADIATION TREATMENTS, +/- VVD065 OR OTHER NRF2 INHIBITORS OR DRUGS/REAGENTS FOR THE TREATMENT OF OSCC IN GEMMS. 2) NORMAL AND TUMOR SAMPLES FOR MOLECULAR ANALYSES. 3) CHARACTERIZATION OF TUMORS FROM GEMMS FOR HISTOLOGY AND IHC. 4) IDENTIFICATION OF OPTIMAL DOSING REGIMEN FOR VVD065 OR OTHER NRF2 INHIBITORS AND DRUGS, +/- RADIATION. AIM 2: DETERMINE IF PYR OR WCDD115 INHIBITS NRF2-DRIVEN RESISTANCE TO RT. PYR AND ITS DERIVATIVE WCDD115 ARE NOT SPECIFIC INHIBITORS OF NRF2, BUT THEY ARE SAFE, POTENT AND EFFICACIOUS IN VIVO. AS OUR PRELIMINARY DATA SHOW, WCDD115 SYNTHETICALLY COMBINES WITH RT TO SUPPRESS IN VITRO GROWTH AND SYNGENEIC MOUSE TUMOR GRAFTS. WE WILL TAKE A SIMILAR APPROACH TO THAT DESCRIBED FOR VVD065 TO DETERMINE IF COMBINATIONS OF WCDD115 OR PYR WITH RT ENHANCE CANCER SUPPRESSION. DOSE AND TREATMENT OPTIMIZATIONS WILL MAXIMIZE THERAPEUTIC EFFICACY AND PROVIDE A DIRECT COMPARISON BETWEEN PYR AND THE MORE POTENT WCDD115. LAST, TOXICITY MEASURES IN NORMAL ORAL CAVITY AND LARYNGEAL TISSUE FOLLOWING PYR, WCDD115 AND VVD065 WITH AND WITHOUT RT WILL BE QUANTIFIED. AS AN FDA APPROVED DRUG, PYR OFFERS FAST ACCESS TO PATIENTS AS A RT SENSITIZING APPROACH. UNC TASKS FOR AIM 2: 1) GENERATE CDKN2AFL/FL;TP53FL/FL;NRF2E79Q (CPNE79Q) MICE, AND TP53FL/FL;CDKN2AFL/FL;NRF2+/+ (CP) MICE. 2) TREAT CPNE79Q, AND CP MICE WITH RADIATION, +/- PYR, WCDD115 OR OTHER NRF2 INHIBITORS AND DRUGS/REAGENTS CHOSEN BY THE MPIS. UNC DELIVERABLES FOR AIM 2: 1) EFFICACY STUDIES OF RADIATION TREATMENTS, +/- PYR, WCDD115 OR OTHER NRF2 INHIBITORS OR DRUGS/REAGENTS FOR THE TREATMENT OF OSCC IN GEMMS. 2) NORMAL AND TUMOR SAMPLES FOR MOLECULAR ANALYSES. 3) CHARACTERIZATION OF TUMORS FROM GEMMS FOR HISTOLOGY AND IHC. 4) IDENTIFICATION OF OPTIMAL DOSING REGIMEN FOR PYR, WCDD115 OR OTHER NRF2 INHIBITORS AND DRUGS, +/- RADIATION. DR. WEISSMAN WILL ALSO PREPARE AND EDIT ALL PUBLICATIONS, INCLUDING RESEARCH REPORTS, REVIEWS, GRANT SUPPLEMENT APPLICATIONS, AND PROGRESS REPORTS IN TANDEM WITH DR. MAJOR.
University Of North Carolina At Chapel Hill
Project Grant R01CA290809
$271.2k 1/30/25