R01AI200543

ATPASE CHROMATIN REMODELING COMPLEXES AS MODULATORS OF HIV-1 LATENCY AND THERAPEUTIC TARGETS - ABSTRACT SIGNIFICANCE: HIV PERSISTS IN LONG-LIVED CD4⁺ T CELL RESERVOIRS DESPITE SUPPRESSIVE ART, AS INTEGRATED PROVIRUSES REMAIN POISED FOR REACTIVATION. CHROMATIN REMODELING IS A CENTRAL BARRIER TO DURABLE SILENCING, YET MOST STUDIES HAVE FOCUSED ON SWI/SNF FAMILY MEMBERS. THE ROLES OF NON- SWI/SNF REMODELERS REMAIN POORLY DEFINED, LIMITING OUR ABILITY TO RATIONALLY DESIGN HOST-DIRECTED “BLOCK-AND-LOCK” CURE STRATEGIES. OUR UNBIASED SHRNA SCREEN OF ALL 16 HUMAN REMODELER ATPASES IDENTIFIED EP400, CHD1, AND CHD9 AS REPRESSORS AND INO80A, SMARCA5, AND CHD2 AS ACTIVATORS, ESTABLISHING CHROMATIN REMODELING AS A KEY DETERMINANT OF HIV LATENCY. INNOVATION: OUR PRIOR STUDIES REVEALED THAT THE P400 COMPLEX REGULATES HIV TRANSCRIPTION THROUGH DUAL MECHANISMS: DIRECTLY, BY ENGAGING TAT VIA THE DMAP1 SUBUNIT TO BLOCK TAT-TAR RNA INTERACTIONS AND RESTRICT P-TEFB RECRUITMENT; AND INDIRECTLY, BY ALTERING HOST TRANSCRIPTIONAL PROGRAMS THAT CONTROL T CELL ACTIVATION STATES. BUILDING ON THIS MECHANISTIC PRECEDENT AND METHODOLOGICAL PLATFORM, WE NOW FOCUS ON INO80A, SMARCA5, CHD1, AND CHD2, REMODELERS FROM DISTINCT ATPASE FAMILIES THAT GOVERN TAT-INDEPENDENT CHECKPOINTS AT INITIATION, PAUSE RELEASE, AND ELONGATION. METHODOLOGICALLY, WE WILL APPLY TURBOID-CHAP-MS (LOCUS-SPECIFIC PROTEOMICS), BEM-SEQ (SINGLE-NUCLEOSOME MAPPING), AND DEGRON-MEDIATED ACUTE DEPLETION WITH ATPASE-DEAD RESCUE TO INTERROGATE REMODELER FUNCTION WITH UNPRECEDENTED RESOLUTION. APPROACH: AIM 1 WILL DEFINE THE ATPASE REQUIREMENT AND TRANSCRIPTIONAL CHECKPOINTS REGULATED BY INO80A, SMARCA5, CHD1, AND CHD2 USING DEGRON/CRISPR PERTURBATIONS, CHIP-SEQ, NASCENT RNA PROFILING, AND NUCLEOSOME MAPPING. AIM 2 WILL CHARACTERIZE REMODELER-SPECIFIC COMPLEXES AND TAT DEPENDENCE AT THE HIV PROMOTER VIA TURBOID PROXIMITY LABELING INTEGRATED WITH CHROMATIN AFFINITY PURIFICATION-MASS SPECTROMETRY. AIM 3 WILL TEST COMBINATORIAL PERTURBATIONS IN JURKAT AND PRIMARY CD4⁺ T CELL LATENCY MODELS, INCLUDING ART-SUPPRESSED DONOR CELLS, TO IDENTIFY SYNERGISTIC “BLOCK-AND-LOCK” STRATEGIES THAT ENFORCE DURABLE PROVIRAL SILENCING. IMPACT: BY DEFINING REMODELER-SPECIFIC MECHANISMS AT DISCRETE TRANSCRIPTIONAL CHECKPOINTS AND LEVERAGING THEIR ENZYMATIC, DRUGGABLE ACTIVITIES, THIS WORK WILL ESTABLISH CHROMATIN REMODELING AS A THERAPEUTIC AXIS FOR DURABLE HIV SUPPRESSION AND FUNCTIONAL CURE.

University Of Florida

<P>THE GOALS ARE:</P><UL><LI>TO FOSTER FUNDAMENTAL CREATIVE DISCOVERIES, INNOVATIVE RESEARCH STRATEGIES, AND THEIR APPLICATIONS AS A BASIS FOR ULTIMATELY PROTECTING AND IMPROVING HEALTH;</LI><LI>TO DEVELOP, MAINTAIN, AND RENEW SCIENTIFIC HUMAN AND PHYSICAL RESOURCES THAT WILL ENSURE THE NATION'S CAPABILITY TO PREVENT DISEASE;</LI><LI>TO EXPAND THE KNOWLEDGE BASE IN MEDICAL AND ASSOCIATED SCIENCES IN ORDER TO ENHANCE THE NATION'S ECONOMIC WELL-BEING AND ENSURE A CONTINUED HIGH RETURN ON THE PUBLIC INVESTMENT IN RESEARCH; AND</LI><LI>TO EXEMPLIFY AND PROMOTE THE HIGHEST LEVEL OF SCIENTIFIC INTEGRITY, PUBLIC ACCOUNTABILITY, AND SOCIAL RESPONSIBILITY IN THE CONDUCT OF SCIENCE.</LI></UL><P>IN REALIZING THESE GOALS, THE NIH PROVIDES LEADERSHIP AND DIRECTION TO PROGRAMS DESIGNED TO IMPROVE THE HEALTH OF THE NATION BY CONDUCTING AND SUPPORTING RESEARCH:</P><UL><LI>IN THE CAUSES, DIAGNOSIS, PREVENTION, AND CURE OF HUMAN DISEASES;</LI><LI>IN THE PROCESSES OF HUMAN GROWTH AND DEVELOPMENT;</LI><LI>IN THE BIOLOGICAL EFFECTS OF ENVIRONMENTAL CONTAMINANTS;</LI><LI>IN THE UNDERSTANDING OF MENTAL, ADDICTIVE AND PHYSICAL DISORDERS; AND</LI><LI>IN DIRECTING PROGRAMS FOR THE COLLECTION, DISSEMINATION, AND EXCHANGE OF INFORMATION IN MEDICINE AND HEALTH, INCLUDING THE DEVELOPMENT AND SUPPORT OF MEDICAL LIBRARIES AND THE TRAINING OF MEDICAL LIBRARIANS AND OTHER HEALTH INFORMATION SPECIALISTS.</LI></UL>

93.855 - Allergy and Infectious Diseases Research

National Institute of Allergy and Infectious Diseases (NIAID) [HHS - NIH]

Jupiter, Florida 33458 United States

Single Zip Code

NIH Research Project Grant (Parent R01 Clinical Trial Not Allowed) (PA-25-301)