Project Grant F31CA318519

SPATIOTEMPORAL REGULATION OF MEMBRANE RECEPTOR ACTIVATION AND CROSSTALK: SINGLE-MOLECULE IMAGING OF RECEPTOR CLUSTERING AND DYNAMICS THAT DRIVE ONCOGENIC SIGNALING. - PROJECT SUMMARY EGFR AND HER2 RECEPTOR TYROSINE KINASES (RTKS) ARE CRITICAL THERAPEUTIC TARGETS IN NUMEROUS CANCERS, YET THE INEVITABLE DEVELOPMENT OF DRUG RESISTANCE SEVERELY LIMITS TREATMENT EFFICACY. RECENT STUDIES SUGGEST THAT THE NANOSCALE CLUSTERING OF THESE RECEPTORS AT THE PLASMA MEMBRANE IS A KEY REGULATOR OF SIGNALING OUTCOMES, BUT THE PRECISE MECHANISMS LINKING THIS SPATIAL ORGANIZATION TO SPECIFIC SIGNALING MECHANISMS AND DRUG RESISTANCE REMAIN POORLY UNDERSTOOD. THIS FELLOWSHIP WILL SUPPORT COMPREHENSIVE TRAINING IN ADVANCED SINGLE-MOLECULE IMAGING TECHNIQUES AND CANCER BIOLOGY, WHILE INVESTIGATING HOW RECEPTOR CLUSTERING DYNAMICS, HETERODIMER INTERACTIONS, AND CROSSTALK BETWEEN DISPARATE RECEPTORS CONTROL DIFFERENTIAL SIGNALING RESPONSES. THE PROPOSED RESEARCH TESTS THE HYPOTHESIS THAT SPECIFIC ACTIVATING CONDITIONS DICTATE RECEPTOR DIFFUSION RATES, DIMERIZATION KINETICS, AND OLIGOMERIZATION STATES, WHICH ENCODE SIGNALING SPECIFICITY. AIM 1 WILL DEFINE HOW EGFR LIGANDS (EGF, EREG, EPGN) AND ONCOGENIC MUTATIONS (L858R) ALTER NANOSCALE CLUSTERING USING DNA-PAINT SUPER-RESOLUTION MICROSCOPY, CORRELATING RECEPTOR CLUSTERING PATTERNS WITH RECRUITMENT OF SIGNALING MEDIATORS GRB2 AND PI3K. AIM 2 WILL QUANTIFY EGFR/HER2 INTERACTION DYNAMICS, INCLUDING THE ONCOGENIC HER2-S310F MUTANT, USING TWO-COLOR QUANTUM DOT TRACKING TO MEASURE HETERODIMER OFF-RATES AND DNA-PAINT TO ASSESS HETERO-CLUSTER FORMATION. AIM 3 WILL EXPLORE EGFR/TGF-B RECEPTOR CROSSTALK, A POTENTIAL MECHANISM FOR THERAPEUTIC ESCAPE. I WILL DETERMINE WHETHER THIS CROSSTALK IS A RESULT OF DIRECT EGFR/TBRII INTERACTIONS AT THE PLASMA MEMBRANE THROUGH SINGLE PARTICLE TRACKING AND CO-CLUSTERING ANALYSIS. TOGETHER, THE OUTCOMES WILL REVEAL MOLECULAR MECHANISMS THAT CONTRIBUTE TO ONCOGENIC SIGNALING AND THERAPEUTIC RESISTANCE. THE FELLOWSHIP TRAINING PLAN ENCOMPASSES FOUR GOALS: GOAL 1 INVOLVES MASTERING SINGLE-MOLECULE IMAGING TECHNIQUES INCLUDING DNA-PAINT AND QUANTUM DOT TRACKING. GOAL 2 FOCUSES ON DEEPENING EXPERTISE THROUGH COMPUTATIONAL AND STRUCTURAL BIOLOGY COURSEWORK AND COLLABORATION WITH STRUCTURAL BIOLOGISTS. GOAL 3 DEVELOPS PROFESSIONAL SKILLS THROUGH GRANT WRITING, REVIEW ARTICLES, AND MENTORING. GOAL 4 BUILDS PEDAGOGICAL EXPERTISE THROUGH THE CERTIFICATE IN UNIVERSITY SCIENCE TEACHING. THE UNIVERSITY OF NEW MEXICO PROVIDES EXCEPTIONAL RESOURCES FOR SINGLE-MOLECULE BIOPHYSICS RESEARCH WITH STATE-OF-THE-ART IMAGING FACILITIES AND ESTABLISHED EXPERTISE IN RECEPTOR BIOLOGY. MY BACKGROUND SPANNING CLINICAL EXPERIENCE, ANALYTICAL CHEMISTRY, AND TRANSLATIONAL RESEARCH PROVIDES STRONG PREPARATION FOR BRIDGING FUNDAMENTAL BIOPHYSICS TO THERAPEUTIC APPLICATIONS. THIS FELLOWSHIP WILL SUPPORT THE CANDIDATE'S GOAL OF LEADING AN INDEPENDENT RESEARCH PROGRAM ADDRESSING THERAPEUTIC RESISTANCE IN CANCER.