Project Grant F30HD123655

DISSECTING THE ROLE OF EPITHELIAL-FIBROBLAST CROSSTALK IN ENDOMETRIOSIS-ASSOCIATED INFERTILITY - PROJECT SUMMARY ENDOMETRIOSIS IS A CHRONIC INFLAMMATORY CONDITION AFFECTING 10-15% OF WOMEN, CAUSING SIGNIFICANT PELVIC PAIN AND INFERTILITY. WHILE THE NORMAL ENDOMETRIUM RELIES ON TIGHTLY REGULATED, SPATIAL WNT-MEDIATED CROSSTALK BETWEEN EPITHELIAL AND FIBROBLAST CELLS TO ESTABLISH A RECEPTIVE LINING, THIS PROCESS IS DISRUPTED IN ENDOMETRIOSIS. IN DISEASED TISSUE, FIBROBLASTS ARE THOUGHT TO DRIVE PATHOLOGY BY INDUCING ABERRANT WNT SIGNALING AND PREVENTING THE ENDOMETRIUM FROM PROPERLY TRANSITIONING INTO A RECEPTIVE STATE. THE GOAL OF THIS PROPOSAL IS TO LEVERAGE A MECHANISTIC UNDERSTANDING OF THIS SPATIAL EPITHELIAL-FIBROBLAST CROSSTALK TO DEVELOP NEW THERAPEUTIC STRATEGIES FOR ENDOMETRIOSIS-ASSOCIATED INFERTILITY. MY PRELIMINARY SPATIAL TRANSCRIPTOMIC DATA REVEAL THAT WNT LIGANDS AND ANTAGONISTS FORM SPECIFIC EXPRESSION GRADIENTS IN FIBROBLASTS, A CELL TYPE DISRUPTED IN ENDOMETRIOSIS. I HYPOTHESIZE THAT THIS DYSREGULATED WNT- MEDIATED CROSSTALK DRIVES ABNORMAL EPITHELIAL DIFFERENTIATION, THEREBY IMPAIRING ENDOMETRIAL RECEPTIVITY. AIM 1 WILL CHARACTERIZE THE EPITHELIAL RESPONSE TO ENDOMETRIOSIS-DERIVED FIBROBLASTS USING PATIENT-DERIVED 3D CO-CULTURES AND SINGLE-CELL RNA SEQUENCING. I WILL SUBSEQUENTLY USE CRISPR-CAS9 GENE EDITING TO SYSTEMATICALLY DEMONSTRATE THE CONTRIBUTION OF SPECIFIC WNT GENES TO THE INFERTILITY PHENOTYPE. AIM 2 WILL SPATIALLY VALIDATE THESE INTERACTIONS IN HUMAN PRE-RECEPTIVE AND RECEPTIVE ENDOMETRIAL BIOPSIES USING HIGH-RESOLUTION SPATIAL TRANSCRIPTOMICS TO PROFILE CELL NEIGHBORHOODS AND SIGNALING NICHES. SUCCESSFUL COMPLETION OF THESE AIMS WILL ELUCIDATE THE PATHOLOGICAL NICHES UNIQUE TO ENDOMETRIOSIS-ASSOCIATED INFERTILITY AND IDENTIFY POTENTIAL THERAPEUTIC TARGETS. HOWEVER, THIS F30 FELLOWSHIP WILL ULTIMATELY PROVIDE ME WITH A UNIQUE RESEARCH FRAMEWORK TO ELUCIDATE MECHANISTIC INSIGHTS ACROSS MANY GYNECOLOGICAL CONDITIONS THROUGH TRANSLATABLE SKILLS IN ADVANCED IN VITRO ORGAN MODELING AND SINGLE-CELL TRANSCRIPTOMICS/SPATIAL OMICS. THEREFORE, THIS TRAINING IS IMPERATIVE FOR MY DEVELOPMENT AS A FUTURE INDEPENDENT PHYSICIAN-SCIENTIST WHO AIMS TO ADDRESS THE BENCH TO BEDSIDE GAPS IN WOMEN'S HEALTH.