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.