Molecular mechanisms of gap junction promotion of lesion formation in Endometriosis

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ABSTRACT This request for a Supplement to RO1HD109027 is in response to RFA NOT-OD-22-208 that addresses several of the priority ORWH Strategic Plan Objectives related to disparities in Women’s Health, specifically: 1.5 (Research on gynecological health, including infertility): Endometriosis is a major issue in gynecological health affecting 10% of women worldwide, of whom as many as 50% suffer problems of infertility 2.2 (Development of biomarkers for women’s health): The current work arises from our development of a much- needed non-surgical diagnostic that is being developed by Hera Biotech (co-founded by the PI and Co-I), who are sponsoring the clinical trial which will provide material for study here to assess if the diagnostic is equally effective across different ethnic groups, and if it reveals different severity of disease. 2.4 (Examination of health disparities stemming from factors such as race and ethnicity). We will expand functional tests on patient endometrial cells proposed in the parent RO1 to definitively test if there are Biological or epigenetic differences that could explain the 3-fold lower rate of diagnosis and 2 fold lower hospitalizations for endometriosis in the Hispanic (and Black) populations compared to non-Hispanic Whites. Absence of such evidence will provide the first hard data to support efforts to address the socioeconomic inequities responsible. The work proposed will utilize our expertise in single cell analysis (Co-I is Director of the Bioanalytics & Single Cell Facility) and ex-vivo functional analyses of cell interactions and invasiveness developed in the Nicholson lab under the parent RO1. We will also utilize our unique S. Texas Ob/Gyn repository of separated endometrial stromal and epithelial cells, which will be trebled in size through the on-going Hera Proof-of-Concept trial. With ~30% of samples being of Hispanic origin, this allows us to conduct the first ever in-depth comparison of gene expression patterns (Aim 1) and cellular phenotypes (Aim 2) between Hispanic and non-Hispanic white control and endometriosis patients. The scope of this short-term study will be kept manageable by focusing in Aim 1 on proteins in the “cell-interactome” – cell components involved in intercellular communication (gap junctions), contact (tight and adhesion junctions), accessory components and regulatory factors (including mediators of epithelial to mesenchymal transition). The Aim 2 functional studies will have a similar focus, concentrating on peritoneal invasiveness in a 3-D ex-vivo model using established and primary mesothelial cells, and measurements of intercellular coupling via gap junctions, as we have demonstrated this to be required for invasion. Should differences be detected, they will provide important guidelines for future personalized therapeutics. Should no differences be observed, it will provide a compelling, data-driven mandate to address the socio-economic or unconscious bias issues that contribute to ethnic disparities in endometriosis treatment.