Molecular mechanisms of gap junction promotion of lesion formation in Endometriosis

摘要 本RO 1HD 109027补充申请是对RFA NOT-OD-22-208的回应，该申请涉及几个问题 * ORWH战略计划中与妇女健康差距有关的优先目标，具体而言： 1.5（研究妇科健康，包括不孕）：子宫内膜异位症是妇科的一个主要问题， 全世界10%的妇女受到健康问题的影响，其中多达50%的妇女患有不孕症 2.2（开发妇女健康生物标志物）：目前的工作源于我们开发了一个多- 需要由Hera Biotech（由PI和Co-I共同创立）开发的非手术诊断， 正在赞助临床试验，该试验将为这里的研究提供材料，以评估诊断是否等同于 在不同的种族群体中有效，如果它揭示了不同的疾病严重程度。 2.4（审查由种族和族裔等因素引起的健康差异）。我们将扩大 在母RO 1中建议对患者子宫内膜细胞进行功能测试，以明确测试是否存在生物学 或表观遗传差异，可以解释3倍的诊断率和2倍的住院率， 与非西班牙裔白人相比，西班牙裔（和黑人）人群的子宫内膜异位症。没有这种 证据将提供第一批硬数据，以支持努力解决造成这种不平等的社会经济问题。 拟议的工作将利用我们在单细胞分析方面的专业知识（Co-I是生物分析和 单细胞设施）和离体功能分析的细胞相互作用和侵袭力， 尼科尔森实验室在母RO 1下。我们还将利用我们独特的S。得克萨斯州妇产科分离 子宫内膜间质和上皮细胞，通过正在进行的Hera概念验证， 审判大约30%的样本是西班牙裔，这使我们能够进行有史以来第一次深入的比较。 西班牙裔和非西班牙裔白色人之间的基因表达模式（目标1）和细胞表型（目标2） 对照组和子宫内膜异位症患者。这项短期研究的范围将保持可控，重点是 目的1：研究细胞间通讯（gap）中的蛋白质 连接）、接触（紧密连接和粘附连接）、辅助成分和调节因子（包括 上皮向间质转化的介质）。Aim 2功能研究将有类似的重点， 集中于使用建立的和原代间皮细胞的3-D离体模型中的腹膜侵袭， 以及通过间隙连接测量细胞间偶联，因为我们已经证明了这是必要的， 入侵如果发现差异，它们将为未来的个性化提供重要的指导方针。 治疗学如果没有观察到差异，它将提供一个令人信服的、数据驱动的任务， 导致子宫内膜异位症治疗中种族差异的社会经济或无意识偏见问题。

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.