CMG2 as a target for safe and effective treatment of endometriosis-associate pain

子宫内膜异位症相关疼痛是女性使用阿片类药物的重要驱动因素。子宫内膜异位症是和 雌激素敏感性、血管生成依赖性疾病，影响约10%的育龄妇女， 在一半的慢性盆腔疼痛女性中发现。子宫内膜异位症增加了长期使用阿片类药物的可能性， 阿片类药物依赖/滥用和阿片类药物过量。子宫内膜异位症目前用NSAIDS、激素 靶向雌激素产生的治疗和手术，但这些选择对约30%的患者不是持久有效的。 患者因此，需要新的目标。 CMG 2是一种整合素样细胞外基质受体，我们已经证明它调节血管生成。是 在子宫内膜异位症组织中相对于正常子宫内膜也过表达。在小鼠模型中，已发表的工作和 我们的初步数据显示，用我们发现的CMG 2拮抗剂（PGG、PGM）治疗， 病变发生率、生长和血管瘤相关疼痛。这些观察结果表明，CMG 2可能是 一种治疗糖尿病相关疼痛的有用靶点。然而，我们对这些问题的理解存在着重大差距， CMG 2生物学会减缓任何以CMG 2为目标的开发计划。第一个关键差距是细胞 在CMG 2拮抗剂治疗后，其靶向导致疼痛减轻的类型尚不清楚。二是 子宫内膜异位症中CMG 2信号转导的分子机制尚不清楚。最后，最大限度地 CMG 2靶向的有效性和重要的安全性尚不清楚。 我们建议通过使用细胞类型特异性CMG 2敲除小鼠来填补这些空白，以鉴定细胞类型， CMG 2表达支持子宫内膜异位症病变生长和疼痛。因为CMG 2缺乏任何催化剂 结构域，我们假设CMG 2信号通过差异蛋白质相互作用。然后，将CMG 2在 将确认人类病变中的适当细胞。接下来，我们将使用邻近蛋白质组学来识别 与CMG 2 ±抑制剂差异性相互作用的下游分子。CMG 2的候选介质 然后将使用CRISPR敲除和基于细胞的CMG 2功能测定来确认信号传导。最后我们 将使用充分表征的、高- 特异性CMG 2抑制剂。在整个离体和体内测定中，RNAseq和scRNAseq将用于 鉴定CMG 2阻断的转录特征，以验证体外测定并产生潜在的 CMG 2抑制剂的药效学标志物。 完成拟议的工作将验证CMG 2作为治疗子宫内膜异位症的靶点- 相关的疼痛。它还将提供子宫内膜异位症病理生理学的关键见解， 产生新的治疗剂，并且可以鉴定用于治疗疾病的另外的靶标。发展 这类药物的使用将改善许多妇女的生活，减少使用阿片类药物治疗这种疾病， 从而改善许多人的生活。

Endometriosis-associated pain is an important driver of opioid use in women. Endometriosis is and estrogen sensitive, angiogenesis dependent disease that affects ~10% of women of childbearing age and is found in half of women with chronic pelvic pain. Endometriosis increases the likelihood of chronic opioid use, opioid dependence/abuse, and opioid overdose. Endometriosis is currently treated with NSAIDS, hormonal therapy targeting estrogen production, and surgery, but these options are not durably effective for ~30% of patients. Thus, new targets are needed. CMG2 is an integrin-like extracellular matrix receptor that we have shown regulates angiogenesis. It is also overexpressed in endometriosis tissue vs. normal endometrium. In mouse models, published work and our preliminary data show that treatment with CMG2 antagonists that we discovered (PGG, PGM) decreases lesion incidence, growth, and endometriosis-associated pain. These observations suggest that CMG2 may be a useful target for the treatment of endometriosis-associated pain. However, key gaps in our understanding of CMG2 biology would slow any development program with CMG2 as a target. The first key gap is that the cell type whose targeting resulted in decreased pain upon CMG2 antagonist treatment is unclear. Second, the molecular mechanism underlying CMG2 signaling in endometriosis is not known. Finally, both the maximum efficacy and, importantly, safety of CMG2 targeting is not known. We propose to fill these gaps by using cell-type-specific CMG2 knockout mice to identify cell types where CMG2 expression supports endometriosis lesion growth and pain. Because CMG2 lacks any catalytic domains, we hypothesize that CMG2 signals via differential protein interaction. Then, expression of CMG2 in appropriate cells in human lesions will be confirmed. Next, we will use proximity proteomics to identify downstream molecules that differentially interact with CMG2 ±inhibitor. Candidate mediators of CMG2 signaling will then be confirmed using CRISPR knockout and cell-based assays of CMG2 function. Finally, we will evaluate safety and efficacy of CMG2 targeting in a mouse model using a well-characterized, high- specificity CMG2 inhibitor. Throughout ex vivo and in vivo assays, RNAseq and scRNAseq will be used to identify transcriptional signatures of CMG2 blockade, both to validate in vitro assays and to generate potential pharmacodynamic markers of CMG2 inhibitors. Completion of the proposed work will validate CMG2 as a target for the treatment of endometriosis- associated pain. It will also provide key insights into endometriosis pathophysiology that will enable the generation of novel therapeutics and may identify additional targets for treatment of the disease. Development of such drugs will improve the lives of many women and decrease the use of opiods to treat this disease, thereby improving many lives.