Developing a nonsteroidal and nonhormonal agent that reverses menopause-related loss of genital epithelial integrity and function

摘要 绝经期泌尿生殖综合征（GSM）影响大多数绝经后（PM）妇女。这种综合症， PM女性中循环雌激素（E）水平较低引起的阴道刺激，包括阴道刺激，性交困难， 复发性尿路感染虽然阴道内施用低剂量的E减轻了症状和体征， 在GSM中，只有大约10%的受影响妇女接受这种治疗。有助于有限地接受这些 受GSM影响的医疗保健提供者和妇女的治疗是他们的黑盒警告， 心血管疾病和子宫内膜癌的风险增加。我们以前的研究表明，生殖器 PM女性和卵巢切除（OVX）小鼠的组织显示出相当的桥粒钙粘蛋白损失 桥粒芯糖蛋白-1（DSG 1）和桥粒芯胶蛋白-1（DSC 1），促进上皮完整性的细胞-细胞粘附分子 屏障功能。最近，我们创建了EFNA 3模拟激动剂肽（EFNA 3-MAP），并观察了其 对OVX小鼠阴道内（ivag）给药恢复生殖器上皮屏障功能， 在通常用于评估致癌性的小鼠品系中的治疗不诱导肿瘤发展。 目前的提案将探讨三个目标，以确定EFNA 3-MAP的作用机制，优化其交付， 和保护生殖器上皮屏障功能的能力，并建立长期治疗的安全性。 第一个目标将使用OVX小鼠来证明ivag EFNA 3-MAP治疗促进生殖器上皮细胞增殖。 增殖和分化，并恢复桥粒钙粘蛋白表达和上皮屏障功能。 其他研究将使用OVX小鼠和市售敲除小鼠来定义EFNA介导的信号传导 ivag EFNA 3-MAP治疗降低生殖器HSV-2感染易感性的途径和能力。的 第二个目的是确定两种独立IVAG递送系统的体外生物相容性和体内功效， 增强生殖器粘膜组织中肽持久性的潜力。这些输送系统是壳聚糖- 增加肽粘膜粘附的基于凝胶和允许水性水凝胶和基于油的递送的bigel。 其他研究将使用OVX小鼠来描述EFNA 3-MAP给药方案，该方案优化了其恢复 生殖器上皮屏障功能第三个目标将使用rasH 2小鼠来探索长期的致癌性。 ivag EFNA 3-MAP治疗作为该小鼠品系有效鉴定促进致癌作用的物质， 提供主要监管机构认可的致癌性测试结果。这些目标将共同提供 开发一种新的非甾体和非激素化合物的基本指导， 生殖器上皮健康与循环E的损失相关，我们预计这项提案将定义EFNA 3- MAP介导的生殖器上皮完整性和屏障功能的调节，确定特定的递送平台 以及优化生殖器上皮健康的给药方案，并建立长期的有利安全性特征， 长期ivag EFNA 3-MAP给药。

ABSTRACT The genitourinary syndrome of menopause (GSM) affects most postmenopausal (PM) women. This syndrome, caused by lower levels of circulating estrogen (E) in PM women, includes vaginal irritation, dyspareunia, and recurrent urinary tract infection. While intravaginal administration of low-dose E alleviates signs and symptoms of GSM, only about 10% of affected women receive this treatment. Contributing to limited acceptance of these therapies by healthcare providers and women affected by the GSM are their black box warnings denoting an increased risk of cardiovascular disease and endometrial cancer. Our research previously showed that genital tissue from PM women and ovariectomized (OVX) mice display comparable loss of the desmosomal cadherins desmoglein-1 (DSG1) and desmocollin-1 (DSC1), cell-cell adhesion molecules that promote epithelial integrity and barrier function. More recently we created an EFNA3-mimetic agonist peptide (EFNA3-MAP) and saw its intravaginal (ivag) administration to OVX mice restore genital epithelial barrier function and that its short-term treatment in a strain of mouse commonly used to assess carcinogenicity does not induce tumor development. The current proposal will explore three aims to define EFNA3-MAP mechanism of action, optimize its delivery and ability to preserve genital epithelial barrier function, and establish the safety profile of long-term treatment. The first aim will use OVX mice to demonstrate that ivag EFNA3-MAP treatment promotes genital epithelial proliferation and differentiation and restores desmosomal cadherin expression and epithelial barrier function. Other studies will use OVX mice and commercially available knockout mice to define EFNA-mediated signaling pathways and capacity of ivag EFNA3-MAP treatment to reduce susceptibility to genital HSV-2 infection. The second aim will define in vitro biocompatibility and in vivo efficacy of two separate ivag delivery systems with the potential to enhance peptide persistence in genital mucosal tissue. These delivery systems are chitosan- based gels that increase peptide mucoadhesion and bigels that allow aqueous hydrogel and oil-based delivery. Other studies will use OVX mice to delineate EFNA3-MAP dosing schedules that optimize its ability to restore genital epithelial barrier function. The third aim will use rasH2 mice to explore the carcinogenicity of long-term ivag EFNA3-MAP treatment as this mouse strain effectively identifies agents that promote carcinogenesis and provides carcinogenicity testing results accepted by key regulatory agencies. Together, these aims will provide essential guidance for developing a novel nonsteroidal and nonhormonal compound that reverses the loss of genital epithelial health associated with loss of circulating E, and we expect this proposal will define EFNA3- MAP-mediated regulation of genital epithelial integrity and barrier function, identify specific delivery platforms and dosing schedules that optimize genital epithelial health, and establish a favorable safety profile for long- term ivag EFNA3-MAP administration.