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

摘要 绝经后泌尿生殖系统综合征(GSM)影响大多数绝经后(PM)女性。这种综合症， PM妇女由于循环雌激素(E)水平较低所致，包括阴道刺激、性交困难和 反复尿路感染。而阴道内注射低剂量的E可以缓解症状和体征 在GSM中，只有大约10%的受影响妇女接受这种治疗。导致人们对这些内容的接受程度有限 医疗保健提供者和受GSM影响的妇女的治疗是他们的黑匣子警告，表明 增加心血管疾病和子宫内膜癌的风险。我们之前的研究表明，生殖器 PM妇女和去卵巢(OVX)小鼠的组织显示桥粒钙粘附素类似的丢失 桥粒芯糖蛋白-1(DSG1)和桥粒芯胶原-1(DSC1)是促进上皮完整性的细胞-细胞黏附分子 和屏障功能。最近，我们创建了一种EFNA3模拟激动肽(EFNA3-MAP)，并看到了它的 OVX小鼠阴道内给药恢复生殖器上皮屏障功能及其短期效应 对一种通常用于评估致癌性的小鼠进行治疗并不会导致肿瘤的发展。 目前的提案将探索三个目标，以定义EFNA3-MAP的行动机制，优化其交付 以及保护生殖器上皮屏障功能的能力，并建立长期治疗的安全概况。 第一个目标将使用OVX小鼠来证明IVAG EFNA3-MAP治疗促进生殖器上皮细胞 促进细胞增殖和分化，恢复桥粒钙粘附素表达和上皮屏障功能。 其他研究将使用OVX小鼠和商用基因敲除小鼠来定义EFNA介导的信号转导 IVAG EFNA3-MAP治疗降低生殖器HSV-2感染易感性的途径和能力。这个 第二个目标是确定两种不同的静脉注射系统的体外生物相容性和体内疗效。 增强多肽在生殖器粘膜组织中持久性的潜力。这些输送系统是壳聚糖- 基于增加多肽粘附性的凝胶和允许水凝胶和油基给药的大凝胶。 其他研究将使用OVX小鼠来描述EFNA3-MAP给药计划，以优化其恢复能力 生殖器上皮屏障功能。第三个目标将使用rash2小鼠来探索长期的致癌性。 IVAG EFNA3-MAP治疗作为该小鼠品系有效地识别促进癌症发生和 提供主要监管机构接受的致癌测试结果。这些目标加在一起将提供 开发一种新的非类固醇和非激素化合物的基本指南，该化合物可以逆转 生殖器上皮健康与循环E丢失相关，我们预计这一提议将定义EFNA3- 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计划，并建立长期安全的有利条件。 术语IVAG EFNA3-MAP管理。

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.