Estrogen reverses progestin-mediated loss of genital mucosal barrier function

雌激素逆转孕激素介导的生殖器粘膜屏障功能丧失

• 批准号: 10024512
• 负责人: Thomas L. Cherpes
• 金额: $ 41.05万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-09-01 至 2023-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10024512
• 关键词: Affect; Biological; Cell Adhesion Molecules; Cell-Cell Adhesion; Cells; Clinical Research; Combined Modality Therapy; Confocal Microscopy; Contraceptive Agents; Contraceptive methods; Cream; Data; Development; Dextrans; Dose; Drug usage; Ephrin-A3; Epithelial; Epithelium; Estrogen Receptors; Estrogens; Foundations; Genes; Genital system; HIV; HIV Infections; HIV-1; HIV-1 load; High Risk Woman; Host Defense; Human; Human Herpesvirus 2; Impairment; Infection; Infectious Agent; Injectable; Left; Life; Macaca; Macaca mulatta; Mediating; Medroxyprogesterone 17-Acetate; Modeling; Molecular; Mucous Membrane; Mus; Pathway interactions; Penetration; Peripheral Blood Mononuclear Cell; Permeability; Pharmaceutical Preparations; Pharmacology; Placebos; Plasma; Population; Predisposition; Progestins; Quantitative Reverse Transcriptase PCR; Regulation; Research; SIV; Serum; Signal Pathway; Systemic infection; Texas red; Tissues; Vagina; Vaginal Ring; Virus; Woman; Work; clinically relevant; desmoglein 1; high risk; hormonal contraception; humanized mouse; lucifer yellow; microscopic imaging; mouse model; prevent; sexually transmitted virus; transmission process

SUMMARY HIV affects more women than any other life-threatening infectious agent. It is most often sexually transmitted, where virus must evade the genital mucosal barrier to cause systemic infection. Clinical studies suggest HIV more easily penetrates this defense among women using the injectable progestin depot-medroxyprogesterone acetate (DMPA). Offering biological plausibility for this possibility, our research group showed that DMPA promoted mouse susceptibility to HSV-2 infection by reducing expression of the cell-cell adhesion molecule desmoglein-1 (DSG1) and increasing genital mucosal permeability. DMPA similarly boosted susceptibility of humanized mice to genital HIV infection. Identifying a potential mechanism for these results, we found DMPA treatment of mice lowered vaginal levels of ephrin A3 (EFNA3); an estrogen (E) receptor target gene shown to promote DSG1 expression in epithelial tissue. Providing clinical relevance for our findings, we showed women initiating DMPA use display changes in ectocervical DSG1 expression and mucosal permeability identical to those seen in mice. Likewise, pharmacologically relevant DMPA doses comparably enhanced genital mucosal permeability in rhesus macaques (RMs). These data newly reveal DMPA impairs mucosal barrier protection. However, using a humanized mouse model of HIV infection, we also uncovered that combined treatment with DMPA and intravaginal (ivag) E blocks virus acquisition by enhancing genital mucosal integrity. These results thus identified an unexpected advantage of hormonal contraceptive strategies that combine use of exogenous progestin and E (i.e., they avert loss of barrier protection caused by progestin use alone). As necessary steps in establishing this approach, this proposal will elucidate mechanisms of E-mediated enhancement of genital mucosal barrier function and define capacity of an E-releasing ivag ring (E-IVR) to protect RMs from genital SIV transmission. Expressly, we will use mice to define E-mediated regulation of EFNA3 pathways that induce DSG1 expression and boost genital mucosal barrier function (Aim 1). We will formulate a RM-sized E-IVR to deliver pharmacologically relevant drug doses, and use these rings to compare EFNA3 and DSG1 expression and mucosal barrier function in RM treated with DMPA, DMPA and placebo IVR, or DMPA and E-IVR (Aim 2). Finally, we will compare genital SIV transmission in RMs administered DMPA, DMPA and placebo IVR, or DMPA and E-IVR with repetitive genital challenges with escalating inoculums of the virus (Aim 3). This work will identify mechanisms by which E induces EFNA3 signaling pathways promoting genital epithelial integrity, and demonstrate that E-IVRs block SIV transmission by abrogating DMPA-mediated weakening of genital mucosal barrier function. These studies will thus deliver important new information in a highly relevant clinical model, and justify exploration of similar contraceptive strategies in populations at high risk for HIV acquisition.

概括 艾滋病毒比其他威胁生命的感染毒剂影响更多的女性。它通常是性传播的， 病毒必须逃避生殖器粘膜屏障以引起全身感染。临床研究表明HIV 使用可注射的孕激素 - 甲状腺酸盐剂，可以更容易地在女性中穿透这种防御 醋酸盐（DMPA）。为这种可能性提供生物学合理性，我们的研究小组表明DMPA 通过降低细胞细胞粘附分子的表达来促进小鼠对HSV-2感染的敏感性 Desmoglein-1（DSG1）和生殖器粘膜渗透性的增加。 DMPA同样增强了 人源化小鼠生殖器HIV感染。确定这些结果的潜在机制，我们发现了DMPA 对小鼠的治疗降低了阴道水平的源代蛋白A3（EFNA3）；雌激素（E）受体靶基因显示为 促进上皮组织中的DSG1表达。为我们的发现提供临床相关性，我们向女性展示了 启动DMPA的使用显示在邻域DSG1表达和粘膜渗透性的变化与 那些在老鼠中看到的。同样，药理学相关的DMPA剂量相当增强的生殖器粘膜 恒河猕猴（RMS）中的渗透性。这些数据新揭示了DMPA会损害粘膜屏障保护。 但是，使用人源化的HIV感染小鼠模型，我们还发现了将治疗与 DMPA和腔内（IVAG）E通过增强生殖器粘膜完整性来阻断病毒的获取。这些结果 因此确定了荷尔蒙避孕策略的意外优势，这些策略结合了外源的使用 孕激素和E（即，它们避免仅使用孕激素造成的屏障保护损失）。作为必要的步骤 在建立这种方法时，该提案将阐明电子介导的生殖器增强的机制 粘膜屏障功能并定义E释放IVAG环（E-IVR）的能力，以保护RMS免受生殖器 SIV传输。明确地，我们将使用小鼠定义诱导EFNA3途径的电子介导的调节 DSG1表达和增强生殖器粘膜屏障功能（AIM 1）。我们将制定RM大小的E-IVR 提供药理学相关的药物剂量，并使用这些环比较EFNA3和DSG1表达 DMPA，DMPA和安慰剂IVR或DMPA和E-IVR处理的RM中的粘膜屏障功能（AIM 2）。 最后，我们将比较施用DMPA，DMPA和安慰剂IVR的RMS生殖器SIV传播或 DMPA和E-IVR具有重复的生殖器挑战，随着病毒的接种升级（AIM 3）。这项工作 将确定E诱导EFNA3信号通路促进生殖器上皮完整性的机制， 并证明E-IVRS通过废除DMPA介导的生殖器削弱来嵌段SIV传播 粘膜屏障功能。因此，这些研究将在高度相关的临床中传递重要的新信息 模型，并证明对艾滋病毒收购高风险的人群中类似的避孕策略的探索是合理的。