Regenerative Therapy for Pelvic Organ Prolapse

描述(申请人提供)：盆腔器官脱垂(POP)降低了美国许多老年女性的生活质量。基于POP病理生理学的非手术治疗的需求尚未得到满足，以预防和/或逆转这种常见的疾病。为了引领这种疗法的发展，我们建议进行一项探索性研究，以(A)评估结构性细胞外基质(ECM)的异常作为POP的病理生理学基础，并(B)建立基于诱导再生细胞外基质修复的创新治疗策略的概念证明。最近在人类和赖氨酰氧化酶样蛋白-1(LOXL1)基因敲除(KO)的数据表明，受损的ECM重塑，特别是弹性基质重塑可能与POP有关。然而，目前尚不清楚是否所有的POP患者都存在LOXL1降低和ECM动态平衡异常，以及它们是否是POP的原因或结果。由于阴道壁ECM对盆底组织的机械支持作用，即使临床病因可能是多样和多因素的，恢复产后盆底组织中阴道ECM的动态平衡可能是预防或逆转POP症状的有效途径。弹性基质通常不会在成人组织中很好地再生，因此将成为我们再生治疗的主要目标，尽管我们也将评估胶原蛋白稳态的恢复。我们建议利用POP的LOXL1基因敲除(KO)小鼠模型进行第一阶段的相关研究，以阐明这些机制方面，在此研究之外，将有助于 评估我们的基质再生方法的体内治疗效果。我们提出了两个目的来检验我们的假设，即LOXL1缺乏导致分娩后阴道ECM组装和修复的异常，与增强的基质降解与POP相关，以及诱导的细胞再生性ECM修复可以减弱或逆转这些异常。目标1a。确定LOXL1KO小鼠阴道壁结构ECM与POP相对应的动态平衡变化的时间进程。目标1b。确定LOXL1缺乏、怀孕、阴道分娩和LOXL1 KO小鼠脱垂如何改变非上皮性阴道细胞(NEVCs)新的ECM沉积。目的2.评价MRF对LOXL1-KO小鼠阴道壁组织器官培养中细胞外基质结构动态平衡指标的恢复能力。拟议的项目建立在合作伙伴PIS、Damaser(动物模型和POP病理生理学)和Ramamurthi(ECM再生)的持续合作和专业知识的基础上。这项研究将在动物模型和培养的人类阴道组织中进行临床前研究，为继续开发这种范式转换、微创再生疗法提供理由、理论基础和可行性数据。这些反过来将使临床试验合理化，以预防和/或治疗POP，从而提高许多女性的生活质量。这项研究的科学影响将是提高对POP发展机制的理解，并测试用于再生弹性基质修复和恢复ECM动态平衡的创新疗法。

DESCRIPTION (provided by applicant): Pelvic organ prolapse (POP) reduces the quality of life of many older women in the U.S. There is an unmet need for non-surgical therapies based on POP pathophysiology, to prevent and/or regress this common condition. Towards pioneering the development of such a therapy, we propose an exploratory study to (a) assess aberrations of the structural extracellular matrix (ECM) as a pathophysiological basis for POP and (b) establish proof of concept of an innovative therapeutic strategy based on induced regenerative extracellular matrix repair. Recent data in humans and lysyl oxidase like protein-1 (LOXL1) knockout (KO) suggest that impaired ECM remodeling, particularly that of elastic matrix may be linked to POP. However, it is not known if decreased LOXL1 and aberrations of ECM homeostasis, occur in all POP patients, and if they are a cause of POP or a result of it. Since vaginal wall ECM mechanically supports pelvic floor tissues, restoring homeostasis of vaginal ECM in post-partum pelvic floor tissues may be an effective approach to prevent or reverse POP symptoms even though the clinical etiology may be varied and multifactorial. Elastic matrix does not usually regenerate well in adult tissues and will therefore be our primary target for regenerative therapy, although we will assess restoration of collagen homeostasis as well. We propose to utilize a LOXL1 knockout (KO) mouse model of POP to perform a first phase correlative study to elucidate these mechanistic aspects that, beyond this study, will be useful to assess in vivo therapeutic efficacy of our matrix regenerative approach. We propose two aims to test our hypotheses that LOXL1 deficiency results in aberrations in vaginal ECM assembly & repair after pup delivery, which together with enhanced matrix degradation are associated with POP, and that these aberrations can be attenuated or reversed by induced cellular regenerative ECM repair. Aim 1a. Determine the time course of changes to the homeostasis of structural ECM of the vaginal wall corresponding to POP in LOXL1 KO mice. Aim 1b. Determine how LOXL1 deficiency, pregnancy, vaginal delivery, and prolapse in LOXL1 KO mice, alter new ECM deposition by Non-Epithelial Vaginal Cells (NEVCs). Aim 2. Assess ability of MRFs to restore indicators of structural ECM homeostasis in organ cultures of vaginal wall tissue isolated from LOXL1 KO mice with and without POP. The proposed project builds on an ongoing collaboration and expertise of the co-PIs, Damaser (animal models and POP pathophysiology) and Ramamurthi (ECM regeneration). The study will provide justification, rationale, and feasibility data to continue development of this paradigm-shifting, minimally invasive regenerative therapy with pre-clinical studies in animal models and cultured human vaginal tissue. These in turn will rationalize clinical trials to prevent and/or treat POP towards improvin quality of life for many women. The scientific impact of this study will be improved understanding of the mechanisms of POP development and testing of an innovative therapy for regenerative elastic matrix repair and restoration of ECM homeostasis.