Regenerative Therapy for Pelvic Organ Prolapse

描述（由申请人提供）：盆腔器官脱垂（POP）降低了美国许多老年女性的生活质量。目前，对基于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/或消退这种常见疾病。为了开创这种疗法的发展，我们提出了一项探索性研究，以（a）评估结构性细胞外基质（ECM）的畸变作为POP的病理生理学基础，（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（LOXL 1）敲除（KO）中的数据表明受损的ECM重塑，特别是弹性基质的重塑可能与POP有关。然而，尚不清楚LOXL 1减少和ECM稳态畸变是否发生在所有POP患者中，以及它们是否是POP的原因或结果。恢复产后骨盆底组织中阴道ECM的稳态可能是预防或逆转POP症状的有效方法，即使临床病因可能是多种多样的和多因素的。弹性基质在成人组织中通常不能很好地再生，因此将是我们再生治疗的主要目标，尽管我们也将评估胶原稳态的恢复。我们建议利用LOXL 1基因敲除（KO）的POP小鼠模型进行第一阶段的相关研究，以阐明这些机制方面，超出本研究，将有助于 评估我们的基质再生方法的体内治疗功效。 我们提出了两个目的来测试我们的假设，即LOXL 1缺陷导致幼鼠分娩后阴道ECM组装和修复中的畸变，这与增强的基质降解一起与POP相关，并且这些畸变可以通过诱导的细胞再生ECM修复来减弱或逆转。目标1a。确定LOXL 1 KO小鼠中对应于POP的阴道壁的结构ECM的稳态变化的时间过程。目标1b。确定LOXL 1缺陷、妊娠、阴道分娩和LOXL 1基因敲除小鼠的脱垂如何改变非上皮阴道细胞（NEVC）的新ECM沉积。目标二。评估MRF恢复从LOXL 1 KO小鼠分离的阴道壁组织器官培养物中结构ECM稳态指标的能力，有和没有POP. The拟议的项目建立在co-PI，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。该研究将提供合理性、基本原理和可行性数据，以继续开发这种范式转变、微创再生疗法，并在动物模型和培养的人类阴道组织中进行临床前研究。这些反过来将使预防和/或治疗POP的临床试验合理化，以改善许多妇女的生活质量。这项研究的科学影响将提高对POP发展机制的理解，并测试再生弹性基质修复和ECM稳态恢复的创新疗法。

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.