Evaluating the role of human cervical smooth muscle cells in normal and premature cervical remodeling

摘要： 自发早产(SPTB)仍然是一个严重的产科困境，给美国带来了巨大的成本。 医疗保健。SPTB的病因各有不同，但最终的共同途径与过早的宫颈重塑有关。 (聚合酶链式反应)、缩短和扩张。尽管这一健康问题很重要，但正常和 人类体内的聚合酶链式反应还不是很清楚。例如，宫颈平滑肌细胞(CSMC)在 宫颈重塑仍不清楚。在过去的五年里，在我杰出的指导下 导师们，我参与了宫颈重塑的研究，并帮助建立了协作式宫颈 哥伦比亚大学研究小组(CCRG)。CCRG的目标是解决以下复杂问题 来自真正多学科方法的聚合酶链式反应。我最初的工作确定了内部操作系统的CSMC是 圆周定向(类似于“括约肌”)，宫颈组织从内口收缩作为反应。 催产素。我认为CSMCs可能有两个关键功能：收缩和/或ECM重塑。AS 妊娠增长时，胎儿生长的压力会伸展到宫颈。我假设CSMC 拉伸导致收缩(以保持括约肌张力)和/或ECM重塑。此外，我假设 患有PCR的女性表现出异常的颈椎伸展反应、收缩力和/或ECM重塑。在……里面 目的1，我将拉伸妊娠宫颈组织活检和来自有(研究组)和无(研究组)妇女的CSMC 聚合酶链式反应(对照，CTL)以确定1)拉伸是否引起宫颈收缩，2)如果宫颈和CSMC 患有PCR症的女性表现出异常的收缩能力和3)潜在的收缩机制，这可能解释 收缩功能缺陷。在目标2中，我将调查患有聚合酶链式反应的妇女的宫颈组织和CSMC是否表现出 异常拉伸反应导致基质金属蛋白酶活性增加，胶原周转/细胞外基质重塑 与CTL相比，以及2)如果炎症进一步增强了拉伸诱导的基质金属蛋白酶的激活/ECM 改建。在目标3中，我将评估在转录组中是否存在拉伸诱导的分子表型变化 患有PCR型和CTL型的女性CSMC水平特别关注收缩和细胞外基质重塑 小路。为了这个目标，我一直在收集有聚合酶链式反应病史的孕妇的组织 来自与胎龄匹配的CTL，使用IRB批准的方案。如果患有聚合酶链式反应的女性有CSMC， 伸展时不能收缩并保持“括约肌样”语调和/或2)表现出异常的ECM 对拉伸的重塑反应导致宫颈机械性变弱，这可能解释了为什么宫颈 最终失败，导致肺结核。在我的导师和CCRG团队的指导下，这个奖项将让我 扩展我的实验知识基础和调查技能，这样我就可以实现我的最终目标 成为一名成功而独立的临床科学家，专注于预防聚合酶链式反应和肺结核。

ABSTRACT: Spontaneous preterm birth (sPTB) remains a significant obstetric dilemma with enormous costs to U.S. healthcare. The etiology of sPTB varies, but the final common pathway involves premature cervical remodeling (PCR), shortening and dilation. Despite the importance of this health issue, the pathophysiology of normal and PCR in humans is not well understood. For instance, the role of cervical smooth muscle cells (CSMCs) in cervical remodeling remains unknown. Over the last five years, under the guidance of my outstanding mentors, I have engaged in studies of cervical remodeling and helped to establish the Collaborative Cervix Research Group (CCRG) at Columbia University. The goal of the CCRG is to tackle the complex problem of PCR from a truly multidisciplinary approach. My initial work established that CSMCs at the internal os are circumferentially oriented (similar to a “sphincter”) and cervical tissue from the internal os contracts in response to oxytocin. I propose that CSMCs may have two key functions: contraction and/or ECM remodeling. As pregnancy grows, the pressure from the growing fetus applies stretch to cervix. I hypothesize that CSMC stretch induces contraction (to maintain sphincter tone) and/or ECM remodeling. Further, I hypothesize that women with PCR exhibit abnormal cervical stretch responses, contractile force and/or ECM remodeling. In Aim 1, I will stretch pregnant cervical tissue biopsies and CSMCs from women with (study group) and without PCR (controls, CTL) to establish 1) if stretch induces cervical contractility, 2) if cervices and CSMCs from women with PCR exhibit aberrant contractility and 3) potential contractile mechanisms which may explain the contractility defect. In Aim 2, I will investigate if cervical tissue and CSMCs from women with PCR 1) exhibit abnormal stretch responses resulting in increased MMP activation, collagen turnover/ECM remodeling compared to CTLs and 2) if inflammation further enhances this stretch-induced MMP activation/ECM remodeling. In Aim 3, I will evaluate if stretch-induced molecular phenotypic changes exist at the transcriptome level in CSMC from women with PCR vs CTL with particular attention to contractility and ECM remodeling pathways. Toward this goal, I have been collecting tissue from pregnant women with a history of PCR and from gestational age-matched CTLs using IRB approved protocols. If women with PCR have 1) CSMCs that cannot contract and maintain “sphincter-like” tone in response to stretch and/or 2) exhibit abnormal ECM remodeling responses to stretch resulting in a mechanically weaker cervix, this may explain why the cervix ultimately fails leading to sPTB. With the guidance of my mentors and CCRG team, this award will allow me to expand my experimental knowledge base and investigational skills so I may achieve my ultimate goal of becoming a successful and independent clinician scientist whose focus is to prevent PCR and PTB.