Project 1 - Lechner

项目总结/摘要 在导致胎儿早产性破裂的机制方面存在着关键的知识空白 膜（PPROM）。这一差距的持续存在是一个重要问题，因为， 尽管如此，预防和治疗PPROM的成功策略的发展仍将是 难以捉摸。该提案的长期目标是减少PPROM引起的早产的发生率。的 目的是确定导致PPROM的细胞外基质机制。中央 该建议的假设是细胞外基质蛋白聚糖双糖链和核心蛋白聚糖有助于 在整个妊娠期保持完整的胎膜。这一假设是基于初步数据 支持这些机制在维持妊娠中的作用。初步研究表明， 小鼠中双糖链蛋白聚糖和核心蛋白聚糖的缺乏导致早产、异常胎膜形态和 改变的信号通路，并且在暴露于炎症时表型增强。此外，委员会认为， PPROM人胎膜显示核心蛋白聚糖调节异常。拟议研究的理由 这些目标的完成将确定关键的大聚糖和核心蛋白聚糖依赖性机制， 的完整性，导致新的和创新的战略，预防和治疗 的PPROM。在强有力的初步数据的指导下，将通过追求三个具体目标来检验这一假设： 1)使用小鼠鉴定蛋白聚糖双糖链和核心蛋白聚糖对成功妊娠的贡献 缺乏这些蛋白聚糖; 2）确定细胞外基质有助于 双糖链蛋白聚糖/核心蛋白聚糖基因敲除小鼠整个妊娠期胎膜的结构完整性; 和3）阐明人类PPROM后胎膜中细胞外基质的状态。下 第一个目标，母胎差异基因型方法将用于评估细胞外基质 对胎膜稳定性的贡献。对于第二个目的，胎膜生物力学测试和 将进行重组双糖链蛋白聚糖拯救试验。第三个目的，人胎膜蛋白聚糖 表达将被评估。每种方法在申请人手中都是可行的。的 所提出的研究的贡献预计将是新的双糖链蛋白聚糖和核心蛋白聚糖的鉴定， 依赖性机制有助于防止PPROM。该研究具有创新性 因为它代表了一种新的和实质性的脱离现状的做法，即 细胞外基质蛋白聚糖对胎膜稳定性的作用 膜机械测试、蛋白多糖治疗和胚胎移植。拟议的研究是 重要的是，因为它有望垂直推进我们对细胞外基质贡献的理解， 在整个怀孕期间稳定胎膜。最终，这些知识有可能 为PPROM新的预防和治疗策略的发展提供信息。

PROJECT SUMMARY/ABSTRACT There is a key gap in knowledge with respect to mechanisms leading to preterm premature rupture of fetal membranes (PPROM). Continued existence of this gap represents an important problem because, until it is filled, the development of successful strategies for the prevention and treatment of PPROM will remain elusive. The long-term goal of this proposal is to reduce the incidence of PPROM-induced preterm birth. The objective here is to determine the extracellular matrix mechanisms that lead to PPROM. The central hypothesis for this proposal is that the extracellular matrix proteoglycans biglycan and decorin contribute to the maintenance of intact fetal membranes throughout gestation. This hypothesis is based on preliminary data that support a role for these mechanisms in maintenance of pregnancy. Preliminary studies show that the absence of biglycan and decorin in mice leads to preterm birth, abnormal fetal membrane morphology and altered signaling pathways, and that the phenotype is enhanced on exposure to inflammation. Furthermore, human fetal membranes with PPROM display decorin dysregulation. The rationale for the proposed research is that completion of these aims will define key biglycan- and decorin-dependent mechanisms that contribute to the integrity of fetal membranes, resulting in new and innovative strategies for the prevention and treatment of PPROM. Guided by strong preliminary data, this hypothesis will be tested by pursuing three specific aims: 1) Identify the contributions of the proteoglycans biglycan and decorin to successful gestation using mice deficient in these proteoglycans; 2) Identify the mechanisms by which the extracellular matrix contributes to the structural integrity of the fetal membranes throughout gestation in the biglycan/decorin knockout mouse; and 3) Elucidate the status of the extracellular matrix in fetal membranes after PPROM in humans. Under the first aim, a maternal-fetal divergent genotype approach will be used to assess the extracellular matrix contribution to fetal membrane stability. For the second aim, fetal membrane biomechanical testing and recombinant biglycan rescue testing will be performed. For the third aim, human fetal membrane proteoglycan expression will be assessed. Each approach has been established as feasible in the applicants' hands. The contribution of the proposed research is expected to be the identification of novel biglycan- and decorin- dependent mechanisms that contribute to protection against PPROM. The proposed research is innovative because it represents a new and substantive departure from the status quo, namely the approach of extracellular matrix proteoglycan contribution to the stability of the fetal membranes using knockout fetal membrane mechanical testing, proteoglycan therapy and embryo transfer. The proposed research is significant because it is expected to vertically advance our understanding of extracellular matrix contribution to the stabilization of the fetal membranes throughout pregnancy. Ultimately, such knowledge has the potential to inform the development of new preventive and therapeutic strategies for PPROM.