Amnion membrane organ-on-chip for modeling intra-amniotic infection

项目摘要 羊膜内感染，也被称为绒毛膜羊膜炎，是早产的一个主要病因。 胎膜破裂(PPROM)，导致早产。尽管它的流行和严重的后果， 由于缺乏易驯服的人类，羊膜内感染的病理机制尚未完全了解。 相关模型。尽管早产的动物模型已经成功地被开发出来用于测试 羊膜内感染的医疗干预，对于动态定量研究来说，它们仍然是次优的 宫腔内细菌-羊膜相互作用。早产人类羊膜样本的稀缺， 特别是从妊娠早期/中期，也阻止了这些人体组织作为实验模型 探讨羊水内感染及其与胎膜早破的功能联系。总而言之，迫切需要 定量的、易处理的、与人类相关的羊膜模型，以促进对基础疾病的理解 羊膜内感染。 NIH R21项目的主要目标是通过以下方式具体满足这一重要的技术需求 开发一种与人类相关的羊膜模型，能够真实地概括 细菌和羊膜组织，同时，允许高分辨率、定量的实验 目的：研究细菌侵入羊膜腔的机制。在我们的初步研究中，我们有 意外发现人类多能干细胞(HPSCs)具有羊膜分化潜能 成功开发了基于hPSC的合成微流控胚胎发生平台，在该平台中 可以概括人类植入后早期发育的发育里程碑 相继以高度可控和可扩展的方式。重要的是，我们还观察到敏感的 HPSC来源的羊水细胞对细菌感染的炎症反应。因此，在这项研究中，我们建议 利用hPSCs的羊膜分化潜力，结合创新的微流体，以 开发首个人羊膜器官片上系统。我们将进一步应用这一易驾驭的 一种定量研究细菌侵入羊膜腔动态的实验系统 阐明炎症诱导的羊膜重塑与羊膜内出血之间的功能联系 羊水细菌贩运。这项拟议研究的成功完成将带来创新 可控、可再生和可扩展的人羊膜制造技术和方法 膜组织，为研究相关妊娠并发症提供了一个易于处理的实验系统， 包括羊膜内感染。人羊膜上器官移植的重复性和可扩展性 芯片系统将使其成为探索人类之间复杂相互作用的一个很有前途的筛选平台 羊膜、细菌病原体、药物和有毒物质。

Project Summary Intra-amniotic infection, also referred to as chorioamnionitis, is a major etiological factor of preterm premature rupture of the membranes (pPROM), leading to preterm birth. Despite its prevalence and grave consequences, the pathology of intra-amniotic infection has yet to be completely understood due to a lack of tractable human- relevant models. Even though animal models of preterm birth have been successfully developed for testing medical interventions of intra-amniotic infection, they remain suboptimal for quantitative studies of dynamic bacterium-amnion interactions in the intrauterine cavity. The scarcity of preterm human amnion samples, especially from early/mid-gestation stages, also prevents these human tissues as experimental models for studying intra-amniotic infection and its functional link to pPROM. Altogether, there is a critical need for quantitative, tractable, human-relevant amnion membrane models for advancing fundamental understanding of intra-amniotic infection. The primary goal of this NIH R21 project is to specifically address this significant technological need, by developing a human-relevant amnion membrane model that can faithfully recapitulate the interaction between bacteria and amnion membrane tissues, and at the same time, allow high-resolution, quantitative experiments to study mechanisms underlying bacterial invasion of the amniotic cavity. In our preliminary study, we have unexpectedly discovered the amniogenic differentiation potency of human pluripotent stem cells (hPSCs) and successfully developed an hPSC-based, synthetic microfluidic embryogenesis platform in which key developmental landmarks during early human post-implantation development can be recapitulated successively in a highly controllable and scalable fashion. Importantly, we also observed sensitive inflammatory response of hPSC-derived amniotic cells to bacterial infection. Thus, in this research we propose to leverage the amnion differentiation potential of hPSCs, in conjunction with innovative microfluidics, to develop the first-of-its-kind human amnion membrane organ-on-chip system. We will further apply this tractable experimental system to quantitatively study the dynamics of bacterial invasion of the amniotic cavity and to elucidate the functional connection between inflammation-induced amniotic membrane remodeling and intra- amniotic bacterial trafficking. Successful accomplishment of this proposed research will lead to innovative technologies and methodologies for controllable, reproducible, and scalable manufacturing of human amnion membrane tissues, offering a tractable experimental system for studying related pregnancy complications, including intra-amniotic infection. The reproducibility and scalability of the human amnion membrane organ-on- chip system will make it a promising screening platform to explore complex interactions between the human amnion membrane, bacterial pathogens, drugs and toxic substances.