Cellular Atlas of the Human Placenta: Structure-Function Relationships and their Implications for Placental Dysfunction

项目摘要/摘要 对人类胎盘的结构-功能关系知之甚少。作为病理学家，我们可以看到 仅在分娩后检查胎盘，并将我们的发现与临床和产前病程以及 新生儿结局。然而，最好的情况是，我们有与疾病相关的损伤，但还没有。 通过分子分析直接验证，或间接通过操作和功能验证 体外模型分析。胎盘功能障碍，临床表现为先兆子痫(PE)伴或 无胎儿生长受限(FGR)与母体血管组织病理损害有关 血流灌注不良(MVM)和胎儿血管灌注不良(FVM)。这些病变不同地影响这三个 滋养细胞隔室：细胞滋养细胞(CTB，推测为干细胞)、合体滋养细胞(STB， 负责气体/营养交换的细胞类型)和绒毛外滋养细胞(EVT，侵袭性细胞类型 在母体-胎儿界面)。然而，我们对这些损伤的理解仅限于形态学。 以及几个标记物的免疫定位。在过去的几年里，包括我们在内的多个组织 使用scRNAseq来表征正常和病变胎盘中的细胞异质性； 然而，这些研究大多是描述性的，缺乏分子数据的空间背景和 对不同的细胞类型进行功能评估。因此，我们建议应用新技术，包括 数字空间轮廓(DSP)和组织脱细胞，随后是细胞外基质(ECM)特异性 质谱学，作为基于发现的方法，更好地描述特定的MVM和FVM病变 在分子水平上。然后，我们将使用主要术语CTB来重现这些表型，并 作为目标评估方法的一部分执行功能验证。我们将检验这一假设 妊娠晚期CTB对特定的细胞和ECM来源的信号作出反应，以增殖和/或分化为 STB或EVT，从而确定这一独特的瞬时器官的潜在再生能力。 我们还将探索PE相关胎盘功能障碍的细胞和细胞外基质来源，测试 假设这种疾病起源于细胞外基质成分和旁分泌信号的改变 胎盘和蜕膜(母体)细胞。成功完成这项提案将建立一个 详细的人体胎盘细胞和基质图谱，具有经过验证的结构-功能 关系，为探讨胎盘再生能力奠定了基础 胎盘功能障碍。

Project Summary/Abstract Little is known about structure-function relationships in the human placenta. As pathologists, we can view the placenta only after delivery, and correlate our findings to clinical and prenatal course, as well as neonatal outcome. At best, however, we have lesions which correlate with disease, but have yet to be validated, either directly, through molecular analysis, or indirectly, through manipulation and functional analysis of in vitro models. Placental dysfunction, manifested clinically as preeclampsia (PE) with or without fetal growth restriction (FGR), is associated with histopathologic lesions of maternal vascular malperfusion (MVM) and fetal vascular malperfusion (FVM). These lesions differentially affect the three trophoblast compartments: cytotrophoblast (CTB, the putative stem cell), syncytiotrophoblast (STB, the cell type responsible for gas/nutrient exchange), and extravillous trophoblast (EVT, the invasive cell type at the maternal-fetal interface). Nevertheless, our understanding of these lesions is limited to morphology and immunolocalization of a few markers. Over the past few years, multiple groups, including ours, have used scRNAseq to characterize cellular heterogeneity within both normal and diseased placentae; however, these studies remain mostly descriptive, lacking both spatial context of the molecular data and functional evaluation of the distinct cell types. We therefore propose to apply novel technologies, including digital spatial profiling (DSP) and tissue decellularization followed by extracellular matrix (ECM)-specific mass spectrometry, as discovery-based approaches to better characterize specific MVM and FVM lesions at the molecular level. We will then use primary term CTB, to reproducibly model these phenotypes and to perform functional validation as part of a targeted evaluation approach. We will test the hypothesis that late gestation CTB respond to specific cell- and ECM-derived signals to proliferate and/or differentiate into either STB or EVT, thus identifying potential regenerative capacity in this unique transient organ. We will also probe the cellular and ECM origins of PE-associated placental dysfunction, testing the hypothesis that this disease originates from alterations in ECM composition and paracrine signaling from both placental and decidual (maternal) cells. Successful completion of this proposal will establish a detailed cellular and matrix atlas of the human placenta, with validated structure-function relationships, laying the groundwork for probing placental regenerative capacity in the setting of placental dysfunction.