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

项目概要/摘要 人们对人类胎盘的结构与功能关系知之甚少。作为病理学家，我们可以看到 仅在分娩后才进行胎盘检查，并将我们的发现与临床和产前过程以及 新生儿结局。然而，充其量，我们有与疾病相关的病变，但尚未被确定。 通过分子分析直接验证，或通过操作和功能间接验证 体外模型分析。胎盘功能障碍，临床表现为先兆子痫 (PE)，伴有或 无胎儿生长受限（FGR），与母体血管的组织病理学病变有关 灌注不良（MVM）和胎儿血管灌注不良（FVM）。这些病变对三个方面有不同的影响 滋养层室：细胞滋养层（CTB，假定的干细胞）、合体滋养层（STB，假定的干细胞） 负责气体/营养物质交换的细胞类型）和绒毛外滋养层（EVT，侵袭性细胞类型） 母胎界面）。然而，我们对这些病变的理解仅限于形态学 和一些标记物的免疫定位。在过去的几年里，包括我们在内的多个团体 使用 scRNAseq 来表征正常胎盘和患病胎盘内的细胞异质性； 然而，这些研究仍然大多是描述性的，缺乏分子数据的空间背景和 不同细胞类型的功能评估。因此，我们建议应用新技术，包括 数字空间分析 (DSP) 和组织脱细胞化，然后是细胞外基质 (ECM) 特异性 质谱分析，作为基于发现的方法，可以更好地表征特定的 MVM 和 FVM 病变 在分子水平上。然后，我们将使用主要术语 CTB 来重复建模这些表型并 执行功能验证作为有针对性的评估方法的一部分。我们将检验以下假设： 妊娠晚期 CTB 对特定细胞和 ECM 衍生的信号作出反应，增殖和/或分化为 STB 或 EVT，从而确定这个独特的短暂器官的潜在再生能力。 我们还将探讨 PE 相关胎盘功能障碍的细胞和 ECM 起源，测试 假设这种疾病起源于 ECM 成分和旁分泌信号的改变 胎盘细胞和蜕膜（母体）细胞。该提案的成功完成将建立一个 人类胎盘的详细细胞和基质图谱，具有经过验证的结构功能 关系，为探索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.