Modeling Human Placentation via Single Cell RNA-Sequencing

通过单细胞 RNA 测序模拟人类胎盘

• 批准号: 10316877
• 负责人: Soumen Paul
• 金额: $ 23.1万
• 依托单位: UNIVERSITY OF KANSAS MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-07-10 至 2023-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10316877
• 关键词: Adult; Affect; Bioinformatics; Biology; Birth; Blood Vessels; Caring; Cell Differentiation process; Cell Lineage; Cell Nucleus; Cells; Chorionic Villi Sampling; Chorionic villi; Clinical Data; Data; Defect; Development; Diagnostic; Disease; Embryo; Ensure; Ethics; Family; Fetal Growth Retardation; Fetal health; First Pregnancy Trimester; Functional disorder; GATA3 gene; Gases; Gene Expression Profile; Gene Expression Profiling; Genomics; Gestational Age; Gestational Diabetes; Healthcare; Hormones; Human; Impairment; Institutes; Kansas; Laboratories; Lead; Maintenance; Maternal Mortality; Mediating; Medical center; Modeling; Modernization; Molecular; Monitor; Mothers; Nutrient; Pathologic; Pathway interactions; Patients; Placenta; Placenta Diseases; Placentation; Pre-Eclampsia; Pregnancy; Pregnancy Complications; Pregnancy Outcome; Pregnancy loss; Premature Birth; Procedures; Process; Proliferating; Public Health; Publishing; Research; Resolution; Risk; Sampling; Small Nuclear RNA; Source; Spontaneous abortion; Syncytiotrophoblast; System; Technology; Testing; Time; Tissues; Undifferentiated; Universities; Villous; cell type; combinatorial; comparative; cytotrophoblast; delivery complications; early pregnancy loss; experimental analysis; experimental study; fetal; follow-up; human model; innovation; insight; knockout gene; longitudinal analysis; mouse genetics; mouse model; mutant mouse model; natural Blastocyst Implantation; postnatal; pregnancy disorder; progenitor; self-renewal; single-cell RNA sequencing; success; transcription factor; transcriptome sequencing; trophoblast; trophoblast stem cell

Abstract Placental dysfunction leads to pregnancy-associated disorders, including intrauterine growth restriction (IUGR) and preeclampsia, and also serves as a developmental cause for postnatal and adult diseases. Often, the causal alterations in the placentation process, which lead to defective pregnancies, occur early in gestation. However, due to ethical barriers and difficulties with the experimental systems, most of the studies on human placental function are performed with samples obtained at term, thereby, limiting our ability to understand placental development and function throughout gestation. Fortunately, the recent advancement of single-nucleus RNA-sequencing (snRNA-seq) strategy and success in deriving true human trophoblast stem cells (human TSCs) from villous cytotrophoblast cells (CTBs) have opened up new strategies for direct assessment of early human placentation process. Chorionic villus sampling (CVS) is a standard health care procedure performed during latter stages of the first trimester of pregnancy. Surplus tissue obtained via CVS represents a valuable source of trophoblast cells for experimental analysis and thus, a unique opportunity to interrogate the early developing placenta. Using gene knockout mouse models, we discovered that depletion of GATA family of transcription factors, GATA2 and GATA3 in trophoblast progenitor cells leads to either loss of pregnancy or fetal growth restriction. As part of our pursuit to define importance of these findings with respect to human placentation, we have successfully performed scRNA-seq study with human chorionic villous samples and found that, within a first-trimester placenta, GATA2 and GATA3 are broadly expressed in distinct cell types, including CTBs, syncytioytrophoblasts (SynTs) and extravillous trophoblasts (EVTs). Furthermore, we have also been able to establish human TSCs from CTBs, isolated from chorionic villi, and found that loss of GATA2 expression in human TSCs impairs both EVT and SynTB differentiation. These observations led us to the central hypothesis of this proposal that single cell gene expression patterns, including expression of GATA factors, in a first-trimester placenta could be predictive of pregnancy associated disorders during late gestation. To test this hypothesis, we will use CVS to perform snRNA-seq and to establish patient-specific human TSCs for subsequent functional studies. We will also monitor those pregnancies for pregnancy- associated disorders, including intrauterine growth restriction and preeclampsia and collect term placental samples for follow up analyses. Together, our proposed study, bolstered by modern technologies, will illuminate previously unknown pathways, which underlie placental adaptation at a single cell level during normative and pathological pregnancies.

抽象的 胎盘功能障碍会导致妊娠相关疾病，包括宫内生长受限 （IUGR）和先兆子痫，也是产后和成人疾病的发育原因。经常， 导致妊娠缺陷的胎盘过程的因果变化发生在早期 妊娠。然而，由于伦理障碍和实验系统的困难，大多数 对人类胎盘功能的研究是用足月获得的样本进行的，因此限制了我们的研究 了解整个妊娠期胎盘发育和功能的能力。幸运的是，最近 单核 RNA 测序 (snRNA-seq) 策略的进步并成功衍生出真正的人类 来自绒毛细胞滋养层细胞（CTB）的滋养层干细胞（人类 TSC）开辟了新的 直接评估人类早期胎盘形成过程的策略。 绒毛膜绒毛取样 (CVS) 是后期阶段执行的标准医疗保健程序 怀孕的前三个月。通过 CVS 获得的剩余组织是宝贵的来源 滋养层细胞进行实验分析，因此是研究早期发育的独特机会 胎盘。 使用基因敲除小鼠模型，我们发现 GATA 转录家族的缺失 滋养层祖细胞中的 GATA2 和 GATA3 因子会导致妊娠失败或胎儿生长 限制。作为我们追求定义这些发现对于人类胎盘的重要性的一部分， 我们已经成功地对人类绒毛膜绒毛样本进行了 scRNA-seq 研究，结果发现， 在妊娠早期胎盘中，GATA2 和 GATA3 广泛表达于不同的细胞类型中，包括 CTB、合体滋养细胞 (SynT) 和绒毛外滋养细胞 (EVT)。此外，我们还 能够从从绒毛膜绒毛中分离出的 CTB 中建立人类 TSC，并发现 GATA2 的缺失 人类 TSC 中的表达会损害 EVT 和 SynTB 分化。这些观察使我们得出 该提案的中心假设是单细胞基因表达模式，包括 GATA 的表达 妊娠早期胎盘中的因素可以预测晚期妊娠相关疾病 妊娠。为了检验这一假设，我们将使用 CVS 进行 snRNA-seq 并建立患者特异性 人类 TSC 用于后续功能研究。我们还将监测这些怀孕情况 - 相关疾病，包括宫内生长受限和先兆子痫，并收集足月胎盘 用于后续分析的样品。总之，我们提出的研究在现代技术的支持下，将 阐明了以前未知的途径，这些途径是胎盘在单细胞水平上适应的基础 正常妊娠和病理妊娠。