Molecular Analysis of the Early Stages of Human Trophoblast Differentiation

人类滋养层分化早期阶段的分子分析

• 批准号: 8435287
• 负责人: SUSAN J. FISHER
• 金额: $ 30.82万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/8435287
• 关键词: Activins; Address; Agreement; Animals; Biological; Cell Culture Techniques; Cell Line; Cell membrane; Cells; Chemicals; Choriocarcinoma; Chorion; Chorionic villi; DNA-Binding Proteins; Data; Data Analyses; Decidua; Defect; Derivation procedure; Development; Developmental Process; Differentiation and Growth; Disease; Edema; Embryonic Development; Endometrial; Endometrium; Event; Fetal Growth Retardation; Fibroblast Growth Factor; Fibroblasts; Geminin; Human; Human Chorionic Gonadotropin; Hypertension; In Vitro; Infertility; Instruction; Link; Methods; MicroRNAs; Modeling; Molecular; Molecular Analysis; Mus; Neuroglia; Nodal; Oocytes; Pathway interactions; Pattern; Pilot Projects; Placenta; Placentation; Plant Roots; Population; Pre-Eclampsia; Pregnancy; Pregnancy Complications; Process; Property; Proteinuria; Reproductive Biology; Research Personnel; Role; Sampling; Seminal; Signal Transduction; Source; Staging; Staining method; Stains; Stem cells; Syncytiotrophoblast; Syndrome; Telomerase; Testing; Training Programs; Translating; Undifferentiated; Villous; Work; base; blastocyst; bone morphogenetic protein 4; cytotrophoblast; endometriosis; established cell line; human embryoid body; human embryonic stem cell; in vitro Model; in vivo; inhibitor/antagonist; insight; nonhuman primate; outreach; pluripotency; progenitor; programs; research study; self-renewal; stem; theories; tool; trophoblast

PROJECT SUMMARY (See Instructions): PROJECT II Very little is known about the formative early stages of human trophoblast (TB) differentiation, in particular, the steps between trophectoderm (TE) specification and formation of the TB populations of the chorionic villi. Recently, we identified the early-gestation human chorion as a niche for cells that coexpressed markers of pluripotency and TB fate determinants. This finding suggested that the chorion is a source of human TB progenitor cells (TBPCs). To test this theory, we established lines of continuously self renewing TBPCs from this membrane and showed that they differentiated into the mature human TB populations¿multi-nucleated syncytiotrophoblasts and invasive cytotrophoblasts. Here we propose using this new cell culture model to analyze TBPC allocation, self-renewal and differentiation. The specific hypotheses to be tested are based on global transcriptional profiling, which showed that TBPCs expressed a combination of DNA binding proteins that have been implicated in human TB development or that regulate the fate of TBs or other stem cells in the mouse. Thus, we will test the hypothesis that these molecules control TB developmental transitions that are key components of human placentation (Aim 1). The results will give us new insights into early steps in TB differentiation that have yet to be studied in humans. Then we will use this information to study the analogous processes in preeclampsia (PE), a human pregnancy complication {e.g., maternal hypertension, proteinuria and edema ¿ intrauterine growth restriction) that is associated with faulty TB differentiation. We will test the hypothesis that the observed deficits can be explained, in part, by perturbations in the molecular circuitry that governs TBPC fate (Aim 2). Thus, these experiments address the molecular underpinnings of faulty placentation, which contributes to a spectrum of disorders ranging from infertility to PE. This project significantly benefits from all the other components of the U-54. With Project III and Core B we will expand our characterization of TBPCs to include microRNA profiling and functional analyses. With Project IV we will explore the dialogue between the placenta and the decidua in terms of decidualized stromal secreted factors (¿ endometriosis). We will contribute to Project I by assisting in the analysis of the oocyte secretome and to Core C by providing expertise in educational outreach that we gained by developing similar programs. Thus, these synergistic interactions expand on the experiments proposed in this project and allow us to study other important aspects of reproductive biology that contribute to infertility.

项目摘要（请参阅说明）：项目II 关于人类滋养细胞（TB）分化的形成早期阶段，尤其是滋养绒毛绒毛绒毛的TB种群之间的步骤，知之甚少。最近，我们确定了早期捕获人绒毛膜是对多能和TB脂肪确定剂共同表达标记的细胞的利基市场。这一发现表明，绒毛膜是人类结核祖细胞（TBPC）的来源。为了检验这一理论，我们从该膜上建立了连续自我更新的TBPC线，并表明它们分化为成熟的人类TB群体。多核核蛋白核细胞和侵入性的细胞增生性细胞细胞。 在这里，我们建议使用这种新的细胞培养模型来分析TBPC分配，自我更新和分化。要测试的特定假设是基于全球转录分析的，这表明TBPC表达了在人类TB发育中隐含的DNA结合蛋白的组合，或者调节小鼠中TBS或其他干细胞的命运。那我们将测试 这些分子控制结核病发育过渡是人类放置的关键组成部分的假设（AIM 1）。结果将使我们对尚未在人类中研究的结核病区分的早期步骤提供新的见解。然后，我们将使用这些信息研究与TB分化有故障有关的人类妊娠并发症（例如，孕妇高血压，蛋白尿和雷氏雷氏生长限制）的类似过程。我们将检验以下假设：可以通过控制TBPC命运的分子电路中的扰动来解释所观察到的定义（AIM 2）。这是这些实验介绍了故障位置的分子基础，这有助于从不育到PE的各种疾病。 该项目从U-54的所有其他组件中都受益匪浅。使用项目III和Core B，我们将扩大TBPC的表征，包括microRNA分析和功能分析。通过项目IV，我们将根据确定的基质分泌因子（子宫内膜异位症）来探讨ploceta和canceua之间的对话。我们将通过协助分析来为项目I做出贡献 卵母细胞分泌型和核心C通过为我们通过制定类似计划获得的教育外展专家提供了专家。这是这些协同的相互作用扩展了该项目中提出的实验，并使我们能够研究有助于不育的生殖生物学的其他重要方面。