Pluripotent Stem Cells: Modeling syncytiotrophoblast development and pathogenesis

多能干细胞：模拟合体滋养层发育和发病机制

• 批准号: 8560788
• 负责人: Toshihiko Ezashi
• 金额: $ 31.85万
• 依托单位: UNIVERSITY OF MISSOURI-COLUMBIA
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-08-15 至 2018-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8560788
• 关键词: Abbreviations; Activins; Affect; Angiogenic Proteins; Anti-Inflammatory Agents; Anti-inflammatory; Arteries; BMP4; Birth; Blood; Capital; Cell Culture Techniques; Cell Death; Cell fusion; Cell model; Cells; Clinical; Complex; Conceptus; Development; Diagnosis; Disease; Embryo; Endocrine; Endogenous Retroviruses; Endometrial; Endothelium; Ensure; Ethics; Event; FGF2 gene; FLT1 gene; Fetal Growth Retardation; Fibroblasts; First Pregnancy Trimester; Gases; Genes; Gestational Age; Goals; HERVs; Human; Human Chorionic Gonadotropin; Hypersensitivity; Immune; In Vitro; Inflammatory; Inflammatory Response; Invaded; Lead; Letters; Longevity; Low Birth Weight Infant; Mammals; Modeling; Molecular; Mothers; Mus; Nutrient; Oxygen; Oxygen measurement, partial pressure, arterial; Pathogenesis; Pathology; Pattern; Perfusion; Phenotype; Placenta; Placenta Diseases; Placentation; Pluripotent Stem Cells; Pre-Eclampsia; Pregnancy; Pregnancy Outcome; Pregnancy loss; Premature Birth; Process; Production; Progesterone; Property; Proteins; Role; Signal Transduction; Small Inducible Cytokine A3; Source; Spiral Artery of the Endometrium; Staging; Stem cells; Structure; Study models; Syncytiotrophoblast; System; Taxon; Testing; Time; Tissues; Umbilical cord structure; Villous; Waste Products; activin A; base; cell type; cytokine; cytotrophoblast; early onset; embryonic stem cell; human embryonic stem cell; in vitro Model; in vivo; induced pluripotent stem cell; inhibitor/antagonist; insight; novel diagnostics; offspring; public health relevance; release factor; surface coating; syncytin; transcription factor; treatment strategy; trophoblast

DESCRIPTION (provided by applicant): A placenta allows eutherian mammals to give birth to progeny that are already developmentally advanced. A wide range of placental phenotypes exist across taxa to ensure adequate nutrient transfer from mother to offspring. The human placenta forms particularly intimate associations with maternal tissues through two types of cells: 1) syncytiotrophoblast (STB) cells, the fetally-derived trophoblast (TB) cell subset that coats the surface of the villous placenta, and 2) extravillous TB (EVCTB), which invade deeply, interact with maternal decidual immune cells, and remodel the spiral arteries. Both TB subsets are in direct contact with maternal blood and maternal immune cells and both are derived from an ill-defined, stem-cell like, cytotrophoblast (cytoTB) cell subpopulation. Although many adverse pregnancy outcomes may result from poor placental development, the study of the earliest stages of placental development cannot be performed in humans for ethical reasons. For example, alterations in STB development and turnover have been implicated in placental disease, including preeclampsia (PE), the leading single cause of premature birth. STB is the source of "placental debris", cytokines, and pro-inflammatory and anti-angiogenic proteins, which become elevated in mothers diagnosed with PE. Appropriate in vitro models of early placental development are essential if we are to better understand and treat diseases of poor placentation. The first goal of this proposal is to generate STB and its mononucleated precursors from human embryonic stem cells (hESC) that have been treated with BMP4 (BMP-hESC) and inhibitors of activin and FGF2 signaling. We will use this system as a model to study STB emergence, especially the process of cell fusion and cell death, as well as intrinsic and extrinsic factors that influence these transitions. This project has high significance because STB forms the major interface with maternal blood perfusing the placenta and is responsible for the production of endocrine factors such as hCG and exchange of gases, nutrients and waste products The second goal is to demonstrate the utility of the BMP-hESC model to examine the molecular events controlling STB formation and lifespan, with an emphasis on the roles of transcription factors, such as GCM1 & GATA2, fusogenic proteins such syncytin-1 & -2 (ERVW1, ERVFRD-1, respectively) and a presently little understood HERV envelope gene product (ERV-Fb1) that inhibits cell-cell fusion. A third goal is to assess whether some features of PE can be unearthed in the BMP-hESC model. Our hypothesis is that TB from a sub-group of conceptuses whose mothers develop PE early in their pregnancies is unusually sensitive to high oxygen tensions. Since STB will only begin to encounter well-oxygenated maternal blood after the uteroplacental arteries "open" towards the end of the first trimester of pregnancy, the oxygen hypersensitivity of cells derived from PE placentas may cause them to turnover at an accelerated rate and shed cell contents and debris into the circulating maternal blood more quickly than normal STB. We will test this hypothesis in vitro using iPSCs derived from the umbilical cords of babies born to mothers with severe preeclampsia and gestational age-matched control iPSCs.

描述(申请人提供)：胎盘允许真兽类哺乳动物生育已经发育成熟的后代。广泛的胎盘表型存在于不同的分类群中，以确保从母亲到后代的足够的营养转移。人类胎盘通过两种类型的细胞与母体组织形成特别密切的联系：1)合体滋养细胞(STB)细胞，即覆盖在绒毛胎盘表面的胎儿来源的滋养细胞(TB)亚群；2)绒毛外TB(EVCTB)，它深入侵袭，与母体蜕膜免疫细胞相互作用，并重塑螺旋动脉。这两个结核病亚型都与母体血液和母体免疫细胞直接接触，都来自定义不清的干细胞样细胞滋养细胞(细胞TB)亚群。虽然许多不良妊娠结局可能是由于胎盘发育不良造成的，但出于伦理原因，对胎盘发育最早阶段的研究不能在人类身上进行。例如，STB发育和周转的改变与胎盘疾病有关，包括先兆子痫(PE)，这是导致早产的主要单一原因。STB是胎盘碎片、细胞因子、促炎和抗血管生成蛋白的来源，在诊断为PE的母亲中，这些蛋白会升高。如果我们要更好地了解和治疗胎盘不良的疾病，合适的早期胎盘发育体外模型是必不可少的。这项提议的第一个目标是从经过BMP4(BMP-hESC)和激活素和FGF2信号抑制剂处理的人胚胎干细胞(HESC)中产生STB及其单核前体细胞。我们将利用这个系统作为模型来研究STB的出现，特别是细胞融合和细胞死亡的过程，以及影响这些转变的内在和外在因素。这个项目具有很高的意义，因为STB形成了母体血液与胎盘灌流的主要界面，并负责产生内分泌因子，如hCG和气体、营养物质和废物的交换。第二个目标是展示BMP-hESC模型的实用性，以研究控制STB形成和寿命的分子事件，重点是转录因子，如GCM1和GATA2，融合蛋白，如Syncytin-1和-2(分别为ERVW1，ERVFRD-1)和目前鲜为人知的抑制细胞-细胞融合的Herv包膜基因产物(ERV-Fb1)。第三个目标是评估是否可以在BMP-hESC模型中发现PE的一些特征。我们的假设是，母亲在怀孕早期患上PE的一组孕妇中的结核病对高氧气紧张异常敏感。由于STB只有在妊娠初期子宫胎盘动脉“开放”后才开始接触到氧气充足的母体血液，来自PE胎盘的细胞对氧气的敏感可能导致它们加速周转，并比正常STB更快地将细胞内容物和碎片排放到循环中的母体血液中。我们将使用来自患有严重先兆子痫的母亲所生婴儿的脐带的IPSCs和与孕龄匹配的对照组IPSCs在体外测试这一假说。