Modeling normal and abnormal trophoblasts

正常和异常滋养层建模

• 批准号: 10188575
• 负责人: Toshihiko Ezashi
• 金额: $ 35.86万
• 依托单位: UNIVERSITY OF MISSOURI-COLUMBIA
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-09-25 至 2023-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10188575
• 关键词: Abbreviations; BMP4; Blood; Cell Line; Cells; Chorionic villi; Conceptus; DNA methylation profiling; Decidual Cell Reactions; Development; Disease; Embryo Loss; Endometrium; Event; Exhibits; Fatty acid glycerol esters; Fetal Growth Retardation; First Pregnancy Trimester; Genes; Giant Cells; Goals; Human; Immunocompromised Host; Immunohistochemistry; Implant; In Vitro; Invaded; Kidney; Lead; Link; Modeling; Molecular; Molecular Profiling; Mothers; Mus; Organoids; Oxidative Stress; Oxygen; Pathologic; Pathway interactions; Perfusion; Perinatal mortality demographics; Phenotype; Physiological; Placenta; Placenta Diseases; Placentation; Pluripotent Stem Cells; Pre-Eclampsia; Pregnancy; Pregnancy Complications; Premature Birth; Process; Regimen; Research Personnel; Second Pregnancy Trimester; Signal Pathway; Site; Spontaneous abortion; Structure; Symptoms; Syncytiotrophoblast; System; Termination of pregnancy; Testing; Third Pregnancy Trimester; Tissues; Transcript; Transgenic Model; Umbilical cord structure; Undifferentiated; Uterus; Villous; Villus; biological adaptation to stress; biological specimen archives; capsule; cytotrophoblast; early onset; early pregnancy; embryonic stem cell; experimental study; genetic association; hazard; human embryonic stem cell; human model; human tissue; implantation; in vivo; induced pluripotent stem cell; innovation; mammary; molecular marker; natural Blastocyst Implantation; stem cell differentiation; stem cell model; stem cells; transcriptome sequencing; trophoblast; trophoblast stem cell

PROJECT SUMMARY Very little is known of human placental development in the period between one and five weeks of gestation when trophoblasts invade the uterus, form the primitive syncytium and cytotrophoblast, and then primary villi. Thus, models are needed to study the molecular and cellular mechanisms controlling early human placental development and what can go wrong with these processes to cause placental disease and early conceptus loss. It is now well established that human embryonic stem cells (ESC) and induced pluripotent stem cells (iPSC) can be driven along the trophoblast lineage by exposing them to BMP4 and inhibiting the signaling pathways that maintain the pluripotent phenotype. The overall premise is that this in vitro system is a valuable model for mimicking placental trophoblast formed early in the first trimester of pregnancy when it is most vulnerable to many of the same hazards that threaten an in vivo pregnancy early in its existence. The project will use this stem cell model of early trophoblast development to understand the development of the placenta in the earliest stages of pregnancy and the immediate pathological basis of early onset preeclampsia (EOPE), a disease linked to shallow placentation and insufficient perfusion with maternal blood. There are three aims: 1) Test the hypothesis that villous TB from the first half of the first trimester represents a transitional state between the primitive placenta encountered at implantation and the mature placenta of the second and third trimester. The goal is to validate the notion that the “primitive” TB generated from ESC and iPSC is an in vitro equivalent of early placental TB. Experiments will also confirm preliminary observations that first trimester villous TB shares many features of its molecular signature with both this primitive TB derived from pluripotent cells and more mature placental TB from the second and third trimesters. 2) Employ the stem cell model of trophoblast differentiation to test the hypothesis that stress response pathways are already aberrant in EOPE placentas upon initial formation of trophoblast. The goal is to employ RNAseq analysis and DNA methylation profiling to compare gene and gene network changes associated with PE and CTL cells when they are cultured under normal and stressful oxygen conditions. 3) Test the hypothesis that a better representation of trophoblast emergence will be gained by conducting the differentiation of the pluripotent stem cells to trophoblast with cultured spheroids rather than in 2D-cultures. The plan is to use such a system to follow the emergence of villous trophoblast within organoids. Additionally, trophoblast stem cells (TSC) will be generated from ESC and iPSC and also use these along with ESC/iPSC to create chimeric organoids. Finally the fate of these organoids will be examined when they are placed under the mammary fat pads and kidney capsules of immunocompromised mice to determine whether they exhibit physiologic and structural interactions and invasive potential within the host mouse. A longer term goal will be to determine whether PE and CTL organoids differ in invasiveness at ectopic sites and in interactions with maternal decidualized endometrium.

项目摘要 在妊娠一到五个星期之间的人类占地发育鲜为人知 当滋养细胞侵入子宫时，形成原始的突触和细胞增生细胞，然后形成原发性绒毛。 这是需要模型来研究控制早期人体位置的分子和细胞机制 开发以及这些过程可能会出现什么问题，以引起局部疾病和早期概念 损失。现在已经很好地确定了人类胚胎干细胞（ESC）和诱导多能干细胞 （IPSC）可以通过将其暴露于BMP4并抑制信号传导来沿滋养细胞谱系驱动 维持多能表型的途径。总体前提是，这种体外系统是有价值的 模仿妊娠早期早期形成的块状滋养细胞的模型 容易受到许多相同的危害，这些危害在其存在早期就威胁着体内怀孕。项目 将使用这种早期滋养细胞发展的干细胞模型来了解 怀孕的最早阶段和早期发作前启动前（EOPE）的直接病理基础， 疾病与遗产血液相关的浅水和灌注不足。有三个目标： 1）检验以下假设：孕期上半年的绒毛结核病代表过渡状态 在植入时遇到的原始plapeta与第二和第三 孕期。目的是验证从ESC和IPSC产生的“原始”结核病是一种体外 等效于早期的结核病。实验还将确认头三个月的初步观察 绒毛结核病具有其分子签名的许多特征，这两个原始结核病都来自多能 细胞和更成熟的斑点结核病，来自第二和第三个三物质。 2）采用干细胞模型 滋养细胞的分化以检验以下假设：应力反应途径在EOPE中已经异常 plapetas在初始形成滋养细胞时。目的是采用RNASEQ分析和DNA甲基化 分析以比较基因和基因网络在培养时与PE和CTL细胞相关的变化 在正常和压力的氧气状态下。 3）检验以下假设，即更好地表示 通过进行多能干细胞的分化为 具有培养的球体而不是2D文化中的滋养细胞。计划是使用这样的系统遵循 类器官中绒毛滋养细胞的出现。此外，将生成滋养细胞干细胞（TSC） 从ESC和IPSC，也将它们与ESC/IPSC一起使用来创建嵌合类器官。最后的命运 这些器官将其放置在乳腺脂肪垫和肾胶囊下时，将被检查 免疫功能低下的小鼠，以确定它们是否暴露了生理和结构相互作用以及 主机鼠标内的侵入性潜力。长期目标是确定PE和CTL是否 器官在生态部位的侵入性和与母体确定的子宫内膜相互作用的侵入性不同。