Modeling Normal and Abnormal Trophoblasts

正常和异常滋养细胞建模

• 批准号: 10660067
• 负责人: R. MICHAEL ROBERTS
• 金额: $ 63.47万
• 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 2028-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10660067
• 关键词: 3-Dimensional; Address; Affect; BMP4; Biological Models; Cell Culture Techniques; Cell Line; Cell Nucleus; Cells; Characteristics; Conceptions; Conceptus; Defect; Development; Disease; Embryo; Environment; Epigenetic Process; Exposure to; Face; Failure; Fibroblasts; First Pregnancy Trimester; Genetic; Germ Cells; Giant Cells; Grouping; HLA G antigen; Heterogeneity; Human; Impairment; Implant; Infant; Invaded; Knowledge; Laboratories; Maternal Mortality; Mitochondria; Modeling; Morbidity - disease rate; Mothers; Organoids; Oxidative Stress; Pathologic; Pathway interactions; Patients; Physiological; Placenta; Placenta Diseases; Placentation; Population; Pre-Eclampsia; Predisposition; Pregnancy; Pregnancy loss; Production; Property; Protocols documentation; Reactive Oxygen Species; Role; Signal Transduction; Site; Source; Spiral Artery of the Endometrium; Stress; Syncytiotrophoblast; Testing; Umbilical cord structure; Uterus; Villous; Villus; blastocyst; cell type; convict; early experience; early onset; experience; fetal; implantation; induced pluripotent stem cell; inhibitor; innovation; migration; mortality; pathophysiology of preeclampsia; stem cell model; stem cells; transcriptome sequencing; transcriptomics; trophoblast; trophoblast stem cell

PROJECT SUMMARY Preeclampsia is a major source of maternal and fetal morbidity and mortality in pregnancy. Multiple studies have documented abnormalities in placental trophoblasts obtained from preeclamptic pregnancies at term, in development and function of both syncytiotrophoblast (STB) and extravillous trophoblast (EVTB). These cell types arise early in gestation, and it is unclear whether the abnormalities contribute to the pathophysiology of preeclampsia or are only a result of the preeclamptic environment. The proposed project will use three innovative models to address this gap in knowledge: (1) Induced pluripotent stem cells (iPSC), derived from control and preeclamptic pregnancies and differentiated to a mixed population of peri-implantation stage trophoblast by exposure to BMP4, and signaling inhibitors; (2) Induced trophoblast stem cells (TSC), obtained by conversion of the control and EOPE iPSC lines, and differentiated to the first trimester, villous stage of placental development, along either the STB or EVTB lineage; (3) 3D trophoblast organoids, derived from the TSC lines, and differentiated to predominantly STB or EVT. There are three specific aims: Aim 1 will test whether STB differentiation and, as a consequence, function, is defective in EOPE in the various models of first trimester placenta. Aim 1A will test the hypothesis that defects in STB assembly and function will be discernible in EOPE in the models of peri-implantation and villous STB developed in this laboratory, particularly under conditions of oxidative stress. In Aim 1B, single nucleus RNAseq will be used to distinguish whether TB types, especially the two previously identified clusters of 8TB, are differentially affected. Aim 2 will characterize EVTB differentiation and invasive properties in normal pregnancy and EOPE by using the TSC models. Aim 2A will determine whether defective TB invasion in EOPE is characteristic of first trimester-like EVTB, as it is in peri-implantation TB. Aim 28 will utilize organoid cultures to assess EVTB differentiation in 3-dimensional space, in homology to anchoring villi, and the role of cell-matrix interactions in acquisition of specific EVTB subtypes. Aim 3 will determine whether mitochondrial defects affect EVT and STB in EOPE Aim 3A will determine whether diminished ATP production reduces invasion in EOPE under stressful conditions. Aim 3B will test whether EOPE TBs are more susceptible to stress-induced production of reactive oxygen species. Accomplishment of these aims will uncover the mechanisms behind impaired STB and EVTB differentiation and function in patient-derived culture models of first trimester placental development.

项目总结 先兆子痫是孕期母婴发病率和死亡率的主要来源。多项研究证实，足月妊娠先兆子痫患者的胎盘滋养层细胞、合体滋养层细胞(STB)和绒毛外滋养层细胞(EVTB)的发育和功能均出现异常。这些细胞类型发生在妊娠早期，目前尚不清楚这些异常是导致先兆子痫的病理生理机制，还是仅仅是先兆子痫环境的结果。拟议的项目将使用三种创新模型来解决这一知识缺口：(1)诱导多能干细胞(IPSC)，来自对照妊娠和子痫前期妊娠，并通过暴露于BMP4和信号抑制剂而分化为围着床期滋养细胞的混合群体；(2)诱导滋养层干细胞(TSC)，通过转化对照和EOPE IPSC系获得，并分化为胎盘发育的早期三个月、绒毛阶段，沿STB或EVTB谱系；(3)3D滋养层有机体，来自TSC系，并主要分化为STB或EVT。有三个具体的目标：目标1将在不同的妊娠早期胎盘模型中测试STB的分化，并因此测试EOPE的功能是否有缺陷。目的在本实验室建立的围植入期和绒毛STB模型中，特别是在氧化应激条件下，验证EOPE中STB的组装和功能缺陷的假设。在目标1B中，将使用单核RNAseq来区分结核病类型，特别是之前确定的两个8TB集群，是否受到不同的影响。目的2利用TSC模型研究EVTB在正常妊娠和EOPE中的分化和侵袭特性。Aim 2A将确定EOPE中有缺陷的结核侵袭是否是早期妊娠样EVTB的特征，就像在种植围术期的结核一样。AIM 28将利用器官培养评估EVTB在三维空间中的分化，与锚定绒毛的同源性，以及细胞-基质相互作用在获得特定EVTB亚型中的作用。目标3将确定线粒体缺陷是否影响EOPE的EVT和STB。目标3A将确定在应激条件下，ATP产量减少是否减少EOPE的侵袭性。Aim 3B将测试EOPE TBS是否更容易受到应激诱导的活性氧产生的影响。这些目标的实现将揭示STB和EVTB在患者来源的早期胎盘发育培养模型中分化和功能受损的机制。