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.

项目摘要 先兆子痫是妊娠期母体和胎儿发病率和死亡率的主要来源。多项研究已经记录了从足月先兆子痫妊娠中获得的胎盘滋养层细胞、合胞体滋养层细胞（STB）和绒毛外滋养层细胞（EVTB）的发育和功能异常。这些细胞类型在妊娠早期出现，目前尚不清楚这些异常是否有助于先兆子痫的病理生理学或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差距：（1）诱导多能干细胞（iPSC），来自对照和先兆子痫妊娠，并通过暴露于BMP 4和信号抑制剂分化为围着床期滋养层的混合群体;（2）诱导的滋养层干细胞（TSC），其通过转化对照和EOPE iPSC系获得，并分化为胎盘发育的前三个月、绒毛阶段，沿着STB或EVTB谱系;（3）3D滋养层类器官，来源于TSC系，并主要分化为STB或EVT。有三个具体目标：目的1将测试STB分化以及因此的功能是否在早期妊娠胎盘的各种模型中在EOPE中存在缺陷。目标1A将测试这样的假设：在本实验室开发的围着床期和绒毛STB模型中，特别是在氧化应激条件下，在EOPE中可以看出STB组装和功能的缺陷。在Aim 1B中，单核RNAseq将用于区分TB类型，特别是先前鉴定的两个8 TB簇是否受到差异影响。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。目的2A将确定EOPE中的缺陷性TB侵袭是否是妊娠早期样EVTB的特征，就像在围着床期TB中一样。目标28将利用类器官培养物来评估EVTB在三维空间中的分化，与锚定绒毛同源，以及细胞-基质相互作用在获得特定EVTB亚型中的作用。目的3将确定线粒体缺陷是否影响EVT和STB在EOPE目的3A将确定是否减少ATP的生产减少侵入EOPE在应激条件下。目标3B将测试EOPE TB是否对应激诱导的活性氧产生更敏感。这些目标的实现将揭示STB和EVTB分化和功能受损背后的机制，在孕早期胎盘发育的患者来源的培养模型。