Trophoblast progenitor heterogeneity and function in normal and Trisomy 21-affected placentae

正常胎盘和 21 三体性胎盘中滋养层祖细胞的异质性和功能

• 批准号: 10804203
• 负责人: Mana M Parast
• 金额: $ 65.99万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN DIEGO
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-09-20 至 2028-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10804203
• 关键词: Address; Affect; Basal Plate; Biological Assay; Biological Models; CDX2 gene; Cell Nucleus; Cells; Chorionic villi; Complement; Data; Data Set; Defect; Derivation procedure; Development; Disease; Down Syndrome; Embryo; Environment; Epithelial Cells; Ethics; Fetal Growth; Fetal Growth Retardation; First Pregnancy Trimester; Functional disorder; Generations; Genetic Models; Goals; Heterogeneity; Human; In Vitro; Karyotype; Knowledge; LGR5 gene; Maintenance; Maternal-Fetal Exchange; Modeling; Molecular; Multiple Pregnancy; Nutrient; Organ; Organoids; Oxygen; Phenotype; Placenta; Placentation; Play; Pluripotent Stem Cells; Pre-Eclampsia; Pregnancy; Pregnancy Complications; Protocols documentation; Reproducibility; Retained Placenta; Risk; Role; Second Pregnancy Trimester; Sorting; Specific qualifier value; Spontaneous abortion; Subgroup; Surface; Syncytiotrophoblast; Third Pregnancy Trimester; Tissues; Villous; adverse pregnancy outcome; blastocyst; cell type; chorionic plate; cytotrophoblast; differentiation protocol; disease phenotype; fetal; high risk; human data; human embryonic stem cell; human pluripotent stem cell; implantation; in vivo; insight; organ growth; overexpression; preference; progenitor; single cell analysis; single-cell RNA sequencing; small hairpin RNA; stem cell differentiation; stem cells; transcriptome; transcriptome sequencing; transcriptomic profiling; trophoblast; trophoblast stem cell

Project Summary/Abstract The human placenta plays a major role in maintaining the proper environment for fetal growth, but remains a poorly-understood organ, particular during early gestation. Over the past few years, advances in single cell analysis and stem cell derivation have finally been applied to this organ, resulting in significant expansion, both of knowledge of cellular heterogeneity as well as of cell-based modeling, of trophoblast, the epithelial cells of the placenta. Specifically, multiple groups have established protocols for derivation of trophoblast stem cells (TSC) and trophoblast organoids from early gestation human placenta, which allow for study of differentiation into functional syncytiotrophoblast (STB) and extravillous trophoblast (EVT). Other groups, including our own, have established reproducible protocols for conversion of human pluripotent stem cells (hPSC) into bona fide TSC, allowing for modeling of both normal and abnormal trophoblast differentiation. However, recent data suggest that primary TSC have a more limited differentiation potential than originally described, with a profile that is most consistent with precursors to EVT, rather than a truly bipotential “TSC.” In fact, while human trophectoderm (TE) cells and early gestation villous CTB (vCTB) co-express TP63 and CDX2, primary TSC lack CDX2, and hPSC-derived TSC lose CDX2 during the transition from TE to TSC. In addition, placentas from Trisomy 21 (T21)-affected pregnancies show a persistent vCTB layer, with a significant proportion retaining CDX2 expression beyond first trimester; this is accompanied by an abnormality of T21-CTB to form STB in vitro, a phenotype which we have recapitulated using T21-affected hPSC. We hypothesize that CDX2+ vCTB represents a distinct trophoblast progenitor state, one that is possibly more primitive, and/or has altered differentiation potential. The goal of this application is to identify the role of CDX2 in establishment of TE, maintenance of a trophoblast progenitor state, and TSC differentiation potential, and characterize the cellular defects associated with its abnormal persistence in T21-affected placentae and the accompanying placental dysfunction. To this end, we propose to use a combination of CTB, TSC, and trophoblast organoids, derived directly from normal and T21-affected placentas or pluripotent stem cells, along with state-of-the-art single cell/single nucleus transcriptome profiling and complementary functional assays to address these goals. Successful completion of this proposal will provide a comprehensive molecular and functional assessment of cytotrophoblast heterogeneity in early gestation placenta, identify capacities and limitations of different in vitro human trophoblast model systems, and offer insights into mechanisms of placental dysfunction in pregnancies affected by Trisomy 21.

项目总结/文摘