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Regulation of Invasive Trophoblast Cell Lineage Development

侵袭性滋养层细胞谱系发育的调控

基本信息

批准号：
10525942
负责人：
Kaela Margaret Varberg
金额：
$ 13.07万
依托单位：
UNIVERSITY OF KANSAS MEDICAL CENTER
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-08-02 至 2024-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10525942
关键词：
Abnormal placentation Adult Affect BHLH Protein Binding Biological Assay Cell Differentiation process Cell Line Cell Lineage Cells Cellular Assay ChIP-seq Child Chromatin Critical Pathways DNA Methylation Data Development Disease Early Diagnosis Embryo Ensure Environment Epigenetic Process Family Fetal Growth Retardation Fetus Foundations Genes Genomics Goals Grant Health Hi-C Homologous Gene Human Impairment Kansas Lead Life Mediating Medical center Mentors Modeling Modification Molecular Molecular Conformation Morbidity - disease rate Mothers Nutrient Phase Physiological Placenta Placentation Pre-Eclampsia Predisposition Pregnancy Pregnancy Complications Pregnancy loss Premature Birth Protocols documentation Publications Rattus Regulation Regulatory Element Research Research Institute Research Project Grants Resistance Resources Series Signal Pathway Spiral Artery of the Endometrium Techniques Testing Training Transposase Universities Uterus Vascular blood supply Vascular remodeling Work Writing adverse outcome adverse pregnancy outcome bisulfite sequencing career chromosome conformation capture early detection biomarkers effective intervention epigenomics fetal gene regulatory network improved in vivo innovation knock-down lentiviral-mediated methylation pattern mortality mutant novel postnatal pregnancy disorder progenitor response single-cell RNA sequencing skill acquisition skills small hairpin RNA transcription factor trophoblast trophoblast stem cell whole genome

项目摘要

Project Summary / Abstract Uterine vascular remodeling occurs during gestation to meet increasing fetal nutrient demands. This remodeling includes modification of the uterine spiral arteries into low resistance vessels for supplying blood to the fetus. Central to uterine spiral artery remodeling are invasive trophoblast cells, known in the human as extravillous trophoblast (EVT). Impaired EVT development leads to suboptimal fetal conditions and adverse pregnancy outcomes including pregnancy loss, preeclampsia, intrauterine growth restriction, and preterm birth. We identified a critical and conserved regulator of EVT lineage development, Achaete-Scute Family Basic Helix-Loop-Helix Transcription Factor 2 (ASCL2). Depletion of ASCL2 in human trophoblast stem (hTS) cells inhibits EVT formation. Similarly, global depletion of ASCL2 in vivo disrupts placental development and causes embryonic lethality in the rat. However, the molecular mechanisms by which ASCL2 directs EVT lineage development are unknown. Our established hTS cell lines and protocols for generating mutant rat models will allow us to directly test our central hypothesis that ASCL2 controls EVT lineage development during placentation. To investigate higher order actions of ASCL2 on the epigenomic landscape of the EVT cell lineage, we will identify how ASCL2 depletion alters DNA methylation, chromatin accessibility and conformation using whole genome bisulfite sequencing (WGBS), assay for transposase-accessible chromatin- sequencing (ATAC-seq), and chromatin capture using Hi-C, respectively (Aim 1A). To identify direct genomic targets of ASCL2 we will perform chromatin immunoprecipitation sequencing (ChIP-seq) in EVT cells (Aim 1B). ASCL2 regulation of trophoblast development and invasion will then be evaluated in vivo using our proven techniques to generate rat hypomorphs (Aim 2A). To examine ASCL2-positive trophoblast cell development in normal and diseased rat placentas we will conduct single cell RNA-sequencing (scRNA-seq) and single cell ATAC-seq (Aim 2B). The proposed research plan will provide the candidate with a body of experimental work necessary for independent publications and preliminary data for R-series grants. The candidate will utilize the expertise of the co-mentoring team as well as resources at the University of Kansas Medical Center and Children’s Mercy Research Institute for cultivation of professional development skills. These skills will be improved through trainee mentoring, data presentation, and scientific writing. During the R00 phase the candidate will develop independence from her mentors by identifying targets of ASCL2 and investigating their contributions to invasive trophoblast lineage development with innovative rat models. The proposed research project serves as the foundation for the candidate’s long-term career goal of identifying how dysregulated spiral artery remodeling leads to a spectrum of diseases ranging from fetal growth restriction to preeclampsia.

项目摘要/摘要