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Regulation of endothelial cell specification

内皮细胞规格的调节

基本信息

批准号：
10569601
负责人：
Juan M Melero-Martin
金额：
$ 60.36万
依托单位：
BOSTON CHILDREN'S HOSPITAL
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-02-15 至 2025-01-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10569601
关键词：
Biological Assay Blood Vessels Cell Communication Cell Differentiation process Cell Line Cell Reprogramming Cells ChIP-seq Chromatin Co-Immunoprecipitations Competence Data Development Endothelial Cells Endothelium Epigenetic Process Exhibits Family Foundations Genes Genetic Transcription Goals Homeostasis Human Impairment In Vitro Knowledge Molecular Mus Mutagenesis Myeloid Cells Natural regeneration Nucleic Acid Regulatory Sequences Pattern Perfusion Phenotype Process Property Proteins Proteomics Regulation Regulatory Element Solid Specific qualifier value Supporting Cell System Technology Time Vascular Diseases Vascular Graft Vascular System Vascular regeneration candidate validation cell type directed differentiation in vitro Assay in vivo induced pluripotent stem cell insight organ regeneration organ repair protein protein interaction repaired single-cell RNA sequencing stem cells transcription factor

项目摘要

SUMMARY Approaches to vascular regeneration and repair require specification of endothelial cells that are competent to form functioning blood vessels. However, the process by which endothelial cells are specified from mesoder- mal precursors remains poorly understood. A key transcriptional regulator of endothelial cell (EC) specification is the ETS-family transcription factor ETV2. Our preliminary data shows that mesodermal progenitor cells (MPCs), differentiated from human induced pluripotent stem cells (iPSCs), are rapidly and efficiently repro- grammed by ETV2 into endothelial cell-like cells (iEC-Ms). These iEC-Ms exhibit properties of endothelial cells in vitro, and assemble into perfused vascular networks in the in vivo microvascular graft assay. In contrast, ETV2 expression directly in iPSCs yielded cells that expressed endothelial cell markers (iEC-Ps) and exhibited a subset of endothelial cell properties in vitro, but did not form perfused vascular networks in microvascular grafts. The overarching goal of this proposal is to use this experimental paradigm to define the mechanisms by which ETV2 drives reprogramming to iECs, and to dissect the mechanisms by which the starting cell type (MPC vs iPSCs) influences the functional properties of the resulting iECs. We propose 3 Specific Aims to achieve these goals: (1) To dissect the transcriptional regulatory landscape of endothelial differentiation. (2) To determine the molecular mechanisms that limit functionality of iECs differ- entiated directly from iPSCs. (3) To characterize the protein-protein interactions required for ETV2 to drive iEC specification. To achieve these aims, we will use cutting edge technologies including single cell RNA-seq, ChIP-seq, and proximity proteomics. Together, these studies will define the molecular mechanisms that underlie the earliest stages of endothe- lial cell specification and that establish endothelial cell competence for interaction with support cells and forma- tion of functional vessels. This fundamental knowledge will form the foundation for strategies to promote vessel development in organ repair and regeneration.

总结