Endothelial Cell Cycle State and Cell Fate

内皮细胞周期状态和细胞命运

基本信息

批准号：
10208947
负责人：
Karen Kemper Hirschi
金额：
$ 52.61万
依托单位：
UNIVERSITY OF VIRGINIA
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-08-15 至 2023-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10208947
关键词：
Arteries Blood Circulation Blood Vessels Blood capillaries Blood flow Cardiovascular system Cell Culture System Cell Cycle Cell Cycle Regulation Cell Proliferation Cells Clinical Data Defect Development Ectoderm Embryonic Development Endoderm Endothelial Cells Endothelium Engineering Ensure Evaluation Exhibits G1 Arrest G1 Phase Gene Expression Genes Growth Human In Vitro Maintenance Mesoderm Metabolic Modeling Molecular Mus Natural regeneration Nutrient Oxygen Pathology Phenotype Process Regenerative Medicine Regulation Research Role Signal Pathway Signal Transduction Structure Testing Tissue Engineering Tissues Tube Up-Regulation Vascular remodeling Vascularization Veins Venous Venous Malformation Waste Products arteriole blood vessel development cell growth embryonic stem cell human stem cells in vivo in vivo Model injured injury and repair insight ischemic injury mechanotransduction migration mouse model neovascularization notch protein novel postnatal prevent repaired replacement tissue response restoration shear stress vascular injury vasculogenesis venule

项目摘要

Establishing a functional vascular network is a rate-limiting step in embryonic development, the repair of injured tissues, and the engineering of tissue replacements. Although we have made progress in identifying factors that promote endothelial cell proliferation and sprouting, we lack understanding of how to properly control endothelial cell growth and phenotypic specialization during vascular remodeling, which has created a significant roadblock for clinical therapies, tissue engineering and regenerative medicine. Although multiple signaling pathways have been implicated in the regulation of arterial-venous network formation, including flow-induced mechanotransduction and Notch signaling, the mechanisms by which these signals coordinately regulate endothelial cell growth suppression and identity were unclear. Our recent studies revealed that remodeling vascular plexi are subject to systemic blood circulation, and that shear stress of different magnitudes promotes differential growth responses and gene expression. That is, arterial/arteriolar shear stress levels promote Notch signaling, and downstream p27-induced late G1 phase arrest that enables arterial gene expression (Fang 2017). Conversely, flow magnitudes typical of veins/venules induce early G1 arrest, and enables upregulation of venous genes. Interestingly, distinct endothelial cell cycle states appear to be maintained in arteries vs. veins postnatally. We know very little about the role of cell cycle control in endothelial cell fate decisions, or the differential signaling pathways induced by vessel-specific flow magnitudes, and how they may coordinately induce and maintain endothelial cell cycle state and identity. The scientific premise of our research is that endothelial cell cycle control is required for proper arterial and venous specification, such that when endothelial cells are in different cell cycle states, they exhibit different propensity for arterial vs. venous gene expression. Support for this idea comes from studies in embryonic stem cells that show cells in early vs. late G1 phase have a propensity for mesoderm/endoderm vs. ectoderm fate, respectively (Paulkin 2014). Thus, our hypothesis is that differential flow forces in arteries and veins induce different intracellular signaling pathways that promote distinct endothelial cell cycle states, creating distinct windows of opportunity for the regulation of arterial vs. venous gene expression. To ensure scientific rigor, we will test this hypothesis in vivo in models of arterial- venous network formation and repair, and in vitro in human endothelial cell culture systems that allow flow manipulation. We will define mechanisms by which vessel-specific flow magnitudes modulate endothelial cell cycle state, determine how distinct endothelial cell cycle states enable differential phenotypic specialization (artery vs. vein), and determine whether manipulation of endothelial cell cycle state can prevent or correct arterial-venous malformations and enhance post-injury vascular repair. Evaluation of this hypothesis will yield novel fundamental insights into blood vessel formation and regeneration that can be used to create human microvasculature ex vivo and treat vascular pathologies.

建立功能性血管网络是胚胎开发的限制步骤受伤的组织和组织置换的工程。尽管我们在识别促进内皮细胞增殖和发芽的因素，我们对如何正确控制血管重塑期间内皮细胞的生长和表型专业化，这产生了重要的用于临床疗法，组织工程和再生医学的障碍。虽然多个信号传导途径已与动脉 - 观网形成的调节有关，包括流动诱导机械传输和置换信号传导，这些信号协调调节的机制内皮细胞生长抑制和身份尚不清楚。我们最近的研究表明，重塑血管丛受到全身性血液循环的影响，而不同幅度的剪切应力会促进差异生长反应和基因表达。也就是说，动脉/动脉剪切应力水平促进了缺口信号传导和下游P27诱导的G1后期停滞，使动脉基因表达（Fang 2017）。相反，静脉/静脉的典型流量大小会引起早期的G1停滞，并可以上调静脉基因。有趣的是，在产后，在动脉与静脉中似乎保持了不同的内皮细胞周期状态。我们对细胞周期控制在内皮细胞命运决策或差异信号的作用知之甚少容器特异性流量诱导的途径，以及它们如何协调诱导和维持内皮细胞周期状态和身份。我们研究的科学前提是内皮细胞周期需要控制适当的动脉和静脉规范，因此当内皮细胞处于不同细胞周期状态，它们表现出动脉与静脉基因表达的不同倾向。支持这个想法来自在胚胎干细胞中的研究，这些干细胞显示早期细胞与G1晚期具有的倾向中胚层/内胚层和外胚层命运（Paulkin 2014）。因此，我们的假设是差异动脉和静脉中的流动诱导不同的细胞内信号通路，促进不同的细胞内信号通路内皮细胞周期状态，为动脉VS的调节创造了不同的机会窗口。静脉基因表达。为了确保科学严谨，我们将在动脉模型中在体内检验这一假设静脉网络形成和修复，以及在人体内皮细胞培养系统中的体外，允许流动操纵。我们将定义机制，该机制通过该机制调节容器特异性流量幅度调节内皮细胞循环状态，确定不同的内皮细胞周期状态如何实现差异表型专业化（动脉与静脉），并确定内皮细胞周期状态的操纵是否可以预防或纠正动脉畸形并增强伤害后血管修复。评估该假设将产生可用于创造人类的血管形成和再生的新型基本见解微脉管系统离体和治疗血管病理。