Elucidating the role of pericytes in angiogenesis in the brain using a tissue-engineered microvessel model

使用组织工程微血管模型阐明周细胞在大脑血管生成中的作用

• 批准号: 10648177
• 负责人: Peter C Searson
• 金额: $ 20.47万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-03-15 至 2025-02-28
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10648177
• 关键词: 3-Dimensional; Acceleration; Address; Adult; Altitude; Angiogenic Factor; Animal Model; Back; Biological Assay; Biological Process; Blood - brain barrier anatomy; Blood Vessels; Blood capillaries; Brain; Cell Therapy; Cells; Central Nervous System; Cerebrovascular system; Complex; Cues; Development; Disease; Endothelial Cells; Endothelium; Engineering; Exercise; Exposure to; FGF2 gene; Fluorescent in Situ Hybridization; Foundations; Genes; Genetic Engineering; Goals; Growth; Growth Factor; Human; Hypoxia; In Situ; Injury; Knowledge; Lead; Malignant neoplasm of brain; Modeling; Mus; Neurodegenerative Disorders; Neurons; Pathologic; Pathologic Processes; Pericytes; Phase; Phenotype; Physiological; Physiological Processes; Play; Process; RNA; Reporting; Retina; Role; Stroke; Testing; Tissue Engineering; Trauma; Traumatic Brain Injury; Vascularization; angiogenesis; cerebral microvasculature; cerebrovascular; experimental study; human model; migration; novel therapeutics; postcapillary venule; repaired; response; response to injury; stem cell technology; therapy design; tissue repair; transcriptome sequencing; transcriptomics; venule; wound healing

Project Summary Angiogenesis is a complex and tightly regulated process essential for tissue repair in response to injury or disease. In the adult human brain, the cerebrovasculature is largely quiescent but angiogenesis is associated with disease (e.g. stroke, neurodegenerative disease, and brain cancer), wound healing (e.g. traumatic brain injury), and physiological factors (e.g. exercise, high altitude adaptation, etc.). In the generally accepted model of brain angiogenesis, when blood vessels are exposed to angiogenic cues, pericytes detach from activated endothelial cells and are passive bystanders during sprout formation and growth, only migrating back to surround the newly formed endothelium in the maturation phase. However, a few studies in humans and animal models have reported that pericytes can play an active role in angiogenesis, leading sprout growth and promoting the migration of stalk cells. The objective of this project is to identify the role of pericytes during angiogenesis in the brain. Our major hypothesis is that pericytes can be selectively activated to become the tip cells and lead the growth of angiogenic sprouts. The objective will be addressed in three aims. In Aim 1, we will use a tissue-engineered post-capillary venule model to assess the role of hypoxia and growth factor gradients on pericyte-guided angiogenesis. In Aim 2, we will evaluate the transcriptomic profile of pericytes during angiogenesis. Immunostaining and in situ RNA hybridization will be used to examine key markers of tip cells as well as to validate key sequencing results. In Aim 3, we will perform a proof-of-concept experiment to test the hypothesis that gene editing can be used to engineer tip cell phenotype. Understanding the role of pericytes in adult brain angiogenesis is important in understanding the response to the cerebrovasculature to a wide range of perturbations and will enable the development of new therapies for cerebrovascular repair.

项目摘要 血管生成是一个复杂的和严格调节的过程，对于组织修复是必要的， 疾病在成年人的大脑中，脑血管系统基本上是静止的，但血管生成是相关的。 疾病（如中风、神经退行性疾病和脑癌）、伤口愈合（如创伤性脑损伤） 损伤）和生理因素（例如，锻炼、高海拔适应等）。在普遍接受的模型中， 在脑血管生成的过程中，当血管暴露于血管生成的信号时，周细胞从激活的 内皮细胞和被动旁观者在芽形成和生长，只有迁移回周围 在成熟阶段新形成的内皮。然而，一些对人类和动物模型的研究 已经报道周细胞可以在血管生成中发挥积极作用，引导芽生长并促进生长。 柄细胞的迁移。 本项目的目的是确定周细胞在脑血管生成过程中的作用。我们的主要 假设周细胞可以被选择性激活成为尖端细胞，并引导血管生成细胞的生长， 芽菜这一目标将在三个目标中实现。在目标1中，我们将使用组织工程后毛细血管 微静脉模型，以评估缺氧和生长因子梯度对周细胞引导的血管生成的作用。在Aim中 2，我们将评估周细胞在血管生成过程中的转录组学谱。免疫染色和原位RNA 杂交将被用于检查尖端细胞的关键标记以及验证关键测序结果。在 目标3，我们将进行概念验证实验，以验证基因编辑可用于 工程化尖端细胞表型。了解周细胞在成人脑血管生成中的作用对于 了解血管系统对各种扰动的反应，并将使 开发脑血管修复的新疗法。