Vascular mural cells in the development of the blood brain barrier

血脑屏障发育中的血管壁细胞

• 批准号: 10687558
• 负责人: Elizabeth Erin Crouch
• 金额: $ 138.89万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-09-01 至 2026-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10687558
• 关键词: Adult; Age; Astrocytes; Atlases; Blood - brain barrier anatomy; Blood Cells; Blood Vessels; Blood flow; Brain; Cell physiology; Cells; Cerebrovascular system; Consumption; Data; Detection; Development; Disease; Effector Cell; Elements; Embryo; Endothelial Cells; Endothelium; Fibroblasts; Fluorescence-Activated Cell Sorting; Future; Genetic; Goals; Heterogeneity; Human; Knowledge; Lead; Microglia; Mitotic; Molecular; Mus; Neonatal; Nervous System Physiology; Neurons; Pathologic; Patients; Pericytes; Physiological; Play; Proteins; RNA; Reporting; Role; Second Pregnancy Trimester; Signal Transduction; Smooth Muscle Myocytes; Time; Tissues; brain cell; experimental study; neurovascular; neurovascular unit; prenatal; progenitor; response; single-cell RNA sequencing; spatiotemporal; stem cells; transcriptomics; treatment strategy

PROJECT ABSTRACT/SUMMARY Brain blood vessel cells play key roles in both physiological and pathological states. Endothelial and mural cells compose the main structural and functional elements of the vasculature. Situated between the endothelial cell lumen and the surrounding astrocytes, microglia, and neurons, mural cells serve as a critical signaling hub of the neurovascular unit. Mural cells are required for the development of the blood brain barrier and regulate blood flow in response to neuronal activity. However, most studies of brain blood vessels use adult mice and therefore little is known about the developing vasculature or human brain blood vessels at any age. These knowledge gaps impact patients. During development, both genetic and sporadic disorders of the vasculature can have a severe impact on long-term neurological function, and we currently have no treatments. To molecularly define the stages and subtypes of mural cells in the developing brain, we have developed an integrated approach to enrich for vascular mural cells, profile them, and functionally characterize their effects. In the prenatal human brain, Fluorescence Activated Cell Sorting (FACS) followed by single cell RNA sequencing (scRNAseq) identified four different types of mural cells during the second trimester: mitotic, classic pericyte, fibroblast, and smooth muscle cells. Trajectory analysis of our transcriptomic data showed smooth muscle cells as potential progenitors that give rise to classic pericytes and fibroblasts. The four mural cell subtypes had recently been reported in the adult human brain, and the presence of their full repertoire at such a young age suggests a highly dynamic and time-consuming maturation process for these cells. In the embryonic mouse brain, previous studies had not been able to define subtypes of mural cells. Our application of FACS followed by scRNAseq at embryonic day 13.5 uncovered the same subtypes as the prenatal human brain. In sum, the above experiments lead me to hypothesize that smooth muscle cells are evolutionarily conserved mural stem cells in the developing brain which give rise to classic pericytes, the differentiated effector cells of the blood brain barrier. My short-term goals for the proposed DP2 are the following: 1. Construct a spatiotemporal atlas of brain mural cells during development using scRNAseq and RNA and protein detection in tissue. 2. Perform lineage tracing with mouse and human cells to interrogate stage-specific roles in blood brain barrier development. As a neuroscientist, vascular biologist, and neonatologist, I am uniquely suited to pioneer our understanding of brain vascular development and the application of this knowledge towards future therapy.

项目摘要/总结 脑血管细胞在生理和病理状态中起着关键作用。 内皮细胞和壁细胞构成了血管的主要结构和功能元件。 脉管系统位于内皮细胞腔和周围的星形胶质细胞之间， 小胶质细胞和神经元，壁细胞作为神经血管的关键信号枢纽， 单位壁细胞是血脑屏障发育和调节血液所必需的 对神经元活动的反应。然而，大多数关于脑血管的研究使用成年小鼠， 因此对于发育中的脉管系统或人脑血管知之甚少， 任何年龄这些知识差距影响患者。在发育过程中，遗传性和散发性 血管系统的紊乱会对长期的神经功能产生严重的影响，我们 目前没有治疗方法。 为了从分子水平上确定发育中大脑壁细胞的阶段和亚型， 开发了一种综合方法来富集血管壁细胞，对其进行分析， 从功能上描述其效果。在产前人类大脑中，荧光激活细胞 分选（FACS）后进行单细胞RNA测序（scRNAseq）鉴定了四种不同的类型 在妊娠中期的壁细胞：有丝分裂，经典的周细胞，成纤维细胞，平滑肌 细胞我们的转录组数据的轨迹分析显示平滑肌细胞是潜在的 产生典型的周细胞和成纤维细胞的祖细胞。四种壁细胞亚型 最近被报道在成年人的大脑中，他们的全部剧目的存在， 年轻的年龄表明这些细胞的成熟过程是高度动态和耗时的。 在胚胎小鼠大脑中，以前的研究未能确定mural的亚型， 细胞我们在胚胎第13.5天应用FACS和scRNAseq，发现了相同的结果。 胎儿期的大脑综上所述，上述实验使我假设 平滑肌细胞是发育中进化上保守的壁干细胞， 产生经典的周细胞，即血脑的分化效应细胞 屏障我对拟议的DP2的短期目标如下：1。构建 使用scRNAseq和RNA的发育期间脑壁细胞的时空图谱， 组织中的蛋白质检测。2.使用小鼠和人类细胞进行谱系追踪以进行询问 在血脑屏障发育中的阶段特异性作用。作为一名神经科学家、血管生物学家， 作为新生儿学家，我非常适合开拓我们对脑血管的理解 开发和应用这些知识对未来的治疗。