Mural stem cells in normal development and germinal matrix hemorrhage in the preterm human brain

正常发育的壁干细胞和早产人脑生发基质出血

基本信息

批准号：
10308716
负责人：
Elizabeth Erin Crouch
金额：
$ 20.29万
依托单位：
UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-12-01 至 2025-11-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10308716
关键词：
3-Dimensional Adult Affect Age Astrocytes Atlases Autopsy Biological Assay Biology Blood - brain barrier anatomy Blood Vessels Blood flow Brain Brain Injuries Brain hemorrhage Brain region CRISPR/Cas technology CSPG4 gene Cell Culture Techniques Cell Lineage Cells Cerebral Palsy Cerebrovascular system Cessation of life Clustered Regularly Interspaced Short Palindromic Repeats Data Development Elements Endothelial Cells Endothelium Fluorescence-Activated Cell Sorting Future Ganglia Genes Gestational Age Goals Hemorrhage Human Immunohistochemistry In Vitro Infant Intellectual functioning disability Knowledge Lead Location Mental Depression Modeling Molecular Mus NOTCH3 gene Neonatal Nerve Degeneration Neurons Organoids Pathogenesis Pathologic Pattern Pericytes Perinatal subependymal hemorrhage Physicians Physiological Platelet-Derived Growth Factor beta Receptor Play Population Pregnancy Premature Infant Proliferating Research Personnel Role Scientist Signal Pathway Smooth Muscle Myocytes Structure Sum Testing Tissues Training Transplantation Tube Ventricular angiogenesis brain tissue capillary bed clinically relevant differential expression experimental study ganglion cell high risk human tissue in vivo lateral ventricle matrigel mouse model neonatal mice nerve stem cell neurogenesis neuroinflammation neuropathology neurovascular novel patient population perinatal medicine premature progenitor relating to nervous system response single-cell RNA sequencing stem cell biomarkers stem cells tool

项目摘要

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. Mural cells, which lie outside of endothelial cells, are critical 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 preferentially affect preterm infants, who are prone to develop hemorrhages specifically located in brain regions called the ganglionic eminences (GE). These hemorrhages confer high risk for permanent brain injury, and we have no treatments. To molecularly define the stages and subtypes of mural cells in the developing human brain, I developed a Fluorescence Activated Cell Sorting (FACS) strategy followed by single cell RNA sequencing (scRNAseq). These data revealed distinct stages of mural cell development from a relatively immature stage at 15GW to the emergence of distinct subtypes at 23 GW. Strikingly, this strategy also uncovered a novel progenitor population that express both mural and neural stem cell markers, suggesting a common neurovascular progenitor that contributes to both angiogenesis and neurogenesis. In the GE, immunohistochemistry with putative neurovascular progenitor markers revealed these cells in a periventricular and perivascular location situated amongst canonical neural stem cells. Consistent with these results, FACS-purified mural cells produce both vascular and neural lineage cells in vitro. In sum, the above experiments lead me to hypothesize that mural cells contribute to both angiogenesis and neurogenesis during development and that a predominance of immature stages of mural cells in the GE contributes to the pathogenesis of hemorrhage. My short-term goals for the proposed K08 are the following: 1. Construct an atlas of human brain mural cells during development using scRNAseq and postmortem human tissue. 2. Develop organoid, mouse transplant, and CRISPR tools to manipulate and interrogate the role of specific signaling pathways of mural cells in human brain development. These results will propel me to an R01. As a physician scientist in Neonatal- Perinatal Medicine, I am uniquely suited to advance 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下都发挥着关键作用。内皮细胞和壁细胞构成了人体的主要结构和功能元素脉管系统。壁细胞位于内皮细胞之外，对于细胞的发育至关重要血脑屏障并根据神经元活动调节血流。然而，大多数脑血管研究使用成年小鼠，因此对发育中的脑血管知之甚少任何年龄的脉管系统或人脑血管。这些知识差距优先影响早产儿，他们容易出现出血，特别是位于称为神经节隆起（GE）。这些出血会带来永久性脑损伤的高风险，我们没有治疗方法。从分子水平上定义发育中的人类壁细胞的阶段和亚型大脑，我开发了一种荧光激活细胞分选（FACS）策略，然后是单细胞 RNA 测序 (scRNAseq)。这些数据揭示了壁细胞发育的不同阶段从 15GW 的相对不成熟阶段到 23GW 不同亚型的出现。引人注目的是，这一策略还发现了一种新的祖先群体，它既表达壁画又表达神经干细胞标记物，表明有一个共同的神经血管祖细胞对两者都有贡献血管生成和神经发生。在 GE 中，免疫组织化学与假定的神经血管祖细胞标记显示这些细胞位于心室周围和血管周围位置典型的神经干细胞之一。与这些结果一致，FACS 纯化的壁细胞在体外产生血管和神经谱系细胞。总之，上述实验引导我假设壁细胞有助于血管生成和神经发生 GE 中壁细胞的未成熟阶段占主导地位有助于出血的发病机制。我对拟议的 K08 的短期目标具体如下： 1. 使用以下方法构建开发过程中的人脑壁细胞图谱： scRNAseq 和死后人体组织。 2. 开发类器官、小鼠移植和 CRISPR 操纵和询问人类壁细胞特定信号通路作用的工具大脑发育。这些结果将推动我达到 R01。作为一名新生儿医学科学家围产期医学，我非常适合增进我们对脑血管的理解这些知识的发展和应用在未来的治疗中。