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/摘要脑血管细胞在生理和病理状态下都起着关键作用。内皮细胞和壁细胞组成的主要结构和功能元素脉管系统。位于内皮细胞之外的壁细胞对于发展至关重要血液脑屏障并调节对神经元活性的血流。但是，大多数对脑血管的研究使用成年小鼠，因此对发展知之甚少在任何年龄的脉管系统或人类脑血管。这些知识差距优先影响早产儿，他们容易发展出在大脑区域的出血神经节恩典（GE）。这些出血给永久性脑损伤带来高风险，而且我们没有治疗。分子定义发展中壁细胞的阶段和亚型大脑，我开发了一种荧光激活的细胞分选（FACS）策略，然后是单细胞 RNA测序（SCRNASEQ）。这些数据揭示了壁细胞开发的不同阶段从相对不成熟的阶段在15GW到23 GW处的不同亚型的出现。令人惊讶的是，该策略还发现了一个新的祖先种群，该祖先既表达壁画又表达神经干细胞标记，提出一种常见的神经血管祖细胞，这两者兼而有之血管生成和神经发生。在GE中，具有假定神经血管的免疫组织化学祖细胞标记揭示了这些细胞位于周围的和血管周的位置在规范神经干细胞中。与这些结果一致，FACS纯化的壁画细胞在体外产生血管和神经谱系细胞。总而言之，以上实验使我假设壁画细胞有助于血管生成和神经发生发育和GE中壁细胞的未成熟阶段的占主导地位有助于出血的发病机理。我针对拟议的K08的短期目标如下：1。使用 scrnaseq和验尸后人体组织。 2。发展器官，鼠标移植和CRISPR 操纵和审问壁细胞在人类中特定信号通路的作用的工具大脑发育。这些结果将使我进入R01。作为新生儿的医师科学家围产期医学，我非常适合促进我们对脑血管的理解发展和将此知识应用于未来的治疗。