Neuroinflammation and vascular development in GMH

GMH 的神经炎症和血管发育

• 批准号: 10685146
• 负责人: Eric J Huang
• 金额: $ 58.07万
• 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: 2023-08-01 至 2028-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10685146
• 关键词: Agonist; Anti-Bacterial Agents; Bioenergetics; Bioinformatics; Biological Assay; Blood Vessels; Brain; Brain region; Cell Communication; Cell Maturation; Cell Nucleus; Cells; Cellular Stress; Cerebral Palsy; Cerebral cortex; Complex; Data; Defensins; Development; Disease; Disease model; Elastases; Endothelial Cells; Enzymes; Erythro; Exhibits; Fluorescence-Activated Cell Sorting; Histologic; Human; In Vitro; Inflammatory; Interneurons; Knowledge; Lead; Macrophage; Mediating; Microglia; Molecular; Morphogenesis; Myelogenous; Neonatal Mortality; Neurodevelopmental Disorder; Neuroglia; Neuroimmune; Organoids; PTPRC gene; Pathogenesis; Perinatal subependymal hemorrhage; Peroxidases; Population; Pregnancy; Premature Birth; Premature Infant; Prevalence; Property; Radial; Research; Research Project Grants; Role; Second Pregnancy Trimester; Signal Transduction; Testing; angiogenesis; antagonist; cytotoxic; improved; insight; matrigel; migration; monocyte; neonatal death; nerve stem cell; nestin protein; neural circuit; neuroblast; neurogenesis; neuroinflammation; neuropathology; neutrophil; prenatal; progenitor; single-cell RNA sequencing; stem cell biology; therapeutic target; transcriptomic profiling; transcriptomics

SUMMARY Preterm infants born between 21 to 30 gestational weeks (GW) have 20-40% chance of developing germinal matrix hemorrhage (GMH), which is a leading cause of neonatal mortality and neurodevelopmental disorders, such as cerebral palsy. Despite decades of research, however, there has been no significant improvement in the prevalence of preterm birth and the mechanism leading to GMH remains unclear. To understand the cause(s) for GMH, we have shown that, during the second trimester, germinal matrix contains enriched populations of Nestin+ radial glia and DCX+ neuroblasts that are fated to become GABAergic interneurons. Furthermore, DCX+ neuroblasts in the germinal matrix are organized as distinct clusters, called DCX-Enriched Nests or DENs, where they expand and migrate to the cerebral cortex and other deep nuclei before becoming mature GABAergic interneurons and integrating into the local neural circuits. To investigate why blood vessels in the germinal matrix are particularly vulnerable to develop GMH, we combined histological and ultrastructural analyses, fluorescence- activated cell sorting (FACS), and single-cell transcriptomics to characterize the properties of nascent blood vessels in the prenatal human brain from 15 to 25 GW. These studies lead to three main conclusions. First, during the second trimester the vascular network in the germinal matrix is much more complex than other brain regions. These nascent blood vessels are tiled by an ensemble of endothelial cells and mural cells, which follow distinct developmental trajectories and use diverse signaling mechanisms to facilitate cell-cell communication and maturation. Second, endothelial cells from younger brain (15-18 GW) exhibit stage-specific transcriptomic and bioenergetic features that are different from those from 20-23 GW. In addition, microglia-vasculature interactions stage-dependently promote angiogenesis in the germinal matrix, but not in the cortical plate. Finally, transcriptomic profiling of CD45+ cells in GMH cases showed that proinflammatory neutrophils and monocytes utilize antibacterial factors and CXCL16-S1PR1 signaling, respectively, to disrupt nascent vasculature in the germinal matrix. Collectively, our results support the overarching hypothesis that proinflammatory neutrophils and monocytes produce cytotoxic factors to disrupt angiogenesis and neurogenesis in the germinal matrix of preterm infants with GMH. To test this hypothesis, we propose to (1) characterize the cytotoxic properties of neutrophil-produced antibacterial factors in disrupting angiogenesis, (2) determine the impacts of CXCL16-S1PR1-mediated signaling in angiogenesis in the germinal matrix, and (3) examine the impacts of GMH on the neurogenesis and migration of GABAergic interneurons. Results from this project will provide important insights into disease mechanism of, and therapeutic targets for, GMH.

总结 在21至30孕周（GW）之间出生的早产儿有20-40%的机会发展为老年痴呆症 基质出血（GMH），是新生儿死亡和神经发育障碍的主要原因， 例如脑瘫。然而，尽管经过几十年的研究， 早产的发生率和导致GMH的机制仍不清楚。了解原因 对于GMH，我们已经表明，在孕中期，germinal矩阵包含丰富的 巢蛋白+放射状胶质细胞和DCX+神经母细胞，它们注定成为GABA能中间神经元。此外，DCX+ 在生发基质中的神经母细胞被组织成不同的簇，称为富含DCX的巢或DEN，其中 它们在变成成熟的GABA能前扩展并迁移到大脑皮层和其他深层核团 中间神经元并整合到局部神经回路中。为了研究为什么血管在生殖基质中 特别容易发展成GMH，我们结合了组织学和超微结构分析，荧光- 活化细胞分选（FACS）和单细胞转录组学来表征新生血液的性质 胎儿大脑中的血管从15到25 GW。这些研究得出三个主要结论。第一、 在孕中期，胚基质中的血管网络比其他脑组织复杂得多， 地区这些新生血管由内皮细胞和壁细胞的集合体平铺， 不同的发育轨迹，并使用不同的信号机制，以促进细胞间的沟通 和成熟。其次，来自年轻大脑（15-18 GW）的内皮细胞表现出阶段特异性转录组学 和不同于20-23 GW的生物能量特征。此外，小胶质血管 相互作用阶段依赖性地促进生发基质中的血管生成，但不促进皮质板中的血管生成。最后， GMH病例中CD 45+细胞的转录组学分析显示，促炎性中性粒细胞和单核细胞 分别利用抗菌因子和CXCL 16-S1 PR 1信号传导来破坏新生血管系统。 几何矩阵总的来说，我们的研究结果支持总体假设，即促炎性 嗜中性粒细胞和单核细胞产生细胞毒性因子，破坏血管生成和神经发生， GMH早产儿的German矩阵。为了验证这一假设，我们建议（1）描述 嗜热菌产生的抗菌因子在破坏血管生成中的细胞毒性特性，（2）确定 CXCL 16-S1 PR 1介导的信号传导对生殖基质中血管生成的影响，以及（3）检查CXCL 16-S1 PR 1介导的信号传导对生殖基质中血管生成的影响。 GMH对GABA能中间神经元的神经发生和迁移的影响。该项目的成果将 为GMH的发病机制和治疗靶点提供了重要的见解。