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.

概括 21 至 30 孕周 (GW) 出生的早产儿有 20-40% 的机会发育成胚芽 基质出血（GMH）是新生儿死亡和神经发育障碍的主要原因， 比如脑瘫。然而，尽管经过数十年的研究，仍没有显着的改善。 早产的发生率以及导致 GMH 的机制仍不清楚。了解原因 对于 GMH，我们已经证明，在妊娠中期，生发基质含有丰富的 Nestin+放射状胶质细胞和DCX+神经母细胞注定会成为GABA能中间神经元。此外，DCX+ 生发基质中的神经母细胞被组织成不同的簇，称为 DCX 富集巢或 DEN，其中 在成为成熟的 GABA 能之前，它们会扩张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。研究为什么生发基质中有血管 特别容易发生 GMH，我们结合了组织学和超微结构分析、荧光- 激活细胞分选 (FACS) 和单细胞转录组学来表征新生血液的特性 产前人类大脑中的血管数量为 15 至 25 GW。这些研究得出三个主要结论。第一的， 在妊娠中期，生发基质中的血管网络比其他大脑复杂得多 地区。这些新生血管由内皮细胞和壁细胞的集合组成，随后 不同的发育轨迹并使用不同的信号机制来促进细胞间的通讯 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 以及与 20-23 GW 不同的生物能特征。此外，小胶质细胞血管系统 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 CD45+ 细胞的转录组学分析表明，促炎性中性粒细胞和单核细胞 分别利用抗菌因子和 CXCL16-S1PR1 信号传导来破坏新生血管 生发基质。总的来说，我们的结果支持了促炎症的总体假设 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 患有 GMH 的早产儿的生发基质。为了检验这个假设，我们建议（1）表征 中性粒细胞产生的抗菌因子在破坏血管生成方面的细胞毒性特性，(2) 确定 CXCL16-S1PR1 介导的信号对生发基质血管生成的影响，以及 (3) 检查 GMH 对 GABA 能中间神经元的神经发生和迁移的影响。该项目的结果将 为 GMH 的疾病机制和治疗靶点提供重要见解。