Fibroblast-derived laminin regulates blood-brain barrier integrity and fibroblast biology in hemorrhagic brain

成纤维细胞衍生的层粘连蛋白调节出血脑中的血脑屏障完整性和成纤维细胞生物学

• 批准号: 10749280
• 负责人: Yao Yao
• 金额: $ 48.98万
• 依托单位: UNIVERSITY OF SOUTH FLORIDA
• 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/10749280
• 关键词: Ablation; Adherens Junction; Affect; Apoptosis; Basal lamina; Biological Process; Biology; Blood - brain barrier anatomy; Blood Vessels; Blood capillaries; Brain; Brain Injuries; Brain hemorrhage; Cause of Death; Cells; Cerebral hemisphere hemorrhage; Cerebrovascular system; Cessation of life; Cicatrix; Death Rate; Development; Disease Outcome; Endothelial Cells; Fibroblasts; Fibronectins; Foundations; Gender; In Vitro; Infarction; Infiltration; Inflammatory; Injury; Innovative Therapy; Knowledge; Laboratories; Laminin; Learning; Mediating; Meninges; Mission; Modeling; Molecular; Molecular Target; Molecular Weight; Morphology; Mus; National Institute of Neurological Disorders and Stroke; Nerve Degeneration; Neurologic; Outcome; Pathogenesis; Physiological; Play; Population; Prevention strategy; Proliferating; Protein Isoforms; Proteins; Research; Role; Secondary to; Sensorimotor functions; Stroke; Testing; Therapeutic; Time; Tracer; Transgenic Mice; blood-brain barrier disruption; blood-brain barrier permeabilization; brain endothelial cell; cell type; central nervous system injury; clinically relevant; cognitive function; density; disability; effective therapy; genetic approach; improved; in vivo; injury and repair; innovation; laminin S; long term memory; middle age; migration; neuroprotection; novel; pharmacologic; receptor; repaired; single-cell RNA sequencing; transcriptome sequencing; transcytosis

Project Summary/Abstract The long-term objective of this application is to develop innovative therapies for intracerebral hemorrhage (ICH), which causes high rates of death and disability worldwide. This is consistent with the mission of NINDS. This proposal aims to investigate the biological functions of fibroblast-derived laminin in blood-brain barrier (BBB) repair and fibroblast biology after ICH and explore the underlying molecular mechanisms. In Aim 1, the function of fibroblast-derived laminin in ICH pathogenesis will be investigated in two clinically relevant ICH models using middle-aged transgenic mice with laminin deficiency in fibroblasts. In Aim 2, the function of fibroblast-derived laminin in BBB repair after ICH will be investigated both in vitro and in vitro. First, how loss of fibroblast-derived laminin affects BBB permeability and inflammatory cell infiltration after ICH will be examined (Aim 2A). Next, whether loss of fibroblast-derived laminin exacerbates BBB disruption via paracellular and/or transcellular mechanisms will be investigated (Aim 2B). Third, the receptors that mediate fibroblast-derived laminin’s “BBB-repairing” effect on endothelial cells will be identified using both pharmacological and genetic approaches (Aim 2C). In Aim 3, the function of fibroblast-derived laminin in fibroblast biology and fibrotic scar composition will be investigated. First, fibroblast biology (proliferation/apoptosis/migration), fibroblast morphology, and fibrotic scar components will be examined in vitro and in vivo (Aim 3A). Next, how exactly fibroblast-derived laminin regulates fibroblast biology and fibrotic scar composition will be explored by bulk and/or single-cell RNAseq analysis (Aim 3B). Third, the receptors that mediate these changes in fibroblasts will be identified using both pharmacological and genetic approaches (Aim 3C). Successful completion of this proposal will elucidate novel functions of fibroblast-derived laminin in BBB repair and fibroblast biology after ICH, identify the receptors that mediate these effects on both endothelial cells and fibroblasts, provide new molecular targets with therapeutic potential, and promote the development of innovative and effective treatments for ICH.

项目总结/摘要 这项应用的长期目标是开发脑出血的创新疗法 (ICH)这导致了世界范围内的高死亡率和残疾率。这与国家土著和民族研究所的使命是一致的。 本研究旨在探讨成纤维细胞源性层粘连蛋白在血脑屏障中的生物学功能 (BBB)修复和成纤维细胞生物学，并探索潜在的分子机制。在目标1中， 将在两个临床相关的ICH中研究成纤维细胞源性层粘连蛋白在ICH发病机制中的功能 使用成纤维细胞层粘连蛋白缺乏的中年转基因小鼠模型。在目标2中， 将在体外和体外研究成纤维细胞衍生的层粘连蛋白在ICH后BBB修复中的作用。第一，如何损失 将检查成纤维细胞衍生的层粘连蛋白影响血脑屏障通透性和ICH后的炎性细胞浸润 (Aim 2A）。接下来，研究成纤维细胞来源的层粘连蛋白的丢失是否通过细胞旁和/或细胞外基质介导而加剧BBB破坏。 将研究跨细胞机制（目的2B）。第三，介导成纤维细胞源性 层粘连蛋白对内皮细胞的“BBB修复”作用将通过药理学和遗传学方法来鉴定 方法（目标2C）。目的3：成纤维细胞源性层粘连蛋白在成纤维细胞生物学和纤维化瘢痕中的作用 组成将进行调查。首先，成纤维细胞生物学（增殖/凋亡/迁移），成纤维细胞 将在体外和体内检查形态和纤维化瘢痕成分（目的3A）。接下来， 成纤维细胞衍生的层粘连蛋白调节成纤维细胞生物学， 和/或单细胞RNAseq分析（Aim 3B）。第三，在成纤维细胞中介导这些变化的受体将 使用药理学和遗传学方法进行鉴定（目标3C）。成功完成本 该提案将阐明成纤维细胞衍生的层粘连蛋白在BBB修复和成纤维细胞生物学中的新功能， ICH，鉴定介导这些作用于内皮细胞和成纤维细胞的受体，提供了新的 具有治疗潜力的分子靶点，促进创新和有效的开发 治疗ICH。