Overcoming the Inhibitory Neurovascular Niche in Preterm Infant Brain Injury

克服早产儿脑损伤中的抑制性神经血管生态位

• 批准号: 10657221
• 负责人: MARK A PETERSEN
• 金额: $ 47.91万
• 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: 2023-04-01 至 2028-02-29
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10657221
• 关键词: Adult; Alzheimer' s Disease; Animal Model; Attenuated; Binding; Binding Sites; Blocking Antibodies; Blood - brain barrier anatomy; Blood Proteins; Blood Vessels; Blood coagulation; Bone Morphogenetic Proteins; Brain; Brain Injuries; Brain hemorrhage; Cell Maturation; Cells; Central Nervous System; Cerebellum; Cicatrix; Coagulation Process; Congenital cerebellar hypoplasia; Cytoplasmic Granules; Data; Deposition; Development; Developmental Disabilities; Disease; Environment; Fibrin; Fibrinogen; Fibrinogen Receptors; Functional disorder; Genetic; Goals; Growth; Hemorrhage; Human; ITGAM gene; ITGB2 gene; Impairment; Inflammation; Inflammatory; Injections; Injury; Innate Immune Response; Integrins; Knock-in Mouse; Link; Lipopolysaccharides; Macrophage; Microglia; Modeling; Molecular; Molecular Profiling; Monoclonal Antibodies; Multiple Sclerosis; Mutant Strains Mice; Mutate; Myelin; Natural regeneration; Neonatal; Nerve Degeneration; Neurodevelopmental Impairment; Neurons; Organoids; Outcome; Oxidative Stress; Pathway interactions; Plasma; Pluripotent Stem Cells; Population; Premature Infant; Proliferating; Proteins; Reactive Oxygen Species; Receptor Signaling; Risk; Role; Signal Transduction; Signaling Protein; Testing; Therapeutic; Trauma; Traumatic injury; Treatment Efficacy; Work; activin A; blood-brain barrier disruption; bone morphogenetic protein receptor type I; bone morphogenetic protein receptors; clinically relevant; defined contribution; disability; efficacy evaluation; human disease; immune activation; improved; in vivo; induced pluripotent stem cell; infant brain injury; inhibitor; neonatal mice; nerve stem cell; nervous system development; nervous system disorder; neurogenesis; neuroinflammation; neurovascular; novel; novel therapeutic intervention; pharmacologic; preservation; prevent; progenitor; receptor; remyelination; repaired; small molecule inhibitor; stem cells; systemic inflammatory response; transcriptomics

PROJECT SUMMARY/ABSTRACT Preterm infants are at risk for central nervous system (CNS) hemorrhage which can disrupt cerebellar maturation and lead to permanent neurodevelopmental impairment. The molecular signals in the disrupted neurovascular niche that block cerebellar development are not known. Thus, no therapeutics are available to prevent the developmental disabilities associated with preterm brain hemorrhage. Fibrinogen, a blood coagulation protein, crosses a leaky blood-brain barrier (BBB) and is a key driver of neuroinflammation, oxidative stress, neurodegeneration, glial scar formation, and inhibition of repair. We hypothesize that fibrinogen is a critical component of the neurovascular niche after BBB disruption that blocks cerebellar development in preterm infants. Our preliminary studies show: 1) Lipopolysaccharide (LPS)-induced systemic inflammation in neonatal mice increases vascular activation, fibrinogen deposition, and neuroinflammation in the cerebellum; 2) Fibrinogen depletion rescues cerebellar growth in systemic neonatal inflammation and plasma injection models of BBB disruption; 3) Fibrinogen inhibits neurogenesis from cerebellar granule neuronal progenitors (CGNPs) and is sufficient to disrupt cerebellar growth in vivo; 4) Fibrinogen activates the bone morphogenetic protein (BMP) receptor activin A receptor type I (ACVR1) in CNS progenitor cells to inhibit remyelination and neurogenesis; 5) Fibrin binds the CD11b/CD18 integrin receptor on microglia/macrophages to induce pro- inflammatory and pro-oxidant pathways that are toxic to CNS progenitor cells and impair regeneration; 6) fggγ390-396A knock-in mice, in which the binding site of fibrin to the CD11b integrin is mutated, have improved cerebellar growth during systemic neonatal inflammation. Our specific aims will test our working model, whereby fibrinogen deposition after BBB disruption alters cerebellar development through: 1) direct inhibitory effects on CGNPs via ACVR1 signaling, and 2) activation of innate immune responses via CD11b. In Aim 1, we will define the contribution of aberrant ACVR1 signaling to fibrinogen-induced cerebellar injury using CGNP-specific ACVR1 mutant mice and clinically relevant ACVR1 small molecule inhibitors. In Aim 2, we will determine the role of fibrin- CD11b-induced innate immune activation to cerebellar injury using fibrinogen mutant mice and a novel monoclonal antibody that blocks the interaction of fibrin with CD11b. In Aim 3, we will define how fibrinogen- ACVR1 signaling alters human cerebellar progenitor cell fate in induced pluripotent stem cell-derived cerebellar organoids using single cell transcriptomics. These studies will reveal molecular mechanisms at the neurovascular interface that link BBB disruption to CNS progenitor cell dysfunction in preterm infant brain injury. Thus, results from this proposal may open new treatment strategies to overcome the inhibitory neurovascular niche in preterm infant brain injury as well as other neurologic diseases with BBB disruption and fibrinogen deposition, such as multiple sclerosis, Alzheimer disease, and traumatic injury.

项目摘要/摘要 早产婴儿有可能破坏小脑成熟的中枢神经系统（CNS）出血 并导致永久性神经发育障碍。神经血管中断的分子信号 该块小脑发育的利基市场尚不清楚。那是没有治疗来防止 与早产脑出血相关的发育障碍。纤维蛋白原，一种血液凝血蛋白， 穿过漏水的血脑屏障（BBB），是神经炎症，氧化应激的关键驱动力， 神经变性，神经胶质疤痕形成和抑制修复。我们假设纤维蛋白原是关键 BBB破坏后神经血管生态位的成分阻碍了早产的小脑发育 婴儿。我们的初步研究表明：1）脂多糖（LPS）诱导的新生儿注射 小鼠会增加小脑血管活化，纤维蛋白原沉积和神经炎症。 2） 纤维蛋白原耗竭挽救了全身新生儿注射和血浆注入模型的小脑生长 BBB中断； 3）纤维蛋白原抑制小脑颗粒神经元祖细胞（CGNP）的神经发生 并且足以破坏体内小脑生长； 4）纤维蛋白原激活骨形态发生蛋白 （BMP）CNS祖细胞中I型受体I型受体A型受体A型受体，以抑制再生和 神经发生； 5）纤维蛋白在小胶质细胞/巨噬细胞上结合CD11b/CD18整联蛋白受体，以诱导促进 对中枢神经系统祖细胞有毒并损害修订的炎性和促氧化途径； 6） FGGγ390-396A敲入小鼠，其中纤维蛋白与CD11b整合素的结合位点已突变，已改善 全身新生儿炎症期间的小脑生长。我们的具体目标将测试我们的工作模型， BBB干扰后的纤维蛋白原沉积通过以下方式改变小脑的发育：1）直接抑制作用对 通过ACVR1信号传导CGNP，以及2）通过CD11b激活先天免疫调查。在AIM 1中，我们将定义 使用CGNP特异性ACVR1，异常ACVR1信号传导对纤维蛋白原诱导的小脑损伤的贡献 突变小鼠和临床相关的ACVR1小分子抑制剂。在AIM 2中，我们将确定纤维蛋白的作用 CD11b诱导的先天免疫对小脑损伤的激活，使用纤维蛋白原突变小鼠和一种新颖 单克隆抗体阻断纤维蛋白与CD11b的相互作用。在AIM 3中，我们将定义纤维蛋白原 - ACVR1信号传导改变了诱导多能干细胞衍生的小脑中的人小脑祖细胞命运 使用单细胞转录组学的器官。这些研究将揭示分子机制 在早产婴儿脑损伤中，将BBB破坏与CNS祖细胞功能障碍联系起来的神经血管界面。 这是该提案的结果可能会开放新的治疗策略，以克服抑制性神经血管 早产婴儿脑损伤以及其他神经系统疾病的利基市场患有BBB破坏和纤维蛋白原 沉积，例如多发性硬化症，阿尔茨海默氏病和创伤性损伤。