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 早产儿有中枢神经系统(CNS)出血的风险，这可能会扰乱小脑成熟 并导致永久性神经发育障碍。受损神经血管中的分子信号 阻碍小脑发育的利基位置尚不清楚。因此，没有可用的治疗方法来预防 与早产儿脑出血相关的发育障碍。纤维蛋白原，一种凝血蛋白， 穿过漏水的血脑屏障(BBB)，是神经炎症、氧化应激、 神经变性、神经胶质瘢痕形成和修复抑制。我们假设纤维蛋白原是一种关键的 早产儿血脑屏障中断后阻碍小脑发育的神经血管壁龛成分 婴儿。我们的初步研究表明：1)脂多糖(LPS)诱导新生儿全身炎症 小鼠增加了小脑的血管激活、纤维蛋白原沉积和神经炎症；2) 在系统性新生儿炎症和血浆注射模型中，纤维蛋白原耗竭挽救小脑生长 3)纤维蛋白原抑制小脑颗粒神经前体细胞的神经发生 4)纤维蛋白原激活骨形态发生蛋白 (BMP)受体激活素A受体I型(ACVR1)在中枢神经系统前体细胞中抑制再髓鞘形成和 5)纤维蛋白与小胶质细胞/巨噬细胞表面的CD11b/CD18整合素受体结合，诱导促进 对中枢神经系统祖细胞有毒性并阻碍再生的炎症和氧化途径；6) FGGCD390-396A敲入鼠中，纤维蛋白与γ整合素的结合位点发生突变，这一点得到了改善 全身性新生儿炎症期间的小脑发育。我们的具体目标将检验我们的工作模式，从而 血脑屏障中断后纤维蛋白原沉积通过以下方式改变小脑发育：1)对 2)通过CD11b激活天然免疫应答。在目标1中，我们将定义 利用CGNP特异性ACVR1研究ACVR1信号在纤维蛋白原诱导的小脑损伤中的作用 突变小鼠和临床相关的ACVR1小分子抑制剂。在目标2，我们将确定纤维蛋白的作用- CD11b诱导的纤维蛋白原突变小鼠对小脑损伤的天然免疫激活 阻断纤维蛋白与CD11b相互作用的单抗。在目标3中，我们将定义纤维蛋白原如何- ACVR1信号改变诱导多能干细胞来源小脑中人小脑祖细胞的命运 使用单细胞转录组的有机化合物。这些研究将揭示分子机制在 早产儿脑损伤中血脑屏障破坏与中枢神经前体细胞功能障碍之间的神经血管接口。 因此，这一建议的结果可能会打开新的治疗策略，以克服抑制神经血管 早产儿脑损伤以及其他伴有血脑屏障紊乱和纤维蛋白原的神经系统疾病的生态位 沉积，如多发性硬化症、阿尔茨海默病和创伤性损伤。