Fibrinogen-mediated mechanisms of preterm infant brain injury

纤维蛋白原介导的早产儿脑损伤机制

基本信息

批准号：
10450658
负责人：
MARK A PETERSEN
金额：
$ 14.61万
依托单位：
J. DAVID GLADSTONE INSTITUTES
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-07-01 至 2023-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10450658
关键词：
Alzheimer&apos s Disease Area Binding Biological Assay Blood - brain barrier anatomy Blood Proteins Blood Vessels Blood coagulation Bone Morphogenetic Proteins Brain Cell Maturation Cells Cellular Assay Central Nervous System Chronic Cicatrix Co-Immunoprecipitations Cytoplasmic Granules Data Deposition Development Developmental Disabilities Environment Event Failure Fibrinogen Fibrinogen Receptors Funding Gene Expression Profile Goals Growth Hemorrhage Hypoxia Impairment In Vitro Integrins Intraventricular Injections Knockout Mice Knowledge Label Ligands Link Mediating Modeling Molecular Multiple Sclerosis Mutant Strains Mice Myelin Neonatal Brain Injury Nerve Degeneration Nervous System Neurons Oligodendroglia Pathologic Pathology Pathway interactions Physicians Premature Birth Premature Infant Preterm brain injury Production Proteins Reporter Research Risk Role Scientist Signal Transduction Signaling Protein Site Slice Testing Therapeutic Time Traumatic injury Work activin A blood-brain barrier disruption bone morphogenetic protein receptors brain dysfunction cell transformation central nervous system injury defined contribution design experience in vivo infant brain injury inhibitor injured intraventricular hemorrhage myelination neonatal brain neonatal mice nerve stem cell nervous system disorder neurodevelopment neurogenesis neuroinflammation neurovascular new therapeutic target newborn brain injury novel novel therapeutic intervention oligodendrocyte progenitor pharmacologic prevent progenitor progenitor system receptor repaired response skills stem cells therapeutic target thrombotic transcriptome transcriptome sequencing two photon microscopy

Alzheimer&apos s Disease Area Binding Biological Assay Blood - brain barrier anatomy Blood Proteins Blood Vessels Blood coagulation Bone Morphogenetic Proteins Brain Cell Maturation Cells Cellular Assay Central Nervous System Chronic Cicatrix Co-Immunoprecipitations Cytoplasmic Granules Data Deposition Development Developmental Disabilities Environment Event Failure Fibrinogen Fibrinogen Receptors Funding Gene Expression Profile Goals Growth Hemorrhage Hypoxia Impairment In Vitro Integrins Intraventricular Injections Knockout Mice Knowledge Label Ligands Link Mediating Modeling Molecular Multiple Sclerosis Mutant Strains Mice Myelin Neonatal Brain Injury Nerve Degeneration Nervous System Neurons Oligodendroglia Pathologic Pathology Pathway interactions Physicians Premature Birth Premature Infant Preterm brain injury Production Proteins Reporter Research Risk Role Scientist Signal Transduction Signaling Protein Site Slice Testing Therapeutic Time Traumatic injury Work activin A blood-brain barrier disruption bone morphogenetic protein receptors brain dysfunction cell transformation central nervous system injury defined contribution design experience in vivo infant brain injury inhibitor injured intraventricular hemorrhage myelination neonatal brain neonatal mice nerve stem cell nervous system disorder neurodevelopment neurogenesis neuroinflammation neurovascular new therapeutic target newborn brain injury novel novel therapeutic intervention oligodendrocyte progenitor pharmacologic prevent progenitor progenitor system receptor repaired response skills stem cells therapeutic target thrombotic transcriptome transcriptome sequencing two photon microscopy

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项目摘要

PROJECT SUMMARY/ABSTRACT Preterm infant brain injury is often associated with blood-brain barrier (BBB) disruption and altered maturation of the central nervous system (CNS) progenitor cells required for normal brain development and myelination. The molecular signals in the injured microenvironment that inhibit CNS progenitor maturation are not fully known. Thus, no therapeutic options are available to prevent the developmental disabilities associated with preterm birth. BBB disruption alters the CNS progenitor niche by allowing blood proteins into the CNS. Fibrinogen, a blood coagulation protein, crosses a leaky BBB and is a key contributor to neuroinflammation, glial scar formation, neurodegeneration, and inhibition of CNS repair. We hypothesize that fibrinogen is a critical component of the microenvironment in preterm infant brain injury that inhibits CNS progenitor cell maturation to impair brain growth and myelination. Our preliminary studies show: (1) neonatal mice subjected to chronic hypoxia display prominent cerebellar pathology that includes fibrinogen deposition, myelination deficits, and impaired cerebellar growth; (2) Intraventricular injection of fibrinogen in neonatal mice disrupts cerebellar development in vivo; (3) Fibrinogen activates the bone morphogenetic protein (BMP) receptor activin A receptor type I (ACVR1) in oligodendrocyte progenitor cells (OPCs) to inhibit OPC maturation and myelination, (4) Fibrinogen inhibits neurogenesis from neuronal progenitor cells in vitro. Our specific aims will test our working model, whereby fibrinogen deposition after BBB disruption induces ACVR1-mediated BMP signaling in CNS progenitor cells to inhibit neurogenesis and myelination leading to abnormal neurodevelopment. In Aim 1, we will define the contribution of fibrinogen to preterm infant brain injury in vivo using fibrinogen mutant mice. In Aim 2, we will identify the cellular mechanisms that contribute to preterm infant brain injury at sites of fibrinogen deposition using in vivo two-photon microscopy (2PM). In Aim 3, we will determine the molecular mechanism of fibrinogen-induced activation of ACVR1 using in vitro binding and cellular assays. These studies will reveal the molecular link between BBB disruption and failure of CNS progenitor cell maturation in preterm infant brain injury. My goal is to become an independent physician- scientist and leader in the field of newborn brain injury. To continue my progress towards this goal, I will build upon the expertise of the Gladstone Institutes and UCSF to expand my research skills in the following areas: (1) in vivo 2PM to study the dynamic cellular responses to BBB disruption and fibrinogen deposition in the neonatal brain, (2) RNA-sequencing and transcriptome analysis to identify fibrinogen-mediated mechanisms of extrinsic inhibition, and (3) binding assays and inhibitor studies to discover novel fibrinogen receptors on CNS progenitor cells. The knowledge and experience gained from this proposal will allow me to compete for R01 funding focused on therapeutics that target the inhibitory microenvironment in preterm infant brain injury.

项目摘要/摘要早产婴儿脑损伤通常与血脑屏障（BBB）破坏和成熟改变有关正常脑发育和髓鞘形成所需的中枢神经系统（CNS）祖细胞。受伤的微环境中抑制CNS祖细胞成熟的分子信号尚未完全已知。因此，没有可用的治疗选择来防止与早产。 BBB破坏通过将血蛋白允许进入中枢神经系统来改变CNS祖细胞生态位。纤维蛋白原（一种血液凝血蛋白）穿过漏水的BBB，是神经炎症的关键因素，神经胶质疤痕形成，神经退行性和中枢神经系统修复的抑制作用。我们假设纤维蛋白原是在早产婴儿脑损伤中抑制CNS祖细胞细胞中微环境的关键成分成熟以损害大脑的生长和髓鞘形成。我们的初步研究表明：（1）受到的新生小鼠慢性缺氧显示出突出的小脑病理学，包括纤维蛋白原沉积，髓鞘化缺陷和小脑生长受损；（2）新生小鼠中脑室室内注射纤维蛋白原体内小脑发育；（3）纤维蛋白原激活骨形态发生蛋白（BMP）受体激活素少突胶质细胞祖细胞（OPC）中的I型I型（ACVR1）以抑制OPC成熟和（4）纤维蛋白原在体外抑制神经元祖细胞的神经发生。我们的具体目标将测试我们的工作模型，该模型通过BBB破坏后的纤维蛋白原沉积诱导ACVR1介导的BMP CNS祖细胞中的信号传导抑制神经发生和髓鞘形成导致异常神经发育。在AIM 1中，我们将定义纤维蛋白原对体内早产婴儿脑损伤的贡献使用纤维蛋白原突变小鼠。在AIM 2中，我们将确定有助于早产的细胞机制使用体内两光子显微镜（下午2点）的纤维蛋白原沉积部位的婴儿脑损伤。在AIM 3中，我们将确定使用体外结合和细胞测定。这些研究将揭示BBB破坏与中枢神经系统失败之间的分子联系早产婴儿脑损伤中的祖细胞成熟。我的目标是成为一名独立的医生 - 新生儿脑损伤领域的科学家和领导者。为了继续我的目标，我将建立借助Gladstone Institutes和UCSF的专业知识，可以在以下领域扩大我的研究技能：（1）在体内2pm研究对BBB破坏和纤维蛋白原沉积的动态细胞反应新生儿大脑（2）RNA测序和转录组分析，以鉴定纤维蛋白原介导的机制外部抑制作用和（3）结合测定和抑制剂研究，发现CNS上的新型纤维蛋白原受体祖细胞。从这一建议中获得的知识和经验将使我能够争夺R01 资金的重点是针对早产婴儿脑损伤的抑制性微环境的治疗剂。