Interrogating the Potential of Ccn1+ Astrocyte Niches to Drive Angiogenesis after Spinal Cord Injury

探讨 Ccn1 星形胶质细胞生态位在脊髓损伤后驱动血管生成的潜力

基本信息

批准号：
10607960
负责人：
Keshav Balaji Suresh
金额：
$ 4.04万
依托单位：
CEDARS-SINAI MEDICAL CENTER
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-02-01 至 2025-10-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10607960
关键词：
Address Adhesions Astrocytes Automobile Driving Axon Behavioral Behavioral Assay Biological Process Blood - brain barrier anatomy Blood Vessels Blood flow Brain Cell Proliferation Cells Central Nervous System Characteristics Complement Data Data Set Development Dimensions Distal Dryness Endothelial Cells Endothelium Environment Evaluation Evolution Family member Gene Expression Genes Genetic Transcription Goals Health Histologic Histology Homeostasis Hybrids Hypoxia ITGB3 gene In Vitro Injury Integrins Ischemia Knockout Mice Lateral Lesion Life Ligation Link Literature Location Locomotor Recovery Maintenance Mediating Methods Modeling Molecular Molecular Evolution Morphology Mus Nervous System Physiology Neurons Paralysed Pathology Pericytes Persons Phenotype Physiological Positioning Attribute Process Proliferating Recovery Recovery of Function Regenerative capacity Regulation Research Research Personnel Research Training Resources Role Signal Transduction Site Specific qualifier value Spinal Spinal Cord Spinal cord injury Stroke Technology Testing Therapeutic Tissues Training Transgenic Mice Up-Regulation Urination Vascular remodeling Vertebral column Walking Work angiogenesis axon regeneration axonal sprouting blood-brain barrier function career central nervous system injury conditional knockout daily functioning design grasp high dimensionality in silico in vivo innovation loss of function motor recovery mouse model nervous system disorder notch protein novel novel therapeutics permissiveness programs regeneration potential regenerative restoration spatiotemporal timeline transcriptomics translational neuroscience white matter

项目摘要

ABSTRACT Spinal Cord Injury (SCI) is a devastating neurological disorder, characterized by disruption to ascending and descending axonal networks, that can leave people paralyzed for life. After SCI, axons in spared white matter (WM) regions attempt to undergo short-range sprouting to restore normal neurological function. However, neuron-extrinsic factors that govern this short-range axon sprouting remain poorly understood. During development and after injury, new axons spatiotemporally follow new blood vessels, hence intimately linking axon sprouting to vasculature. Central Nervous System (CNS) vasculature is orchestrated by astrocytes, which physically interact with endothelia and pericytes to form the gliovascular unit- a tissue niche with indispensable roles in modulating blood flow and Blood Brain Barrier (BBB) maintenance. Astrocytes are also chief responders to any CNS insult and undergo highly context dependent changes in morphology, gene expression, and function in a process collectively referred to as “astrocyte reactivity”. Recent work in stroke and hypoxia have uncovered necessary roles for reactive astrocytes in restorative angiogenesis, but specific astrocyte-secreted molecules mediating these effects remain an outstanding question. I have recently identified a novel, spatially restricted subpopulation of reactive astrocytes defined by persistent upregulation of the powerful pro-angiogenic molecule CCN Family Member 1(CCN1). In my proposal I will utilize two independent, yet complimentary, aims to test the hypothesis that Ccn1+ astrocytes demarcate an evolving pro-angiogenic tissue niche, that promotes functional recovery after SCI by directly governing endothelial cell phenotype including cell proliferation, maturation, or Notch signaling. In Aim 1 I will analyze a first of its kind longitudinal Spatial Transcriptomics dataset of spared tissue regions in a mouse hemisection model of SCI (mhSCI) to A) interrogate the molecular evolution of intraspinal tissue niches harboring Ccn1+ astrocytes, and B) establish a powerful resource for the SCI field. From this aim I will understand the unique molecular features of Ccn1+ astrocyte niches and computationally infer the evolution of associated biological processes, signaling cascades, and transcriptional regulators. In Aim 2 I will complement the computational data from Aim 1 by utilizing a newly generated astrocyte specific CCN1 knockout mouse for A) behavioral assays of locomotor recovery and B) histological assessment of endothelial cell phenotype after mhSCI. From this aim I will interrogate the therapeutic potential of CCN1 for SCI and the direct effect it has on endothelial cell specification. Taken together, this study will uncover the angiogenic potential of tissue niches harboring Ccn1+ astrocytes and start to provide a glimpse into a potential mechanism of action. Such findings may have important implications in the development of new therapeutics that are aimed at providing a more permissive environment for regenerative axon sprouting after SCI.

抽象的脊髓损伤 (SCI) 是一种毁灭性的神经系统疾病，其特征是上行和上行神经功能受到破坏。脊髓损伤后，白质中的轴突会幸免于难，导致患者终生瘫痪。（WM）区域尝试进行短程发芽以恢复正常的神经功能。控制这种短程轴突萌发的神经元外在因素仍然知之甚少。发育和受伤后，新的轴突在时空上跟随新的血管，因此紧密连接轴突发芽到脉管系统是由星形胶质细胞精心策划的。与内皮细胞和周细胞发生物理相互作用，形成胶质血管单元——一个具有不可或缺的组织生态位星形胶质细胞在调节血流和维持血脑屏障（BBB）方面的作用也是主要的反应者。任何中枢神经系统损伤，并在形态、基因表达和功能方面经历高度依赖的变化最近在中风和缺氧方面的研究发现了一个统称为“星形胶质细胞反应性”的过程。反应性星形胶质细胞在恢复性血管生成中的必要作用，但特定的星形胶质细胞分泌分子调节这些影响仍然是一个悬而未决的问题，我最近发现了一个新颖的、空间受限的问题。由强大的促血管生成分子持续上调定义的反应性星形胶质细胞亚群 CCN 家庭成员 1（CCN1）。在我的提案中，我将利用两个独立但互补的目标来测试假设 Ccn1+ 星形胶质细胞划定一个不断进化的促血管生成组织生态位，促进功能性通过直接控制内皮细胞表型（包括细胞增殖、成熟或在目标 1 中，我将分析第一个纵向空间转录组学数据集。 SCI (mhSCI) 小鼠半切片模型中的组织区域 A) 探究 SCI 的分子进化脊髓内组织微环境中含有 Ccn1+ 星形胶质细胞，B) 为 SCI 领域建立了强大的资源。为了这个目标，我将了解 Ccn1+ 星形胶质细胞生态位的独特分子特征，并通过计算推断在目标 2 中，我将研究相关生物过程、信号级联和转录调节因子的进化。利用新生成的星形胶质细胞特异性 CCN1 敲除来补充 Aim 1 的计算数据小鼠 A) 运动恢复的行为测定和 B) 内皮细胞的组织学评估 mhSCI 后的表型我将探讨 CCN1 对 SCI 的治疗潜力及其直接作用。总而言之，这项研究将揭示其对内皮细胞规范的影响。含有 Ccn1+ 星形胶质细胞的组织壁龛，开始让我们了解潜在的作用机制。这些发现可能对开发针对以下疾病的新疗法具有重要意义：为 SCI 后再生轴突萌发提供更宽松的环境。