Mechanisms of oligodendroglial ciliary function in white matter injury repair

少突胶质细胞纤毛功能在白质损伤修复中的机制

• 批准号: 10659990
• 负责人: Stephen Philip James Fancy
• 金额: $ 40.38万
• 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: 2023-04-15 至 2028-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10659990
• 关键词: Acute; Adult; Affect; Agonist; Area; Biological; Blood Vessels; CREB1 gene; Catalogs; Cell Communication; Cell Culture Techniques; Cell Proliferation; Cells; Cellular biology; Central Nervous System Diseases; Cerebral Palsy; Cilia; Collaborations; Cues; Cyclic AMP; Data; Demyelinations; Development; Disease; Disease Progression; Enzymes; Erinaceidae; Failure; G-Protein-Coupled Receptors; Gene Expression; Gene Targeting; Genes; Grant; Homeostasis; Human; Image; In Vitro; Injury; Label; Lesion; Letters; LoxP-flanked allele; Mammalian Cell; Maps; Mediating; Methods; Microtubules; Multiple Sclerosis; Multiple Sclerosis Lesions; Mus; Myelin; Myelin Sheath; Natural regeneration; Nature; Neurologic Dysfunctions; Organelles; Pathway interactions; Pericytes; Population; Proliferating; Proteins; Proteome; Regulation; Rest; Role; Signal Pathway; Signal Transduction; Signaling Molecule; Site; Slice; Spinal Cord; Surveys; Techniques; Tissues; Transducers; Transgenic Organisms; Visualization; cell behavior; cell type; hypoxia neonatorum; in vivo; injury and repair; insight; mRNA sequencing; migration; neonatal human; nervous system disorder; newborn brain injury; oligodendrocyte precursor; oligodendrocyte progenitor; pharmacologic; precursor cell; progenitor; programs; recruit; remyelination; repaired; response; response to injury; stem cells; therapeutic target; tool; white matter; white matter injury

PROJECT ABSTRACT After damage to white matter tracts (WMI) in CNS diseases such as multiple sclerosis (MS) in adults and newborn brain injuries that cause cerebral palsy (CP), myelin sheaths can be regenerated by activated oligodendrocyte precursor cells (OPCs). Failure of this remyelination program often occurs due to the improper recruitment of OPCs into injury sites, contributing significantly to ongoing neurological dysfunction and disease progression. Understanding the mechanisms controlling OPC biology during remyelination will provide insights as to why myelin repair fails in human cases. Importantly, OPCs dynamically produce primary cilia, microtubule- based organelles that transduce intercellular cues in a specialized signaling compartment. The role of primary cilia in regulating developmental pathways in OPCs remains poorly understood. Here, we show that OPCs require primary cilia to respond properly to WMI. First, this grant will demonstrate that genetically removing primary cilia from OPCs results in inadequate WMI repair, identifying the primary cilium as a critical effector of biological change in OPCs necessary for the WMI response. Furthermore, as there remains little mechanistic understanding of ciliary signaling pathways in OPCs, we will use a combination of approaches that ultimately define a GPCR/cAMP/CREB signaling axis beginning at the primary cilium as a crucial regulator of OPC biology. Finally, with recent advances in proximity-labeling, we can now catalogue the proteins that survey OPC primary cilia using a technique termed cilia-APEX. This grant will utilize cilia-APEX to identify signaling molecules that localize to OPC primary cilia in vitro and during remyelination in vivo. This will demonstrate dynamic changes in the protein content of OPC primary cilia during different stages of remyelination, while also adding significant insight into the extent of ciliary functions in OPCs. Together, these studies will show that primary cilia are a critical signaling module in OPCs for the regulation of remyelination, and will reveal potential therapeutic target for conditions such as MS and CP, where the OPC response to injury can be dysfunctional. 1

项目摘要 成人多发性硬化症 (MS) 等中枢神经系统疾病导致白质束 (WMI) 受损后 新生儿脑损伤导致脑瘫（CP），髓鞘可以通过激活来再生 少突胶质细胞前体细胞（OPC）。这种髓鞘再生计划的失败常常是由于不当的操作而发生的。 OPCs 募集到损伤部位，对持续的神经功能障碍和疾病有显着影响 进展。了解髓鞘再生过程中控制 OPC 生物学的机制将提供见解 为什么人类病例中髓磷脂修复失败。重要的是，OPC 动态产生初级纤毛、微管- 基于细胞器，在专门的信号室中转导细胞间信号。初级的作用 纤毛在调节 OPC 发育途径中的作用仍知之甚少。在这里，我们展示了 OPC 需要初级纤毛对 WMI 做出正确反应。首先，这笔赠款将证明基因去除 OPC 的初级纤毛导致 WMI 修复不充分，将初级纤毛确定为关键 WMI 反应所必需的 OPC 生物变化的效应器。此外，由于所剩无几 为了对 OPC 中纤毛信号通路的机制有所了解，我们将结合使用以下方法： 最终定义了从初级纤毛开始的 GPCR/cAMP/CREB ​​信号轴作为关键的调节因子 OPC 生物学。最后，随着邻近标记的最新进展，我们现在可以对以下蛋白质进行编目： 使用称为纤毛-APEX 的技术调查 OPC 初级纤毛。这笔赠款将利用 cilia-APEX 来识别 体外和体内髓鞘再生过程中定位于 OPC 初级纤毛的信号分子。这将 证明 OPC 初级纤毛蛋白质含量在不同阶段的动态变化 髓鞘再生，同时还可以深入了解 OPC 中纤毛功能的程度。在一起，这些 研究表明，初级纤毛是 OPC 中调节髓鞘再生的关键信号模块， 并将揭示 MS 和 CP 等疾病的潜在治疗靶点，其中 OPC 对损伤的反应 可能会出现功能障碍。 1