Mechanisms of oligodendroglial ciliary function in white matter injury repair

项目摘要 中枢神经系统疾病（如成人多发性硬化症（MS））中的白色物质束（白质束）受损后， 新生儿脑损伤导致脑瘫（CP），髓鞘可以通过激活再生 少突胶质前体细胞（OPCs）。这种髓鞘再生计划的失败往往是由于不适当的 OPCs募集到损伤部位，显著促进持续的神经功能障碍和疾病 进展了解在髓鞘再生过程中控制OPC生物学的机制将提供深入的见解 为什么人类的髓鞘修复失败重要的是，OPCs动态地产生初级纤毛，微管- 在一个专门的信号室中传递细胞间信号的细胞器。小学的作用 纤毛在调节OPCs发育途径中的作用仍然知之甚少。在这里，我们表明， 需要初级纤毛来正确地响应振动。首先，这项拨款将证明， 来自OPCs的初级纤毛导致不充分的纤毛修复，确定初级纤毛是一个关键的 OPCs的生物学变化的效应子，这是OPCs应答所必需的。此外，由于几乎没有 在OPCs的纤毛信号通路的机制的理解，我们将使用的方法， 最终确定GPCR/cAMP/CREB信号轴开始于初级纤毛作为一个重要的调节器 OPC生物学最后，随着邻近标记的最新进展，我们现在可以对 使用称为cilia-APEX的技术调查OPC初级纤毛。这项赠款将利用纤毛APEX，以确定 在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 证明在不同阶段的OPC初级纤毛蛋白质含量的动态变化， 髓鞘再生，同时也增加了对OPCs中纤毛功能程度的重要了解。所有这些 研究将表明初级纤毛是OPC中调节髓鞘再生的关键信号模块， 并将揭示潜在的治疗目标的条件，如MS和CP，其中OPC响应损伤 可能会功能失调 1

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