Defining mechanisms of disease and repair in a viral model of multiple sclerosis

翻译后摘要：脱髓鞘疾病多发性硬化症（MS）患者的一个重要的未满足的临床需求 是一种有效的促进髓鞘再生的方法， 免疫抑制剂可以抑制脱髓鞘并恢复运动功能，同时限制免疫细胞浸润到CNS中。长期 本研究的目的是i）确定趋化因子信号传导如何控制神经炎症， 疾病进展，ii）评估趋化因子信号传导在调节少突胶质细胞祖细胞中的作用 (OPC)iii）进一步表征移植的人和小鼠神经细胞如何在体外成熟和髓鞘再生， 祖细胞增强轴突的完整性，促进髓鞘再生和影响 神经炎症/脱髓鞘，iv）定义小胶质细胞限制神经炎症/脱髓鞘的严重性的机制， 脱髓鞘并影响髓鞘再生。为了实现这些目标，我们将使用一个公认的临床前 20多年来，本实验室一直使用敏感的C57 BL/6颅内感染， 小鼠与神经适应的小鼠肝炎病毒（JHMV）的JHM株作为病毒诱导的模型 脱髓鞘研究控制神经炎症，脱髓鞘， 髓鞘再生建议的实验程序，将有助于实现我们的研究目标，将包括 通过产生小鼠的遗传方法，其中靶基因被选择性诱导/消除 评估对疾病进展和修复的影响，CRISPR技术消除NPC中的特定靶基因 免疫细胞和驻留CNS细胞上的单细胞和核RNA测序以及2-光子 (2P)在显微镜下观察轴突损伤/修复和髓鞘再生。总的来说，我们相信 本提案中概述的实验目标将为MS的发病机制提供新的见解， 确定治疗干预的新靶点，以阻止疾病进展并促进髓鞘再生。

Abstract: An important unmet clinical need for patients with the demyelinating disease multiple sclerosis (MS) is an effective method for promoting remyelination that can ameliorate clinical symptoms associated with demyelination and restore motor function while limiting immune cell infiltration into the CNS. The long-term objectives of this research proposal are to i) define how chemokine signaling controls neuroinflammation and disease progression, ii) assess the effects of chemokine signaling in regulating oligodendrocyte progenitor cell (OPC) maturation and remyelination, iii) further characterize how engrafted human and mouse neural progenitor cells enhance axonal integrity, promote remyelination and influence neuroinflammation/demyelination, iv) define mechanisms by which microglia restrict the severity of demyelination and influence remyelination. To accomplish these goals, we will use a well-accepted pre-clinical animal models of MS. For over 20 years, my laboratory has used intracranial infection of susceptible C57BL/6 mice with the neuroadapted JHM strain of mouse hepatitis virus (JHMV) as a model of viral-induced demyelination to study molecular and cellular events controlling neurioinflammation, demyelination, and remyelination. Proposed experimental procedures that will aid in accomplishing our research goals will include genetic approaches through generation of mice in which targeted genes are either selectively induced/ablated to assess effect on disease progression and repair, CRISPR technology to ablate specific target genes in NPC cultures, single cell and nuclear RNA sequencing on immune cells and resident CNS cells and use of 2-photon (2P) microscopy to visualize axonal damage/repair and remyelination. Collectively, we believe our experimental goals outlined in this proposal will provide new insight into the pathogenesis of MS as well as identify new targets for therapeutic intervention to impede disease progression and promote remyelination.