Targeting SNO-GAPDH in inflammatory neurodegeneration and mitochondrial injury

项目摘要/摘要 尽管多发性硬化症(MS)被经典地认为是一种脱髓鞘疾病，但神经轴突丢失发生在 疾病的复发期和进展期，并代表了DISA的主要病理相关性- 保佑。现有的疗法主要针对外周免疫系统，防止临床复发，但在很大程度上 未能预防进行性多发性硬化治疗中的神经变性，其主要神经保护模式为Ac- 运动是目前研究的一个主要目标，既可以减缓进展性多发性硬化症的残疾，也可以限制再次发生多发性硬化症时的伤害 尽管有目前的治疗方法，失误还是会发生。一氧化氮(NO)是一种由中枢神经系统产生的有毒自由基。 中枢神经系统巨噬细胞和小胶质细胞在复发性和进行性神经轴索损伤中的作用 MS和神经炎症模型中，NO诱导的线粒体功能障碍起主要作用。SPE- 介导这种损伤的确切的、可用药的信号通路尚未确定。我们建议研究一项 候选信号通路涉及蛋白质GAPDH的亚硝化。硝基GAPDH(SNO-GAPDH) 转位到细胞核和线粒体，在细胞死亡和核靶向等方面具有既定的作用 作为sirt1和pgc-1α，对线粒体生物能量学至关重要。此外，SNO-GAPDH信令可以 被CGP3466阻断，CGP3466是一种高度特异的口服中枢神经系统渗透药物，在人类中已建立了安全性 临床翻译门槛低。我们有初步证据表明SNO-GAPDH信令是活跃的 在实验性自身免疫性脑脊髓炎(EAE)小鼠的神经炎症模型中， 从多发性硬化症患者的死后获取物质组织。我们发现系统性地给药 CGP3466减轻C57BL/6 MOG35-55/CFA EAE的神经功能障碍并防止神经元损伤 一氧化氮对培养神经元线粒体呼吸的影响。在拟议的研究中，我们计划充分发挥以下作用- 鉴定MOG35-55/CFA EAE和死后人类MS组织中SNO-GAPDH通路的活性。我们会阻止- 探讨CGP3466在EAE中的保护作用是否来自于主要的神经保护机制。 依赖外周免疫效应，与现有治疗方法的区别及其临床前的建立 潜力。最后，我们将通过评估SNO-GAPDH对神经元有丝分裂的影响来寻求机制上的见解。 软骨功能。这一机制和临床前治疗研究的积极结果将建立一个 治疗多发性硬化症和更广泛的神经炎性疾病的新方法。公安人员的职业发展 该计划将在免疫学、神经炎症模型和线粒体方面提供新的培训机会 生物能量学，为未来炎症性神经变性的独立研究提供了基础。

PROJECT SUMMARY/ABSTRACT Although multiple sclerosis (MS) is classically considered a demyelinating disease, neuro-axonal loss occurs in both relapsing and progressive phases of the disease and represents the primary pathologic correlate of disa- bility. Existing therapies primarily target the peripheral immune system, preventing clinical relapses but largely failing to prevent neurodegeneration in progressive MS. Therapies with a primary neuroprotective mode of ac- tion are a major goal of current research, both to slow disability in progressive MS and to limit injury when re- lapses occur despite current therapies. Nitric oxide (NO), a toxic free radical produced by central nervous sys- tem (CNS) macrophages and microglia, contributes to neuro-axonal injury in both relapsing and progressive MS and in models of neuroinflammation, with NO-induced mitochondrial dysfunction playing a major role. Spe- cific, druggable signaling pathways that mediate this injury have not been identified. We propose to study a candidate signaling pathway involving nitrosylation of the protein GAPDH. Nitrosylated GAPDH (SNO-GAPDH) translocates to both nucleus and mitochondria, with an established role in cell death and nuclear targets, such as SIRT1 and PGC-1α, critical for mitochondrial bioenergetics. Moreover, SNO-GAPDH signaling can be blocked by CGP3466, a highly specific, oral CNS-penetrant drug with an established safety profile in humans and a low threshold for clinical translation. We have preliminary evidence that SNO-GAPDH signaling is active in an experimental autoimmune encephalomyelitis (EAE) mouse model of neuroinflammation, and in white matter tissue obtained post-mortem from MS patients. We have found that systemic administration of CGP3466 attenuates neurologic disability in C57BL/6 MOG35-55/CFA EAE and prevents impairment of neuronal mitochondrial respiration in cultured neurons exposed to NO. In the proposed studies, we plan to fully charac- terize SNO-GAPDH pathway activity in MOG35-55/CFA EAE and post-mortem human MS tissue. We will deter- mine whether the protective effects of CGP3466 in EAE derive from a primary neuroprotective mechanism in- dependent of peripheral immune effects, differentiating it from current therapies and establishing its pre-clinical potential. Finally, we will seek mechanistic insights by evaluating the effects of SNO-GAPDH on neuronal mito- chondrial function. Positive results from this mechanistic and pre-clinical therapeutic research will establish a new therapeutic approach for MS and neuroinflammatory disease more broadly. The PI's career development plan will provide new training opportunities in immunology, models of neuroinflammation, and mitochondrial bioenergetics, providing a foundation for future independent studies of inflammatory neurodegeneration.