Rho GTPase siRNA as a Therapeutic Strategy in Experimental Autoimmune Neuritis

Rho GTPase siRNA 作为实验性自身免疫性神经炎的治疗策略

• 批准号: 8730419
• 负责人: Evan B. Stubbs
• 金额: --
• 依托单位: EDWARD HINES JR VA HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-04-01 至 2016-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8730419
• 关键词: Acute; Affect; Attenuated; Autoimmune Diseases; Blood-Nerve Barrier; CCL2 gene; Caring; Clinical; Development; Disease; Dose; Economic Burden; Electrophysiology (science); Endothelial Cells; Etiology; Experimental Autoimmune Neuritis; Goals; Guanosine Triphosphate Phosphohydrolases; Immune; Immunohistochemistry; Inflammatory; Laboratories; Leukocytes; Mainstreaming; Mediating; Modeling; Neuropathy; Paralysed; Paresis; Peripheral Nerves; Peripheral Nervous System; Peripheral Nervous System Diseases; Polyradiculopathy; Prevalence; Publishing Peer Reviews; Rattus; Rehabilitation therapy; Reverse Transcriptase Polymerase Chain Reaction; Severities; Small Interfering RNA; Technology; Testing; Therapeutic; Translations; Vascular Endothelium; Veterans; cytokine; health administration; in vivo; macrophage; nervous system disorder; neuromuscular; novel; novel therapeutics; palliative; patient population; prophylactic; public health relevance; response; rho GTP-Binding Proteins; sciatic nerve; social; socioeconomics; trafficking; translational study

Acquired Inflammatory neuropathies are a considerable social and economic burden to our Veterans and to the Veterans Health Administration. Encompassing both known infectious or possibly infectious etiologies, inflammatory neuropathies constitute one of the largest and least understood spectrums of neurologic disorders. Inclusive among these disorders is acute inflammatory demyelinating polyradiculopathy (AIDP), a highly disabling inflammatory autoimmune disease of the peripheral nervous system that is characterized by acute/subacute symmetrical paresis with areflexia progressing to neuromuscular paralysis. Despite its overwhelming prevalence and socioeconomic impact, the treatment of Veterans with inflammatory peripheral neuropathies, including AIDP, remains palliative. Cytokine-mediated recruitment and trafficking of autoreactive leukocytes across the blood-nerve barrier and into peripheral nerves is a well-established early pathological hallmark of inflammatory peripheral neuropathies, including AIDP. Localized GTPase-dependent activation of the peripheral nerve vascular endothelium in response to proinflammatory cytokines represents an initiating pathological insult. Peer- reviewed and published preliminary studies from our laboratory strongly support that trafficking of autoreactive leukocytes into peripheral nerves during AIDP may proceed by a mechanism that involves Cdc42 GTPase- dependent secretion of CCL2. Novel translational studies are critically needed to develop therapeutic and rehabilitative strategies for the advanced care of Veterans debilitated by acquired inflammatory neuropathies. In this two-year SPiRE study, we will determine whether siRNA-mediated GTPase knockdown therapeutically protects against the development and progression of inflammatory neuropathy. Hypothesis: Therapeutically administered siRNA directed against key monomeric GTPases will attenuate the development and progression of experimental autoimmune neuritis by inhibiting endothelial cell CCL2 expression. This hypothesis will be tested in vivo with the following two Specific Objectives using an established clinically- translatable rat model of AIDP (experimental autoimmune neuritis, EAN). Specific Objective 1 will determine whether prophylactic or therapeutically administered siRNAs targeting key monomeric GTPases protects against the development and progression of EAN. We will determine the dose- dependent effect of siRNAs targeting Cdc42 or RalA GTPases on (a) the clinical severity and course of EAN and on (b) EAN-induced changes in peripheral nerve function, using evoked-response electrophysiology. Specific Objective 2 will determine whether prophylactic or therapeutically administered siRNAs targeting key monomeric GTPases attenuates trafficking of autoreactive leukocytes into peripheral nerves during EAN. We will (a) validate siRNA-mediated knockdown of key monomeric GTPases (Cdc42 or RalA) within sciatic nerves of siRNA-treated EAN rats, compared with scrambled siRNA-treated EAN controls, and (b) quantify the content and distribution of CCL2, CCR2, and immune infiltrates (macrophages and leukocytes) within sciatic nerves of siRNA-treated rats, compared with scrambled siRNA-treated EAN controls, with immunohistochemistry. The goal of this study is to establish siRNA-mediated Rho GTPase knockdown as a viable therapeutic strategy for the management of inflammatory peripheral neuropathies. We argue that successful completion of this study will expedite the translation of siRNA technology into mainstream management of Veterans with acquired inflammatory neuropathies, including AIDP.

获得性炎症性神经病是我们退伍军人的一个相当大的社会和经济负担， 退伍军人健康管理局包括已知的传染性或可能的传染性病因， 炎性神经病是神经系统疾病中最大和最不了解的疾病之一， 紊乱这些疾病包括急性炎性脱髓鞘性多神经根病（AIDP）， 一种高度致残的周围神经系统炎性自身免疫性疾病，其特征在于 急性/亚急性对称性轻瘫伴无反射进展为神经肌肉麻痹。尽管 压倒性的患病率和社会经济影响，治疗退伍军人与炎症性外周 包括AIDP在内的神经病仍然是姑息性的。 细胞因子介导的自身反应性白细胞的募集和运输穿过血-神经屏障， 周围神经炎是周围神经炎的一个公认的早期病理标志， 神经病变，包括AIDP。外周神经血管的局部GTP酶依赖性激活 内皮细胞对促炎细胞因子的反应代表了起始的病理损伤。同行- 我们实验室的回顾和发表的初步研究强烈支持自体反应性细胞的运输， AIDP期间白细胞进入外周神经可能通过涉及Cdc 42 GT3的机制进行， CCL 2的依赖性分泌。 迫切需要新的转化研究来制定治疗和康复策略， 对因获得性炎性神经病而衰弱的退伍军人的高级护理。在这两年中， SPiRE研究，我们将确定siRNA介导的GT3基因敲低是否能在治疗上保护 炎症性神经病的发展和进展。 假设：治疗性施用的针对关键单体GTP酶的siRNA将减弱 抑制内皮细胞CCL 2诱导实验性自身免疫性神经炎的发生发展 表情 该假设将使用已建立的临床- 实验性自身免疫性神经炎（Experimental autoimmune neuritis，EAN） 具体目标1将确定是否预防性或治疗性施用靶向关键的siRNA。 单体GTP酶保护EAN的发展和进展。我们会确定剂量- 靶向Cdc 42或RalA GTP酶的siRNA对（a）EAN的临床严重性和病程的依赖性作用 和（B）EAN诱导的外周神经功能变化，使用诱发反应电生理学。 具体目标2将确定是否预防性或治疗性施用靶向关键的siRNA， 单体GTP酶减弱EAN期间自身反应性白细胞向外周神经的运输。我们 将（a）验证坐骨神经内关键单体GTP酶（Cdc 42或RalA）的siRNA介导的敲低 和（B）定量siRNA处理的EAN大鼠中与乱序siRNA处理的EAN对照相比， 以及CCL 2、CCR 2和免疫浸润细胞（巨噬细胞和白细胞）在坐骨神经内的分布。 siRNA处理的大鼠，与乱序siRNA处理的EAN对照相比，用免疫组织化学。 本研究的目的是建立siRNA介导的Rho GT3基因敲低作为一种可行的治疗策略 用于炎性周围神经病变的管理。我们认为，成功完成这一任务 这项研究将加快siRNA技术转化为退伍军人的主流管理， 获得性炎性神经病，包括AIDP。