Treatment of multiple sclerosis model with RNA padlocks

描述（由申请人提供）：本提案的目的是检验以下假设：阻断TNF-α翻译的RNA套索（TM）可以减少实验性自身免疫性脑脊髓炎（EAE）（一种多发性硬化模型）的临床体征和组织损伤。RNA套索是含有酶特征的反义RNA，该酶特征使它们在杂交时与靶mRNA拓扑连接。在以前的工作中，我们确定了一个针对TNF-α的前导套索，它有效地抑制了巨噬细胞系和小鼠中TNF-α的分泌。最初的实验将比较几种策略，包括电穿孔，阳离子脂质和病毒，以优化将RNA反义套索递送到髓鞘碱性蛋白（MBP）特异性T细胞系中。然后，这些靶细胞将接受刺激，以确定RNA套索对TNF-α产生的影响。如果RNA套索有效地降低TNF-α，则将细胞注射到幼稚受体中以确定TNF-α RNA套索是否降低EAE的严重程度。此外，我们将测试RNA套索的体内施用是否阻断EAE的发作或改善EAE的严重性。如果肿瘤坏死因子不是最好的靶点，我们将测试套索对抗干扰素γ。还将进行急性毒性研究。 商业潜力：在临床前研究中证明疗效将导致单独使用RNA套索和/或与目前批准的治疗剂（例如Betaseron、Avonex、Copaxone）联合使用的临床试验。 这项技术也可能在TNF-α导致炎症和病理学的其他疾病中具有潜在的商业应用，包括类风湿性关节炎和1型糖尿病。 如果我们证明RNA套索能有效地传递到T细胞，那么也有可能传递阻断HIV基因表达和干扰病毒复制的套索。 最终的产品将是用于治疗自身免疫的套索药物，估计市场每年超过60亿美元

DESCRIPTION (provided by the applicant): The goal of this proposal is to test the hypothesis that RNA lassos(TM) that block the translation of TNF-alpha can decrease the clinical signs and tissue damage in experimental autoimmune encephalomyelitis (EAE), a model of multiple sclerosis. RNA lassos are antisense RNAs containing an enzymatic feature that causes them to become topologically linked to a target mRNA upon hybridization. In previous work, we identified a lead lasso directed against TNF-alpha that effectively inhibits TNF-alpha secretion in a macrophage cell line and in mice. Initial experiments will compare several strategies, including electroporation, cationic lipids, and viruses, to optimize the delivery of RNA antisense lassos into myelin basic protein (MBP)-specific T cell lines. Those targeted cells will then receive stimulation to determine the effect of RNA lassos on TNF-alpha production. If the RNA lassos effectively decrease TNF-alpha, the cells will be injected into naive recipients to determine whether TNF-alpha RNA lassos decrease the severity of EAE. In addition, we will test whether in vivo administration of RNA lassos blocks the onset or ameliorates the severity of EAE. If TNF turns out not to be the best target, we will test lassos against interferon gamma. An acute toxicity study will also be performed. Commercial potential: A demonstration of efficacy in preclinical studies would lead to a clinical trail of RNA lassos alone and/or in combination with a currently approved therapeutic (e.g. Betaseron, Avonex, Copaxone). This technology may also have potential commercial applications in other conditions in which TNF-alpha contributes to inflammation and pathology, including rheumatoid arthritis and type-1 diabetes. If we demonstrate effective delivery of RNA lassos to T cells, it may also be possible to deliver lassos that block HIV gene expression and interfere with viral replication. The ultimate product would be lasso drugs for treatment of autoimmunity, an estimated market of over $6 billion per year