TRANS-SPLICING TO REPAIR CYSTIC FIBROSIS MRNA

反式剪接修复囊性纤维化 mRNA

• 批准号: 6430807
• 负责人: LLOYD G MITCHELL
• 金额: $ 56.22万
• 依托单位: INTRONN, INC.
• 依托单位国家: 美国
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-09-15 至 2002-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6430807
• 关键词: RNA splicing; adeno associated virus group; biotherapeutic agent; chloride channels; cystic fibrosis; drug design /synthesis /production; gene expression; gene targeting; gene therapy; laboratory mouse; laboratory rat; polymerase chain reaction; precursor mRNA; spliceosomes; tissue /cell culture; transfection /expression vector

The ultimate objective of this proposal is to develop a new general mechanism to reprogram any human or eukaryotic gene at the pre- messenger RNA level. Specifically, this proposal will develop our patented technology, spliceosome mediated RNA trans-splicing (SMaRT[TM]), towards the production of a practical gene therapy for the treatment of Cystic Fibrosis. In Phase 1, we produced a series of pretherapeutic RNA molecules (PTMs) that are capable of initiating SMaRT reactions with mutant Cystic Fibrosis Transmembrane Regulator (CFTR) pre-mRNA, repairing the mRNA and expressing full length CFTR protein in cultured cells. The goal in Phase 2 is to identify an optimal PTM that can repair mutant CFTR mRNA, appropriately express normal CFTR protein, and restore chloride channel function in the most clinically relevant models of Cystic Fibrosis. This lead PTM therapeutic candidate will enter full preclinical testing in Phase 3 and ultimately, human clinical trials if results warrant. Many other genetic diseases may also be amenable to SMaRT[TM] therapeutics. Trans-splicing PTMs could improve many aspects of gene therapies by conferring intra-cellular specificity, decreasing the size of the delivered gene, and acquiring the regulated expression of the endogenous target gene. PROPOSED COMMERCIAL APPLICATIONS: Development of RNA molecules capable of repairing mutant CFTR by trans-splicing could lead to a therapeutic for Cystic Fibrosis that could slow or halt disease progression. There is a clinical need to serve and treat this market of approximately 30,000 affected individuals in the U.S. alone. Additionally, the development of effective spliceosome mediated RNA trans-splicing technology would be of utility in many gene transfer applications including the treatment of other genetic diseases, infections by splicing viruses ( HIV, EBV, papilloma, etc) and cancer.

该提案的最终目标是开发一种新的通用机制，在前信使 RNA 水平上重新编程任何人类或真核基因。具体来说，该提案将开发我们的专利技术，即剪接体介导的 RNA 反式剪接 (SMaRT[TM])，以生产用于治疗囊性纤维化的实用基因疗法。在第一阶段，我们生产了一系列治疗前 RNA 分子 (PTM)，它们能够与突变的囊性纤维化跨膜调节因子 (CFTR) 前 mRNA 启动 SMaRT 反应，修复 mRNA 并在培养细胞中表达全长 CFTR 蛋白。第 2 阶段的目标是确定一种最佳 PTM，可以修复突变​​的 CFTR mRNA，适当表达正常的 CFTR 蛋白，并在临床最相关的囊性纤维化模型中恢复氯离子通道功能。该领先的 PTM 治疗候选药物将进入第 3 阶段的全面临床前测试，如果结果证明的话，最终将进入人体临床试验。许多其他遗传疾病也可能适合 SMaRT[TM] 治疗。 反式剪接 PTM 可以通过赋予细胞内特异性、减小传递基因的大小以及获得内源靶基因的调节表达来改善基因治疗的许多方面。拟议的商业应用：开发能够通过反式剪接修复突变 CFTR 的 RNA 分子可能会产生一种治疗囊性纤维化的方法，从而减缓或阻止疾病的进展。仅在美国就有大约 30,000 名受影响个体的市场需要服务和治疗。此外，有效剪接体介导的RNA反式剪接技术的开发将在许多基因转移应用中发挥作用，包括治疗其他遗传疾病、剪接病毒（HIV、EBV、乳头状瘤等）感染和癌症。