Nonsense Readthrough: a Therapeutic Approach to Inherited Vascular Disorders

无意义的通读：遗传性血管疾病的治疗方法

• 批准号: 9616979
• 负责人: Micheala A Aldred
• 金额: $ 20.95万
• 依托单位: INDIANA UNIV-PURDUE UNIV AT INDIANAPOLIS
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-01-01 至 2018-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9616979
• 关键词: Nonsense; Readthrough; Therapeutic; Approach; Inherited

 DESCRIPTION (provided by applicant) The goal of this study is a pre-clinical evaluation of nonsense readthrough as a potential therapy for inherited vascular disorders caused by mutations in the bone morphogenetic protein signaling pathway. We will evaluate two drugs with complementary properties. Ataluren is a FDA-designated orphan drug in phase 3 trials for two genetic disorders. It promotes readthrough of nonsense mutations without affecting the level of mRNA transcript and is most effective against UGA stop codons. Amlexanox is an anti-allergy drug that also promotes nonsense readthrough, but does so by inhibiting nonsense- mediated mRNA decay. Information on its codon specificity is limited, but it may show higher activity than ataluren against UAG codons. We will test the comparative effects of these two compounds on mutations that underlie two distinct but related vascular disorders: hereditary hemorrhagic telangiectasia (HHT) and pulmonary arterial hypertension (PAH). The mutation spectrum in these two disorders in complementary, with UGA codons predominating in PAH, whereas UAG/UAA mutations are more common in HHT. PAH is a life-threatening and incurable disease affecting the lung vasculature that is in desperate need of new therapies to tackle the underlying molecular changes that drive vascular remodeling. HHT affects blood vessels throughout the body and has high morbidity associated with arteriovenous malformations in the lungs and brain. Rarely, HHT patients may also develop PAH. We will utilize a combination of (a) a novel dual-tag in vitro reporter construct that we have developed to measure readthrough efficiency and sequence the resulting protein product, (b) patient-derived endothelial cells with defined genetic mutations to study functional restoration of cell signaling and (c) a novel knock-in mouse model with a nonsense mutation for in vivo testing. In the US alone, correction of nonsense mutations could benefit 6000-12,000 people affected with HHT and up to 1000 with PAH. Successful completion of this study will establish the effectiveness of this approach in vivo and define the sub-groups o patients most likely to respond. In addition, the novel dual-tag reporter that we have developed to measure both the efficiency of readthrough and the nature of the amino acid that is incorporated will be generalizable to many other genetic disorders The results of this study could translate rapidly to interventional human trials and pave the way for personalized approaches to therapy for genetic vascular diseases.

 描述（由申请人提供） 本研究的目的是对无义通读作为骨形态发生蛋白信号通路突变引起的遗传性血管疾病的潜在治疗方法进行临床前评价。我们将评估两种具有互补特性的药物。Ataluren是FDA指定的孤儿药，用于治疗两种遗传性疾病的3期试验。它促进无义突变的通读而不影响mRNA转录水平，并且对UGA终止密码子最有效。氨来赞是一种抗过敏药物，也促进无义通读，但通过抑制无义介导的mRNA衰变来实现。关于其密码子特异性的信息是有限的，但它可能显示出比阿他卢仑对UAG密码子更高的活性。我们将测试这两种化合物对两种不同但相关的血管疾病（遗传性出血性毛细血管扩张症（HHT）和肺动脉高压（PAH））突变的比较作用。这两种疾病的突变谱是互补的，在PAH中以UGA密码子为主，而UAG/UAA突变在HHT中更常见。PAH是一种影响肺血管系统的危及生命且无法治愈的疾病，迫切需要新的治疗方法来解决驱动血管重塑的潜在分子变化。HHT影响全身的血管，并且具有与肺和脑中的动静脉畸形相关的高发病率。HHT患者也可能发生PAH，这种情况很少见。我们将利用以下的组合：（a）我们开发的新型双标签体外报告基因构建体，用于测量通读效率并对所得蛋白质产物进行测序，（B）具有确定的基因突变的患者来源的内皮细胞，用于研究细胞信号传导的功能恢复，以及（c）具有无义突变的新型敲入小鼠模型，用于体内测试。仅在美国，无义突变的纠正可能使6000- 12，000名HHT患者和多达1000名PAH患者受益。本研究的成功完成将确定该方法在体内的有效性，并确定最有可能应答的患者亚组。此外，我们开发的用于测量通读效率和并入的氨基酸性质的新型双标签报告基因将可推广到许多其他遗传性疾病。这项研究的结果可以迅速转化为干预性人体试验，并为遗传性血管疾病的个性化治疗方法铺平道路。