Novel Approaches for Read-through of Nonsense Mutations in COL7A1

通读 COL7A1 无义突变的新方法

• 批准号: 9314378
• 负责人: Qiujie Jiang
• 金额: $ 7.8万
• 依托单位: THOMAS JEFFERSON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-08-07 至 2018-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9314378
• 关键词: Affect; Alleles; Amino Acids; Biological Assay; Biological Models; Bulla; Cell Culture Techniques; Cell physiology; Cells; Chronic; Cicatrix; Clinical Trials; Collagen; Collagen Type VII; Complication; Connective Tissue Diseases; Development; Disease; Drug Kinetics; Embryo; Epidermolysis Bullosa; Epidermolysis Bullosa Dystrophica; Fibroblasts; Genes; Heritability; Human; In Vitro; Incidence; Individual; Inherited; Intervention; Lead; Length; Mediating; Medical; Membrane; Messenger RNA; Modeling; Mutation; Nonsense Codon; Nonsense Mutation; Patients; Pharmaceutical Preparations; Pharmacologic Substance; Pharmacology; Phenotype; Property; Proteins; Reporting; Risk; Signs and Symptoms; Skin; Squamous cell carcinoma; Suppressor Mutations; System; Terminator Codon; Testing; Toxicology; Transcript; Transfection; Translating; Ulcer; United States; Validation; Zebrafish; amlexanox; design; effective therapy; efficacy testing; experimental study; in vitro testing; in vivo; keratinocyte; knock-down; mRNA Decay; mutant; novel; novel strategies; patient subsets; polypeptide; pre-clinical; premature; public health relevance

 DESCRIPTION (provided by applicant): Dystrophic epidermolysis bullosa (DEB), an inherited connective tissue disease, manifests with blisters, erosions and chronic ulcers in the skin and mucosal membranes. The signs and symptoms of this condition vary widely among affected individuals; the most severe cases involve widespread blistering that can lead to scarring disfigurement and other serious medical problems. Furthermore, as a complication of the chronic skin damage, patients with DEB have an increased risk in development of highly aggressive squamous cell carcinomas. This disease is caused by mutations in the COL7A1 gene, ~10% of them being nonsense mutations. So far there is no effective treatment. The incidence of EB is estimated to be ~1:20,000 implying that there are 30,000-40,000 affected individuals in the United States. Assuming the same global incidence, there are over half a million EB patients in the world. Since approximately 20 percent of the patients have the dystrophic forms, even with ~10% of patients having nonsense mutations in COL7A1, thousands of patients would benefit from a successful strategy of PTC read-through worldwide. This study will test novel pharmaceutical compounds that facilitate read-through of nonsense mutations and counteract nonsense-mediated mRNA decay, thus allowing synthesis of the full-length polypeptide from the mutant allele, with the hypothesis that correction of COL7A1 mutations will counteract their phenotypic consequences. The read-through compounds with antagonistic properties against nonsense-mediated mRNA decay will be tested in vitro in cell cultures with mutant keratinocytes and fibroblasts derived from the skin of patients with DEB as well as in vivo using zebrafish as a platform. Preclinical demonstration of the efficacy of compounds, such as PTC124 and Amlexanox, can be translated quickly to the clinical trials due to the fact that the pharmacokinetics and toxicology profiles of these drugs have already been established.

 描述（由适用提供）：营养不良的表皮分层鳞茎（DEB），一种遗传的结缔组织疾病，在皮肤和粘膜膜上表现出水泡，侵蚀和慢性溃疡。在受影响的个体中，这种情况的体征和症状差异很大；最严重的病例涉及宽度的起泡，这可能导致畸形和其他严重的医疗问题。此外，作为慢性皮肤损伤的并发症，DEB患者在高度侵袭性的鳞状细胞癌的发展中具有增加的风险。该疾病是由COL7A1基因突变引起的，其中约有10％是胡说八道突变。到目前为止，还没有有效的治疗方法。 EB的事件估计约为1：20,000，这意味着在美国有30,000-40,000名受影响的人。假设同样的全球事件，世界上有超过半百万的EB患者。由于大约20％的患者患有营养不良形式，即使约有10％的患者在COL7A1中患有胡说八道突变，因此，成千上万的患者将受益于全球PTC的成功策略。这项研究将测试新型的药物化合物，以促进无义突变的读取并抵消废话介导的mRNA衰变，从而允许从突变等位基因中合成全长多肽，并假设Col7a1突变的校正将与他们的表型后果相抵触。具有拮抗特性的读取化合物对无义介导的mRNA衰变的介导化合物将在体外在具有突变体角质形成细胞的细胞培养物中进行测试，并且使用Zebrafish作为平台的DEB和INVIVO患者的皮肤产生的成纤维细胞和成纤维细胞。由于已经建立了这些药物的药代动力学和毒理学概况，因此可以快速转化化合物（例如PTC124和Ammlexanox）效率的临床前证明。