NINDS Exploratory/Developmental Projects in Translational Research

NINDS 转化研究探索/开发项目

• 批准号: 7574330
• 负责人: Nancy Elise Braverman
• 金额: $ 22.77万
• 依托单位: UNIVERSITY OF SOUTHERN CALIFORNIA
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-09-15 至 2010-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7574330
• 关键词: Acetyl-CoA C-Acetyltransferase; Affect; Alleles; Animals; Biochemical; Biogenesis; Biological Assay; Biological Availability; Cells; Chemicals; Clinical; Clinical Treatment; Clinical Trials; Code; Cultured Cells; Cystic Fibrosis; Cystic Fibrosis Transmembrane Conductance Regulator; Development; Disease; Duchenne muscular dystrophy; Engineering; Evaluation; Fibroblasts; Future; Gene Expression; Gene Expression Profiling; Generations; Genes; Goals; Immunoblot Analysis; Immunoblotting; Individual; Inherited; Length; Maintenance; Mammalian Cell; Monitor; Morphology; Muscular Dystrophies; Mutation; Neurodegenerative Disorders; Nonsense Codon; Nonsense Mutation; Nucleotides; Numbers; Parents; Patients; Peroxisomal Disorders; Pharmaceutical Preparations; Phytanic Acid; Plasmalogens; Portraits; Positioning Attribute; Process; Protein Import; Proteins; Public Health; Reading; Relative (related person); Reporter; Reporter Genes; Rodent; Sampling; Series; Skin; Spinal Muscular Atrophy; Structure; Terminator Codon; Testing; Therapeutic Agents; Therapeutic Effect; Toxic effect; Transcript; Translational Research; Translations; Very Long Chain Fatty Acid; analog; base; interest; nervous system disorder; novel; oxidation; peroxisome; protein function; research clinical testing; response

DESCRIPTION (provided by applicant): Peroxisomal biogenesis disorders (PBDs) are a group of autosomal recessive neurodegenerative disorders caused by mutations in PEX genes. Our goal is to evaluate the efficacy of a set of newly developed drugs capable of selectively promoting the read-through of premature stop codons (nonsense suppressor compounds) in PEX genes as therapeutic agents for a subset of individuals with PBDs. One such nonsense suppressor drug, PTC-124 is in clinical trials for the treatment of cystic fibrosis and Duchenne's muscular dystrophy cases caused by nonsense mutations in CFTR and DMD, respectively. Another series of analogs based on the parent compound indoprofen have shown nonsense suppressor activity in cell culture assays and have favorable toxicity and bioavailability profiles in rodents. A necessary step in achieving this goal is to determine the ability of these drugs to rescue peroxisome structure and function in cultured cells derived from PBD patients with nonsense mutations in peroxisome assembly (PEX) genes. In this proposal, we will conduct mutational analyses of PEX genes in thirty PBD patients to expand upon a known subset of PBD patients with disease-causing nonsense mutations (Specific Aim 1). Fibroblasts exist from each of these patients which will be used in downstream analyses of peroxisome function. In parallel, we will conduct a series of functional assays on drug-treated cultured fibroblasts derived from eighteen PBD patients with previously identified nonsense mutations in peroxin genes (Specific Aim 2). This includes biochemical, immunolocalization, immunoblot, and reporter gene-based characterization of peroxisome protein function and assembly. Should any of these nonsense suppressor compounds prove effective in rescuing peroxisome function in cultured fibroblasts, we will analyze their off-target effects through microarray-based gene expression profiling of drug-treated cells. Overall, these studies will provide a necessary initial evaluation of the potential efficacy of nonsense mutation suppressor therapies for PBDs prior to our long-term goal of developing therapeutic agents for PBDs that are used in clinical settings. PUBLIC HEALTH RELEVANCE: Project Narrative We are interested in developing therapies for patients with peroxisome biogenesis disorders (PBD), which are a group of inherited, often fatal, neurological diseases in which there is no current therapy. We will evaluate a series of drugs for their ability to rescue peroxisome functions in skin cells cultured from patients. These drugs have been shown to read through nonsense mutations in two other diseases, and preliminary chemical refinement and animal toxicity studies are in progress. Cellular rescue is a necessary preliminary step for the potential future application of these drugs to treat PBD caused by nonsense mutations.

描述（由申请人提供）：过氧化物酶体生物发生障碍（PBD）是由PEX基因突变引起的一组常染色体隐性神经退行性疾病。我们的目标是评估一组新开发的药物的功效，能够选择性地促进PEX基因中的过早停止密码子（废话抑制剂化合物）作为治疗剂的读入读物，以促进PBD的一部分。 PTC-124的一种无意义的抑制药物是临床试验，用于治疗囊性纤维化和Duchenne的肌肉营养不良病例，分别由CFTR和DMD中的胡说八道引起。基于母体化合物的另一个类似物在细胞培养分析中表现出废话的抑制作用，并且在啮齿动物中具有有利的毒性和生物利用度。实现这一目标的必要步骤是确定这些药物在过氧化物酶体组装（PEX）基因中衍生出的PBD患者衍生出的培养细胞中挽救过氧化物酶体结构和功能的能力。在此提案中，我们将对30名PBD患者的PEX基因进行突变分析，以扩大患有疾病的无义突变的PBD患者的一部分（特定的目标1）。这些患者中的每一个都存在成纤维细胞，这些患者将用于过氧化物酶体功能的下游分析。同时，我们将对从十八位PBD患者得出的药物处理的成纤维细胞进行一系列功能测定，这些PBD患者先前鉴定出了过氧蛋白基因中的废话突变（特定的AIM 2）。这包括生化，免疫定位，免疫印迹和基于报告基因过氧化物酶体蛋白功能和组装的表征。如果这些废话抑制剂化合物在培养的成纤维细胞中挽救过氧化物酶体功能有效，我们将通过基于微阵列的基因基因表达分析药物处理的细胞来分析其脱靶作用。总体而言，这些研究将提供必要的初步评估，以了解PBD的无义突变抑制疗法的潜在疗效，然后我们长期开发用于临床环境中使用的PBD的治疗剂。 公共卫生相关性：我们有兴趣针对过氧化物酶体生物发生障碍（PBD）的患者开发疗法的项目叙述，这些患者是一组遗传性的，通常是致命的，致命的，神经系统疾病，没有当前治疗。我们将评估一系列药物，以挽救从患者培养的皮肤细胞中过氧化物酶体功能的能力。这些药物已被证明可以通过另外两种疾病中的胡说八道突变读取，并且正在进行初步的化学细化和动物毒性研究。蜂窝救援是这些药物将来可能应用来治疗无义突变引起的PBD的必要初步步骤。