DRUG-INDUCED ATM READTHROUGH OF PTC MUTATIONS

药物诱导的 ATM 读取 PTC 突变

基本信息

批准号：
7488560
负责人：
RICHARD A GATTI
金额：
$ 33.69万
依托单位：
UNIVERSITY OF CALIFORNIA LOS ANGELES
依托单位国家：
美国
项目类别：
财政年份：
2007
资助国家：
美国
起止时间：
2007-09-01 至 2012-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7488560
关键词：
A Mouse ATM Gene Mutation ATM function ATM gene ATM promoter Aminoglycoside Antibiotics Aminoglycosides Animal Model Animal Testing Animals Ataxia Telangiectasia Biological Assay Biological Markers Cancer Patient Cell Line Cells Characteristics Chemical Structure Child Class Clinical Clinical Trials DNA biosynthesis DNA chemical synthesis Development Diagnostic tests Disease Drug or chemical Tissue Distribution Geneticin Gentamicins Goals Hereditary Disease Heterozygote Laboratories Life Measurement Monitor Mutation Neurodegenerative Disorders Nonsense Codon Nonsense Mutation Nucleotides Numbers Patients Pharmaceutical Preparations Phase Protein Truncation Radiation Tolerance Range Reading Research Residual state Terminator Codon Testing Tissue Sample Toxic effect antimicrobial ataxia telangiectasia mutated protein base design high throughput screening improved in vivo mouse model response restoration small molecule libraries stem tool

项目摘要

DESCRIPTION (provided by applicant): This project stems from our long-standing goal to treat ATM deficiency, whether this be in cancer patients or in those suffering from ataxia-telangiectasia (A-T), a rare neurodegenerative disorder of children for which no treatment exists. Recently, we have obtained exciting evidence that certain antibiotic aminoglycosides, such as gentamicin and geneticin, induce the readthrough of premature termination codon (PTC) mutations in the ATM gene. The resulting ATM protein is functional, in that it corrects the radiosensitivity of A-T cells, phosphorylates ATM targets, and partially restores the S phase checkpoint, as demonstrated by measurement of radioresistant DNA synthesis. There is reason to believe that during the development of antibiotic aminoglycosides, over the past 40 years, many compounds were developed and screened for antimicrobial activity but not for PTC readthrough ability. High throughput screening will be developed, using assays based on protein truncation testing, to screen for new readthrough drugs. Promising compounds will then be further evaluated in secondary cell-based assays for ATM function, using mutations and cell lines that correspond to patient mutations. Tertiary testing will investigate the readthrough efficiency of various stop codons and the effect of the +4 nucleotide of each termination codon. Aminoglycoside-induced readthrough will be tested in combination with ATM promoter inducers in an effort to further boost intracellular ATM levels. A mouse model carrying a PTC mutation in the ATM gene will be generated for animal testing of selected drugs. Biomarkers will be developed for following in vivo responses to aminoglycoside treatment. These efforts are encouraged by the rationale that ATM heterozygotes live essentially normal lives with less than 50 percent of normal ATM protein levels. Furthermore, a subset of A-T patients with <15 percent of normal ATM protein tends to manifest less severe disease. These studies may impact upon other genetic disorders as well, and upon cancer patients with low levels of ATM protein.

描述（由申请人提供）：该项目源于我们治疗 ATM 缺陷的长期目标，无论是癌症患者还是患有共济失调毛细血管扩张症 (A-T) 的患者，这是一种罕见的儿童神经退行性疾病，目前尚无治疗方法。最近，我们获得了令人兴奋的证据，表明某些抗生素氨基糖苷类药物，例如庆大霉素和遗传霉素，会诱导 ATM 基因中提前终止密码子 (PTC) 突变的通读。由此产生的 ATM 蛋白具有功能性，因为它可以纠正 A-T 细胞的放射敏感性，磷酸化 ATM 靶标，并部分恢复 S 期检查点，如抗辐射 DNA 合成的测量所证明的。有理由相信，在过去的 40 年里，在抗生素氨基糖苷类抗生素的开发过程中，开发和筛选了许多化合物的抗菌活性，但没有针对 PTC 通读能力。将开发高通量筛选，使用基于蛋白质截断测试的测定来筛选新的通读药物。然后，将使用与患者突变相对应的突变和细胞系，在基于细胞的二次 ATM 功能测定中进一步评估有前景的化合物。第三次测试将研究各种终止密码子的通读效率以及每个终止密码子的+4核苷酸的影响。氨基糖苷诱导的通读将与 ATM 启动子诱导剂结合进行测试，以进一步提高细胞内 ATM 水平。将生成 ATM 基因中携带 PTC 突变的小鼠模型，用于选定药物的动物测试。将开发生物标志物用于跟踪氨基糖苷类治疗的体内反应。这些努力受到以下基本原理的鼓励：ATM 杂合子基本上过着正常的生活，其 ATM 蛋白水平低于正常的 50%。此外，A-T 患者中 ATM 蛋白正常值低于 15% 的亚群往往表现出较不严重的疾病。这些研究也可能对其他遗传性疾病以及 ATM 蛋白水平较低的癌症患者产生影响。