HIGH THROUGHPUT DETECTION OF GENETIC MUTATIONS AS BIOMARKERS OF FUT

作为 FUT 生物标志物的基因突变的高通量检测

• 批准号: 7342283
• 负责人: RICHARD A MATHIES
• 金额: $ 39.12万
• 依托单位: UNIVERSITY OF CALIFORNIA BERKELEY
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-08-15 至 2011-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7342283
• 关键词: 5q31; 7q22; Alkylating Agents; Aneuploidy; Benzene; Biological Assay; Biological Markers; Blood Cells; Cell Line; Cell Separation; Cells; Chemicals; Chromosome Structures; Chromosome abnormality; Chromosomes; Cytogenetics; Detection; Development; Disease; Disease Pathway; Early Diagnosis; Emulsions; Epidemiologic Studies; Event; Flow Cytometry; Fluorescence; Frequencies; Future; Gene Mutation; Genes; Genetic; Genomics; Genotype; Goals; Hematopoietic Neoplasms; Hereditary Disease; Human; Hypoxanthine Phosphoribosyltransferase; Individual; Label; Link; Malignant Neoplasms; Measures; Metaphase Spread; Methods; Microfluidics; Modeling; Monitor; Mutation; N-ras Genes; NPM1 gene; NRAS gene; Normal Cell; Numbers; Plasmids; Point Mutation; Polymerase Chain Reaction; Production; Range; Risk; Solutions; Somatic Mutation; Specimen; Stem cells; Technology; Testing; Time; Work; cancer risk; epidemiology study; genetic analysis; human population study; leukemia; leukemia/lymphoma; micro-total analysis system; multiplex detection; nanolitre; nucleophosmin; p21 N-Ras Protein; prospective; research study; single molecule; size; tool

Cancer is a genetic disease involving the sequential accumulation of somatic mutations. Genetic damage leading to these mutations can occur at the level of the gene, (e.g. point mutations) or the chromosome (e.g. aneuploidy, translocations). Numerous biomarkers have been developed to detect early mutational and chromosomal effects of carcinogenic exposure in humans. Historically, biomarkers have tended to measure mutations in surrogate genes, such as HPRT, or use cytogenetics to assess chromosomal aberrations (CA). These biomarkers are elevated by a wide range of carcinogenic exposures and CA have been shown to be predictive of future cancer risk in prospective epidemiology studies. They do, however, have several drawbacks in that surrogate genes are not on the causal pathway of disease and cytogenetic markers require metaphase spreads to be prepared and their analysis is time consuming and expensive. New biomarkers of early effect for cancer are therefore urgently needed. We propose to develop biomarkers of early effect for blood cancers by generating high-throughput single cell PCR assays for mutation in key genes or regions linked to leukemia and lymphoma. This approach will take advantage of recent advances in microfluidics and lab-on-a-chip technologies that make single cell genetic analysis (SCGA) feasible on a high-throughput scale. Specifically we plan to develop methods for performing high throughput single template copy PCR amplification in nanoliter emulsion droplets and then apply these methods to develop high-throughput SCGA methods to detect low frequencies of mutations of importance in blood cancers, such as translocation t(14;18), deletion of 5q31 and mutations in NFtAS and NPM1. We will test if the new SCGA methods can detect mutations in cells exposed to alkylating agents and benzene metabolites and humans exposed to benzene. The studies proposed will be a first step towards providing revolutionary new assays for the detection of genetic mutations of relevance to blood cancer risk in healthy individuals. Such biomarkers will be useful in epidemiological studies of leukemia and lymphoma, which have long latency periods, as well as providing tools for early detection for those individuals at risk. The high-throughput detection methods we propose to develop should be applicable to large human population studies and will work with existing biobanked specimens

癌症是一种遗传疾病，涉及体细胞突变的顺序积累。遗传损伤