Power calculation and design issues in next-generation sequencing

下一代测序中的功率计算和设计问题

• 批准号: 9278965
• 负责人: George C. Tseng
• 金额: $ 21.57万
• 依托单位: UNIVERSITY OF PITTSBURGH AT PITTSBURGH
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-07-07 至 2019-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9278965
• 关键词: Alleles; Binomial Model; Bioconductor; Bioinformatics; Biological Markers; Budgets; Computer software; Cost-Benefit Analysis; DNA Methylation; Data; Detection; Diagnosis; Dimensions; Drops; Equilibrium; Event; Expenditure; Experimental Designs; Future; Galaxy; Gene Expression; Gene Fusion; Genes; Genomics; Goals; Health; Java; Low Prevalence; Malignant Neoplasms; Malignant neoplasm of prostate; Massive Parallel Sequencing; Methods; Methylation; Modeling; Online Systems; Patient-Focused Outcomes; Prevalence; Prostate; Research; Research Design; Research Personnel; Resolution; Sample Size; Sampling; Software Tools; Somatic Mutation; Specific qualifier value; System; Technology; Transcript; base; candidate marker; collaborative environment; cost; cost effective; data modeling; deep sequencing; design; differential expression; experimental study; fusion gene; genome-wide; genome-wide analysis; human disease; improved; next generation sequencing; novel; open source; outcome forecast; public health relevance; sample collection; software development; statistics; tool; transcriptome sequencing; tumor; two-dimensional

 DESCRIPTION (provided by applicant): Next-generation sequencing has brought revolutionary genome-wide, dense resolution and high-throughput capability to perform various types of omics analyses, including gene expression, methylation, fusion gene, somatic mutation and many others. With the dropping costs, the technology is gaining popularity. The experimental expenses, however, remain significant and power calculation tools are essential to adequately design and guide an NGS analysis. Unlike power calculation in traditional experiments or microarrays, power calculation in NGS require simultaneous consideration of sample size and sequencing depth and count-data also bring statistical challenges. We propose the following aims in this proposal: (1a) Develop power calculation tools for differential expression analysis from RNA-seq experiments. Optimal sample size and sequencing depth are jointly determined by power function and budget constraints. (1b) Develop power calculation tools for differential methylation in methyl-seq experiments. (2a) Develop power calculation tools for fusion gene detection in cancer using RNA-seq. Identify sample size and sequencing depth needed for fusion genes with low prevalence and low allelic-fraction. (2b) Perform additional ultra-deep sequencing in the preliminary prostate study to identify additional low-allelic-fraction and prognosis predictive fusion genes. Successful completion of these aims will provide state-of-the-art power calculation tools for the fast growing projects using NGS technology for candidate marker and fusion gene detection.

 描述（由申请人提供）：下一代测序带来了革命性的全基因组、高密度分辨率和高通量能力，可进行各种类型的组学分析，包括基因表达、甲基化、融合基因、体细胞突变等。随着成本的下降，该技术越来越受欢迎，然而，实验费用仍然很高，功效计算工具对于充分设计和指导 NGS 分析至关重要，与传统实验或微阵列中的功效计算不同，NGS 中的功效计算需要同时考虑样本。大小和测序深度以及计数数据也带来了统计挑战：（1a）开发用于RNA-seq实验差异表达分析的功效计算工具最佳样本量和测序深度由功效共同决定。 (1b) 开发甲基测序实验中差异甲基化的功效计算工具 (2a) 开发使用 RNA 测序进行癌症融合基因检测的功效计算工具。患病率低(2b) 在初步前列腺研究中进行额外的超深度测序，以确定额外的低等位基因部分。 成功完成这些目标将为使用 NGS 技术进行候选标记和融合基因检测的快速发展项目提供最先进的功率计算工具。