Standardized Gene Expression Core Facility Development

描述（由申请人提供）： 需要灵敏、高通量、标准化的基因表达 俄亥俄医学院专门针对癌症进行的分析 一般研究界。基因表达之间的经验关联 模式和临床上重要的表型将提供改进的标记 预测癌症行为和对特定药物治疗的反应。在 此外，此类协会将为新的机制提供知情基础 旨在改善癌症预防、诊断和治疗的研究。 微阵列（cDNA 和/或寡核苷酸）基因表达分析是一种 有价值的筛查工具，因此，俄亥俄医学院 目前正在建立微阵列核心设施。 然而，很明显微阵列分析不够灵敏， 对于许多临床应用来说是可重复的或灵活的。因此， 微阵列数据一般必须在后续定量基因中进行验证 表达分析。由六名 NCI 赞助的研究人员组成的小组 俄亥俄医学院加入支持这项申请，建立一个 高通量、标准化的基因表达分析设施基于 标准化 RT (StarRT)-PCR。这将是一个全新的资源，这 其他机构的研究人员将加入目标 4 的小组，他们 使用微阵列并支持使用 StaRT-PCR 作为“黄金标准” 他们可以优化微阵列分析方案。该设施将 以低成本提供给一般研究界。高 StaRT-PCR 的重现性、灵敏度和低检测阈值 基因表达方法已在多项独立研究中得到验证。在 在一项盲法研究中，四个基因在一个共同的 cDNA 中测量了十个基因的表达。 实验室。平均实验室间相关方差 (CV) 为 0.48，证实基因表达可能存在 2-3 倍的差异 充满信心地检测到。使用微通道毛细管电泳进行 CV 减少到0.26。该提案的具体目标如下。我; 建立可容纳4,000个基因的自动化StaRT-PCR核心设施 表达测定/24小时。目标2；以低成本向 NCI 资助的 MCO 调查人员以及其他感兴趣的调查人员 机构，3；开发一个带注释的基因表达数据库来服务 NCI 资助的 MCO 以及其他机构的研究人员，AIM：4 开发 标准化微阵列数据分析的可接受的方法。

DESCRIPTION (provided by applicant): There is a need for sensitive, high throughput, standardized gene expression analysis at the Medical College of Ohio specifically and for the cancer research community in general. Empiric association between gene expression patterns and clinically important phenotypes will provide improved markers to predict cancer behavior and response to treatment with specific agents. In addition, such associations will provide an informed basis for new mechanistic studies aimed at improving cancer prevention, diagnosis and treatment. Microarray (cDNA and/or oligonucleotide) gene expression analysis is a valuable screening tool and for this reason, the Medical College of Ohio currently is in the process of establishing a microarray core facility. However, it is clear that microarray analysis is not sufficiently sensitive, reproducible, or flexible for many clinical applications. Consequently, microarray data generally must be validated in follow-up quantitative gene expression analyses. The group of six NCI sponsored investigators at the Medical College of Ohio join in supporting this application to establish a high throughput, standardized gene expression analysis facility based on standardized RT (StaRT)-PCR. This will be a completely new resource, This group will be joined in Aim 4 by investigators at other institutions who are using microarrays and support the use of StaRT-PCR as a "gold standard" by which they may optimize microarray analysis protocols. This facility will be available to the general research community at low cost. The high reproducibility, sensitivity, and low detection threshold of the StaRT-PCR gene expression method have been validated in multiple independent studies. In a blinded study, expression of ten genes was measured in a common cDNA by four laboratories. The average inter-laboratory correlation of variance (CV) was 0.48, confirming that 2-3 fold differences in gene expression could be detected with confidence. Using microchannel capillary electrophoresis the CV was reduced to 0.26. The Specific Aims of this proposal are as follows. I; establish an automated StaRT-PCR core facility capable of 4,000 gene expression assays/24 hours. AIM 2; Provide this service at low cost to the NCI-funded investigators at MCO as well as interested investigators at other institutions, 3; Develop an annotated gene expression database to service both NCI-funded investigators at MCO as well as other institutions, ,AIM: 4 develop an acceptable method for standardizing microarray data analysis.