Alternate Splicing and Alcohol Exposure: TransExpress Hematome 1.0

交替剪接和酒精暴露：TransExpress Hematome 1.0

• 批准号: 7105995
• 负责人: SUBHA SRINIVASAN
• 金额: $ 11.15万
• 依托单位: JIVAN BIOLOGICS, INC.
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-09-01 至 2008-02-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7105995
• 关键词: Macaca; RNA splicing; alcoholism /alcohol abuse; biomarker; biotechnology; blood; ethanol; high throughput technology; microarray technology; nucleic acid hybridization; nucleic acid purification; technology /technique development

DESCRIPTION (provided by applicant): Despite a critical need for biomarkers for alcohol exposure, the literature is unusually sparse in the applications of high-throughput genomics technologies in this area. The limited reports on biomarkers of alcohol exposure mostly confined to biomarkers identified in mouse or rat brains in response to ethanol. The inaccessibility of brain samples from human subjects renders them untranslatable to clinical settings. There is a dire need for an aggressive approach for high-throughput applications in alcohol research that can be ready tested in clinical settings. High-throughput experiments with blood on genome-wide microarrays can only monitor 15-20% of genes while missing majority of the functionally diverse mRNA spliced products. Considering evolutionary implications of alternate splicing as a mechanism used by metazoans to create functional diversity from small number of genes parallel to gene duplication, the limitations of gene microarrays are very limiting in biomarker research. Here, by utilizing a blood-wide splice microarray technology developed by Jivan, Hematome 1.0, we propose a research plan that bridges the gap between genome research and clinical application. Furthermore, by allowing for measurement of regulated splicing, Hematome 1.0 provide 3 to 4 fold more biomarkers for alcohol exposure from blood. Jivan's proprietary TransExpress(tm) Hematome 1.0 microarray platform has the capability to simultaneously discover and measure splice regulation of genes. The discovery aspect of TransExpress(tm) is critical to this endeavor because a majority of splice variants of human genes in blood are yet uncharacterized. The key personnel on this project will be Dr. Subha Srinivasan (Principal Investigator) and Jonathan Bingham, both of whom have extensive industry experience in genomics, microarray technologies, and bioinformatics. The specific aim of the SBIR Phase I project is to compile a comprehensive list of candidate genes that display regulated splicing in monkey blood in response to alcohol exposure. The monkey model is used both to make our findings as relevant to humans as possible and to avoid bias in human samples from selfreporting alcohol/food/drug consumption. Splice regulation in blood from 5 animals at various time points before and after exposure to alcohol will be profiled in Phase I for candidate genes. In Phase II, these gene candidates will be validated for predictive significance in hundreds of human blood samples.

描述（由申请人提供）：尽管迫切需要酒精暴露的生物标志物，但在该领域应用高通量基因组学技术的文献异常稀少。关于酒精暴露的生物标志物的有限报告大多局限于小鼠或大鼠大脑中响应于乙醇的生物标志物。来自人类受试者的大脑样本的不可接近性使得它们无法转化为临床环境。迫切需要一种积极的方法，用于酒精研究中的高通量应用，可以在临床环境中进行测试。在全基因组微阵列上使用血液进行的高通量实验只能监测15-20%的基因，而错过了大多数功能多样的mRNA剪接产物。考虑到选择性剪接作为后生动物使用的一种机制，从少量基因平行于基因复制产生功能多样性的进化意义，基因微阵列的局限性在生物标志物研究中是非常有限的。在这里，通过利用由Jivan开发的全血液剪接微阵列技术，Hematome 1.0，我们提出了一个研究计划，弥合基因组研究和临床应用之间的差距。此外，通过允许测量调节剪接，Hematome 1.0提供了3至4倍的血液酒精暴露生物标志物。Jivan公司专有的TransExpress（TM）Hematome 1.0微阵列平台具有同时发现和测量基因剪接调控的能力。TransExpress（tm）的发现方面对这一奋进至关重要，因为血液中人类基因的大多数剪接变体尚未被表征。该项目的主要人员将是Subha Srinivasan博士（首席研究员）和Jonathan Bingham，他们都在基因组学，微阵列技术和生物信息学方面拥有丰富的行业经验。SBIR第一阶段项目的具体目标是编制一份全面的候选基因清单，这些基因在猴子血液中显示出对酒精暴露的反应。使用猴子模型既可以使我们的研究结果尽可能与人类相关，也可以避免人类样本中自我报告的酒精/食物/药物消费的偏倚。将在I期分析5只动物在暴露于酒精之前和之后的不同时间点的血液中的剪接调节，以获得候选基因。在第二阶段，这些候选基因将在数百个人类血液样本中验证其预测意义。