Alternate Splicing and Alcohol Exposure: TransExpress Hematome 1.0

描述(由申请人提供)：尽管酒精暴露对生物标记物的需求非常迫切，但高通量基因组学技术在这一领域的应用文献异常稀少。关于酒精暴露的生物标志物的有限报道大多局限于在小鼠或大鼠大脑中发现的对乙醇的反应的生物标志物。人类受试者的大脑样本无法获取，因此无法翻译到临床环境中。对于酒精研究中的高通量应用，迫切需要一种积极的方法，这种方法可以在临床环境中随时进行测试。在全基因组微阵列上对血液进行的高通量实验只能监测15%-20%的基因，而丢失了大多数功能多样的mRNA剪接产物。考虑到后生动物使用交替剪接作为一种机制来创造从少量平行于基因复制的功能多样性的进化意义，基因芯片在生物标志物研究中的局限性是非常有限的。在这里，通过利用Jivan，hematome 1.0开发的全血剪接微阵列技术，我们提出了一个研究计划，弥合了基因组研究和临床应用之间的差距。此外，通过允许测量受调节的剪接，血液组1.0提供了3到4倍于血液中酒精暴露的生物标记物。Jivan的专利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)的发现方面对这一努力至关重要，因为血液中人类基因的大多数剪接变体尚未确定特征。该项目的关键人员将是Subha Srinivesan博士(首席调查员)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汇编一份全面的候选基因清单，这些基因显示猴子血液中受调控的剪接对酒精暴露的反应。猴子模型的使用既是为了让我们的发现尽可能与人类相关，也是为了避免人类样本因自我报告酒精/食物/药物消费而产生的偏见。在第一阶段中，将对5只动物在酒精暴露前后不同时间点的血液剪接调节进行分析，寻找候选基因。在第二阶段，这些候选基因将在数百个人类血液样本中进行预测意义的验证。

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.