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. ? ? ?
