HLS Detection of RNA biomarkers in individual platelets

Name of Applicant Last First Middle GNATENKO Dmitri V ABSTRACT This SBIR Phase I application is submitted in response to Small Business Topics of Special Interest TOSI initiative HLS The overall goal of the proposal is to develop and commercialize an assay for detection of RNA biomarkers in circulating blood platelets developed as a novel diagnostic approach with potential applicability to wide ranging disorders including cancer cardiovascular or autoimmune disorders Platelets are anucleate blood cells that have a crucial role in the maintenance of hemostasis thrombosis and wound healing They contain a broad variety of miRNAs and mRNAs In addition to their canonical functions platelets mediate intercellular RNA transfer and are known to have altered genetic signatures that allow for disease tracking and or early diagnostics We have successfully adapted and compared several RNA profiling platforms to transcript profiling of platelets and developed a class prediction model that discriminates clonal from non clonal phenotypes of thrombocytosis with accuracy We will now extend these advances to combine fluorescent in situ hybridization and flow cytometry for multiplexed mRNA and miRNA biomarkers in individual platelets The high sensitivity and specificity of this technology bypasses the need for technically rigorous RNA isolation and allows for routine genetic biomarker detection and quantification using standard flow cytometric analyses In Phase I we propose to develop a sensitive assay for detection and quantification of miRNA and mRNA transcripts in platelets at a single cell level and to compare this technology to quantitative RT PCR Phase II will be focused on refinement analytical and clinical validation of the technology for diagnostics and disease tracking human disorders encompassing genetically altered biomarker subsets Since platelets are easily accessible this technology represents a significant step forward towards minimally invasive detection and quantification of RNA biomarkers in circulating blood Name of Applicant Last First Middle GNATENKO Dmitri V We propose to develop a sensitive technology to detect and quantify biomarkers of various diseases in human blood This technology will work on the single cell level and detect biomarkers in blood cells thrombocytes using fluorescent signals Today accumulating evidences indicate that human blood thrombocytes contain biomarkers of many human diseases including cancer We propose to develop a sensitive diagnostic assay based on nanotechnology fluorescent probes and routine blood test Once fully developed this technology can be adapted to diagnostics of many types of cancer and other common diseases