HLS-Detection of RNA biomarkers in individual platelets

HLS-检测个体血小板中的 RNA 生物标志物

• 批准号: 9341731
• 负责人: Dmitri V GNATENKO
• 金额: $ 24.31万
• 依托单位: BLOOD CELL TECHNOLOGIES, LLC
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-09-15 至 2019-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9341731
• 关键词: Address; Affect; Area; Autoimmune Diseases; Biological Assay; Biological Markers; Biological Process; Blood; Blood Cells; Blood Platelets; Blood Tests; Businesses; Bypass; Cancer Diagnostics; Cancer Patient; Cardiovascular Diseases; Cardiovascular system; Cells; Chemistry; Chronic; Clinical; Colorectal; Complex; DNA; Data; Detection; Development; Diagnostic; Disease; Early Diagnosis; Flow Cytometry; Fluorescent Probes; Fluorescent in Situ Hybridization; Foundations; Generations; Genetic Markers; Goals; Half-Life; Hemostatic function; Human; Immune; In Situ; Individual; Inflammation; Kidney; Maintenance; Malignant Neoplasms; Mediating; Messenger RNA; Methods; MicroRNAs; Modeling; Modification; Myocardial; Names; Nanotechnology; Nucleic Acids; Nucleotides; Organism; Phase; Phenotype; Physiology; Plasma; Population; Procedures; RNA; Recurrent disease; Reproducibility; Role; Sensitivity and Specificity; Sickle Cell; Signal Transduction; Small Business Innovation Research Grant; Source; Specificity; System; Technology; Testing; Thrombosis; Transcript; Transfer RNA; Untranslated RNA; Validation; Whole Blood; Work; Wound Healing; assay development; base; cancer type; cell type; clinically relevant; design; diagnostic assay; genetic signature; human disease; improved; interest; microRNA biomarkers; minimally invasive; novel; novel diagnostics; response; technology validation; thrombocytosis; transcriptome sequencing; tumor

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 HLS17-03. 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 93.6% 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.

申请人姓名（姓、名、中间名）：GNATENKO，Dmitri V. 摘要 此SBIR第一阶段申请是为了响应小企业专题特别 利益（TOSI）倡议HLS 17 -03。该提案的总体目标是发展和 将开发的用于检测循环血小板中RNA生物标志物的测定商业化 作为一种新的诊断方法，可能适用于广泛的疾病， 癌症、心血管疾病或自身免疫性疾病。血小板是一种无核血细胞， 在维持止血、血栓形成和伤口愈合中起关键作用。它们含有 各种各样的miRNAs和mRNAs。除了它们的典型功能外，血小板还介导 细胞间RNA转移，并且已知具有允许疾病发生的改变的遗传标记 跟踪和/或早期诊断。我们已经成功地适应和比较了几种RNA 分析平台，以转录血小板谱，并开发了一个类预测模型， 区分血小板增多症的克隆和非克隆表型，准确率为93.6%。我们将 现在将这些进展扩展到结合联合收割机荧光原位杂交和流式细胞术， 单个血小板中的多重mRNA和miRNA生物标志物。的高灵敏度和 这种技术的特异性绕过了对技术上严格的RNA分离的需要， 用于使用标准流式细胞术进行常规遗传生物标志物检测和定量 分析。在第一阶段，我们建议开发一种灵敏的检测和定量分析， 在单细胞水平检测血小板中的miRNA和mRNA转录物，并将该技术与 定量RT-PCR。第二阶段将侧重于改进，分析和临床验证 用于诊断和疾病跟踪的人类疾病技术，包括遗传学上的- 改变的生物标志物子集。由于血小板很容易获得，这项技术代表了 向RNA微创检测和定量迈出重要一步 循环血液中的生物标志物。