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.

申请人的名称（最后，第一个，中间）：Gnatenko，Dmitri V. 抽象的 此SBIR I期申请是针对特殊小企业主题提交的 利息（TOSI）倡议HLS17-03。该提案的总体目标是发展和 商业化在循环血小板中检测RNA生物标志物的测定，开发的 作为一种新颖的诊断方法，可能适用于广泛的疾病 癌症，心血管或自身免疫性疾病。血小板是具有Anuclete的血细胞，具有 在止血，血栓形成和伤口愈合的维持中至关重要的作用。他们包含一个 各种各样的miRNA和mRNA。除了它们的规范功能外，血小板还介导 细胞间RNA转移，已知遗传特征改变了疾病 跟踪和/或早期诊断。我们已经成功适应并比较了几个RNA 将平台分析到血小板的成绩单分析，并开发了一个类预测模型 将克隆与血小板病的非克隆表型区分开，精度为93.6％。我们将 现在扩展这些进步以结合原位杂交和流式细胞仪的结合 单个血小板中的多路复用mRNA和miRNA生物标志物。高灵敏度和 该技术的特异性绕过了对技术严格的RNA隔离的需求，并允许 用于常规的遗传生物标志物检测和使用标准流式细胞术 分析。在第一阶段，我们建议开发一种敏感测定法以检测和定量 小血小板中的miRNA和mRNA转录本在单细胞水平上，并将这种技术与 定量RT-PCR。第二阶段将侧重于改进，分析和临床验证 诊断和疾病的技术跟踪人类疾病，包括基因 - 改变生物标志物子集。由于血小板很容易进入，因此该技术代表 朝着最小的侵入性检测和RNA定量迈出的显着步骤 循环血液中的生物标志物。