Rapid, low-cost mRNA analysis system for cancer companion diagnostics

用于癌症伴随诊断的快速、低成本 mRNA 分析系统

• 批准号: 8394718
• 负责人: Aaron R. Hawkins
• 金额: $ 27.21万
• 依托单位: LIQUILUME DIAGNOSTICS, INC.
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-09-01 至 2014-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8394718
• 关键词: Algorithms; Antineoplastic Agents; Area; Biological Assay; Biological Markers; Cancer Detection; Cancer Diagnostics; Cancer Patient; Cells; Clinical; Communicable Diseases; Companions; Detection; Development; Diagnostic; Disease; Evaluation; FDA approved; Genetic Screening; Genetic Transcription; Genome; Genomics; Goals; Gold; Government; Growth; Immune response; Immunotherapy; In Vitro; Interferon Type II; Intervention; Malignant Neoplasms; Medicine; Messenger RNA; Methods; Microfluidics; Molecular; Molecular Analysis; Monitor; Noise; Nucleic Acid Amplification Tests; Nucleic Acids; Optics; Organ Transplantation; Outcome; Peripheral Blood Mononuclear Cell; Persons; Pharmaceutical Preparations; Phase; Play; Polymerase Chain Reaction; Principal Investigator; Public Health; RNA; Regulatory T-Lymphocyte; Research; Role; Sampling; Sensitivity and Specificity; Signal Transduction; Small Business Innovation Research Grant; Solutions; Specimen; Spottings; Structure; System; Systems Analysis; Technology; Testing; Time; Translating; Viral Load result; Work; base; cancer immunotherapy; cost; genetic analysis; innovation; instrument; melanoma; mind body interaction; multiplex detection; novel; novel strategies; nucleic acid detection; oncology; operation; pathogen; programs; single molecule; synthetic nucleic acid

DESCRIPTION (provided by applicant): Nucleic acid tests comprise a large and growing segment of emerging molecular diagnostics ("MDx"). Companion diagnostics is an extremely important aspect of MDx due its potential to inform and accompany treatment of diseases using a genome-based, personalized approach. Taking the next step in molecular diagnostics will require the development of sensitive and specific instruments that provide repeated testing of disease biomarkers at low cost and low complexity. The long-term goal of LiquiLume Diagnostics Inc. is to develop solutions to these challenges based on optofluidics - the combination of integrated optics and microfluidics on a single chip-scale system. The objective of this application is to demonstrate key capabilities of LiquiLume's envisioned companion diagnostic instrument, Optofluidic RNA Biomarker Testing System (ORBITS). The ORBITS is a platform technology introducing optofluidic waveguide- based detection on the single molecule level for amplification-free nucleic acid detection. As a first, representative example, we will demonstrate amplification-free monitoring of cancer biomarkers in clinical specimens. The objectives of this application will be accomplished by completing the following specific aims: (1) Nucleic acid (mRNA) detection in clinical samples; and (2) Multi-spot excitation nucleic acid detection, both using ORBITS optofluidic chips. Under the first Aim, we will show that mRNA cancer biomarkers from clinical sample material can be identified and quantitated. Specifically, we will demonstrate detection of five mRNA biomarkers in peripheral blood mononuclear cells (PBMCs) from melanoma cancer patients treated with the anti-cancer drug ipilimumab. The second Aim will introduce multi-spot optical excitation via interferometric waveguide structures for enhancing signal-to-background ratios. A 100x improvement over single-spot excitation will be demonstrated. The project's innovative aspects lie in the first demonstration of integrated optofluidic technology for cancer immunotherapy, and the introduction of interferometric excitation for optical signal-to-noise enhancement. The proposed work is significant because it will introduce a new type of instrument that features the specificity and sensitivity of gold standard polymerase chain reaction methods at much lower complexity and cost. Demonstration of the ORBITS principle will not only be applicable to companion diagnostics of cancer, but also serve as the blueprint for a broader class of molecular diagnostic instruments for rapid pathogen and biomarker detection. PUBLIC HEALTH RELEVANCE: Quantitative monitoring of tens of biomarkers is an essential component of cancer immunotherapy and other forms of emerging companion diagnostics. This application describes a novel approach to this problem using amplification-free detection of cancer mRNA biomarkers on an optofluidic chip. The proposed integrated diagnostic platform would optimize public health interventions by providing sensitive and accurate molecular diagnostics with low cost and complexity.

描述（由申请人提供）：核酸检测构成了新兴分子诊断（“MDx”）的一个庞大且不断增长的部分。伴随诊断是 MDx 的一个极其重要的方面，因为它有可能使用基于基因组的个性化方法为疾病提供信息并伴随治疗。分子诊断的下一步需要开发灵敏且特异的仪器，以低成本和低复杂性提供疾病生物标志物的重复测试。 LiquiLume Diagnostics Inc. 的长期目标是开发基于光流控技术（在单芯片级系统上集成光学和微流控技术的组合）来应对这些挑战的解决方案。该应用的目的是展示 LiquiLume 设想的配套诊断仪器 Optofluidic RNA 生物标志物测试系统 (ORBITS) 的关键功能。 ORBITS 是一种平台技术，在单分子水平上引入基于光流控波导的检测，用于无扩增核酸检测。作为第一个代表性示例，我们将演示对临床样本中癌症生物标志物的无扩增监测。本应用的目的将通过完成以下具体目标来实现：（1）临床样本中的核酸（mRNA）检测； (2)多点激发核酸检测，均使用ORBITS光流控芯片。在第一个目标下，我们将证明可以识别和定量临床样本材料中的 mRNA 癌症生物标志物。具体来说，我们将展示在接受抗癌药物 ipilimumab 治疗的黑色素瘤癌症患者的外周血单核细胞 (PBMC) 中检测 5 种 mRNA 生物标志物。第二个目标将通过干涉波导结构引入多点光学激发，以提高信号背景比。将展示比单点激励 100 倍的改进。该项目的创新之处在于首次展示了用于癌症免疫治疗的集成光流控技术，以及引入了用于光学信噪比增强的干涉激发。拟议的工作意义重大，因为它将引入一种新型仪器，该仪器具有金标准聚合酶链式反应方法的特异性和灵敏度，且复杂性和成本低得多。 ORBITS 原理的演示不仅适用于癌症的伴随诊断，而且还可以作为更广泛的分子诊断仪器的蓝图，用于快速病原体和生物标志物检测。 公共卫生相关性：数十种生物标志物的定量监测是癌症免疫治疗和其他形式的新兴伴随诊断的重要组成部分。该应用描述了一种解决该问题的新方法，使用光流控芯片上的癌症 mRNA 生物标志物的无扩增检测。拟议的综合诊断平台将通过以低成本和复杂性提供灵敏且准确的分子诊断来优化公共卫生干预措施。