Multiplex Gene Expression in Single Cells for Circulating Tumor Cell Detection

描述(申请人提供)：循环肿瘤细胞(CTCs)的检测和计数具有成为癌症诊断和治疗的非常强大的工具的潜力。然而，目前的CTC检测技术还没有建立在常规的临床实践中。其中一个原因是，CTCs在血液中的浓度极低，每1000万个白细胞中就有一个。现有技术缺乏可靠检测这些稀有CTCs所需的灵敏度和/或特异性。另一个原因是，目前的技术没有提供关于四氯化碳分子状态的信息。该项目的总体目标是开发一种名为QMAGEX的检测技术，用于单细胞基因表达的定量复合分析，并利用该技术对体液中CTCs进行灵敏检测、准确计数和功能分析。这项工作的长期目标是利用这项检测技术开发一套高价值的癌症诊断产品。这些产品将满足癌症诊断中许多尚未得到满足的重要需求，特别是在早期发现、治疗指导和复发监测方面。在第一阶段，我们开发了一个原型QMAGEX方法，证明了它能够以单拷贝灵敏度在单个细胞中原位检测RNA转录本，并证明了其用于血液中癌细胞检测的可行性。在第二阶段，我们的目标是将QMAGEX从目前的概念验证阶段转移到一个强大的和高信息含量的分析阶段。第二阶段的具体目标包括：1)开发高信息含量4-plex QMAGEX分析的基本成分；2)开发和验证用于检测血液中稀有细胞的流式细胞仪4-plex QMAGEX分析；以及3)评估高信息含量4-plex QMAGEX检测转移性乳腺癌患者CTCs的临床应用价值。第三阶段将开发专注于特定癌症和特定诊断应用的商业诊断产品。与公共卫生相关：众所周知，循环肿瘤细胞(CTCs)可以提供肿瘤生长和转移的早期迹象。因此，使用QMAGEX检测系统对CTC的检测和分子分析将在癌症的早期发现、确定癌症患者的靶向治疗和确定治疗反应、监测癌症进展和复发方面具有重要价值。

DESCRIPTION (provided by applicant): The detection and enumeration of circulating tumor cells (CTCs) has the potential to become a very powerful tool in cancer diagnosis and treatment. However, current technologies for CTC detection are not yet established in routine clinical practice. One reason is that CTCs are present in blood at an extremely low concentration of one in 10 million white blood cells. Existing technologies lack the sensitivity and/or specificity required for reliable detection of these rare CTCs. Another reason is that current technology does not offer information about the molecular status of CTCs. The overall goal of this project is to develop an assay technology named QMAGEX for Quantitative Multiplex Analysis of Gene Expression in Single Cells and to utilize the technology for sensitive detection, accurate enumeration, and functional analysis of CTCs in body fluids. The long-term goal of this effort is to leverage this assay technology to develop a suite of high value cancer diagnostic products. These products would satisfy many significant unmet needs in cancer diagnostics, especially in the area of early detection, therapy guidance and recurrence monitoring. In Phase I, we have developed a prototype QMAGEX assay, demonstrated its capability for detecting RNA transcripts in situ in individual cells with single copy sensitivity, and proven its feasibility for cancer cell detection in blood. In Phase II, we aim to move QMAGEX from the current proof-of-concept stage to a robust and high information content assay. The specific aims in Phase II include: 1) Develop essential components for high information content 4-plex QMAGEX assay; 2) Develop and validate a flow cytometric 4-plex QMAGEX assay for rare cell detection in blood; and 3) Evaluate the clinical utility of the high information content 4-plex QMAGEX assay for detection of CTCs in metastatic breast cancer patients. Commercial diagnostic products focusing on specific cancers and specific diagnostic applications will be developed in Phase III. PUBLIC HEALTH RELEVANCE: It has been known that circulating tumor cells (CTCs) can provide an early indication of tumor growth and metastasis. Thus, detection and molecular analysis of CTC using the QMAGEX assay system will be valuable in early detection of cancer, in identifying cancer patients for targeted therapies and determining response to treatment, in monitoring cancer progression and recurrence.