Multiplex Gene Expression in Single Cells for Circulating Tumor Cell Detection

描述（由申请人提供）：循环肿瘤细胞（CTC）的检测和计数有可能成为癌症诊断和治疗中非常强大的工具。然而，目前的CTC检测技术尚未在常规临床实践中建立。一个原因是CTC以极低的浓度存在于血液中，为1千万分之一的白色血细胞。现有技术缺乏可靠检测这些罕见CTC所需的灵敏度和/或特异性。另一个原因是目前的技术不能提供关于CTC分子状态的信息。本项目的总体目标是开发一种用于单细胞基因表达定量多重分析的检测技术QMAGEX，并利用该技术对体液中的CTC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种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满足的重要需求，特别是在早期检测、治疗指导和复发监测领域。在第一阶段，我们已经开发了一个原型QMAGEX检测，证明了其在单个细胞中以单拷贝灵敏度原位检测RNA转录本的能力，并证明了其在血液中检测癌细胞的可行性。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高信息含量检测。II期的具体目标包括：1）开发用于高信息含量4重QMAGEX测定的基本组件; 2）开发并验证用于血液中稀有细胞检测的流式细胞术4重QMAGEX测定;以及3）评价高信息含量4重QMAGEX测定用于检测转移性乳腺癌患者中CTC的临床效用。第三阶段将开发针对特定癌症和特定诊断应用的商业诊断产品。公共卫生相关性：众所周知，循环肿瘤细胞（CTC）可以提供肿瘤生长和转移的早期指征。因此，使用QMAGEX测定系统的CTC的检测和分子分析将在癌症的早期检测、鉴定癌症患者以进行靶向治疗和确定对治疗的反应、监测癌症进展和复发中有价值。

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.