Automated Systems for Detection and Molecular Characterization of Circulating Tum

循环肿瘤检测和分子表征的自动化系统

• 批准号: 8145206
• 负责人: YULING LUO
• 金额: $ 100.63万
• 依托单位: ADVANCED CELL DIAGNOSTICS, INC.
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-09-05 至 2013-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8145206
• 关键词: Applications Grants; Area; Biological Assay; Biological Markers; Biological Preservation; Blood; Blood specimen; Cancer Diagnostics; Cancer Patient; Cell Line; Cellular Morphology; Collection; Data; Detection; Development; Diagnostic; ERBB2 gene; Early Diagnosis; Epidermal Growth Factor Receptor; FDA approved; Foundations; Goals; Grant; Image; In Situ; Label; Lead; Leukocytes; Malignant Neoplasms; Manuals; Methods; Modeling; Molecular; Molecular Analysis; Monitor; Monitoring for Recurrence; Outcomes Research; Patients; Performance; Phase; Phenotype; Primary Neoplasm; Procedures; RNA; RNA analysis; RNA marker; Recovery; Reproducibility; Research; Screening for cancer; Sensitivity and Specificity; Signal Transduction; Small Business Innovation Research Grant; Specificity; System; Technology; Validation; Work; base; cancer cell; cancer diagnosis; cancer recurrence; cancer therapy; clinical practice; clinical research site; fluorescence microscope; improved; malignant breast neoplasm; molecular phenotype; multiplex detection; neoplastic cell; prototype; public health relevance; response; single molecule; tool; tumor progression

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 technologies do not offer information about the molecular status of CTCs, which can be used to select appropriate targeted therapy and monitor therapy response. The overall goal of this proposal is to develop an automated system called CTCscope" for detection and molecular phenotyping of circulating tumor cells (CTCs) in blood. This automated system is based on the successful development of ACD's RNAscope" technology and prototype CTCscope"system, both of which are supported in part by a SBIR Phase II grant R44CA122444. Our aim in this proposal is to develop and validate the automated CTCscope" system for sensitive detection, accurate enumeration, and molecular characterization of CTCs. The long term goal of this effort is to leverage CTCscope" to develop a suite of high value cancer diagnostic products. These products would satisfy many significant unmet needs in cancer management, especially in the areas of early detection, therapy selection and guidance, and recurrence monitoring. With the support of the SBIR phase II grant, we have successfully developed the RNAscope"assay capable of in situ multiplex detection and quantification of RNA markers with single molecule sensitivity and exquisite specificity. We have also developed a manual prototype system for CTC detection and analysis. The manual system includes three components: 1) CTC preservation: CTCs are preserved and RNA stabilized in the blood sample; 2) CTC labeling: CTCs are labeled for multiple RNA markers in situ by the RNAscope"assay; and 3) CTC detection and analysis: CTCs are detected, enumerated and characterized based on RNA marker signals observed under the fluorescent microscope. Using this system, we have demonstrated highly specific detection of circulating tumor cells and characterized their EGFR expression status in breast cancer patients. These data demonstrated that CTCscope" has several unique capabilities which could potentially offer significant advantages over existing CTC detection methods. In this proposal, we will extend our current work to develop and validate an automated CTCscope" system for reliable detection and enumeration of CTCs, and for quantitative analysis of RNA markers reflective of CTCs' molecular phenotype. In addition, as part of the CTCscope" system validation, the HER2 and EGFR status of the CTCs and their concordance/discordance with their status in primary tumor will be determined. The specific aims include: 1) Develop an automated CTCscope" system for CTC detection and molecular characterization; 2) Validate CTCscope" by comparing its performance with the CellSearch(R) system using blood samples from ~230 metastatic breast cancer patients; and 3) Determine HER2 and EGFR status on CTCs and compare their status on primary tumors. The specific outcomes of this research will include an automated CTCscope" platform that is successfully developed and validated, enabling a suite of high value diagnostic products to be developed for greatly improved cancer management. This research will also lay the foundation for a valuable diagnostic product that can guide targeted therapy selection and monitor therapy response for metastatic breast cancer patients. With the CTCscope" platform successfully developed and validated, a suite of high value diagnostic products can be developed for greatly improved cancer management. PUBLIC HEALTH RELEVANCE: This is a SBIR Phase II Bridge Grant application. The automated CTCscope" system developed under this proposal will enable reliable detection, accurate enumeration, and molecular analysis of circulating tumor cells, which will lead to a suite of high value cancer diagnostic products for early detection of cancer, for selecting right cancer patients for targeted therapies, for determining their response to treatment, and for monitoring cancer progression and recurrence.

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有关CTC分子状态的信息，这些信息可用于选择适当的靶向治疗和监测治疗反应。该提案的总体目标是开发一种名为“CTC scope”的自动化系统，用于检测血液中循环肿瘤细胞（CTC）的分子表型。该自动化系统基于ACD的RNAscope”技术和原型CTCscope“系统的成功开发，两者都得到SBIR第二阶段资助R44 CA 122444的部分支持。本提案的目的是开发和验证自动化CTCscope系统，用于灵敏检测、准确计数和CTC分子表征。这项工作的长期目标是利用CTCscope”开发一套高价值的癌症诊断产品。这些产品将满足癌症管理中许多尚未满足的重要需求，特别是在早期检测、治疗选择和指导以及复发监测等领域。在SBIR II期资助的支持下，我们成功开发了RNAscope“检测试剂盒，能够原位多重检测和定量RNA标记物，具有单分子灵敏度和精确的特异性。我们还开发了一个手动CTC检测和分析原型系统。手动系统包括三个组件：1）CTC保存：CTC被保存并且RNA在血液样品中稳定化; 2）CTC标记：通过RNAscope测定原位标记CTC的多个RNA标志物;和3）CTC检测和分析：基于在荧光显微镜下观察到的RNA标志物信号检测、计数和表征CTC。使用该系统，我们已经证明了循环肿瘤细胞的高度特异性检测，并在乳腺癌患者中表征其EGFR表达状态。这些数据表明，CTCscope“具有几种独特的能力，可能比现有的CTC检测方法具有显着的优势。在本提案中，我们将扩展我们目前的工作，以开发和验证自动化CTCscope系统，用于可靠地检测和计数CTC，并用于定量分析反映CTC分子表型的RNA标记物。此外，作为CTCscope系统验证的一部分，将确定CTC的HER 2和EGFR状态及其与原发性肿瘤状态的一致性/不一致性。具体目标包括：1）开发用于CTC检测和分子表征的自动化CTCscope”系统; 2）使用来自约230名转移性乳腺癌患者的血液样本，通过将其性能与CellSearch（R）系统进行比较来验证CTCscope”;以及3）确定CTC上的HER 2和EGFR状态，并比较它们在原发性肿瘤上的状态。这项研究的具体成果将包括一个成功开发和验证的自动化CTCscope平台，从而能够开发一套高价值的诊断产品，大大改善癌症管理。这项研究还将为一种有价值的诊断产品奠定基础，该产品可以指导转移性乳腺癌患者的靶向治疗选择和监测治疗反应。随着CTCscope平台的成功开发和验证，可以开发一套高价值的诊断产品，大大改善癌症管理。 公共卫生相关性：这是SBIR第二阶段桥梁补助金申请。根据该提案开发的自动化CTCscope系统将能够对循环肿瘤细胞进行可靠的检测、准确的计数和分子分析，这将产生一套高价值的癌症诊断产品，用于早期检测癌症、选择合适的癌症患者进行靶向治疗、确定他们对治疗的反应以及监测癌症进展和复发。