Ading color to cancer, adenovirus and flow cytometry to identify and capture CTCs

为癌症、腺病毒和流式细胞术着色以识别和捕获 CTC

• 批准号: 8223340
• 负责人: Lyudmila Bazhenova
• 金额: $ 14.89万
• 依托单位: NANOCELLECT BIOMEDICAL, INC.
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-09-21 至 2013-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8223340
• 关键词: Adenovirus Vector; Adenoviruses; Aerosols; Affinity Chromatography; Antibodies; Antigens; Biological Assay; Biological Markers; Biology; Biomedical Engineering; Biopsy; Blood; Blood Cells; Blood Circulation; Blood specimen; Cancer Patient; Cardiovascular system; Cell Nucleus; Cell Separation; Cells; Cessation of life; Characteristics; Chimeric Proteins; Clinic; Clinical; Color; Cytosol; Data; Detection; Devices; Diagnosis; Diagnostic; Disseminated Malignant Neoplasm; Doctor of Philosophy; Early identification; Engineering; Epithelial; Evaluation; Flow Cytometry; Fluorescent in Situ Hybridization; Frequencies; Gene Expression Profile; Genetic; Genome; Goals; Growth; Hour; Individual; Invaded; Label; Left; Legal patent; Lesion; Libraries; Malignant - descriptor; Malignant Neoplasms; Malignant neoplasm of lung; Measures; Mediating; Microfluidics; Microscopy; Molecular; Mutation; Neoplasm Metastasis; Non-Small-Cell Lung Carcinoma; Nucleic Acids; Oncologist; Organ; Pathway interactions; Patients; Phenotype; Primary Neoplasm; Process; Proliferating; Proteins; Reporter; Reporting; Research Personnel; Reverse Transcriptase Polymerase Chain Reaction; Risk; Sampling; Science; Series; Serotyping; Signal Pathway; Site; Solid Neoplasm; Solutions; Sorting - Cell Movement; Staging; System; Techniques; Technology; Testing; Tissues; Translating; Viral; Viral Vector; Virus; base; cancer cell; cancer therapy; cost; defective adenoviral vector; design; detector; malignant phenotype; micro-total analysis system; microchip; neoplastic cell; novel; novel diagnostics; peripheral blood; point-of-care diagnostics; prognostic; response; selective expression; therapeutic target; tool; tumor; viral detection; voltage

DESCRIPTION (provided by applicant): We propose a novel technique to identify and capture circulating tumor cells (CTCs) using engineered adenoviruses and sophisticated flow cytometry. Current techniques for detection of CTCs include reverse transcriptase-polymerase chain reaction (RT-PCR), flow cytometry, fluorescence in situ hybridization, and, more recently, microfluidics. Unfortunately, RT-PCR does not distinguish between viable metastatic CTC versus nucleic acids or cellular fragments originating from the primary tumor. ! Antibody-based techniques cannot be used for detection of all cancers, but only those cancers that express the most common and well- characterized markers. As such, there is a desperate need to develop new diagnostic agents and tools that not only detect and capture CTCs but also quantify their malignant potential and identify 'up-front' the therapies that are most effective in ablating an individual patient's tumor. Despite the complexity and variability of cancers at a genome scale, a unifying theme is their growth deregulation phenotypes, the so-called hallmarks of cancer, which are conferred by mutations in a relatively small number of key pathways. Rather than focus on detecting individual genetic lesions that are numerous and highly variable between tumors, we propose to create diagnostic viruses that incorporate multiple transcriptional and molecular modules in their genomes to infect and detect a patient's tumor, report its molecular 'hallmarks' and its response to different therapies 'up- front'. Using these agents, the molecular lesions and malignant characteristics of any given tumor will be rapidly discerned (within 24 hours) and scored via a standardized automated platform. Furthermore, these agents could also be used as reporters to determine rapidly and directly if a patient's tumor is likely to respond to a particular therapy. Our goal is to develop a standardized automated platform that provides point-of-care diagnostics to inform clinical decisions at a level of molecular sophistication and prognostic power that is not possible with any other detection system, biomarkers or correlative gene expression signatures. To achieve this, we will combine transformative new technological platforms developed at the Salk, UCSD, and NanoSort that label tumor cells in different colors based on their acquisition of molecular lesions that dictate malignant progression and response to therapy, facilitating their detection, quantification and isolation using an integrated 'lab-on a chip' flow cytometer. PUBLIC HEALTH RELEVANCE: To aid in the diagnosis and treatment of cancer, we will specially design adenoviruses that preferentially infect circulating tumor cells (CTCs, cells that have left a tumor and entered the bloodstream), drastically distinguishing these rare cells from the billions of blood cells found in a routine blood sample. This technology also detects the status of the cancer, giving doctors information about how aggressive or treatable a cancer might be. We combine this adenoviral system with a sophisticated new lab-on-a-chip cell-sorting device that passes the virally infected cells along microfluidic channels to be measured and sorted from the blood, and examined for characteristic hallmarks of cancer.

描述（由申请人提供）：我们提出了一种利用工程腺病毒和复杂的流式细胞术识别和捕获循环肿瘤细胞（ctc）的新技术。目前检测ctc的技术包括逆转录聚合酶链反应（RT-PCR）、流式细胞术、荧光原位杂交，以及最近的微流体技术。不幸的是，RT-PCR不能区分活的转移性CTC与源自原发肿瘤的核酸或细胞片段。！ 基于抗体的技术不能用于检测所有的癌症，而只能用于检测那些表达最常见和表征良好的标志物的癌症。因此，迫切需要开发新的诊断试剂和工具，不仅可以检测和捕获ctc，还可以量化其恶性潜能，并“预先”确定最有效的消融单个患者肿瘤的治疗方法。尽管癌症在基因组尺度上具有复杂性和可变性，但一个统一的主题是它们的生长失调表型，即所谓的癌症标志，这是由相对少数关键途径的突变所赋予的。与其专注于检测肿瘤之间数量众多且高度可变的单个遗传病变，我们建议创建诊断病毒，该病毒在其基因组中包含多个转录和分子模块，以感染和检测患者的肿瘤，报告其分子“特征”及其对不同治疗的反应。使用这些药物，任何给定肿瘤的分子病变和恶性特征将被快速识别（在24小时内），并通过标准化的自动化平台进行评分。此外，这些药物还可以作为报告者，快速而直接地确定患者的肿瘤是否可能对特定治疗有反应。我们的目标是开发一个标准化的自动化平台，提供即时诊断，在分子复杂性和预后能力的水平上为临床决策提供信息，这是任何其他检测系统、生物标志物或相关基因表达特征所不可能实现的。为了实现这一目标，我们将结合Salk， UCSD和NanoSort开发的变革性新技术平台，根据肿瘤细胞获得的指示恶性进展和对治疗反应的分子病变，以不同的颜色标记肿瘤细胞，促进它们的检测，定量和分离，使用集成的“实验室-芯片”流式细胞仪。