Culturing viably captured circulating tumor cells using conditional reprogramming

使用条件重编程培养活捕获的循环肿瘤细胞

• 批准号: 8622303
• 负责人: RICHARD JAMES COTE
• 金额: $ 18.16万
• 依托单位: UNIVERSITY OF MIAMI SCHOOL OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-12-01 至 2015-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8622303
• 关键词: Biological; Biological Assay; Biological Models; Blood; Blood Cells; Blood specimen; CDX2 gene; Cancer Model; Cancer Patient; Cell Count; Cell Culture System; Cell Culture Techniques; Cells; Cessation of life; Clinical; Colon Carcinoma; Colorectal Cancer; Cultured Cells; Cultured Tumor Cells; Data; Development; Devices; Disease; Disease Progression; Drug Targeting; Engineering; Ensure; Epithelial Cells; Generations; Genetic; Hematopoietic Neoplasms; Human; Immunofluorescence Immunologic; Individual; Label; Lead; Malignant Neoplasms; Malignant neoplasm of lung; Malignant neoplasm of prostate; Metastatic Prostate Cancer; Methods; Molecular; Mutation; Neoplasm Circulating Cells; Neoplasm Metastasis; Patients; Pharmaceutical Preparations; Phenotype; Play; Population; Process; Prognostic Marker; Research Personnel; Rho-associated kinase; Role; Sampling; System; Techniques; Technology; Therapeutic; Time; Tissues; Whole Blood; abstracting; anticancer research; base; cancer cell; cell type; clinically significant; cost; drug sensitivity; evidence base; inhibitor/antagonist; insight; kinase inhibitor; malignant breast neoplasm; metastatic process; mouse model; neoplastic cell; next generation; novel; outcome forecast; peripheral blood; programs; response; success; treatment planning; treatment response; tumor

Abstract The presence or absence of metastasis is the most important determinant of prognosis and management of cancer. Spread of circulating tumor cells (CTC) via the peripheral blood is shown to be prognostic indicator of later overt metastases. Currently we have a very limited overall fundamental biological understanding of CTCs, and of the clinical significance of CTCs in individual patients' disease progression and treatment response. Several approaches have been described for capture or isolation of CTC from the blood, most have limitations and a very few allow culture of the CTC upon isolation. We are thus forced to study a finite, small number of CTC captured at static time points to yield limited cellular and molecular data, precluding our ability to study CTC dynamically for their functional aspects. We have developed a unique precisely engineered microfilter platform that effectively separates larger CTC from smaller blood cells, and have modified it for viable CTC capture. In parallel, we have also developed a novel method for tumor cell culture, which we refer to as conditionally reprogrammed cell (CRC) system, using a ROCK-inhibitor-treated feeder layer. We propose to integrate the two novel platforms to achieve viable CTC capture and culture, following two Specific Aims: 1) To combine the microfilter and CRC technologies to capture and reliably grow cancer cells from blood in a model system, and 2) To use combined technologies to capture and grow CTC from patients with metastatic prostate, breast, colon and lung cancer. We have substantial preliminary data which shows that our capture technology can efficiently capture viable tumor cells from blood in a syngeneic mouse model which can be cultured such that the cultured cells preserve the original phenotype. In a key advance, we have also demonstrated the ability to efficiently isolate viable CTCs and successfully establish cultures that can be available for downstream functional and molecular characterization. Our CRC technology can efficiently and reliably grow tumor cells, starting even from small numbers of founder cells, and that the CRC show morphologic, phenotypic and genetic fidelity to the tumor of origin. Our data also shows that the CRC established from clinical samples show functional utility through their use to predict response to drugs in actual patients. Thus, the combination of the microfilter and CRC systems proposed here for capture and culture of rare CTC has a high likelihood of success, and can transform the way we evaluate and manage patients with cancer. We also believe that successful CTC cultures will lead to an important and necessary insight into metastatic process that the present cancer model systems cannot provide.

抽象的 转移的存在或不存在是预后和 癌症的管理。通过外周血传播循环肿瘤细胞（CTC） 被证明是后来明显转移的预后指标。目前我们有一个非常有限的 对CTC的总体基本生物学理解以及CTC在CTC中的临床意义 个别患者的疾病进展和治疗反应。有几种方法 描述了用于从血液中捕获或隔离CTC，大多数都有局限性，并且非常 很少有隔离后CTC的培养。因此，我们被迫研究有限的数量 在静态时间点捕获的CTC以产生有限的细胞和分子数据，排除了我们的 能够动态研究其功能方面的能力。我们已经开发了一个独特的 精确设计的微滤器平台，可有效地将较大的CTC与较小的CTC分开 血细胞并已将其修改为可行的CTC捕获。同时，我们还开发了 肿瘤细胞培养的新方法，我们称之为有条件重编程的细胞（CRC） 系统，使用岩石抑制剂处理的馈线层。 我们建议整合两个新型平台，以实现可行的CTC捕获和文化， 以下两个具体目的：1）结合微滤器和CRC技术以捕获和 可靠地在模型系统中从血液中生长癌细胞，2）使用合并技术 从转移性前列腺，乳腺癌，结肠癌和肺癌捕获和生长CTC。 我们有大量的初步数据，这表明我们的捕获技术可以有效 在合成小鼠模型中，从血液中捕获可行的肿瘤细胞，该模型可以培养 培养的细胞保留原始表型。在关键的进步中，我们也有 证明了有效隔离可行CTC并成功建立培养物的能力 这可以用于下游功能和分子表征。我们的CRC 技术可以有效，可靠地生长肿瘤细胞，甚至从少量 创始人细胞，CRC表现出对肿瘤的形态，表型和遗传保真度 起源。我们的数据还表明，从临床样品中建立的CRC显示功能 通过使用它们预测实际患者对药物反应的实用性。因此，结合 在此提议的捕获和培养的稀有CTC的微滤器和CRC系统具有很高的 成功的可能性，可以改变我们评估和管理癌症患者的方式。 我们还认为，成功的CTC文化将带来重要和必要的见解 进入当前癌症模型系统无法提供的转移过程。