Magnetorotation: a Rapid Assay for Single Cell Drug Sensitivity of Cancer Cells

磁旋转：癌细胞单细胞药物敏感性的快速测定

• 批准号: 8154084
• 负责人: Raoul Kopelman
• 金额: $ 17.7万
• 依托单位: UNIVERSITY OF MICHIGAN AT ANN ARBOR
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-09-14 至 2013-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8154084
• 关键词: Achievement; Agreement; Antineoplastic Agents; Apoptosis; Bacteria; Biological Assay; Biopsy; Blood; Blood specimen; CD44 gene; Cancer Cell Growth; Cell Count; Cell Cycle; Cell Death; Cell Size; Cell division; Cells; Cessation of life; Clinical; Collaborations; Cytotoxic agent; DU145; Detection; Development; Devices; Diagnostic; Dose; Frequencies; Goals; Growth; Health; Human; Immunomagnetic Separation; In Vitro; Individual; Lead; Lethal Dose 50; Life; Liquid substance; Magnetism; Malignant Neoplasms; Malignant neoplasm of prostate; Manuals; Measurement; Measures; Medical; Medicine; Methods; Microscope; Microscopic; Mission; Monitor; Nanotechnology; Optics; Patients; Population; Population Heterogeneity; Protocols documentation; Public Health; Research; Resolution; Rotation; Shapes; Specimen; Speed; Staining method; Stains; Stem cells; Surface; System; Techniques; Technology; Testing; Therapeutic Agents; Time; Translations; Validation; Work; anticancer research; base; cancer cell; cancer stem cell; cell growth; chemotherapy; clinical application; cytotoxic; cytotoxicity; docetaxel; dosage; drug development; drug sensitivity; high throughput screening; improved; magnetic beads; magnetic field; meter; monitoring device; nanomedicine; nanoscale; neoplastic cell; novel; novel strategies; particle; rapid growth; response; stem cell differentiation

DESCRIPTION (provided by applicant): The current state of the art of many high throughput cell-based assays, used for cancer research, drug development and determination of precision therapy, often requires growth on 2D surfaces and requires the time-consuming and labor-intensive culturing step. Furthermore, current assays use large cell populations, which can pose a challenge when working with the critically important, but small in number, cancer stem cells (CSCs). When investigating CSCs, differentiation can occur in as little as 1-2 cell divisions. Currently, no high throughput method exists to measure cytotoxic effects on suspended cancer stem cells. So it is desirable to have a high throughput assay that is sensitive to volume and shape changes in single suspended cells and small spheroids. In response to the limitation of current protocols, we are developing a cell magnetorotation method, based on asynchronous magnetic bead rotation (AMBR), that is sensitive to single-cell changes, and that will allow for rapid growth and cytotoxicity analysis of individual suspended cells and spheroids. Due to its submicroscopic resolution, high throughput, and short observation time, we anticipate that this AMBR assay will drastically reduce the culture step time, the overall time to results, and the number of cells needed for such an assay. For the preliminary validation of this new approach we selected to investigate single cells, single stem cells and single spheroids relating to prostate cancer. We set up a collaboration using the nanotechnology and nanomedicine expertise of the Kopelman Lab and the prostate cancer and drug sensitivity expertise of the Pienta Lab. Based on preliminary single cancer cell results, we hypothesize that the volumetric changes, resulting from the response to chemotherapy agents by cancer cells and spheroids, can be accurately and sensitively monitored. Furthermore, due to the combined sensitivity, speed and simplicity of the proposed approach, the typically very small number of stem cells can be tested rapidly, before their turning into a heterogeneous population. This will enable rapid drug sensitivity tests on cancer stem cells and on spheroids containing cancer stem cells, leading to a fast and smart selection protocol for therapeutic agents that can be tailored to a specific patient. We plan to achieve growth and cytotoxicity demonstration measurements on individual prostate cancer and cancer stem cells. We also plan to demonstrate that the AMBR method can be used to measure the growth of prostate cancer spheroids and their response to the cytotoxic agent docetaxel. Our long-term objective is to develop an assay device that will detect target cells directly in liquid clinical specimens (circulating tumor cells from patient blood) and provide automated results, with no manual steps, for personalized identification and drug sensitivity testing, for any form of cancer. PUBLIC HEALTH RELEVANCE: This proposal is relevant to public health because determining the optimum treatment for the individual patient is of prime importance in medicine, and particularly when it comes to cancer cases. A new approach, based on cell magnetorotation, enables drug sensitivity tests with higher sensitivity, throughput, speed and fidelity. Therefore, this proposal is relevant to the part of NIH's mission for developing medical discoveries that improve human health.

描述（由申请人提供）：用于癌症研究，药物开发和精确治疗的许多高吞吐细胞测定法的现行状态，通常需要在2D表面上增长，并且需要耗时且劳动密集型的培养步骤。此外，当前的测定法使用较大的细胞群体，在使用至关重要的癌症干细胞（CSC）时，可能会构成挑战。在研究CSC时，分化可能仅在1-2个细胞分裂中发生。目前，尚无高通量方法来测量对悬浮癌干细胞的细胞毒性作用。因此，需要具有对体积敏感并形成单个悬浮细胞和小球体的变化的高吞吐量测定。响应当前方案的局限性，我们正在基于异步磁珠旋转（AMBR）开发一种细胞磁化方法，该方法对单细胞变化很敏感，这将允许对单个悬浮细胞和球体的快速生长和细胞毒性分析。由于其亚镜下的分辨率，高吞吐量和短期观察时间，我们预计该AMBR分析将大大减少培养时间的阶段时间，总体结果以及这种测定所需的细胞数量。对于这种新方法的初步验证，我们选择研究与前列腺癌有关的单个细胞，单个干细胞和单个球体。我们使用Kopelman实验室的纳米技术和纳米医学专业知识以及Prostate癌症和PROSTATE癌症和药物敏感性专业知识建立了合作。基于初步的单个癌细胞结果，我们假设可以准确且敏感地监测癌细胞和球体对化学疗法剂的反应而产生的体积变化。此外，由于提出方法的敏感性，速度和简单性的组合，通常可以在变成异质种群之前迅速测试干细胞的数量很少。这将在癌症干细胞和含有癌症干细胞的球体上进行快速的药物敏感性测试，从而为治疗剂提供快速，智能选择方案，可以针对特定患者定制。我们计划在单个前列腺癌和癌症干细胞上实现生长和细胞毒性示范测量。我们还计划证明，AMBR方法可用于测量前列腺癌球体的生长及其对细胞毒性剂多西他赛的反应。我们的长期目标是开发一种测定装置，该设备将直接检测液体临床标本中的靶细胞（从患者血液中循环肿瘤细胞），并为任何形式的癌症提供自动化结果，没有手动步骤，用于个性化鉴定和药物敏感性测试。 公共卫生相关性：该提案与公共卫生有关，因为确定个人患者的最佳治疗在医学中至关重要，尤其是在癌症病例方面。一种基于细胞磁化的新方法可实现具有更高敏感性，吞吐量，速度和忠诚度的药物敏感性测试。因此，该提案与NIH的一部分有关改善人类健康的医疗发现的任务。