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: 8332762
• 负责人: Raoul Kopelman
• 金额: $ 9.89万
• 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/8332762
• 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.

描述（由申请人提供）：目前许多用于癌症研究、药物开发和精确治疗测定的高通量细胞检测技术，通常需要在二维表面上生长，并且需要耗时和劳动密集型的培养步骤。此外，目前的检测方法使用大量细胞群，这在处理至关重要但数量很少的癌症干细胞（CSCs）时可能会带来挑战。当研究CSCs时，分化可以发生在1-2个细胞分裂中。目前，尚无高通量的方法来测量悬浮肿瘤干细胞的细胞毒性作用。因此，需要一种对单个悬浮细胞和小球体的体积和形状变化敏感的高通量测定方法。为了应对当前方案的局限性，我们正在开发一种基于异步磁珠旋转（AMBR）的细胞磁旋转方法，该方法对单细胞变化敏感，并且可以对单个悬浮细胞和球体进行快速生长和细胞毒性分析。由于其亚显微分辨率、高通量和短观察时间，我们预计这种AMBR试验将大大减少培养步骤时间、总体结果时间和这种试验所需的细胞数量。为了初步验证这种新方法，我们选择研究单个细胞、单个干细胞和单个球体与前列腺癌的关系。我们利用Kopelman实验室的纳米技术和纳米医学专业知识以及Pienta实验室的前列腺癌和药物敏感性专业知识建立了合作关系。基于单个癌细胞的初步结果，我们假设癌细胞和球体对化疗药物的反应引起的体积变化可以准确而灵敏地监测。此外，由于该方法的灵敏度、速度和简单性的结合，通常非常少量的干细胞可以在它们变成异质群体之前进行快速测试。这将使对癌症干细胞和含有癌症干细胞的球体进行快速药物敏感性测试成为可能，从而导致针对特定患者量身定制的治疗剂的快速和智能选择方案。我们计划实现单个前列腺癌和癌症干细胞的生长和细胞毒性演示测量。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药物多西他赛的反应。我们的长期目标是开发一种检测设备，可以直接检测液体临床标本（来自患者血液的循环肿瘤细胞）中的靶细胞，并提供自动化结果，无需手动步骤，用于任何形式的癌症的个性化识别和药物敏感性测试。

项目成果

Fractal dimension of microbead assemblies used for protein detection.

用于蛋白质检测的微珠组件的分形维数。

• DOI: 10.1002/cphc.201402048
• 发表时间: 2014
• 期刊: Chemphyschem : a European journal of chemical physics and physical chemistry
• 作者: Hecht,Ariel;Commiskey,Patrick;Lazaridis,Filippos;Argyrakis,Panos;Kopelman,Raoul
• 通讯作者: Kopelman,Raoul

Bead assembly magnetorotation as a signal transduction method for protein detection.

珠组装磁旋转作为蛋白质检测的信号转导方法。

• DOI: 10.1016/j.bios.2013.03.073
• 发表时间: 2013
• 期刊: Biosensors & bioelectronics
• 影响因子: 12.6
• 作者: Hecht,Ariel;Commiskey,Patrick;Shah,Nicholas;Kopelman,Raoul
• 通讯作者: Kopelman,Raoul