Development of a Flexible Micro Spring Array device for viable circulating tumor

开发用于活循环肿瘤的柔性微弹簧阵列装置

• 批准号: 8303786
• 负责人: Siyang Zheng
• 金额: $ 14.88万
• 依托单位: PENNSYLVANIA STATE UNIVERSITY, THE
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-08-06 至 2014-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8303786
• 关键词: Accounting; Antineoplastic Agents; Automobile Driving; Blood; Blood Cells; Blood Circulation; Blood specimen; Cancer Patient; Cell Culture Techniques; Cell Size; Cells; Cessation of life; Clinical; Cultured Tumor Cells; Detection; Development; Device Designs; Devices; Diagnosis; Diagnostic; Disease; Disseminated Malignant Neoplasm; Distant; ERBB2 gene; Edetic Acid; Epithelial; FDA approved; Filtration; Goals; Growth; Health; Invaded; Leukocytes; Lymphatic System; Malignant - descriptor; Malignant Neoplasms; Mammalian Cell; Methods; Microfabrication; Modeling; Monitor; Neoplasm Metastasis; Organ; Patients; Performance; Pilot Projects; Population; Porosity; Primary Cell Cultures; Primary Neoplasm; Process; Proliferating; Protocols documentation; Recovery; Research Personnel; Sampling; Selection for Treatments; Solid Neoplasm; Stress; Structure; System; Technology; Testing; Therapeutic; Toxic effect; Travel; Treatment Efficacy; Tube; Work; base; cancer therapy; cell injury; chemotherapy; commercialization; cost; drug efficacy; drug sensitivity; efficacy testing; experience; flexibility; immunocytochemistry; improved; instrument; malignant breast neoplasm; meetings; neoplastic cell; novel; operation; outcome forecast; peripheral blood; pressure; product development; prototype; research clinical testing; response; sample collection; scale up; tool; treatment planning; tumor

DESCRIPTION (provided by applicant): Development of a flexible micro spring array device for viable circulating tumor cell enrichment, analysis, primary culture and ex vivo therapeutic response test Metastatic cancer accounts for over 90% of cancer related deaths. Cancer metastasis is the process by which malignant circulating tumor cells (CTCs) detach from a primary tumor, spread through peripheral blood, then invade and proliferate in distant organs. The fundamental challenge of detecting CTCs in blood samples is the fact that they are so rare, requiring detection sensitivity of only a few tumors cells in billions of blood cells. Since the majority of solid tumor cells are of epithelial origin, and are almost always significantly larger and more rigid than normal blood cells, size based separation has been demonstrated as an effective method for CTC capture. The first goal of this project is to characterize and optimize a new flexible micro spring array (FMSA) device for the enrichment of CTCs from peripheral blood samples. The novel spring structures incorporated into the device design minimize concentrated stresses experienced by the CTCs and encourage their health, survival and proliferation. We have optimized the gap size of the device to reliably achieve over 85% recovery, 104 enrichment against leukocytes, 80% viability, and throughput of 5mL fresh blood in EDTA tube in 5 minutes. This technology is portable, cheap, fast, and more effective than CellSearchTM, which is the only current FDA approved system for CTC enumeration in a limited variety of cancers. In our preliminary study, we also found the driving differential pressure is critical for ell viability and proliferability. The safe operation pressure is orders of magnitude lower than the pressure normally used for conventional cell filtration. Thus our second goal is to develop a MEMS enabled automatic low pressure control system to facilitate the FMSA device. Together with the FMSA device, the whole system can solve the over 40 years' problem that widely used conventional filtration can't enrich viable mammalian cells because of cell damage and device clogging. More importantly, the FMSA system is capable of the capture of viable tumor cells that are available for primary culture and ex vivo therapeutic drug efficacy test, which is the third goal of the proposed study. The successful capture of viable CTCs will prompt the establishment of systems and protocols for their growth into a primary culture. This will provide the basis for testing the tumor cells' response to a variety of anticancer drugs ex vivo. If proven to be effective, a more specific and personalized treatment plan may be developed without having to unnecessarily expose a patient to the cost and toxic effects of chemotherapy. With further optimization for device flexibility and primary cell culture, and establishment of an automated low pressure control prototype, the FMSA enrichment system can be readily adapted for clinical usage. A pilot study of clinical samples obtained from our clinical collaborators will evaluate CTC detection, primary culture, and ex vivo drug efficacy tests with the FMSA system. PUBLIC HEALTH RELEVANCE: The most deadly forms of cancer can release aggressive cells that travel through the bloodstream and spread throughout the body. This project will explore the use of a new microfabricated device to capture these invading cells and isolate them from a patient blood sample. The information obtained by growing and analyzing these cells can be used for improved detection, diagnosis, and development of personalized treatment plans.

描述（由申请人提供）：开发用于活循环肿瘤细胞富集、分析、原代培养和离体治疗反应测试的柔性微弹簧阵列装置转移性癌症占癌症相关死亡的 90% 以上。癌症转移是恶性循环肿瘤细胞（CTC）从原发肿瘤脱落，通过外周血扩散，然后侵入远处器官并增殖的过程。检测血液样本中的 CTC 的根本挑战是它们非常罕见，只需要检测数十亿血细胞中的少数肿瘤细胞的灵敏度。由于大多数实体瘤细胞都是上皮细胞来源，并且几乎总是比正常血细胞明显更大且更坚硬，因此基于尺寸的分离已被证明是捕获 CTC 的有效方法。该项目的首要目标是表征和优化一种新型柔性微弹簧阵列 (FMSA) 装置，用于从外周血样本中富集 CTC。装置设计中采用的新颖弹簧结构最大限度地减少了 CTC 所承受的集中应力，并促进它们的健康、生存和增殖。我们优化了设备的间隙尺寸，以可靠地实现超过 85% 的回收率、104 个白细胞富集、80% 的活力以及 5 分钟内 EDTA 管中 5mL 新鲜血液的吞吐量。该技术便携、便宜、快速且比 CellSearchTM 更有效，CellSearchTM 是目前 FDA 批准的唯一用于有限种类癌症 CTC 计数的系统。在我们的初步研究中，我们还发现驱动压差对于细胞活力和增殖能力至关重要。安全操作压力比常规细胞过滤通常使用的压力低几个数量级。因此，我们的第二个目标是开发一种支持 MEMS 的自动低压控制系统，以促进 FMSA 设备的发展。与FMSA设备一起，整个系统可以解决40多年来广泛使用的传统过滤由于细胞损伤和设备堵塞而无法富集活哺乳动物细胞的问题。更重要的是，FMSA系统能够捕获活的肿瘤细胞，可用于原代培养和离体治疗药效测试，这是该研究的第三个目标。成功捕获可行的 CTC 将促进其生长成原代培养的系统和方案的建立。这将为离体测试肿瘤细胞对多种抗癌药物的反应提供基础。如果证明 为了有效，可以制定更具体和个性化的治疗计划，而不必让患者承受化疗的成本和毒性作用。通过进一步优化设备灵活性和原代细胞培养，以及建立自动化低压控制原型，FMSA 富集系统可以轻松适应临床使用。对从我们的临床合作者处获得的临床样本进行的试点研究将 使用 FMSA 系统评估 CTC 检测、原代培养和离体药效测试。 公共卫生相关性：最致命的癌症会释放侵袭性细胞，这些细胞会通过血流传播到全身。该项目将探索使用新型微型设备来捕获这些入侵细胞并将其从患者血液样本中分离出来。通过培养和分析这些细胞获得的信息可用于改进检测、诊断和制定个性化治疗计划。