Integration of Flexible Micro Spring Array and High Throughput Microfluidics for

柔性微弹簧阵列和高通量微流体的集成

• 批准号: 8358451
• 负责人: Siyang Zheng
• 金额: $ 223.5万
• 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: 2012-09-17 至 2017-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8358451
• 关键词: Accounting; Antineoplastic Agents; Biological Assay; Biopsy; Blood; Blood Cells; Blood Circulation; Blood Volume; Blood specimen; Body part; Cancer Patient; Cancer Prognosis; Cell Size; Cells; Cessation of life; Clinic; Detection; Development; Diagnosis; Disseminated Malignant Neoplasm; FDA approved; Filtration; Genetic; Invaded; Lead; Leukocytes; Liquid substance; Malignant Neoplasms; Mechanical Stress; Methods; Microfabrication; Microfluidics; Molecular Profiling; Monitor; Mutation Detection; Neoplasm Metastasis; Outcome; Patient Isolation; Patients; Pharmaceutical Preparations; Primary Neoplasm; Process; Reaction; Risk; Staging; System; Techniques; Technology; Testing; Therapeutic; Time; Toxic effect; Travel; abstracting; base; cancer diagnosis; cancer type; chemotherapy; clinically relevant; cost; cost effective; drug efficacy; efficacy testing; experience; flexibility; improved; minimally invasive; neoplastic cell; novel strategies; pressure; public health relevance; treatment planning; tumor progression

DESCRIPTION (Provided by the applicant) Abstract: The ability of metastatic cancer to release circulating tumor cells (CTCs) that travel through the blood and invade different parts of the body accounts for over 90% of cancer related deaths. New techniques for improved diagnosis and therapeutic strategies are desperately needed to improve patient outcomes in late stage metastatic cancer patients. One such technique involves the analysis of these CTCs through isolation from patient blood samples. Since molecular profiles of CTCs can be quite different from those of the primary tumor and more similar to the metastatic tumors, CTCs are better suited for metastatic cancer prognosis and diagnosis. Current CTC technologies have serious difficulties and limitations for clinic applications due to poor sensitivity and selectivity, high cost, and long processing times. CellSearchTM, the only current FDA approved system for CTC analysis, is only used for detection of CTCs and is not capable of preserving viable cells. The fundamental challenge with obtaining CTCs from blood samples is the fact that they are so rare, with only a few tumors cells occurring among billions of blood cells. Since tumor cells are almost always significantly larger than normal blood cells, size based separation has been demonstrated as an effective method for CTC capture. We have taken a novel approach to established microfiltration technology by implementing an array of flexible microsprings and using a regulated low pressure flow system to minimize the mechanical stresses experienced by cells during the filtration process. This is an efficient and cost effective system that is capable of the enrichment of viable CTCs from a clinically relevant blood volume of 7.5 mL in only 10 minutes. Despite achieving greater than 104 enrichment against leukocytes, the purity of the enriched cells is not sufficient for obtaining clinically relevant genetic information. We propose the incorporation of a high throughput microfluidic system that will physically partition these cells for analysis on a single cell level.By exploiting this inherent advantage of microtechnology, a large volume of parallel reactions can be used to overcome the issues with purity. Genetic expression profiles and mutation detections may be used for improved diagnosis, and lead to the development of highly personalized therapy plans that are optimized for each patient. Furthermore, the use of microfluidics for multiwell partitioning of CTCs will be used to attempt the establishment of favorable conditions for the culture of CTCs, even at an initially low seeding number. Successful primary culture of CTCs will allow drug efficacy tests that may be used to assay potential drugs ex vivo without exposing a patient to the unnecessary cost or toxic effects of chemotherapy. These new approaches based on the analysis of viable CTCs represent a different approach that has not been proven. However, since this technological platform is applicable to almost every type of cancer, it could fairly quickly revolutionize the way that therapies are derived for metastatic cancer patients. Public Health Relevance: The most deadly forms of cancer can release aggressive cells that circulate through the bloodstream and spread throughout the body. Microfabrication technology has been developed to effectively isolate these cells from a patient blood sample, allowing a minimally invasive "liquid biopsy" that may be performed often for monitoring tumor progression. This project explores the integration of a high throughput approach to analysis that will make it possible to test various anticancer drugs on these cells at no risk to the patient, and obtain genetic information that will be crucial to developing a highly personalized treatment plan.

描述(由申请人提供) 摘要：转移癌释放循环肿瘤细胞(CTCs)的能力占癌症相关死亡的90%以上。CTCs通过血液传播并侵袭身体不同部位。迫切需要改进诊断和治疗策略的新技术来改善晚期转移性癌症患者的预后。一种这样的技术涉及通过从患者血液样本中分离来分析这些CTC。由于CTCs的分子特征与原发癌有很大不同，而与转移性肿瘤更相似，因此CTCs更适合于转移性肿瘤的预后和诊断。由于灵敏度和选择性差、成本高、处理时间长，目前的CTC技术在临床应用中存在严重的困难和限制。CellSearchTM是目前FDA批准的唯一用于CTC分析的系统，仅用于CTC的检测，不能保存活细胞。从血液样本中获取CTCs的根本挑战是，它们非常罕见，在数十亿个血细胞中只有几个肿瘤细胞。由于肿瘤细胞几乎总是比正常血细胞大得多，基于大小的分离已被证明是一种有效的CTC捕获方法。我们采用了一种新的方法来建立微过滤技术，通过实施一系列灵活的微弹簧和使用可调节的低压流动系统来最大限度地减少细胞在过滤过程中遇到的机械压力。这是一种高效且具有成本效益的系统，能够在10分钟内从临床相关的7.5毫升血液中浓缩出可存活的CTCs。尽管对白细胞达到了104以上的浓缩，但浓缩后的细胞的纯度不足以获得 临床相关的遗传信息。我们建议加入一个高通量的微流控系统，将这些细胞进行物理分割，以便在单个细胞水平上进行分析。通过利用微技术的这一固有优势，可以使用大量的平行反应来克服纯度问题。基因表达谱和突变检测可用于改进诊断，并导致针对每个患者优化的高度个性化治疗计划的开发。此外，将使用微流控技术进行CTCs的多孔分配，以尝试为CTCs的培养创造有利条件，即使在最初的低播种量情况下也是如此。CTCs的成功原代培养将允许进行药效测试，这些测试可用于在体外测试潜在的药物，而不会使患者暴露在化疗的不必要成本或毒副作用中。这些基于对可行的四氯化碳的分析的新方法代表了一种尚未得到证实的不同方法。然而，由于这一技术平台适用于几乎所有类型的癌症，它可能很快就会彻底改变转移性癌症患者的治疗方法。 与公共卫生相关：最致命的癌症可以释放侵袭性细胞，这些细胞在血液中循环并扩散到全身。微制造技术已经被开发出来，可以有效地从患者血液样本中分离出这些细胞，从而允许进行微创的“液体活组织检查”，这可能经常用于监测肿瘤的进展。该项目探索了高通量分析方法的集成，使在这些细胞上测试各种抗癌药物成为可能，而不会对患者造成风险，并获得对开发高度个性化的治疗计划至关重要的遗传信息。

项目成果

Separable Bilayer Microfiltration Device for Label-Free Enrichment of Viable Circulating Tumor Cells.

可分离双层微滤装置，用于无标记富集活循环肿瘤细胞。

• DOI: 10.1007/978-1-4939-7144-2_6
• 发表时间: 2017
• 期刊: Methods in molecular biology (Clifton, N.J.)
• 作者: Hao,Sijie;Nisic,Merisa;He,Hongzhang;Tai,Yu-Chong;Zheng,Si-Yang
• 通讯作者: Zheng,Si-Yang

Highly sensitive DNA detection using cascade amplification strategy based on hybridization chain reaction and enzyme-induced metallization.

• DOI: 10.1016/j.bios.2014.11.035
• 发表时间: 2015-04-15
• 期刊: BIOSENSORS & BIOELECTRONICS
• 影响因子: 12.6
• 作者: Yu, Xu;Zhang, Zhi-Ling;Zheng, Si-Yang
• 通讯作者: Zheng, Si-Yang

Synthesis of Self-Assembled Multifunctional Nanocomposite Catalysts with Highly Stabilized Reactivity and Magnetic Recyclability.

• DOI: 10.1038/srep25459
• 发表时间: 2016-05-05
• 期刊: Scientific reports
• 影响因子: 4.6
• 作者: Yu X;Cheng G;Zheng SY
• 通讯作者: Zheng SY

Point-of-care microdevices for blood plasma analysis in viral infectious diseases.

• DOI: 10.1007/s10439-014-1044-2
• 发表时间: 2014-11
• 期刊: Annals of biomedical engineering
• 影响因子: 3.8
• 作者: Yeh YT;Nisic M;Yu X;Xia Y;Zheng SY
• 通讯作者: Zheng SY

Separable bilayer microfiltration device for viable label-free enrichment of circulating tumour cells.

• DOI: 10.1038/srep07392
• 发表时间: 2014-12-09
• 期刊: Scientific reports
• 影响因子: 4.6
• 作者: Zhou MD;Hao S;Williams AJ;Harouaka RA;Schrand B;Rawal S;Ao Z;Brenneman R;Gilboa E;Lu B;Wang S;Zhu J;Datar R;Cote R;Tai YC;Zheng SY
• 通讯作者: Zheng SY