Fractionation and Profiling of Heterogeneous Circulating Tumor Cells Using a Hyperuniform- Structured Microchip

使用超均匀结构微芯片对异质循环肿瘤细胞进行分级和分析

• 批准号: 10025872
• 负责人: Wei Li
• 金额: $ 55.31万
• 依托单位: TEXAS TECH UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-08-07 至 2024-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10025872
• 关键词: Address; Adherence; Adhesions; Adhesives; Adopted; Affinity; Blood; Blood Cells; Blood specimen; CD44 gene; CD47 gene; Cancer Biology; Cell Adhesion; Cell Cycle; Cell Line; Cell Separation; Cell Size; Cell Survival; Cells; Characteristics; Clinical; Clinical Oncology; Collaborations; Correlation Studies; Devices; Diagnostic; Disease model; Ensure; Fractionation; Genomics; Goals; Health Sciences; Heterogeneity; Immunofluorescence Immunologic; In Situ; Individual; LNCaP; Label; Laboratory Research; Location; Malignant Neoplasms; Metastatic Neoplasm to the Bone; Metastatic Neoplasm to the Liver; Metastatic Neoplasm to the Lung; Methods; Microfluidic Microchips; Microscope; Monitor; Morphologic artifacts; Neoplasm Circulating Cells; Neoplasm Metastasis; Optics; PC3 cell line; Pattern; Phenotype; Population; Population Heterogeneity; Process; Proteomics; Protocols documentation; Pump; Recovery; Reporting; Research; Research Personnel; Resolution; Sampling; Sensitivity and Specificity; Structure; Surface; Syringes; TACSTD1 gene; Technology; Time; Treatment Efficacy; Work; Yang; assay development; base; biophysical properties; cancer cell; cancer diagnosis; cancer therapy; cell type; clinical application; clinical care; density; design; innovation; insight; manufacturing process; microchip; microfluidic technology; prevent; sample fixation; screening; tool; tumor; tumor progression

Title: Fractionation and Profiling of Heterogeneous Circulating Tumor Cells Using a Hyperuniform-structured Microchip Project Summary/Abstract Circulating tumor cells (CTCs) are highly heterogeneous, and specific CTC subpopulations, rather than the whole, are responsible for cancer metastasis. Current CTC technologies simply isolate all CTCs in a blood sample without resolving them into distinct subpopulations, preventing researchers from acquiring true insights into the metastatic potential of CTCs. As a consequence, correlation between CTC heterogeneity and tumor progression is largely unknown. In addition, existing CTC characterization methods often involve destructive fixation and permeabilization protocols, which limit the potential for subsequent phenotypic analysis of CTCs and other downstream applications. The goal of this proposal is to understand the metastatic potential of CTCs through effective fractionation and profiling of CTC subpopulations. I propose to isolate, in-situ identify, and selectively recover CTCs using a microchip with hyperuniform structure. Hyperuniformity (HU) is an emerging concept of a packing pattern which contains local heterogeneity or randomness and global regularity or homogeneity. My work, for the first time, will integrate the concept of hyperuniformity into affinity-based microfluidic devices for CTC isolation. I hypothesize that due to the controlled differences in local flow patterns induced by the hyperuniform structure, cell arrest in different locations on the microchip will require different adhesive strengths. Further, this adhesive strength is anticipated to be related to the types and densities of surface markers on the captured CTCs and therefore, their metastatic character. Specific aims include (1) Design and characterize HU structured microchip for CTC capture and analyze flow pattern and adhesion force in the device; (2) Capture and identify subpopulations of CTCs with variable expression of the surface marker using a HU microchip; and (3) Explore several key factors for potential incorporation of the HU microchip platform into clinical oncology settings. The HU microchip offers a simple and unique resolution for fractionation of CTCs, as its global homogeneity provides equal possibility of CTC adherence; and local heterogeneity allows simultaneous differentiation of subpopulations by analyzing adhesive strength required for individual CTCs. As a result, subpopulations of CTCs can be identified using only their capture locations on the HU chip without requiring additional post-capture immunofluorescence characterization. The most significant quantitative milestones are to achieve 80% accuracy on statistical correlation on: 1) the locations on a HU chip with strength of cell-post interaction, 2) predictions of location vs. cell type in a cancer cell mixture (PC3 and LNCaP), and 3) achieving 80% accuracy on identification of released EMT and non-EMT cells from their locations on a HU microchip, validated by immunostaining. If successfully developed, this HU microchip can be easily integrated into research laboratories to study fundamental cancer biology related to CTC heterogeneity, as well as into clinical settings to profile CTC subpopulations to assist cancer diagnosis, predict tumor progression, and monitor therapeutic efficacy.

标题：使用A的异质循环肿瘤细胞的分馏和分析 超一结构的微芯片 项目摘要/摘要 循环肿瘤细胞（CTC）是高度异质的，特定的CTC亚群，而不是整体 负责癌症转移。当前的CTC技术简单地隔离了血液样本中的所有CTC 不用将它们分解为不同的亚群，阻止研究人员获得真正的见解 CTC的转移潜力。结果，CTC异质性与肿瘤进展之间的相关性 在很大程度上未知。此外，现有的CTC表征方法通常涉及破坏性固定和 通透方案，这限制了对CTC和其他表型分析的潜力 下游应用程序。该提议的目的是了解CTC的转移潜力 CTC亚群的有效分馏和分析。我建议隔离，原位识别和选择性地识别 使用具有超平均结构的微芯片恢复CTC。超均匀性（HU）是一个新兴的概念 包装模式，其中包含局部异质性或随机性以及全球规律性或同质性。我的 工作首次将超均匀性的概念整合到基于亲和力的微流体设备中 CTC隔离。我假设，由于由局部流动模式受控的差异 超平均结构，微芯片上不同位置的细胞停滞将需要不同的粘合强度。 此外，这种粘合强度预计与表面标记的类型和密度有关 捕获了CTC，因此是其转移性特征。具体目的包括（1）设计和表征HU 用于CTC捕获和分析设备中的流动模式和粘附力的结构化微芯片； （2）捕获和 使用HU微芯片确定具有表面标记的CTC的亚群； （3） 探索将HU微芯片平台纳入临床肿瘤学环境中的几个关键因素。 Hu Microchip为CTC分馏提供了简单而独特的分辨率，作为其全球同质性 提供了CTC依从性的同等可能性；和局部异质性允许同时分化 通过分析单个CTC所需的粘合强度来亚群。结果，CTC的亚群 只能使用HU芯片上的捕获位置来识别，而无需额外的捕获后 免疫荧光表征。最重要的定量里程碑是实现80％的精度 关于以下统计相关性：1）HU芯片上的位置具有细胞 - 柱相互作用的强度，2） 癌细胞混合物中的位置与细胞类型（PC3和LNCAP），3）在鉴定方面达到80％的精度 通过免疫染色验证的HU微芯片上释放的EMT和非EMT细胞的释放。如果 成功开发了此HU微芯片可以轻松地整合到研究实验室中以研究 与CTC异质性以及临床环境有关的基本癌症生物学 亚群以帮助癌症诊断，预测肿瘤进展并监测治疗功效。