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 超均匀结构微芯片 项目概要/摘要 循环肿瘤细胞 (CTC) 具有高度异质性，并且是特定的 CTC 亚群，而不是整个细胞， 负责癌症转移。目前的 CTC 技术只是简单地分离血液样本中的所有 CTC 没有将它们分解为不同的亚群，从而阻止研究人员获得对 CTC 的转移潜力。因此，CTC 异质性与肿瘤进展之间的相关性 很大程度上是未知的。此外，现有的 CTC 表征方法通常涉及破坏性固定和 透化方案，限制了 CTC 和其他物质后续表型分析的潜力 下游应用。该提案的目标是通过以下方式了解 CTC 的转移潜力： CTC 亚群的有效分级和分析。我建议隔离、就地识别、有选择地 使用具有超均匀结构的微芯片回收 CTC。超均匀性（HU）是一个新兴的概念 包含局部异质性或随机性和全局规则性或同质性的堆积模式。我的 这项工作首次将超均匀性的概念整合到基于亲和力的微流体装置中 CTC 隔离。我假设由于局部流动模式的受控差异 由于超均匀结构，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，预计这种粘合强度与表面标记的类型和密度有关。 捕获了 CTC，因此捕获了它们的转移特征。具体目标包括 (1) 设计和表征 HU 用于捕获 CTC 并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 微芯片识别具有不同表面标记表达的 CTC 亚群；和（3） 探索将 HU 微芯片平台纳入临床肿瘤学环境的几个关键因素。 HU 微芯片为 CTC 的分离提供了简单而独特的分辨率，因为其全球同质性 提供 CTC 依从性的平等可能性；和局部异质性允许同时分化 通过分析单个 CTC 所需的粘合强度来确定亚群。因此，CTC 亚群 仅使用 HU 芯片上的捕获位置即可识别，无需额外的后捕获 免疫荧光表征。最重要的定量里程碑是达到 80% 的准确率 统计相关性：1) HU 芯片上的位置与细胞后相互作用的强度，2) 的预测 癌细胞混合物（PC3 和 LNCaP）中的位置与细胞类型，以及 3) 实现 80% 的识别准确度 HU 微芯片上释放的 EMT 和非 EMT 细胞，并通过免疫染色进行验证。如果 研发成功，这款HU微芯片可以轻松集成到研究实验室进行研究 与 CTC 异质性相关的基础癌症生物学，以及临床环境以分析 CTC 亚群以协助癌症诊断、预测肿瘤进展和监测治疗效果。