Microfluidic Apheresis to Isolate Circulating Tumor Clusters

微流体血浆分离术分离循环肿瘤簇

• 批准号: 10380192
• 负责人: Mehmet Toner
• 金额: $ 23.56万
• 依托单位: MASSACHUSETTS GENERAL HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-01-14 至 2023-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10380192
• 关键词: Anoikis; Antitumor Drug Screening Assays; Apoptotic; Architecture; Back; Biological Markers; Biomechanics; Biopsy; Blood; Blood Cells; Blood Chemical Analysis; Blood Component Removal; Blood Platelets; Blood Transfusion; Blood Volume; Cancer Patient; Cell Separation; Cells; Cellular Morphology; Centrifugation; Clinic; Clinical; Coagulation Process; Complete Blood Count; Detection; Diagnostic; Embolism; Ensure; Event; Filtration; Genome; Geometry; Goals; Hematopoietic Neoplasms; Hour; Intercellular Junctions; Lateral; Leukocytes; Link; Liquid substance; Microfluidic Microchips; Microfluidics; Molecular; Neoplasm Circulating Cells; Neoplasm Metastasis; Operative Surgical Procedures; Patients; Peripheral Blood Mononuclear Cell; Play; Population; Procedures; Process; Property; Proteome; Reporting; Role; Running; Sampling; Screening for cancer; Seeds; Sorting - Cell Movement; Speed; Structure; Surface; System; Technology; Testing; Therapeutic; Time; TimeLine; Tissues; Travel; Whole Blood; base; blood rheology; cancer cell; clinical decision-making; epigenetic marker; high risk; high throughput technology; innovation; liquid biopsy; metastatic process; microfluidic technology; neoplastic cell; parallelization; peripheral blood; precision oncology; prevent; quantum; safety study; stem cells; technology development; transcriptome; tumor; tumor progression

ABSTRACT Circulating tumor cells clusters (CTC-clusters), also called tumor emboli in 1970s, are known to play a major role in the metastatic process. For example, CTC-clusters have been inferred to be 50-fold as metastatic as one CTC. They also differ in many ways from single CTCs, including having distinct epigenetic markers. Due in part to their intact cell-cell junctions, the evidence also suggests they avoid anoikis, hence unlike single CTCs they are less apoptotic. However, CTC-clusters are even rarer than single CTCs, with in many cases an average of just one cluster-associated tumor cell in 10 mL of blood, meaning most samples of 10-20 mL do not contain a CTC-cluster. Isolating CTC-clusters from larger blood volumes is thus required to unlock the unique reservoir of biomarkers linked to this most metastatic population of tumor cells. Although there has been a number of approaches to isolate CTC-clusters, mostly using microfluidics, they primarily use a small amount of blood up to 10-20 mL and hence are not able to sort CTC-clusters reliably to impact clinical-decision making. To overcome the shortcomings of current approaches, we propose to develop core technology enabling a microfluidic CTC-cluster apheresis (CTApheresis) system that can interrogate 20-100% (i.e., 1-5 L) of the entire blood volume within an hour to isolate enough CTC-clusters in a majority of patients. We will reach our goal of CTApheresis by building an innovative microfluidic technology to gently and selectively sort CTC-clusters from undiluted blood in a high-throughput system. Here, we will create the “functional modular unit” for scalable CTApheresis. We will split the project in two distinct, integrated, aims. In aim 1, we will create a microfluidic chip to concentrate CTC-clusters within large volumes of whole blood. In aim 2, we will integrate this sorter into a functional modular unit sorter to process 120 mL of whole blood in 1 hour (scalable by nine-fold by parallelization to 1 L/h). Throughout and as a comprehensive safety study after aim 2 completes, we will run a battery of tests comparing processed and unprocessed blood, to examine if CTApheresis blood processed using the proposed microfluidic device will be safe to return back to the patient. Gentle sorting of CTC-clusters from liters of whole blood has the potential to unlock some of the mysteries of the metastatic process but also to enable ex-vivo culture of CTCs for “real time” functional drug testing for cancer patients. Routine sampling of the CTC-cluster genome, transcriptome and proteome could become a replacement for invasive surgical biopsies, and ultimately provide a powerful approach for early detection of cancer in high-risk patients.

抽象的 循环肿瘤细胞簇（CTC 簇），在 20 世纪 70 年代也称为肿瘤栓子，已知在 转移过程。例如，CTC 簇的转移性被推断为一个 CTC 的 50 倍。他们也有不同 在很多方面都来自于单一的 CTC，包括具有独特的表观遗传标记。部分原因是它们的细胞间完整 连接处，证据还表明它们避免失巢凋亡，因此与单个 CTC 不同，它们的细胞凋亡较少。然而， CTC 簇比单个 CTC 更为罕见，在许多情况下，平均只有一个簇相关的肿瘤细胞 10 mL 血液，意味着大多数 10-20 mL 样本不包含 CTC 簇。将 CTC 簇与较大的 CTC 簇隔离 因此，需要血量来解锁与这一最具转移性的群体相关的独特生物标志物库。 肿瘤细胞。尽管已经有许多方法来分离 CTC 簇，其中大多数使用微流体，但它们 主要使用最多 10-20 mL 的少量血液，因此无法可靠地对 CTC 簇进行分类以产生影响 临床决策。为了克服现有方法的缺点，我们建议开发核心技术 启用微流控 CTC 簇血浆分离术 (CTApheresis) 系统，该系统可以询问整个细胞的 20-100%（即 1-5 L） 一小时内的血容量足以在大多数患者中分离出足够的 CTC 簇。我们将实现我们的目标 CTApheresis 通过构建创新的微流体技术来温和地、选择性地从未稀释的 CTC 簇中分类 高通量系统中的血液。在这里，我们将为可扩展的 CTApheresis 创建“功能模块化单元”。我们将 将项目分为两个不同的、综合的目标。在目标 1 中，我们将创建一种微流控芯片来浓缩 CTC 簇 在大量全血中。在目标 2 中，我们将把这个分拣机集成到功能模块化单元分拣机中来处理 1 小时内获得 120 mL 全血（可通过并行化扩展至 1 L/h 九倍）。贯穿始终并作为一个综合性的 目标 2 完成后的安全性研究，我们将进行一系列测试，比较经过处理和未经处理的血液，以检查 如果使用所提议的微流体装置处理的 CTA 血液可以安全地返回患者体内。温和的 从几升全血中分选 CTC 簇有可能解开转移性癌症的一些谜团 不仅如此，还可以对 CTC 进行离体培养，以便对癌症患者进行“实时”功能药物测试。常规 CTC 簇基因组、转录组和蛋白质组采样可能成为侵入性手术的替代品 活组织检查，并最终为高危患者的癌症早期检测提供强大的方法。