Microfluidic sorting of lung cancer cells from leukapheresis product as an alternative to metastatic tumor biopsy

从白细胞分离术产品中对肺癌细胞进行微流体分选，作为转移性肿瘤活检的替代方法

• 批准号: 10455704
• 负责人: Daniel A. Haber
• 金额: $ 44.33万
• 依托单位: MASSACHUSETTS GENERAL HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-08-01 至 2026-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10455704
• 关键词: Active Sites; Address; Antibodies; Biological; Biomedical Engineering; Biopsy; Blood; Blood Cells; Blood Platelets; Blood Volume; Blood specimen; Cancer Patient; Cell Separation; Cell Surface Proteins; Cells; Chemotherapy-Oncologic Procedure; Cirrhosis; Clinical; Complex; DNA; Detection; Diagnostic; Disease Progression; Drug Kinetics; Drug resistance; Environmental Risk Factor; Epithelial; Evolution; Excision; Explosion; Genetic; Goals; Hereditary Breast Carcinoma; Hour; Human; Immune response; Immunomodulators; Immunotherapy; Individual; Label; Lesion; Leukapheresis; Location; Lung; Lung nodule; Magnetism; Malignant Neoplasms; Malignant neoplasm of liver; Malignant neoplasm of lung; Measurement; Measures; Mesenchymal; Metastatic to; Microfluidics; Modality; Monitor; Mononuclear; Neoplasm Circulating Cells; Neoplasm Metastasis; Non-Small-Cell Lung Carcinoma; Operative Surgical Procedures; Outcome; Patients; Performance; Persons; Pharmacotherapy; Pilot Projects; Population; Procedures; Property; Proteins; RNA; Resistance; Resources; Risk; Sampling; Signal Pathway; Signal Transduction; Site; Smoker; Solid Neoplasm; Sorting - Cell Movement; Source; Speed; Technology; Testing; Therapeutic; Therapeutic Intervention; Time; Tissues; Tube; Tumor Antigens; Tumor Debulking; Tumor-Derived; Validation; Whole Blood; acquired drug resistance; base; cancer cell; cancer immunotherapy; cancer invasiveness; cancer therapy; clinical application; clinically relevant; design; diagnostic accuracy; experimental study; first-in-human; innovation; liquid biopsy; lung cancer cell; magnetic beads; molecular marker; mouse model; multidisciplinary; multiple omics; neoantigens; neoplastic cell; new technology; peripheral blood; pre-clinical; precision oncology; predictive marker; pressure; prevent; radiological imaging; real time monitoring; response; small molecule inhibitor; standard of care; success; targeted treatment; temporal measurement; tool; tumor; tumor progression; validation studies; virtual

With the advent of precision oncology, including both small molecule inhibitors targeting specific genetic drivers of cancer and modulators of the immune response to cancer neoantigens, there is a pressing need for advanced diagnostics to direct and monitor therapeutic interventions. Currently, such real time monitoring is performed through repeat, or resistance, biopsies; however, these surgical biopsies are invasive, may have significant complications including insufficient tissue for the intended analyses, and only sample one specific site of tumor, which may not be representative of the entire tumor cell population within a patient. Liquid biopsies are poised to revolutionize cancer therapies, by enabling frequent blood-based monitoring of tumor-derived materials, as cancers evolve in response to therapeutic interventions. The technological hurdle in isolating sufficient numbers of rare CTCs from routine blood specimens has been the single major limitation preventing the clinical deployment of CTC-based diagnostic opportunities. Enabled by the technology proposed here, our shift from processing routine 10 mL blood tubes (0.2% of whole blood volume) to making use of clinical leukapheresis to sample near-whole blood volumes (40-100%) addresses this challenge. The fundamental basis of the technology is the highly efficient depletion of antibody-tagged blood cells away from unmanipulated CTCs (“negative depletion” as opposed to “positive selection”), thereby enabling tumor antigen– independent enrichment of unperturbed, viable CTCs. This strategy is applicable to cells disseminated from virtually any solid tumor type without making any a priori assumption on the physical or biological properties of tumor cells. Our hypothesis is that liquid biopsy of large number of CTCs using leukopheresis is equivalent to the invasive biopsies of metastatic tumor lesions, currently performed at the time of on-treatment disease progression in lung cancer. To address our hypothesis, we formulated 3 Aims. In Aim 1, we will develop a large-volume CTC isolation technology based on microfluidic negative selection. In Aim 2, we will integrate microfluidic components designed in Aim 1 into a monolithic chip for sorting of CTCs from leukopaks. In Aim 3, we will test our hypothesis by a direct comparison of tumor biopsy and liquid biopsy, performed within a few weeks of each other, assessing both their success rate and diagnostic accuracy. We believe that the convergence of resources and multidisciplinary expertise available in our team will lead to a transformative bioengineering technology, paired with a highly clinically-relevant clinical challenge, providing a new tool for preclinical lung cancer therapeutics. A positive outcome in this pilot study would set the stage for testing more complex clinical applications, ranging from measuring cancer cell signaling effects of drug therapy (“noninvasive pharmacokinetics”) and quantitation of cell surface protein targets for cancer immunotherapy (protein- based predictive markers), and ultimately even enable detection and tissue localization of early invasive cancers in at-risk individuals such as familial breast cancer, people with environmental risks (e.g., lung nodules in smokers, liver cancer in cirrhosis), abnormal radiographic findings of unknown significance or inaccessible biopsy.

随着精确肿瘤学的冒险，包括针对特定遗传驱动因素的小分子抑制剂 癌症和对癌症新抗原的免疫反应的调节剂，迫切需要先进的诊断 直接和监视热干预措施。目前，通过重复或 阻力，活检；但是，这些手术活检是侵入性的，可能有明显的并发症 预期分析的组织不足，仅采样一个特定的肿瘤部位，这可能不是代表性的 患者内的整个肿瘤细胞种群液体活检被中毒以彻底改变癌症疗法 随着癌症响应治疗而进化，可以对肿瘤衍生的材料进行经常基于血液的监测 干预措施。从常规血液标本中隔离足够数量的稀有CTC的技术障碍具有 是阻止基于CTC的诊断机会的临床部署的唯一主要局限性。启用 在这里提出的技术，我们从加工日常的10毫升血管（全血量的0.2％）转变为 利用临床白细胞术来采样近乎全体血量的（40-100％），以解决这一挑战。这 该技术的基本依据是抗体标记的血细胞的高效部署 无操纵的CTC（与“阳性选择”相反），从而实现肿瘤抗原 - 独立的不受干扰，可行的CTC的富集。此策略适用于几乎任何任何分布的细胞 实体瘤类型，而没有对肿瘤细胞的物理或生物学特性进行任何先验假设。我们的 假设是使用白细胞植物进行大量CTC的液体活检等同于入侵活检 转移性肿瘤病变，目前在肺癌中进行治疗疾病进展时进行。 为了解决我们的假设，我们提出了3个目标。在AIM 1中，我们将开发大量的CTC隔离技术 基于微流体负选择。在AIM 2中，我们将将AIM 1中设计的微流体组件集成到一个 从列科帕克人分类CTC的整体芯片。在AIM 3中，我们将通过直接比较来检验我们的假设 肿瘤活检和液体活检在几周内进行，评估其成功率和 诊断准确性。我们认为，我们团队中提供的资源和多学科专业知识的融合 将导致一种变革性的生物工程技术，并与高度临床相关的临床挑战搭配 为临床前肺癌治疗提供了一种新工具。这项试验研究的积极结果将为 测试更复杂的临床应用，从测量药物治疗的癌细胞信号传导效应 （“无创药代动力学”）以及细胞表面蛋白靶标的癌症免疫疗法（蛋白质 - 基于预测标记），甚至最终使早期侵入性癌症的检测和组织定位 诸如家庭乳腺癌，有环境风险的人（例如吸烟者的肺淋巴结，肝癌 肝硬化），异常的射线照相发现，具有未知的意义或无法访问的活检。