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: 10199185
• 负责人: Daniel A. Haber
• 金额: $ 45.23万
• 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/10199185
• 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; Diagnostic radiologic examination; 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; 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; 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.

随着精准肿瘤学的出现，包括针对特定基因驱动因素的小分子抑制剂 癌症和癌症新抗原免疫反应的调节剂，迫切需要先进的诊断技术 目前，这种实时监测是通过重复或监测来进行的。 阻力、活检；然而，这些手术活检是侵入性的，可能会产生严重的并发症，包括 组织不足以进行预期分析，并且仅对肿瘤的一个特定部位进行采样，这可能不具有代表性 液体活检有望彻底改变癌症治疗方法。 随着癌症响应治疗而进化，能够对肿瘤衍生物质进行频繁的血液监测 从常规血液样本中分离足够数量的稀有 CTC 存在技术障碍。 是阻碍基于 CTC 的诊断机会的临床部署的单一主要限制。 此处提出的技术，我们从处理常规 10 mL 血管（全血量的 0.2%）转变为 利用临床白细胞分离术来采集接近全血量（40-100%）的样本解决了这一挑战。 该技术的基本原理是高效地去除带有抗体标记的血细胞 未经操作的 CTC（“负耗竭”而不是“正选择”），从而使肿瘤抗原– 独立富集未受干扰的、存活的 CTC，该策略适用于几乎任何来源的细胞。 实体瘤类型，无需对肿瘤细胞的物理或生物学特性做出任何先验假设。 假设使用白细胞去除术对大量 CTC 进行液体活检相当于侵入性活检 转移性肿瘤病灶，目前在肺癌治疗疾病进展时进行。 为了解决我们的假设，我们制定了 3 个目标 在目标 1 中，我们将开发一种大容量 CTC 分离技术。 基于微流控负选择，在目标 2 中，我们将把目标 1 中设计的微流控组件集成到一个模型中。 用于从 leukopaks 中分选 CTC 的单片芯片 在目标 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，评估它们的成功率和 我们相信我们团队的资源和多学科专业知识的融合。 将带来变革性的生物工程技术，并伴随着与临床高度相关的临床挑战， 该试点研究的积极成果将为临床前肺癌治疗提供新的工具。 测试更复杂的临床应用，包括测量药物治疗的癌细胞信号传导作用 （“无创药代动力学”）和癌症免疫治疗的细胞表面蛋白靶标的定量（蛋白- 基于预测标记），最终甚至能够检测和组织定位处于危险中的早期浸润性癌症 个人，例如家族性乳腺癌、有环境风险的人（例如吸烟者的肺结节、患有肝癌的人） 肝硬化）、意义不明的异常放射学发现或无法进行活检。