Determining treatment sensitivity in B cell lymphoma by novel microfluidics-based NK cell immunogenicity platform

通过基于微流体的新型 NK 细胞免疫原性平台确定 B 细胞淋巴瘤的治疗敏感性

• 批准号: 9919540
• 负责人: Andrew M Evens
• 金额: $ 37.93万
• 依托单位: NORTHEASTERN UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-05-01 至 2022-10-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9919540
• 关键词: Accounting; Address; Antibodies; Antibody Therapy; B lymphoid malignancy; B-Cell Lymphomas; B-Cell NonHodgkins Lymphoma; Basal Cell; Binding; Biochemical; Biological; Biological Assay; Biological Availability; Biomedical Engineering; Biomedical Research; Categories; Cell Communication; Cells; Characteristics; Classification; Clinical; Clinical Sensitivity; Collaborations; Comb animal structure; Communities; Complex; Development; Devices; Drops; Drug Screening; Event; FDA approved; Fluorescence; Generations; Heterogeneity; Hyperactive behavior; Image; Immune; Immune Targeting; Immune response; Immunoassay; Immunomodulators; Immunotherapeutic agent; Immunotherapy; Individual; Kinetics; Link; Lymphocyte; Lymphoma; MS4A1 gene; Malignant Neoplasms; Measurement; Mediating; Methods; Microfluidic Microchips; Microfluidics; Molecular; Monitor; Monoclonal Antibodies; Natural Killer Cells; Non-Hodgkin' s Lymphoma; Pharmaceutical Preparations; Pharmacologic Substance; Phenotype; Population; Research; Research Personnel; Resistance; Resolution; Site; Sorting - Cell Movement; Speed; System; Systems Biology; Techniques; Technology; Testing; Therapeutic; Therapeutic Human Experimentation; Time; Transducers; Tumor-infiltrating immune cells; base; behavioral phenotyping; biological heterogeneity; biomaterial compatibility; cell killing; chemotherapy; clinical application; clinical assay development; experience; high throughput technology; immune function; immunogenicity; innovation; multidisciplinary; nanosystems; neoplastic cell; novel; novel drug combination; novel strategies; personalized medicine; predictive test; response; rituximab; screening; single cell analysis; success; targeted treatment; tositumomab; transcriptomics; tumor; tumor immunology

Abstract B cell non-Hodgkin lymphomas (bNHL) are the most common lymphoma subtype representing >85% of all NHLs. bNHL are typically treated the anti-CD20 antibody (e.g., rituximab) alone or in combination with chemotherapy. There are currently, however, no biological methods or markers to predict the sensitivity or resistance to rituximab (or any other) antibody therapy. A key feature of antibody activity occurs through natural killer (NK) cell-mediated killing of antibody-coated target tumor cells, however, antitumor activity and subsequent resistance, is poorly understood. In this application, we propose to develop and validate a high throughput droplet based microfluidic platform to investigate the key features of NK cells associated with rapid, slow or inactive tumor killing kinetics in NHL. We will first adapt a novel approach and integrate the biocompatible acoustofluidic droplet sorter during the droplet microarray formation to determine the phenotypes of immune-target cell interaction in microfluidic droplets. We will validate a droplet-based microfluidic device to interrogate single-cell dynamic responses and cell-cell interactions within intact droplets. Next, we will demonstrate a high-purity (>95%), high-throughput (>10,000 events/s), four-channel acoustofluidic droplet sorter to integrate with droplet analysis array. The downstream 4-channel sorting will allow, after establishing the kinetic profiles of interactions, to identify and sort droplets containing active lymphocytes into a distinctive pool; separate basal lymphocytes into another pool based on fluorescence. A unique function of selecting sorting criteria based on imaging analysis can be provided by the combination of droplet imaging array and acoustofluidic droplet sorters, which is unachievable for conventional fluorescence activated droplet sorters (FADS) since imaging tracking is inherently tricky in high-speed flow. Thus, our approach serves as a “bottom-up” method of classification, by first identifying distinct functional categories and then probing the content of the individual cell category to determine the key factors for the molecular classification of heterogeneous immune functions of NK cells related to target cell kill. In addition, we will identify NK cell heterogeneity and bio-functional characteristics to discover novel drug combinations for NK cell dependent immunotherapy via an integrated acoustofluidic droplet sorting platform. We will demonstrate that the accuracy of phenotype identification of our device and its suitability for clinical applications by monitoring and classifying NK/NHL single cell interactions in the presence of monoclonal antibodies and performing biochemical secretome assay from ‘hyperactive’, ‘basal’ and non-responsive pools. By combing these findings with drug screening and identification of phenotype altering drugs, we will demonstrate the applicability of this technology for personalized medicine and rational clinical immunotherapeutic applications. We envision our platform may be leveraged in a variety of single-cell analysis applications in immunotherapy and it will provide high value to the bioengineering, biomedical, and therapeutic research communities.

抽象的 B 细胞非霍奇金淋巴瘤 (bNHL) 是最常见的淋巴瘤亚型，占 85% 以上 所有 NHL 通常单独使用抗 CD20 抗体（例如利妥昔单抗）或与抗 CD20 抗体联合治疗。 然而，目前没有生物学方法或标记来预测敏感性或化疗。 对利妥昔单抗（或任何其他）抗体疗法的耐药性 抗体活性的一个关键特征是通过自然发生的。 杀伤细胞（NK）介导的抗体包被靶肿瘤细胞的杀伤作用，然而，抗肿瘤活性和随后的 阻力，对此知之甚少。在此应用中，我们建议开发并验证高通量液滴。 基于微流控平台来研究 NK 细胞与快速、缓慢或不活动相关的关键特征 NHL 中的肿瘤杀伤动力学我们将首先采用一种新方法并整合生物相容性声流体。 液滴分选仪在液滴微阵列形成过程中确定免疫靶细胞的表型 我们将验证基于液滴的微流体装置来询问单细胞。 接下来，我们将演示高纯度的液滴内的动态响应和细胞间相互作用。 (>95%)、高通量（>10,000 个事件/秒）、四通道声流控液滴分选器，可与液滴集成 下游 4 通道排序将允许在建立相互作用的动力学曲线后， 识别含有活性淋巴细胞的液滴并将其分类到独特的单独的基础淋巴细胞池中； 进入另一个基于荧光的池基于成像分析选择排序标准的独特功能。 可以通过液滴成像阵列和声流控液滴分选器的组合来提供，这是 传统的荧光激活液滴分选器 (FADS) 无法实现，因为成像跟踪本质上是 因此，我们的方法通过首先识别来充当“自下而上”的分类方法。 区分功能类别，然后探究单个细胞类别的内容以确定关键 与靶细胞杀伤相关的 NK 细胞异质免疫功能的分子分类因素。 此外，我们将鉴定NK细胞异质性和生物功能特征，以发现新药 通过集成声流体液滴分选平台进行 NK 细胞依赖性免疫治疗的组合。 将证明我们的设备表型识别的准确性及其临床适用性 通过在单克隆抗体存在的情况下监测和分类 NK/NHL 单细胞相互作用来应用 抗体并对“过度活跃”、“基础”和无反应池进行生化分泌组测定。 通过将这些发现与药物筛选和表型改变药物的鉴定相结合，我们将 展示该技术对于个性化医疗和合理临床的适用性 我们预计我们的平台可用于各种单细胞分析。 其在免疫治疗中的应用将为生物工程、生物医学和治疗提供很高的价值 研究社区。