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，代表> 85％ 所有NHLS。 BNHL通常单独处理抗CD20抗体（例如利妥昔单抗）或与 化学疗法。但是，目前没有生物学方法或标记来预测灵敏度或 对利妥昔单抗（或任何其他）抗体疗法的抗性。抗体活性的关键特征是通过自然发生的 然而 抵抗，知之甚少。在此应用程序中，我们建议开发和验证高吞吐量液滴 基于微流体平台，研究与快速，缓慢或不活跃相关的NK细胞的关键特征 NHL肿瘤杀死动力学。我们将首先适应一种新颖的方法，并整合生物相容性的声流液 液滴微阵列形成期间的液滴分辨率，以确定免疫目标细胞的表型 微流体液滴中的相互作用。我们将验证基于液滴的微流体设备来询问单细胞 完整液滴中的动态响应和细胞 - 细胞相互作用。接下来，我们将展示高纯度 （> 95％），高通量（> 10,000事件/s），四通道声流体液滴分散器与液滴整合 分析阵列。在建立相互作用的动力学轮廓后，下游的4通道分类将允许 识别和排序含有活性淋巴细胞的液滴到独特的池中；单独的碱性淋巴细胞 基于荧光进入另一个池。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动液滴分辨率的组合提供 对于常规荧光激活的液滴划分角（FADS），由于成像跟踪本质上是无法实现的 高速流棘手。这是通过首先识别的，我们的方法是一种“自下而上”的分类方法 不同的功能类别，然后探测单个单元格类别的内容以确定键 与靶细胞杀死有关的NK细胞异质免疫功能的分子分类的因素。 此外，我们将确定NK细胞异质性和生物功能特征以发现新的药物 通过集成的声流液滴分选平台的NK细胞依赖免疫疗法的组合。我们 将证明我们设备表型识别的准确性及其对临床的适用性 通过在单克隆存在的情况下监视和分类NK/NHL单细胞相互作用来应用 来自“多动”，“基础”和无反应池的抗体和生化分泌组测定。 通过将这些发现与药物筛查和鉴定进行表型改变药物的鉴定，我们将 证明该技术在个性化医学和合理的临床上的适用性 免疫治疗应用。我们设想我们的平台可能会在各种单细胞分析中利用 免疫疗法的应用，它将为生物工程，生物医学和治疗提供高价值 研究社区。