Automated Patient-Specific Dendritic Cell Generation for Transcriptomics-Driven Vaccinology

用于转录组驱动的疫苗学的自动患者特异性树突状细胞生成

• 批准号: 9275355
• 负责人: Shashi Murthy
• 金额: $ 38.49万
• 依托单位: NORTHEASTERN UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-06-20 至 2019-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9275355
• 关键词: Address; Arts; Autoimmunity; Autologous; Autologous Dendritic Cells; BCG Vaccine; Basic Science; Benchmarking; Biological Assay; Biomedical Engineering; Biomedical Technology; Blood; Blood Cells; Blood Volume; Blood specimen; CD14 gene; CD4 Positive T Lymphocytes; Cell Culture System; Cell Therapy; Cells; Clinical Research; Coculture Techniques; Communicable Diseases; Data; Dendritic Cell Vaccine; Dendritic Cells; Development; Dinoprostone; Disease; Elements; Environment; Epitopes; Flow Cytometry; Generations; Genetic Transcription; Goals; Graft Rejection; Granulocyte-Macrophage Colony-Stimulating Factor; Homing; Hour; Human; Human Resources; IL4 gene; IL6 gene; Immune; Immunity; Immunology; Individual; Infection; Interleukin-1; Knowledge; Malignant Neoplasms; Manuals; Metabolic Pathway; Methods; Microfluidics; Patients; Pattern; Performance; Perfusion; Peripheral Blood Mononuclear Cell; Phase; Phenotype; Prevention; Process; Protocols documentation; Regenerative Medicine; Regimen; Reproducibility; Research; Resources; Role; Sampling; Savings; Series; Standardization; System; T cell response; T cell therapy; T memory cell; T-Lymphocyte; TNF gene; Techniques; Technology; Testing; Time; Tube; Vaccination; Vaccine Clinical Trial; Vaccines; Whole Blood; blood perfusion; cohort; commercialization; comparative; cost; cost effective; cytokine; design; drug discovery; efficacy study; experience; experimental study; genome-wide; head-to-head comparison; improved; individual patient; infectious disease treatment; insight; memory CD4 T lymphocyte; monocyte; new technology; novel vaccines; pathogen; programs; prototype; public health relevance; response; self-renewal; tool; transcriptomics; vaccine candidate; vaccine development; vaccine discovery; vaccine trial; vaccinology; whole genome

 DESCRIPTION (provided by applicant): Dendritic cells (DCs) are an indispensable part of studying human responses that are important for protective immunity against cancer and infectious diseases as well as prevention of autoimmunity and transplant rejection. These cells are also key elements of personalized vaccines which are a major research focus in cancer and infectious diseases. Despite the vital role of DCs in both clinical and basic research contexts, methods for obtaining these cells from individuals remains a comparatively under-developed and inefficient process. Because DCs are present in very low concentrations (<1%) in blood, these cells must be generated from monocytes and the state of the art in such generation involves a laborious process of static culture and stimulation with cytokines contained in culture medium. Numerous manual steps are required to go from a sample of patient-derived blood or peripheral blood mononuclear cells (PBMCs) to sufficient numbers of DCs that can be utilized for vaccine development, T cell therapy, or mechanistic studies. When scaled even to the level of tens of samples for a study involving one or two conditions or separate blood draws, the resource requirement in terms of personnel hours and number of manual steps becomes significant. Considering the existing and projected use of these cells at much larger scale, such as in Phase II or III clinical trials of vaccines and personalized cell therapy regimens, the curret approach to DC generation poses an unusually large burden, most significantly in terms of cost, but also in terms of the time required to perform comprehensive studies and trials. This proposal aims to address the unmet need for effective DC generation technologies by designing of a fully-automated microfluidic system (microDEN) that accepts a blood or PBMC sample and directly delivers DCs following a period of perfusion with cytokines. This system will combine monocyte isolation from blood and perfusion culture into individual, patient-specific chips. The elimination of manual steps associated with monocyte purification and culturing in two different media types containing cytokines will, in itself, represent a major advance relative to start of th art DC generation. We further hypothesize that the perfusion technique employed in our microfluidic method will allow reduction in the time required for DC generation (currently ~ 6 days), thereby offering significant additional savings in cost and resources. The proposed microDEN system will be built and rigorously benchmarked against the conventional DC generation technique using a range of functional assays, the most critical of which will be transcriptomal profiling of CD4+ T cells stimulated with autologous, BCG-infected DCs.

 描述（由适用提供）：树突状细胞（DC）是研究人类反应的必不可少的一部分，这些反应对于保护癌症和传染病的保护性免疫学很重要，并且预防自身免疫性和移植抑制。这些细胞也是个性化疫苗的关键要素，这是癌症和传染病的主要研究重点。尽管DC在临床和基础研究环境中都起着至关重要的作用，但从个体获得这些细胞的方法仍然是相对较低的发达和效率低下的过程。由于DC的血液中存在非常低的浓度（<1％），因此这些细胞必须由单核细胞产生，并且在这一生成中的最新细胞涉及静态培养的实验室过程，并用培养基中包含的细胞因子进行刺激。需要进行许多手动步骤，从患者衍生的血液或外周血单核细胞（PBMC）的样本到可用于疫苗开发，T细胞疗法或机械研究的兼容数量的DC。当将研究范围缩放到涉及一个或两个条件或单独抽血的研究水平时，就人员小时和手动步骤数量而言，资源要求变得很大。考虑到这些细胞在更大范围内的现有和预测使用，例如在II期或III期临床试验中，疫苗和个性化细胞疗法方案的临床试验，DC生成的库雷方法在成本方面具有异常大的Burnen，但在进行全面研究和试验的时间方面也很重要。该提案旨在通过设计完全自动化的微流体系统（微型）来满足对有效DC生成技术的未满足需求，该系统接受血液或PBMC样品，并在用细胞因子灌注后直接提供DC。该系统将将从血液和灌注培养物中的单核细胞分离为单个，患者特异性芯片。在包含细胞因子的两种不同培养基类型中与单核细胞纯化和培养相关的手动步骤的演变本身就是相对于TH ART DC生成的开始的主要进步。我们进一步假设，在微流体方法中采用的灌注技术将允许DC生成所需的时间（目前约6天）减少，从而为成本和资源提供了可观的节省。拟议的微源系统将使用一系列功能测定法对传统的DC生成技术进行严格测试，其中最关键的是用自体内BCG感染的DC刺激的CD4+ T细胞的转录组分析。