Microfluidic Precision Engineered Artificial Antigen Presenting Cells for Cancer Immunotherapy

用于癌症免疫治疗的微流控精密工程人工抗原呈递细胞

• 批准号: 10696138
• 负责人: Anshu Agrawal
• 金额: $ 37.37万
• 依托单位: UNIVERSITY OF CALIFORNIA-IRVINE
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-02 至 2025-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10696138
• 关键词: APC Vaccine; Adhesions; Adjuvant; Affect; Animal Testing; Antibodies; Antigen Presentation; Antigen-Presenting Cells; Antigens; Area; Autoantigens; Autologous; Back; Binding; Biological; Blood; Breast Cancer Patient; CD28 gene; CD8-Positive T-Lymphocytes; CD8B1 gene; Cancer Patient; Cell Adhesion Molecules; Cell Size; Cell membrane; Cells; Complex; Consumption; Cytoskeleton; Cytotoxic T-Lymphocytes; Data; Dendritic Cells; Emulsions; Engineering; Engraftment; Evaluation; Formulation; Goals; Grant; Hydrogels; Immune system; Immunologic Memory; In Vitro; Incubated; Individual; Influenza; Integral Membrane Protein; Integrins; Interferon Type II; Ligands; Lipid Bilayers; Malignant Neoplasms; Mechanics; Membrane; Methods; Microfluidic Microchips; Microfluidics; Modeling; Molecular; Mus; Patients; Peripheral Blood Mononuclear Cell; Process; Production; Proliferating; Proteins; Specificity; Synapses; T cell response; T cell therapy; T-Cell Activation; T-Lymphocyte; TNF gene; Testing; Time; Tumor Antigens; Tumor Expansion; Vaccination; antigen-specific T cells; cancer immunotherapy; cell preparation; cytotoxic CD8 T cells; efficacy evaluation; fighting; immunological synapse; in vivo; in vivo evaluation; malignant breast neoplasm; manufacturing scale-up; mechanical properties; melanoma; neoplastic cell; response; scale up; tumor; tumor eradication; tumor microenvironment; tumor progression; unilamellar vesicle

Abstract The goal of cancer immunotherapy is to build long-lasting tumor-specific immunologic ‘memory’ in patients that enables the lifelong rejection of tumors. The two prominent types of antigen-specific cancer immunotherapy, adoptive T cell therapy and APC-based vaccination, both require expansion of anti-tumor T cells via APCs. However, for the purpose of effective adoptive T cell therapy, the critical question is how to generate, within a short period of time, large numbers of antitumor T cells. Furthermore, in vitro-expanded T cells must also possess the capacity to engraft, proliferate, and persist in vivo with sufficient antitumor function to induce sustained antitumor activity. Autologous antigen-presenting cells (APCs) such as DCs also have several serious limitations. The necessity to access large amounts of cancer patients’ blood to prepare autologous APC from each patient in a timely manner is cumbersome. To overcome these problems, we developed the microfluidic process to generate cell-sized unilamellar vesicles (CUVs) and decorated them with antigen presenting ligands for artificial APCs (or aAPCs). Preliminary results show that aAPCs are able to bind and interact with T cells and cause their expansion. The objective of the present proposal is to further optimize the aAPCs preparation and test its capacity to induce tumor specific responses in vitro and in vivo. The hypothesis is that the optimized aAPC functionalization will result in enhanced expansion of cytotoxic CD8 T cells and a reduction in tumor progression over the present one (original). The Specific Aims are- 1) Bioinspired optimization of artificial antigen presenting cell (aAPC) production via microfluidic engineering. We will insert the antigen presenting ligands in the membrane to mimic cells. The aAPCs will also be produced with hydrogel cytoskeletons to optimize its mechanical properties for maximum T cell expansion. 2) Evaluation of the capacity of aAPCs to induce tumor specific T cell responses in vitro. Using PBMCs from healthy donors and breast cancer patients we will evaluate the capacity of aAPCs to induce cytotoxic T cells. 3) Evaluation of the capacity of aAPCs to induce T cell responses and tumor killing with an in vivo mice tumor model. Methods to scale up the production of aAPCs for in vivo use will be developed. The capacity of aAPCs to kill tumor in vivo in mice will also be determined using a melanoma model. The goal is to produce an aAPC preparation that mimics cells, is stable, easy to produce in large quantities and capable of expanding tumor specific CD8 T cells for immunotherapy of cancer.

抽象的 癌症免疫疗法的目的是在患者中建立持久的肿瘤特异性免疫学“记忆”。 使肿瘤的终身拒绝。两种突出类型的抗原特异性癌症免疫疗法， 养子T细胞疗法和基于APC的疫苗接种都需要通过APC扩大抗肿瘤T细胞。 但是，出于有效的自适应T细胞疗法的目的，关键的问题是如何在 短时间，大量抗肿瘤T细胞。此外，体外扩展的T细胞还必须拥有 具有足够的抗肿瘤功能的体内植入，增殖和持续的能力，以诱导持续 抗肿瘤活性。自体抗原细胞（APC）（例如DC）也有几个严重的局限性。 获得大量癌症患者血液以从每位患者准备自体APC的必要条件 及时的方式很麻烦。为了克服这些问题，我们开发了微流体过程 产生细胞大小的单层蔬菜（CUV），并用抗原呈现艺术配体进行装饰 APC（或AAPC）。初步结果表明，AAPC能够结合并与T细胞相互作用并引起其 扩张。本提案的目的是进一步优化AAPC的准备并测试其 在体外和体内诱导肿瘤特异性反应的能力。假设是优化的AAPC 功能化将导致细胞毒性CD8 T细胞的膨胀增强和肿瘤进展的减少 在目前的（原始）上。具体目的是-1）呈现人造抗原的生物启发优化 通过微流体工程生产细胞（AAPC）。我们将在 膜到模拟细胞。 AAPC还将用水凝胶细胞骨架生产，以优化其 最大T细胞扩展的机械性能。 2）评估AAPC诱导肿瘤的能力 特定的T细胞反应体外。使用来自健康供体和乳腺癌患者的PBMC，我们将评估 AAPC诱导细胞毒性T细胞的能力。 3）评估AAPC诱导T细胞的能力 用体内小鼠肿瘤模型的反应和肿瘤杀死。扩大AAPC生产的方法 将开发体内使用。 AAPC在小鼠体内杀死肿瘤的能力也将使用A确定 黑色素瘤模型。目的是生产AAPC制备，以模拟细胞稳定，易于生产 大量，能够扩大肿瘤特异性CD8 T细胞进行癌症的免疫疗法。