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.

摘要 癌症免疫治疗的目标是在符合以下条件的患者中建立持久的肿瘤特异性免疫“记忆” 使肿瘤能够终生排斥。两种主要的抗原特异性癌症免疫疗法， 过继T细胞疗法和基于APC的疫苗接种都需要通过APC扩增抗肿瘤T细胞。 然而，为了有效地采用T细胞疗法，关键问题是如何在一个 短时间内，大量的抗肿瘤T细胞。此外，体外扩增的T细胞还必须具备 在体内植入、增殖和存活的能力，并具有足够的抗肿瘤功能以诱导持续 抗肿瘤活性。树突状细胞等自体抗原提呈细胞(APC)也有一些严重的局限性。 获取大量癌症患者血液以制备每个患者自体APC的必要性 及时的是繁琐的。为了克服这些问题，我们开发了微流控工艺来 产生细胞大小的单层囊泡(CUV)，并用抗原提呈配体修饰它们以用于人工 APC(或aAPC)。初步结果表明，aAPC能够与T细胞结合和相互作用，并使其 扩张。本建议的目标是进一步优化AAPC的制备并测试其 在体外和体内诱导肿瘤特异性反应的能力。假设是最优的AAPC 功能化将导致细胞毒性CD8 T细胞的增强扩张和肿瘤进展的减缓 而不是现在的(原件)。具体目标是：1)人工抗原呈递的生物启发优化 通过微流控工程生产细胞(AAPC)。我们将把抗原提呈配体插入到 膜来模拟细胞。AAPC还将与水凝胶细胞骨架一起生产，以优化其 最大限度地扩大T细胞的机械性能。2)评价aAPC的致瘤能力 体外特异性T细胞反应。使用来自健康捐赠者和乳腺癌患者的PBMC，我们将评估 AAPC诱导细胞毒性T细胞的能力。3)aAPC诱导T细胞能力的评价 体内小鼠肿瘤模型的反应和肿瘤杀伤。扩大AAPC生产规模的方法 将开发活体使用。AAPC在小鼠体内杀伤肿瘤的能力也将通过使用 黑色素瘤模型。我们的目标是生产一种模拟细胞、稳定、易于生产的AAPC制剂 大量扩增肿瘤特异性CD8T细胞，用于肿瘤免疫治疗。