Scaffolds mimicking antigen presenting cells

模拟抗原呈递细胞的支架

• 批准号: 10001355
• 负责人: David J Mooney
• 金额: $ 60万
• 依托单位: HARVARD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-09-20 至 2021-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10001355
• 关键词: Scaffolds; mimicking; antigen; presenting; cells

Current approaches to expand T cells ex vivo for therapeutic applications are limited by low expansion rates and T-cell products of limited functionality. To address these issues, we have recently described a system that mimics natural antigen-presenting cells (APCs). These APC-mimetic scaffolds (APC-ms) consist of a fluid lipid bilayer supported by mesoporous silica micro-rods (MSRs). The lipid bilayer presents membrane-bound cues for T-cell receptor stimulation and costimulation at predefined densities, while the micro-rods enable sustained release of soluble paracrine cues. Using anti-CD3, anti-CD28 and controlled release of interleukin-2, we have shown that the APC-ms promotes ten-fold greater polyclonal expansion of primary mouse and human T cells than commercial beads (Dynabeads), and can be used to tune the phenotypic attributes of expanded T-cell products. APC-ms also support over 5-fold greater expansion of CD19 CAR-T cells than Dynabeads, with increased efficacy in a clinically-relevant xenograft lymphoma model. While APC-ms is a promising T cell expansion system, there are several development activities that would aid its clinical and commercial translation. The current MSR synthesis process has not been standardized for this application, and lacks established SOPs to yield material products with consistent properties. Currently, surface cues are presented on APC-ms using biotin-streptavidin, but the use of click chemistry in place of streptavidin could provide a number of advantages. While APC-ms is designed to completely degrade during the process of T cell culture period, obviating the need for its removal before cell delivery, we have not yet thoroughly explored the impact of any potential residual materials on the infused T cell products. These needs lead to the following specific objectives for this project (1) Establish SOPs for APC-ms synthesis. This will include identifying MSR critical quality attributes (CQAs) for functional APC-ms and understanding how critical process parameters (CPPs) in MSR synthesis affect those CQAs (2) Develop a process to directly and selectively conjugate surface cues onto lipid bilayers, via click chemistry, to simplify and modularity of APC-ms assembly and function. (3) Characterize residual APC-ms materials during T cell processing, and perform a thorough in vivo safety assessment. The successful achievement of these aims will immediately address key issues related to using APC-ms as an ex vivo T-cell expansion platform.

目前用于治疗应用的离体扩增 T 细胞的方法受到低扩增率的限制 以及功能有限的 T 细胞产品。为了解决这些问题，我们最近描述了一个系统 模仿天然抗原呈递细胞（APC）。这些 APC 模拟支架 (APC-ms) 由液体脂质组成 由介孔二氧化硅微棒（MSR）支撑的双层。脂质双层呈现膜结合信号 用于以预定密度刺激 T 细胞受体和共刺激，而微棒能够持续 释放可溶性旁分泌信号。使用抗 CD3、抗 CD28 和白细胞介素 2 的控释，我们 显示 APC-ms 促进原代小鼠和人类 T 细胞多克隆扩增十倍 比商业珠 (Dynabeads) 更有效，可用于调整扩增 T 细胞的表型属性 产品。 APC-ms 还支持比 Dynabeads 多 5 倍的 CD19 CAR-T 细胞扩增， 提高临床相关异种移植淋巴瘤模型的疗效。虽然APC-ms是一种有前途的T细胞 扩展系统，有几项开发活动将有助于其临床和商业 翻译。目前的 MSR 合成工艺尚未针对该应用进行标准化，并且缺乏 建立标准操作程序以生产具有一致特性的材料产品。目前，表面线索已呈现 在 APC-MS 上使用生物素-链霉亲和素，但使用点击化学代替链霉亲和素可以提供 数量优势。而APC-ms被设计为在T细胞培养过程中完全降解 期间，避免了在细胞交付之前将其移除的需要，我们尚未彻底探讨其影响 输注 T 细胞产品上任何潜在残留物质。这些需求导致了以下具体需求 该项目的目标 (1) 建立 APC-ms 合成的 SOP。这将包括确定 MSR 关键 功能性 APC-MS 的质量属性 (CQA) 并了解关键过程参数 (CPP) 在 MSR 合成影响这些 CQA (2) 开发直接且选择性地缀合表面线索的流程 通过点击化学将其连接到脂质双层上，以简化 APC-ms 组装和功能并使其模块化。 (3) 表征 T 细胞处理过程中残留的 APC-ms 材料，并执行彻底的体内安全性 评估。这些目标的成功实现将立即解决与使用相关的关键问题 APC-ms 作为离体 T 细胞扩增平台。