High-efficiency microfluidic cell fusion for dendritic cell/tumor cell vaccine production

项目摘要 使用树突状细胞-肿瘤(DC/肿瘤)杂交细胞制成的癌症疫苗进行个性化免疫治疗 被开发用于一系列癌症，从急性髓系白血病(AML)开始，现在是 适用于其他血癌(淋巴瘤、多发性骨髓瘤)和实体瘤(肾细胞癌、 乳腺癌)。这些癌症疫苗显示出持久的缓解、持久的保护作用， 毒性最小。在加工过程中，基于DC的疫苗使用数百万患者来源的融合 肿瘤细胞和自体DC以创建呈现广泛的肿瘤抗原的杂交细胞， 包括由个人肿瘤的独特突变特征产生的新抗原，在上下文中 直流电介导的共刺激。尽管他们有希望，但在疫苗接种过程中仍存在一些技术挑战 产生的原因是DC-肿瘤融合效率低且可变。其中一个挑战涉及 获取足够的肿瘤组织，这可能会限制疫苗在早期肿瘤和实体瘤中的适用性 癌症。更重要的是，低融合效率意味着许多肿瘤细胞不会对 疫苗，这可能会限制新抗原库和疫苗的最终效力。要达到高度 融合效率，我们建议将细胞融合的微流控方法改造为离心式， 使用微流控旋转圆盘融合数十万个细胞。在本R21中，我们建议 解决在进一步发展之前必须解决的两个高风险问题：1)我们能否融合 临床上相关数量的高效率细胞，以及2)提高融合效率是否提供 任何生物学(因此，潜在的临床)益处。因此，我们提出两个具体目标： 目的1：研制高效融合DC/AML细胞的离心式微流控圆盘平台。 我们将把微流控细胞融合平台改造成旋转圆盘离心式，它可以处理 一大片区域上的细胞。 目的2：对盘状DC/AML融合肿瘤疫苗进行体内外评价。我们将利用EX 活体人患者来源和小鼠AML模型评估盘状DC/AML融合细胞 与常规处理的融合细胞相比。

Project Summary Personalized immunotherapy using cancer vaccines made of dendritic-tumor (DC/Tumor) hybrid cells is being developed for a range of cancers, starting with acute myeloid leukemia (AML) and now being adapted to other blood cancers (lymphoma, multiple myeloma) and solid tumors (renal cell carcinoma, breast cancer). These cancer vaccines have demonstrated durable remissions, long-lasting protection, and minimal toxicity. During processing, DC-based vaccines use fusion of millions of patient-derived tumor cells and autologous DCs to create hybrid cells that present a broad array of tumor antigens, including neoantigens generated by the unique mutational profile of an individual's tumor, in the context of DC-mediated co-stimulation. Despite their promise, several technical challenges exist during vaccine production stemming from the low and variable DC-tumor fusion efficiency. One challenge involves obtaining adequate tumor tissue, which can restrict vaccine applicability in early-stage tumors and solid cancers. More significantly, low fusion efficiency means that many tumor cells do not contribute to the vaccine, which may limit the neoantigen repertoire and the ultimate efficacy of the vaccine. To attain high fusion efficiency, we propose to adapt a microfluidic method for cell fusion to a centrifugal format that uses a microfluidic spinning disc to fuse hundreds of thousands of cells. In this R21 we propose to address two high-risk questions necessary before further development can take place: 1) can we fuse clinically relevant quantities of cells at high efficiency, and 2) does increased fusion efficiency provide any biological (and, thus, potentially clinical) benefits. We thus propose two specific aims: Aim 1: Development of a centrifugal microfluidic disc platform for high-efficiency fusion of DC/AML cells. We will adapt a microfluidic cell fusion platform to a spinning disc centrifugal format, which processes cells over a large area. Aim 2: Ex vivo and in vivo evaluation of disc-made DC/AML fusion cancer vaccines. We will utilize ex vivo human patient-derived and murine AML models to evaluate disc-made DC/AML fusion cells in comparison to conventionally processed fusion cells.