Microfluidic technology to isolate tumoricidal T-cells from peripheral blood

项目摘要 使用离体扩增的抗肿瘤T细胞的连续性细胞转移（ACT）由于以下原因而引起了极大的兴趣： 在治疗黑色素瘤和其他癌症方面取得了成功。这是一种高度个性化的疗法，其中自体 需要能够靶向肿瘤的T细胞。然而，寻找特异性靶向肿瘤的细胞仍然是一个难题。 这是广泛应用基于T细胞的ACT疗法的主要障碍。目前淋巴细胞的检测方法 富集导致杀肿瘤T细胞的适度增加，对克隆异质性几乎没有控制。一 克服这些挑战的技术将显著降低障碍（例如，降低成本 脱靶效应）用于广泛传播ACT疗法。该项目的主要目标是开发一个 分离技术，以基于它们的能力富集具有杀肿瘤T细胞的淋巴细胞群， 识别自体肿瘤抗原。我们微流体技术的前提是， 当暴露于肿瘤衍生肽-主要组织相容性时， 最佳流动条件下的复合体I。具体目标是：1）开发一种微流控装置，以富集 具有抗原特异性T细胞的淋巴细胞群，以及2）证明微流体免疫的能力。 平台，以使用患者的细胞培养物富集具有杀肿瘤T细胞的患者来源的外周血单核细胞。 匹配的肿瘤细胞实现我们的主要目标将创造一种潜在的颠覆性技术， 为基于T细胞的ACT疗法的广泛应用铺平了道路，并且不可知特征（即，没有先验 需要肿瘤抗原的知识）的技术将使其广泛适用于 针对一系列癌症的个性化医疗方法。

PROJECT SUMMARY Adoptive cell transfer (ACT) using ex vivo expanded anti-tumor T-cells has garnered significant interest due to successes in treating melanoma and other cancers. This is a highly personalized therapy, in which autologous T-cells that can target the tumors are required. However, finding cells that specifically target tumors remains a major hurdle for the widespread application of T-cell based ACT therapies. The current methods of lymphocyte enrichment result in modest increases in tumoricidal T-cells with little control over the clonal heterogeneity. A technology that overcomes these challenges would significantly lower the barriers (e.g., reduce cost, reduce off-target effects) for broad dissemination of ACT therapies. The primary goal of this project is to develop a separation technology to enrich a population of lymphocytes with tumoricidal T-cells based on their capacity to recognize autologous tumor antigens. The premise of our microfluidic technology is that tumoricidal T-cells can be separated from a bulk leukocyte population when exposed to tumor-derived peptide-major histocompatibility complex I under optimal flow conditions. The specific aims are to: 1) Develop a microfluidic device to enrich a population of lymphocytes with antigen specific T-cells, and 2) Demonstrate the capacity of the microfluidic platform to enrich patient-derived Peripheral Blood Mononuclear Cells with tumoricidal T-cells using patient- matched tumor cells. Accomplishing our primary goal will create a potentially disruptive technology that could pave way for wide-spread application of T-cell based ACT therapies, and the agnostic feature (i.e., no a priori knowledge of tumor antigen(s) is required) of the technology would make it broadly applicable for a personalized medicine approach to a range of cancers.