Engineering injectable T cell stimulating microparticles for cancer immunotherapy

项目摘要 免疫性T细胞疗法（ACT）是一种基于T细胞的癌症疗法，其中自体T细胞被用于治疗癌症。 从患者分离细胞，离体活化和扩增，然后再输注到细胞中。 病人虽然ACT已显示出巨大的临床成功，但成功仅限于 黑色素瘤和复杂的制造考虑造成了巨大的代价。创新在 用于T细胞活化的可扩展的非细胞系统，例如人工抗原呈递细胞 （aAPCs），通过缩短培养时间改善了CD 8 + T细胞的离体扩增， 提供对所得T细胞表型和功能的更严格控制。然而，T细胞培养 使用aAPC仍然需要几周时间并且需要制造劳动力和成本。平台为在 抗原特异性T细胞的体内活化将降低T细胞免疫的成本和复杂性， 疗法该项目的目标是创建第一个生物材料支架， 用于癌症免疫治疗的CD 8 + T细胞的体内抗原特异性活化。平台， 称为人工淋巴结（aLN），是一种透明质酸水凝胶与信号1 （肽-MHC）、2（抗-CD 28）和3（细胞因子支持物），其可以皮下注射以 创造一个T细胞激活的微环境。我们将研究3D支架的作用 参数的T细胞活化，以及深入了解体内抗原的动态， 在免疫活性宿主中特异性T细胞活化。 我们将开发可注射的aLN微粒（MP），其将在体内压缩以形成T 细胞活化支架。我们将首先研究aLN MP的物理性质，例如 信号密度、刚度和尺寸。我们还将研究细胞粘附蛋白的加入 以促进T细胞在支架内的迁移。第二，我们将在当地和 持续信号3（细胞因子）信号传导组分。我们将测试各种细胞因子，包括 IL-2、IL-7、IL-15和IL-21，因为它们产生效应细胞和记忆细胞的能力。与 我们将测试两种整合方法，抗体呈递和 封装这些aLN参数将针对鼠和人抗原两者进行优化。 特异性T细胞扩增。最后，我们将测试aLN MP用于体内活化的功效。 和CD 8 + T细胞的扩增及其抗癌功效，使用B16-OVA、B16-SIY和 小鼠T细胞的MC 38-OVA和人SK-MEL-37（A2+/NY-ESO-1+）黑素瘤细胞 人T细胞的细胞系。如果成功的话，这一提议将产生一种新的非细胞方法， 原位产生抗原特异性T细胞应答，扩大了 为更多患者提供免疫治疗。

Project Summary Adoptive T cell therapy (ACT) is a T cell based cancer therapy in which autologous T cells are isolated from the patient, activated and expanded ex vivo, then reinfused into the patient. While ACT has shown great clinical success, the success has been limited to melanoma, and complex manufacturing considerations create a large price tag. Innovations in in scalable, acellular systems for T cell activation, such as artificial antigen presenting cells (aAPCs), has improved the ex vivo expansion of CD8+ T cells by shortening culture times and providing tighter control of the resulting T cell phenotype and function. However, T cell culture with aAPCs still takes several weeks and requires manufacturing labor and cost. Platforms for in vivo activation of antigen-specific T cells would decrease the cost and complexity of T cell therapy. The goal of the proposed project is to create the first biomaterial scaffold for direct, in vivo, antigen-specific activation of CD8+ T cells for cancer immunotherapy. The platform, termed the artificial lymph node (aLN), is a hyaluronic acid hydrogel conjugated with signals 1 (peptide-MHC), 2 (anti-CD28) and 3 (cytokine support) that can be injected subcutaneously to create a T cell activating microenvironment. We will investigate the effects of 3D scaffold parameters on T cell activation as well as gain insight into the dynamics of in vivo antigen- specific T cell activation in an immune competent host. We will develop injectable aLN microparticles (MPs) that will compact in vivo to form a T cell activating scaffold. We will first investigate physical properties of the aLN MPs such as signal density, stiffness, and size. We will also investigate the addition of cell adhesion proteins to facilitate migration of the T cells within the scaffold. Second, we will incorporate a local and sustained signal 3 (cytokine) signaling component. We will test a variety of cytokines, including IL-2, IL-7, IL-15, and IL-21, for their ability to generate both effector and memory cells. With the lead cytokine cocktail, we will test two methods of integration, antibody presentation and encapsulation. These aLN parameters will be optimized for both murine and human antigen- specific T cell expansion. Finally, we will test the efficacy of the aLN MPs for in vivo activation and expansion of CD8+ T cells and their anti-cancer efficacy, using B16-OVA, B16-SIY, and MC38-OVA for mouse T cells, and the human SK-MEL-37 (A2+/NY-ESO-1+) melanoma cell line for human T cells. If successful, this proposal will produce a novel acellular approach for the in situ generation of an antigen-specific T cell response, expanding the access of immunotherapy to more patients.