T cell receptor signaling-responsive protein nanogels for the T cell-mediated treatment of solid liver tumors

肝细胞癌（HCC）是世界上第五大最常见的癌症形式，也是导致癌症相关死亡的第三大原因。发生与预后不良有关，治疗选择有限。免疫疗法在其他类型的癌症，如黑色素瘤或B细胞恶性肿瘤中显示出令人鼓舞的效果。然而，HCC免疫治疗方法的有限成功至少部分是由于肝脏固有的免疫抑制微环境，在实体肝肿瘤中更为明显。因此，开发新的策略来克服肝脏的耐受状态并刺激肿瘤特异性T细胞是非常重要的。因此，该研究的重点将是开发和表征新型T细胞受体（TCR）信号应答纳米凝胶制剂，用于在小鼠肝癌模型中使用过继T细胞转移进行免疫治疗。纳米凝胶将基于不同的免疫刺激细胞因子或抗体，如抗pd -1、抗ctla -4、抗il -10、抗tgf β、OX40配体或IL-12，通过蛋白质分子交联和引入氧化还原敏感的二硫键来合成。所生成的纳米凝胶制剂靶向和结合T细胞质膜的能力以及它们在TCR连接后释放蛋白质货物的潜力将被研究。下一步，纳米凝胶将在体外对抗原特异性CD8+ T细胞的激活以及随后对表达抗原的肝细胞的杀伤方面进行评估。在最后的研究中，纳米凝胶负载的CD8+ T细胞将被过继地转移到具有抗原表达肝细胞的受体小鼠或具有肝细胞癌肿瘤模型的小鼠中。治疗成功将通过测量肝脏归巢和过继转移T细胞的肝内扩张、抗原特异性肝细胞裂解、肿瘤生长和荷瘤小鼠的存活来分析。

Hepatocellular carcinoma (HCC) is the fifth most common form of cancer and the third leading cause of cancer-related deaths worldwide. Occurrence is associated with poor prognosis and the treatment options are limited. Immunotherapy has shown promising results in other cancer types, such as melanoma or B cell malignancies. However, the limited success of immunotherapeutic treatment approaches for HCC is at least partly due to the immunosuppressive microenvironment inherent in the liver and even more pronounced in solid liver tumors. As such, the development of novel strategies to overcome the state of tolerance in the liver and to stimulate tumor-specific T cells is of great importance. Therefore, the focus of the proposed study will be the development and characterization of novel T cell receptor (TCR) signaling-responsive nanogel formulations for immunotherapeutic treatment using adoptive T cell transfer in a mouse model of hepatocellular carcinoma. Nanogels will be synthesized based on different immunostimulatory cytokines or antibodies, such as anti-PD-1, anti-CTLA-4, anti-IL-10, anti-TGFβ, OX40 ligand or IL-12, by crosslinking of protein molecules and introducing redox-sensitive disulphide bonds. The capability of the generated nanogel formulations to target and bind to the plasma membrane of T cells and their potential to release protein cargo upon TCR ligation will be investigated. In a next step, nanogels will be evaluated in terms of activation of antigen-specific CD8+ T cells in vitro and the subsequent killing of antigen-expressing hepatocytes. In final investigations, the nanogel-loaded CD8+ T cells will be adoptively transferred in recipient mice with antigen-expressing hepatocytes or in mice bearing a tumor model of hepatocellular carcinoma. Treatment success will be analyzed measuring liver homing and intrahepatic expansion of adoptively transferred T cells, the antigen-specific lysis of hepatocytes, tumor growth, and the survival of tumor-bearing mice.