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

肝细胞癌（HCC）是第五大常见癌症，也是全球癌症相关死亡的第三大原因。发生与预后不良相关，治疗选择有限。免疫疗法在其他癌症类型，如黑色素瘤或B细胞恶性肿瘤中显示出有希望的结果。然而，HCC的免疫抑制治疗方法的有限成功至少部分是由于肝脏中固有的免疫抑制微环境，并且在实体肝肿瘤中甚至更明显。因此，开发克服肝脏耐受状态并刺激肿瘤特异性T细胞的新策略至关重要。因此，拟议研究的重点将是开发和表征新型T细胞受体（TCR）信号传导响应性纳米凝胶制剂，用于在肝细胞癌小鼠模型中使用过继性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性细胞因子或抗体（例如抗PD-1、抗CTLA-4、抗IL-10、抗TGF β、OX 40配体或IL-12）通过蛋白质分子的交联和引入氧化还原敏感性二硫键来合成。将研究所产生的纳米凝胶制剂靶向和结合T细胞质膜的能力及其在TCR连接后释放蛋白质货物的潜力。在下一步中，纳米凝胶将在体外抗原特异性CD 8 + T细胞的活化和随后对抗原表达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CD 8 + T细胞将过继转移到具有抗原表达肝细胞的受体小鼠中或携带肝细胞癌肿瘤模型的小鼠中。将通过测量过继转移的T细胞的肝归巢和肝内扩增、肝细胞的抗原特异性裂解、肿瘤生长和荷瘤小鼠的存活率来分析治疗成功。

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.