Experimental development of strategies that effectively combine T cell immunotherapy with the inhibition of tumor-promoting signal transduction pathways for the treatment of melanoma

发展有效结合T细胞定向免疫治疗和抑制促肿瘤信号转导通路治疗黑色素瘤的策略是当前最重要的临床挑战之一。这项建议的主要目标是使用最先进的临床前基因工程小鼠模型对这些策略进行实验评估。在我们的工作中，我们将调查一般假设，即干扰素驱动的细胞毒性CD8+T细胞免疫在黑色素瘤微环境中受到生理保护反应和黑色素瘤细胞免疫抑制活性的局部限制。这些反调节机制包括髓系免疫细胞在受损的肿瘤组织中的募集，刺激PD1/PDL1免疫抑制受体的相互作用，以及通过肿瘤细胞中的致癌信号通路的活性而产生免疫抑制环境。实验工作旨在干扰这些机制，分为三个部分，目的如下：(I)鉴定I型IFN在调节抗肿瘤CD8+T细胞反应中的作用；(Ii)建立体内生物发光成像技术，以非侵入性地评估过继转移的细胞毒性CD8+T细胞调节黑色素瘤组织渗透以及随后髓系免疫细胞的招募的治疗策略的有效性；(Iii)扩展我们的模型系统，探索过继转移的CD8+T细胞治疗与BRAF(V600E)信号转导抑制剂相结合的治疗方案。拟议中的实验将对T细胞免疫疗法与促进肿瘤信号转导途径的抑制剂相结合的可能性产生基本的见解。这将为正在进行的和未来转移性黑色素瘤患者的临床翻译研究提供有价值的信息。

The development of strategies that effectively combine T cell directed immunotherapy with the inhibition of tumor-promoting signal transduction pathways for the treatment of melanoma represents one of the most important current clinical challenges. The principle goal of this proposal is to experimentally evaluate such strategies using state-of-the-art preclinical genetically engineered mouse models. In our work we will investigate the general hypothesis that IFN-driven cytotoxic CD8+ T cell immunity is limited locally in the melanoma microenvironment by both physiologic protective responses and by the immunosuppressive activity of melanoma cells. These counter-regulatory mechanisms include the recruitment of myeloid immune cells into injured tumor tissue, the stimulation of PD1/PDL1 immune-inhibitory receptor interactions, and the generation of an immunosuppressive milieu by the activity of oncogenic signalling pathways in tumor cells. The experimental work is directed at interfering with these mechanisms and is divided in three parts with the following aims: (i) to characterize the role of type I IFNs for the regulation of anti-tumoral CD8+ T cell responses; (ii) to establish in vivo bioluminescence imaging techniques to non-invasively evaluate the efficacy of therapeutic strategies that modulate the infiltration of melanoma tissue with adoptively transferred cytotoxic CD8+ T cells and the subsequent recruitment of myeloid immune cells; and (iii) to expand our model system and explore treatment protocols that combine adoptive CD8+ T-cell therapies with BRAF(V600E) signal transduction inhibitors. The proposed experiments will yield fundamental insights into the possibilities to combine T cell immunotherapies with the inhibitors of tumor-promoting signal transduction pathways. This will provide valuable information for ongoing and future clinical translational studies in patients with metastatic melanoma.