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

有效地将联合收割机T细胞定向免疫疗法与抑制肿瘤促进信号转导途径结合起来治疗黑素瘤的策略的开发代表了当前最重要的临床挑战之一。本提案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，我们将调查的一般假设，IFN-驱动的细胞毒性CD 8 + T细胞免疫是有限的局部在黑色素瘤微环境中的生理保护反应和黑色素瘤细胞的免疫抑制活性。这些反调节机制包括将髓样免疫细胞募集到受损的肿瘤组织中，刺激PD 1/PDL 1免疫抑制受体相互作用，以及通过肿瘤细胞中致癌信号传导途径的活性产生免疫抑制环境。实验工作旨在干扰这些机制，并分为三个部分，其目的如下：（i）表征I型IFN在调节抗肿瘤CD 8 + T细胞应答中的作用;（ii）建立体内生物发光成像技术，以非侵入性地评估用过继转移的细胞毒性CD 8+调节黑色素瘤组织浸润的治疗策略的功效T细胞和随后的骨髓免疫细胞的募集;和（iii）扩展我们的模型系统并探索将联合收割机过继性CD 8 + T细胞疗法与BRAF（V600 E）信号转导抑制剂相结合的治疗方案。所提出的实验将产生对将联合收割机T细胞免疫疗法与肿瘤促进信号转导通路的抑制剂相结合的可能性的基本见解。这将为正在进行的和未来的转移性黑色素瘤患者的临床转化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.