Genetically engineering T cells to overcome the immunosuppressive melanoma microenvironment

摘要 虽然检查点免疫疗法在过去改变了转移性黑色素瘤的治疗 十年来，这种疗法未能为大多数患者提供持久的疗效。免疫治疗耐药 实体瘤的发生通常是由免疫抑制的肿瘤微环境(TME)驱动的，TME抑制了 传入T细胞的抗肿瘤能力。在这项提案中，我的目标是通过基因工程改造T细胞，以克服一种关键 免疫抑制的途径，我用一个无偏见的生物信息学筛选确定的。我的目标是提高 CRISPR基因敲除T细胞过继转移对其表达的影响 抑制性受体表达于T细胞表面。我已经开发出一种方法来敲除这种基因 在小鼠和人类T细胞中有效，通过这个提议，我将确定基因敲除的效果 关于黑色素瘤中T细胞的内在适合性和功能(目标1)、T细胞的聚集和功能 微环境(目标2)和T细胞抗肿瘤效果(目标3)。我预计这项研究将产生一个 转化法改善过继转移T细胞在免疫抑制中的功能 黑色素瘤TME。我们的团队已经展示了将新的T细胞疗法从实验室应用到临床的能力， 我们已建立的用基因工程T细胞治疗患者的计划将使翻译变得容易。

ABSTRACT While checkpoint immunotherapy has transformed the treatment of metastatic melanoma over the past decade, this therapy fails to provide durable responses for the majority of patients. Immunotherapy resistance in solid tumors is often driven by the immunosuppressive tumor microenvironment (TME), which dampens the anti-tumor capacity of incoming T cells. In this proposal, I aim to genetically engineer T cells to overcome a key pathway of immunosuppression which I identified using an unbiased bioinformatics screen. I aim to improve the function of adoptively transferred T cells by using CRISPR to knockout the expression of this key immune inhibitory receptor expressed on the T cell surface. I have already developed a method to knockout this gene efficiently in murine and human T cells, and through this proposal, I will determine the effect of gene knockout on intrinsic T cell fitness and function (Aim 1), T cell accumulation and function in the melanoma microenvironment (Aim 2), and T cell anti-tumor efficacy (Aim 3). I expect that this research will yield a translational method to improve the function of adoptively transferred T cells in the immunosuppressive melanoma TME. Our team has demonstrated the ability to take novel T cell therapies from the lab to the clinic, and our established program to treat patients with genetically-engineered T cells will ease translation.