Functional Activation of Low Affinity TILs

项目摘要/摘要 在理解如何激活和增强低亲和力肿瘤的杀瘤活性方面存在根本性的差距 浸润性淋巴细胞(TIL)，识别来自失调的未突变蛋白的抗原。 这种间隙的持续存在是一个重要的问题，因为有许多肿瘤具有低的 突变负荷对目前的治疗不起作用，但引发低亲和力T细胞反应。我们的长期合作 目标是确定在TIL和表观遗传靶点中功能丰富的独特的抑制性受体 帮助建立有助于开发这些癌症未来治疗方法的机制。这个 这项研究的首要目标是阐明可用于提高抗肿瘤T细胞的途径 活性，特别是低亲和力TIL的活性。中心假说是低分子化合物的抗肿瘤活性。 亲和力T细胞通过阻断抑制性受体和靶向表观遗传调节而对增强敏感 地区。这一假设是基于申请者在 实验室。这项拟议研究的基本原理是，理解 低亲和力T细胞有可能转化为更好地理解癌症的更好治疗方法 现在这个国家每两到三个人中就有一个人受到这种疾病的困扰。在强劲的初步数据的指导下，这一假设 将通过追求两个特定的目标来进行测试：1)建立调节低分子药物抗肿瘤活性的机制。 亲和力TIL通过定义其抑制受体，以及2)建立表观遗传机制 在低亲和力TIL中编程。这两个目标都将使用回溯基因技术来产生大量的 高亲和力或低亲和力的肿瘤特异性T细胞。在第一个目标中，将从列表中识别抑制受体 研究人员发现的含有ITIM的基因在TIL中上调，并使用生化方法 利用SH2结构域从低亲和力T细胞中识别抑制性受体的方法。人类的基因 在这个目标中确定的兴趣将在其他系统中得到确认，并在人类结直肠样本上进行探测，以 确定它们在人类TIL上的表达。在第二个目标中，基因表达和染色质可及性 外周和肿瘤中的低亲和力和高亲和力T细胞将被检测。T细胞发育对人类免疫功能的影响 肿瘤抗原缺陷小鼠也将接受检查。在申请人看来，这种方法是创新的， 因为它不同于使用新奇技术查看针对肿瘤的最高亲和力反应的现状 方法：研究方法。拟议的研究的贡献将是重大的，因为识别抑制 有望激活内源性低亲和力T细胞的受体和表观遗传靶点毒性可能较小 并激活一些尚未体验到免疫疗法好处的肿瘤中的T细胞。最终， 这些知识有可能为目前还不起作用的癌症的免疫疗法的发展提供信息。 接受治疗。

PROJECT SUMMARY/ABSTRACT There is a fundamental gap in understanding how to activate and increase tumoricidal activity low-affinity tumor infiltrating lymphocytes (TILs) that recognize antigens derived from dysregulated unmutated proteins. Continued existence of this gap represents an important problem because there are many tumors with a low mutation load that do not respond to current treatments, but elicit low-affinity T cell responses. Our long-term goal is to identify unique inhibitory receptors that are functionally enriched in TILs and epigenetic targets to help establish mechanisms that contribute to development of future therapies for these cancers. The overarching goal of this research is to elucidate pathways that can be harnessed to improve antitumor T cell activity, particularly the activity of low-affinity TILs. The central hypothesis is that the antitumor activity of low- affinity T cells is sensitive to enhancement by blocking inhibitory receptors and targeting epigenetic regulatory regions. This hypothesis has been formulated on the basis of preliminary data produced in the applicants' laboratories. The rationale for the proposed research is that understanding the activation and differentiation of low-affinity T cells has the potential to translate into better understanding of better therapies for cancer that now afflicts one of every two to three people in this country. Guided by strong preliminary data, this hypothesis will be tested by pursuing two specific aims: 1) Establish mechanisms that modulate antitumor activity of low- affinity TILs by defining their inhibitory receptor repertoire, and 2) Establish mechanisms of epigenetic programming in low-affinity TILs. Both of these aims will use retrogenic technology to generate large pools of either high or low-affinity tumor-specific T cells. In the first aim, inhibitory receptors will be identified from a list of ITIM-containing genes that were found by the investigators to be upregulated in TILs and using biochemical approaches that identifies inhibitory receptors from low-affinity T cells using SH2 domains. The genes of interest identified in this aim will be confirmed in other systems and probed on human colorectal samples to determine their expression on human TILs. In the second aim, gene expression and chromatin accessibility of low and high affinity T cells in the periphery and tumor will be examined. The effects of T cells development in tumor antigen-deficient mice will also be examined. The approach is innovative, in the applicant's opinion, because it departs from the status quo of looking at the highest affinity reactions against tumors using novel methods. The contribution of the proposed research will be significant because identification of inhibitory receptors and epigenetic targets expected to activate endogenous low-affinity T cells may have fewer toxicities and activate T cells in some tumors that have not yet experienced the benefits of immunotherapies. Ultimately, such knowledge has the potential to inform the development of immunotherapies for cancers that do not as yet have treatments.