Characterizing and Targeting the Hypoxic T Cell Surfaceome to Promote Immune Function in Cancer

项目摘要 最近的努力集中在开发基于T细胞的免疫疗法来治疗癌症。然而，这些 治疗有几个局限性，包括对实体肿瘤的最小疗效。的一个特点是 实体瘤微环境为低氧可利用性，或低氧。人们普遍认为，肿瘤缺氧 抑制抗肿瘤免疫反应。该项目的目标是提高我们对T如何 细胞对低氧作出反应，并揭示了提高低氧T细胞抗肿瘤功能的新策略。 这些发现将最终指导T细胞疗法的工程学，以克服缺氧性肿瘤 微环境。调节抗肿瘤免疫功能的两个关键T细胞亚型是促炎性T细胞 效应器(Tef)和抗炎T调节(Treg)细胞。之前的研究考察了 低氧对这些细胞类型的影响是高度不一致的。阐明低氧对TREF和Treg功能的影响 扩大我们对基本T细胞生物学的理解，并有可能找到增强抗肿瘤T细胞的新方法 细胞在低氧条件下的活性。在这里，我们假设低氧改变T细胞表面蛋白(“表面体”)和 T细胞的功能与净免疫抑制效应一致。为了检验这一假设，我们将 利用对原代人类T细胞、动物模型和患者样本的研究相结合。我们会 利用我们已建立的基于表面组学和噬菌体展示的重组抗体工程策略 研究低氧对可疑的低氧诱导抗原(HIA)的影响，并鉴定新的HIA。 我们将首先将这些技术应用于在常氧或低氧条件下培养的原代人胸腺和Treg细胞。 体外低氧条件。然后我们将检查来自人源化的低氧肿瘤浸润性T细胞 小鼠胰腺癌模型以及患者胰腺癌标本。这种表面上的轮廓， 结合T细胞增殖、激活和迁移研究，将提供一个全面的图景 低氧诱导T细胞表面蛋白和功能的变化。此外，我们将设计新的抑制物 针对可疑和新发现的HIA的抗体或双特异性构建物。这些都是精心设计的 蛋白质将在低氧培养和人源化胰腺中测试TJeff/Treg的调节功能 肿瘤小鼠模型，最终目标是确定促进低氧Tef功能的策略和 免疫介导的肿瘤杀伤。从长远来看，基本的生物学见解和抗体工具从 这项拟议的研究将为正在进行的使用T细胞治疗癌症的努力提供信息，并将为 提高缺氧性肿瘤免疫靶向性的未来疗法。

Project Summary Recent efforts have focused on developing T cell-based immune therapies to treat cancer. However, these therapies have several limitations, including minimal efficacy against solid tumors. One characteristic of the solid tumor microenvironment is low oxygen availability, or hypoxia. It is generally thought that tumor hypoxia suppresses the anti-tumor immune response. The goal of this project is to improve our understanding of how T cells respond to hypoxia and reveal new strategies for increasing the anti-tumor function of hypoxic T cells. These findings will ultimately inform the engineering of T cell therapies to overcome the hypoxic tumor microenvironment. Two key T cell subtypes that modulate anti-tumor immune function are pro-inflammatory T effector (Teff) and anti-inflammatory T regulatory (Treg) cells. Previous studies examining the effects of hypoxia on these cell types are highly discordant. Clarifying the effects of hypoxia on Teff and Treg function will expand our understanding of basic T cell biology and potentially identify new means of enhancing anti-tumor T cell activity in hypoxia. Here, we hypothesize that hypoxia alters T cell surface proteins (the “surfaceome”) and T cell function in a manner consistent with a net immunosuppressive effect. To test this hypothesis, we will utilize a combination of studies with primary human T cells, animal models, and patient samples. We will employ our established surfaceomic and phage display-based recombinant antibody engineering strategies to examine the effect of hypoxia on suspected hypoxia-induced antigens (HIAs) as well as to identify novel HIAs. We will first apply these techniques to isolated primary human Teff and Treg cells cultured in normoxic or hypoxic conditions in vitro. We will then examine hypoxic tumor-infiltrating T cells derived from humanized mouse models of pancreatic cancer as well as patient pancreatic tumor samples. This surfaceomic profiling, combined with T cell proliferation, activation, and migration studies, will provide a comprehensive picture of hypoxia-induced T cell surface protein and functional changes. Furthermore, we will engineer novel inhibitory antibodies or bispecific constructs targeting both suspected and newly-identified HIAs. These engineered proteins will be tested for Teff/Treg modulatory function in hypoxic culture and in a humanized pancreatic cancer mouse model with the ultimate goal of identifying strategies to promote hypoxic Teff function and immune-mediated tumor killing. In the long term, the basic biological insights and antibody tools gained from the proposed studies will inform ongoing efforts to treat cancer using T cells and will lay the foundation for future therapeutics to boost immune targeting of hypoxic tumors.