Analyzing the Efficacy of Galectin-1 Ligand Inhibition in Adoptive T cell Therapy

描述（由申请人提供）：半乳糖凝集素-1（Gal-1）是一种对b-半乳糖具有亲和力的碳水化合物结合蛋白。Gal-1参与细胞粘附、迁移和凋亡。此外，已发现Gal-1与多种癌症相关。几个研究小组已经证明，黑色素瘤细胞分泌丰富的Gal-1。这一观察结果与Gal-1对辅助T细胞的众所周知的促凋亡和免疫调节作用一起导致了Gal-1通过诱导对黑素瘤抗原特异性CD 8 + T细胞的免疫抑制来增强肿瘤免疫逃逸机制的假设。 该项目的长期目标是研究在过继性T细胞治疗环境中消除Gal-1配体对黑色素瘤抗原特异性CD 8+细胞毒性T细胞（CTL）的功效。目前，过继性CD 8 + T细胞疗法在晚期黑色素瘤患者中显示出适度的改善。因此，在过继免疫治疗之前阻断抗原特异性CTL上的Gal-1-Gal-1配体相互作用可以帮助避免当前的缺点，例如T细胞对肿瘤来源的Gal-1的易感性，从而增强抗肿瘤功效。为此，我的目标是确定缺乏Gal-1配体的CTL是否可以在自体过继性T细胞转移环境中有效地减弱肿瘤生长。这些研究的结果将极大地影响改善过继免疫疗法治疗癌症的努力。 具体目标1。已发表的研究表明，Gal-1对T辅助细胞具有免疫调节作用。然而，关于Gal-1对CD 8 + T细胞的作用的研究很少。我的初步数据确定，CTL表达Gal-1配体，并进行Gal-1介导的调节。此外，我们发现Gal-1表达与疾病进展和无效的CD 8 + T细胞应答相关。这些数据导致了Gal-1抑制有助于肿瘤免疫逃逸的CTL的假设。为了验证这一假设，我将分析Gal-1介导的细胞因子表达，增殖潜力和凋亡活性对抗原特异性CTL的影响。 具体目标2。我实验室的研究表明，用Gal-1配体抑制剂4-F-GlcNAc治疗B16荷瘤小鼠，可产生抗肿瘤活性2。4-F-GlcNAc对许多类型的免疫细胞具有抗碳水化合物作用，因此我计划设计一种更有针对性的治疗方法。我将用4-F-GlcNAc体外处理CTL，然后将细胞转移到荷瘤小鼠体内。我推测用4-F-GlcNAc处理的CTL将免于Gal-1介导的细胞死亡和抑制，从而增加CTL水平并增强抗肿瘤活性。为了检验这一假设，我将使用FACS和ELISA在4-F-GlcNAc处理和未处理的CTL中分析细胞活力和细胞因子表达。我还将研究转移的CD 8 + T细胞在荷黑色素瘤小鼠中的抗肿瘤功效。总之，我的数据将探索用4-F-GlcNAc处理的CTL在过继性T细胞免疫疗法中的治疗效用。 公共卫生相关性：该项目的长期目标是研究在过继性T细胞治疗环境中消除半乳糖凝集素-1（Gal-1）配体对黑色素瘤抗原特异性CD 8+细胞毒性T细胞（CTL）的疗效。在过继性免疫疗法之前阻断抗原特异性CTL上的Gal-1 - Gal-1配体相互作用可以帮助规避当前的缺点，例如T细胞对肿瘤来源的Gal-1的易感性和体内寿命，从而增强抗肿瘤功效。

DESCRIPTION (provided by applicant): Galectin-1 (Gal-1), is a carbohydrate-binding protein with affinity for b-galactose. Gal-1 is involved in cell adhesion, migration, and apoptosis. In addition, Gal-1 has been found to be associated with a variety of cancers. Several groups have demonstrated that melanoma cells secrete an abundance of Gal-1. This observation taken together with Gal-1's well-known pro-apoptotic and immunoregulatory effects on helper T cells has led to the hypothesis that Gal-1 elicits tumor-immune escape mechanisms by inducing immunosuppression on melanoma antigen-specific CD8+ T cells. The long-term goal of this project is to study the efficacy of eliminating Gal-1 ligands on melanoma antigen-specific CD8+ cytotoxic T cells (CTLs) in an adoptive T cell therapy setting. Currently, adoptive CD8+ T cell therapy has shown modest improvements in patients with advanced melanoma. Thus, blocking Gal-1-Gal-1 ligand interactions on antigen-specific CTLs, prior to adoptive immunotherapy, could help avoid current shortcomings, such as T cell susceptibility to tumor-derived Gal-1, thereby enhancing anti-tumor efficacy. To this end, my goals are to determine whether CTLs lacking Gal-1 ligands can effectively attenuate tumor growth in an autologous adoptive T cell transfer setting. Results from these studies will greatly impact efforts to improve adoptive immunotherapy for treating cancer. Specific Aim 1. Published studies demonstrate that Gal-1 has immunoregulatory effects on T helper cells. However, few studies have been published about the role of Gal-1 on CD8+ T cells. My preliminary data establish that CTLs express Gal-1 ligands and undergo Gal-1-mediated modulation. Furthermore, we show that Gal-1 expression is correlated to disease progression and ineffective CD8+ T cell responses. These data has led to the hypothesis that Gal-1 suppresses CTLs contributing to tumor immune escape. To test this hypothesis, I will analyze the effects of Gal-1-mediated cytokine expression, proliferative potential and apoptotic activity on antigen-specific CTLs. Specific Aim 2. Studies in my lab have demonstrated that treatment of B16 tumor-bearing mice with Gal-1 ligand inhibitor, 4-F-GlcNAc, and results in anti-tumor activity2. 4-F-GlcNAc has anti-carbohydrate effects on many types of immune cells, so I plan to devise a more focused treatment. I will treat CTLs ex vivo with 4-F-GlcNAc and then transfer the cells into tumor-bearing mice. I hypothesize that CTLs treated with 4-F-GlcNAc will be spared from Gal-1-mediated cell death and suppression, thus increasing CTL levels and enhancing anti-tumor activity. To test this hypothesis, I will analyze for cell viability and cytokine expression using FACS and ELISA in 4-F-GlcNAc treated and untreated CTLs. I will also study the anti-tumor efficacy of the transferred CD8+ T cells in melanoma-bearing mice. Overall, my data will explore the therapeutic utility of CTLs treated with 4-F-GlcNAc in adoptive T cell immunotherapy. PUBLIC HEALTH RELEVANCE: The long-term goal of this project is to study the efficacy of eliminating Galectin-1 (Gal-1) ligands on melanoma antigen-specific CD8+ cytotoxic T cells (CTLs) in an adoptive T cell therapy setting. Blocking Gal-1 - Gal-1 ligand interactions on antigen-specific CTLs, prior to adoptive immunotherapy, could help circumvent current shortcomings, such as T cell susceptibility to tumor- derived Gal-1 and longevity in vivo, thereby enhancing anti-tumor efficacy.