Grail-targeting breaks the immune tolerance to melanoma

圣杯靶向打破了对黑色素瘤的免疫耐受

• 批准号: 9379209
• 负责人: Roza Insafetdinovna Nurieva
• 金额: $ 8万
• 依托单位: UNIVERSITY OF TX MD ANDERSON CAN CTR
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-07-01 至 2019-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9379209
• 关键词: Biological Response Modifiers; CD4 Positive T Lymphocytes; CD8-Positive T-Lymphocytes; Cancer Etiology; Cause of Death; Cell physiology; Cells; Clinical; Data; Development; Future; Gene Expression Microarray Analysis; Growth; Human; Immune; Immune Tolerance; Immune response; Immune system; Immunosuppressive Agents; Immunotherapy; Implant; In Vitro; Incidence; Laboratories; Lead; Lymphoma; Malignant Neoplasms; Melanoma Cell; Methods; Molecular; Mus; Neoplasm Metastasis; Obstruction; Outcome; Patients; Property; Reagent; Regulation; Regulatory T-Lymphocyte; Relapse; Research; Resistance; Risk Factors; Role; Sampling; Small-Cell Lymphoma; System; T-Lymphocyte; Technology; Testing; Therapeutic; Time; Tumor Antigens; Tumor Tissue; Work; base; cancer immunotherapy; cancer type; cytotoxic; cytotoxicity; improved; in vivo; innovation; insight; knockout gene; melanoma; memory recall; mouse model; neoplastic cell; novel; novel therapeutic intervention; pre-clinical; prevent; response; screening; tool; tumor; ubiquitin-protein ligase; yeast two hybrid system

Abstract Efficacy of anti-cancer immunotherapy is limited due to various tumor evasion mechanisms including the tolerance induction of tumor-reactive T cells. Therefore, strategies to break T cell tolerance and thereafter enhance T-cell cytotoxicity towards tumor cells are needed. One such innovative and rational approach is to manipulate the intrinsic properties of T cells by suppressing the activity of tolerance-associated factors. We have acknowledged the E3 ubiquitin ligase, Grail as an essential component of T-cell tolerance and showed that Grail deficiency resulted in CD4+ T cell hyper-responsiveness and defective regulatory T cell suppressive function. Thus, targeting Grail may help to break tumor immune tolerance; however, the role of Grail in tumor development and in the function of cytotoxic CD8+ T lymphocytes (CTLs) remains unknown. Our preliminary data show that loss of Grail enhanced cytolytic functionality of CTLs. Furthermore, utilizing lymphoma tumors, we have determined that Grail-deficient CTLs are essential for control of established tumor. Remarkably, Grail expression level was significantly higher in CTLs from lymphoma patients, suggesting the novel role of Grail in immune tolerance to lymphoma and providing a rationale for exploring the role of Grail in melanoma that is often observed in a lymphoma setting. Further understanding the function of Grail in different cancer types will be beneficial for development of common therapeutic approach for these malignancies. We have evidence that Grail deficiency in mice confers control of implanted B16 melanoma tumor. Moreover, in melanoma patient samples, we found high Grail expression in CD8+ T cells from the tumor tissues, suggesting that Grail expression in the immune cells could be a big obstruction for controlling melanoma as well. Therefore, we propose to investigate the therapeutic potential of Grail-targeted CTLs for melanoma as well as their regulatory mechanisms using both mouse and human systems. In Aim 1, we will investigate the role of Grail KO CTLs in melanoma growth control by utilizing gene knockout approaches. We will also assess the mechanism(s) whereby Grail controls anti-tumor function and persistence of CTLs. Moreover, we will try to identify targets through which Grail facilitates control of tumor-reactive CD8+ T cell function by employing cutting edge technologies, including two-hybrid screening and microarray analysis of gene expression. In Aim 2, we will establish an efficient Grail targeting approach for human and mouse CD8+ T cells in order to improve the efficiency of adoptive therapy for established malignancies. Overall the proposed research will help to validate Grail as a functional mediator of immune tolerance in melanoma as well as evaluate the therapeutic potential of Grail deficient CD8+ T cells for melanoma treatment. The implications from this work are significant since the results will provide mechanistic insights into the understanding of the function of Grail in melanoma and will potentially lead to the development of novel targeted treatments to reverse immune tolerance in melanoma and other malignancies.

摘要 抗癌免疫疗法的功效由于各种肿瘤逃避机制而受到限制，所述肿瘤逃避机制包括免疫抑制剂。 肿瘤反应性T细胞的耐受诱导。因此，打破T细胞耐受性的策略以及此后 需要增强T细胞对肿瘤细胞的细胞毒性。其中一个创新和合理的方法是 通过抑制耐受相关因子的活性来操纵T细胞的内在特性。 我们已经认识到E3泛素连接酶，Grail作为T细胞耐受性的重要组成部分， 结果表明，Grail缺陷导致CD 4 + T细胞高反应性和调节性T细胞缺陷， 抑制功能因此，靶向Grail可能有助于打破肿瘤免疫耐受;然而， 圣杯在肿瘤发展和细胞毒性CD 8 + T淋巴细胞（CTL）的功能仍然未知。 我们的初步数据显示，Grail的丢失增强了CTL的细胞溶解功能。此外，利用 在淋巴瘤肿瘤中，我们已经确定Grail缺陷型CTL对于控制已建立的肿瘤是必需的。 值得注意的是，Grail表达水平在来自淋巴瘤患者的CTL中显著更高，这表明， Grail在淋巴瘤免疫耐受中的新作用，并为探索Grail在淋巴瘤免疫耐受中的作用提供了理论基础。 黑色素瘤，通常在淋巴瘤环境中观察到。进一步了解圣杯在不同领域的作用 癌症类型将有利于开发这些恶性肿瘤的共同治疗方法。我们 有证据表明，小鼠中的Grail缺陷可以控制植入的B16黑色素瘤。而且在 在黑色素瘤患者样本中，我们发现Grail在来自肿瘤组织的CD 8 + T细胞中的高表达，表明 Grail在免疫细胞中的表达也可能是控制黑色素瘤的一大障碍。 因此，我们建议研究Grail靶向CTL对黑色素瘤的治疗潜力， 以及使用小鼠和人类系统的调节机制。 在目的1中，我们将通过利用基因敲除来研究Grail KO CTL在黑色素瘤生长控制中的作用 接近。我们还将评估Grail控制抗肿瘤功能和持久性的机制 的CTL。此外，我们将尝试确定Grail促进肿瘤反应性CD 8+细胞控制的靶点。 通过采用尖端技术，包括双杂交筛选和微阵列分析， 的基因表达。在目标2中，我们将建立一种有效的人和小鼠的Grail靶向方法， CD 8 + T细胞，以提高过继治疗已确立的恶性肿瘤的效率。 总的来说，拟议的研究将有助于验证Grail作为免疫耐受的功能介质， 本发明还提供了用于治疗黑色素瘤的方法，以及评估Grail缺陷型CD 8 + T细胞用于黑色素瘤治疗的治疗潜力。 这项工作的意义是重大的，因为结果将提供机械的见解， 了解Grail在黑色素瘤中的功能，并可能导致开发新的 逆转黑色素瘤和其他恶性肿瘤免疫耐受的靶向治疗。