Metabolic Engineering of Bacteria for Cancer Immunotherapy by Gamma Delta T Cells

Gamma Delta T 细胞用于癌症免疫治疗的细菌代谢工程

• 批准号: 10412920
• 负责人: CRAIG T MORITA
• 金额: --
• 依托单位: IOWA CITY VA MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-10-01 至 2023-09-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10412920
• 关键词: Adoption; Adoptive Immunotherapy; Adoptive Transfer; Anabolism; Antibodies; Antigens; Attenuated; B lymphoid malignancy; Bacteria; Bacterial Infections; Bacterial Vaccines; Bladder; Blood Cells; CAR T cell therapy; CTLA4 gene; Cancer Etiology; Cells; Cessation of life; Colon Carcinoma; Colorectal; Colorectal Cancer; Developed Countries; Developing Countries; Diphosphates; Disease remission; Dose; Effectiveness; Environmental Hazards; Exposure to; Goals; Growth Factor; Gulf War; Human; Immunity; Immunization; Immunize; Immunodeficient Mouse; Immunoglobulins; Immunotherapy; In complete remission; Incidence; Infection; Inflammatory; Interleukin-2; Kidney; Koreans; Listeria; Listeria monocytogenes; Lung; Lymphoma; Malignant Neoplasms; Malignant neoplasm of lung; Malignant neoplasm of prostate; Metabolism; Methods; Microbe; Military Personnel; Minority; Molecular; Monitor; Monkeys; Mutation; Natural Immunity; Partial Remission; Pathway interactions; Patients; Pattern recognition receptor; Persian Gulf; Persons; Play; Primates; Prodrugs; Prostatic Neoplasms; Proteins; Protozoan Infections; Renal carcinoma; Risk; Role; Site; Solid Neoplasm; Stable Disease; T cell response; T cell therapy; T-Lymphocyte; Techniques; Testing; Tissues; Tuberculosis; Tularemia; Tumor Immunity; Typhoid Fever; United States; Vaccines; Veterans; Vietnam; World War II; Zoledronic Acid; analog; anergy; bisphosphonate; butyrophilin; cancer immunotherapy; cancer type; cell killing; checkpoint receptors; chemokine; chimeric antigen receptor; cytokine; effectiveness evaluation; experience; high risk; immune checkpoint blockade; improved; in vivo Model; inorganic phosphate; intravenous injection; isopentenyl pyrophosphate; isoprenoid; malignant breast neoplasm; melanoma; metabolic engineering; mevalonate; microbial; mortality; mouse model; neoantigens; neoplastic cell; novel; pamidronate; partial response; prenyl; prevent; programmed cell death ligand 1; programmed cell death protein 1; tumor; γδ T cells

In the United States and throughout the world, cancer incidence and mortality has increased dramatically in both developed and developing nations. Cancer causes ~13% of human deaths with 7.6 million people dying from cancer in 2007. More people in the US die of lung cancer than breast, colon, kidney, and prostate cancers combined. Recent studies show that veterans are 25 to 75 percent more likely to develop lung cancer than people who did not serve in the military. Advances in cancer immunotherapy are leading to breakthroughs in treatment. Adoptive transfer of T cells expressing chimeric antigen receptors (CAR-T) results in durable remis- sions for B cell malignancies. Checkpoint blockade with antibodies against PD-1, PD-L1, and CTLA-4 results in partial and complete responses in patients with a variety of malignancies. Yet, significant limitations exist. With the exception of patients with melanoma, only a minority of patients respond to checkpoint blockage. Common cancers such as prostate and colorectal cancer generally do not respond. Thus, additional approaches are needed to realize the full potential of cancer immunotherapy. Treatment with γδ T cells expressing Vγ2Vδ2 TCRs is one such approach. Unlike αβ T cells, the response of Vγ2Vδ2 T cells is not MHC restricted but instead requires the Ig superfamily protein, butyrophilin 3A1, to sense the foreign-microbial isoprenoid metabolite, HMBPP, and the self-metabolite, IPP. This sensing allows tumor cells to be recognized and killed by Vγ2Vδ2 T cells independent of their mutational burden. Vγ2Vδ2 T cells safely expand to very high numbers during many infections (up to 1 in 2 circulating T cells) where they kill infected cells and secrete inflammatory Th1 cytokines, chemokines, and growth factors. Two approaches are being used to treat cancer with Vγ2Vδ2 T cells. The first is to immunize with stimula- tors such as the bromohydrin analog of HMBPP or the aminobisphosphonate zoledronic acid with low-dose IL- 2. Although treatment has resulted in partial remissions, these vaccines eventually cause anergy and deletion of the Vγ2Vδ2 T cells. The second is to adoptively transfer Vγ2Vδ2 T cells. This approach is safe and has in- duced complete remissions in three patients with solid tumors, and induced partial remissions or stable dis- ease in others. However, for widespread adoption, Vγ2Vδ2 T cell therapy needs to be more effective. Live bacterial vaccines have been used to prevent tuberculosis, typhoid fever, and tularemia. The bacteria produce compounds that activate innate immunity and antigens that stimulate αβ T cells to provide help to the Vγ2Vδ2 T cells as they expand. We have now identified an attenuated Listeria strain that consistently ex- pands Vγ2Vδ2 T cells. Listeria preferentially accumulate in tumors which should allow the specific tar- geting of adoptively-transferred Vγ2Vδ2 T cells to tumors as well as TCR stimulation at the tumor site by HMBPP. We also find that Vγ2Vδ2 T cells rapidly express checkpoint receptors such as CTLA-4, PD- 1, TIM-3, and LAG-3 upon stimulation and that adding PD-1 checkpoint blockade markedly enhances prostate tumor immunity by Vγ2Vδ2 T cells in a mouse model. We have also identified a novel bisphosphonate prodrug. To accomplish these goals, we will: delete inlB in ΔactA prfA* (G155S) Listeria vaccine and assess dosing and timing of immunization, assess the ability of Listeria bacteria to target and acti- vate adoptively transferred Vγ2Vδ2 T cells to control tumors, and assess effectiveness of combining checkpoint blockade and a new PTA bisphosphonate prodrug with adoptively transferred Vγ2Vδ2 T cells. We have an outstanding team with an excellent track record and have extensive experience working with γδ T cells and isoprenoid metabolism. We have established in vivo models and techniques. An effective Liste- ria vaccine have been identified. The molecular methods to create more vaccines are well developed. In con- clusion, immunotherapy using bacterial vaccines for Vγ2Vδ2 T cells has the potential to be broadly applicable for the treatment of many different tumors both by direct activation and through potentiating adoptive transfer.

在美国和世界各地，癌症发病率和死亡率急剧增加 发达国家和发展中国家约有 13% 的人死于癌症，其中 760 万人死于癌症。 2007 年死于癌症。美国死于肺癌的人数多于乳腺癌、结肠癌、肾癌和前列腺癌 最近的研究表明，退伍军人患肺癌的可能性比退伍军人高 25% 至 75%。 未曾参军的人在癌症免疫治疗方面取得了突破。 表达嵌合抗原受体 (CAR-T) 的 T 细胞的过继转移可导致持久缓解。 使用针对 PD-1、PD-L1 和 CTLA-4 的抗体阻断检查点会导致 B 细胞恶性肿瘤的发生。 然而，对多种恶性肿瘤患者的部分和完全缓解存在显着的局限性。 除黑色素瘤患者外，只有少数患者对常见检查点阻断有反应。 前列腺癌和结直肠癌等癌症通常没有反应，因此需要采取其他方法。 需要充分发挥癌症免疫疗法的潜力。 使用表达 Vγ2Vδ2 TCR 的 γδ T 细胞进行治疗就是一种此类方法，其反应与 αβ T 细胞不同。 Vγ2Vδ2 T 细胞的功能不受 MHC 限制，而是需要 Ig 超家族蛋白、嗜丁酸蛋白 3A1 感测外来微生物类异戊二烯代谢物 HMBPP 和自身代谢物 IPP 这种感测允许。 肿瘤细胞被 Vγ2Vδ2 T 细胞识别并杀死，与其突变负荷无关。 在许多感染期间，细胞安全地扩增到非常高的数量（高达二分之一的循环 T 细胞），并杀死它们 感染细胞并分泌炎症性 Th1 细胞因子、趋化因子和生长因子。 有两种方法用于使用 Vγ2Vδ2 T 细胞治疗癌症，第一种方法是通过刺激进行免疫。 诸如 HMBPP 的溴醇类似物或氨基二膦酸唑来膦酸与低剂量 IL- 2. 虽然治疗已导致部分缓解，但这些疫苗最终会导致无反应和缺失 第二种是过继转移 Vγ2Vδ2 T 细胞。这种方法安全且具有以下优点。 诱导三名实体瘤患者完全缓解，并诱导部分缓解或稳定缓解 然而，为了广泛采用，Vγ2Vδ2 T 细胞疗法需要更有效。 活细菌疫苗已用于预防结核病、伤寒病和兔热病。 产生激活先天免疫的化合物和刺激 αβ T 细胞的抗原，为 当 Vγ2Vδ2 T 细胞扩增时，我们现在已经鉴定出一种减毒的李斯特菌菌株，该菌株始终会消失。 Pands Vγ2Vδ2 T 细胞优先在肿瘤中积聚，这应该允许特定的目标。 将 Vγ2Vδ2 T 细胞过继转移至肿瘤并在肿瘤部位进行 TCR 刺激 我们还发现 Vγ2Vδ2 T 细胞快速表达检查点受体，例如 CTLA-4、PD- 1、TIM-3 和 LAG-3 受到刺激后，添加 PD-1 检查点阻断可显着增强 我们还在小鼠模型中发现了一种新的 Vγ2Vδ2 T 细胞免疫前列腺肿瘤的方法。 为了实现这些目标，我们将： 删除 ΔactA prfA* (G155S) 李斯特菌中的 inlB。 疫苗并评估免疫接种的剂量和时间，评估李斯特菌细菌靶向和激活的能力 过继转移Vγ2Vδ2 T细胞来控制肿瘤，并评估联合检查点的有效性 阻断和一种新的 PTA 双膦酸盐前药与过继转移的 Vγ2Vδ2 T 细胞。 我们拥有一支出色的团队，拥有出色的业绩记录，并拥有丰富的合作经验 γδ T 细胞和类异戊二烯代谢我们已经建立了有效的 Liste- 体内模型和技术。 ria 疫苗已被确定，用于制造更多疫苗的分子方法已得到很好的开发。 结论是，使用细菌疫苗针对 Vγ2Vδ2 T 细胞的免疫疗法具有广泛适用的潜力 通过直接激活和增强过继转移来治疗许多不同的肿瘤。