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: 10516094
• 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 2024-09-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10516094
• 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; 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; Populations at Risk; Primates; Prodrugs; Prostatic Neoplasms; Proteins; Protozoan Infections; Renal carcinoma; 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; 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; response; 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.

在美国和全世界，癌症发病率和死亡率在#年急剧上升。 无论是发达国家还是发展中国家。癌症占人类死亡的13%，760万人死于癌症 在2007年死于癌症。在美国，死于肺癌的人比死于乳腺癌、结肠癌、肾癌和前列腺癌的人多 加在一起。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%到75% 那些没有在军队服役的人。癌症免疫治疗的进展正在导致在 治疗。过继转移表达嵌合抗原受体(CAR-T)的T细胞可获得持久的再治疗。 B细胞恶性肿瘤的诊断。使用PD-1、PD-L1和CTLA-4抗体阻断检查点可导致 多种恶性肿瘤患者的部分和完全反应。然而，存在着重大的限制。使用 除了黑色素瘤患者，只有少数患者对检查点阻断有反应。普普通通 前列腺癌和结直肠癌等癌症通常不起作用。因此，其他方法有 需要实现癌症免疫疗法的全部潜力。 用γδT细胞表达V-γ-2V-δ-2TCR就是这样一种方法。与αβT细胞不同，这种反应 Vγ2Vδ2T细胞的数量不受MHC的限制，而是需要Ig超家族蛋白BuyroPhilin 3A1来 感觉外来微生物的异戊二烯代谢物HMBPP和自身代谢物IPP。这种感测允许 肿瘤细胞被Vγ2Vδ2T细胞识别和杀伤，而不受其突变负荷的影响。Vγ2Vδ2 T 在许多感染期间，细胞安全地膨胀到非常高的数量(高达2个循环T细胞中的1个)，在那里它们会杀死 感染细胞并分泌炎性Th1细胞因子、趋化因子和生长因子。 使用Vγ2Vδ2T细胞治疗癌症有两种方法。第一种是用刺激免疫-- 如HMBPP的溴代醇类似物或带有低剂量IL-1的氨基双膦酸唑来膦酸。 2.虽然治疗导致部分缓解，但这些疫苗最终导致无效性和缺失性 Vγ2Vδ2T细胞。二是过继转移Vγ2Vδ2T细胞。这种方法是安全的，并且在- 3例实体瘤患者获完全缓解，部分缓解或稳定消失。 安逸地对待别人。然而，要得到广泛采用，Vγ2Vδ2T细胞疗法需要更有效。 活细菌疫苗已被用于预防结核病、伤寒和兔热病。细菌 产生激活先天免疫的化合物和刺激αβT细胞的抗原，以帮助 Vγ2Vδ2 T细胞扩增。我们现在已经鉴定出一种减毒李斯特菌菌株，它一直在- 扩增Vγ2Vδ2T细胞。李斯特菌优先聚集在肿瘤中，这应该允许特定的焦油- 过继转移的V-γ-2V-δ-2T细胞在肿瘤中的获得及肿瘤部位的TcR刺激 由HMBPP提供。我们还发现，Vγ2Vδ2T细胞快速表达检查点受体，如CTLA-4，PD-4。 1、TIM-3和LAG-3在刺激时，加入PD-1检查点阻断剂显著增强 V-γ-2V-δ-2T细胞免疫小鼠前列腺癌的实验研究我们还确认了一部小说 双膦酸类前体药物。为了实现这些目标，我们将：删除ΔActa PRFA*(G155S)Listeria中的inlB 疫苗接种和评估免疫剂量和时间，评估李斯特菌靶向和活性的能力 VATE过继转移V-γ-2V-δ-2T细胞控制肿瘤并评估联合检查点的效果 Blockade和过继转移Vγ2Vδ2T细胞的新的PTA双膦酸前药。 我们有一支优秀的团队，有着出色的业绩记录，并拥有丰富的合作经验 γδT细胞与类异戊二烯代谢。我们已经建立了体内模型和技术。一个有效的列表- RIA疫苗已经确定。制造更多疫苗的分子方法已经开发得很好了。在狱中- 结论：V-γ-2V-δ-2T细胞细菌疫苗免疫治疗具有广阔的应用前景 用于多种不同肿瘤的治疗，包括直接激活和增强过继转移。

项目成果

Expansion of human γδ T cells for adoptive immunotherapy using a bisphosphonate prodrug.

• DOI: 10.1111/cas.13491
• 发表时间: 2018-03
• 期刊: Cancer science
• 影响因子: 5.7
• 作者: Tanaka Y;Murata-Hirai K;Iwasaki M;Matsumoto K;Hayashi K;Kumagai A;Nada MH;Wang H;Kobayashi H;Kamitakahara H;Okamura H;Sugie T;Minato N;Toi M;Morita CT
• 通讯作者: Morita CT