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: 8922337
• 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: 2010
• 资助国家: 美国
• 起止时间: 2010-10-01 至 2019-09-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8922337
• 关键词: Accounting; Adoptive Immunotherapy; Adoptive Transfer; Age; Antigens; Attenuated; Bacteria; Bacterial Vaccines; Biochemical Pathway; Blood Cells; Cancer Etiology; Cancer Vaccines; Cells; Cessation of life; Colon Carcinoma; Cytotoxic T-Lymphocyte-Associated Protein 4; Developing Countries; Diphosphates; Disease; Disease remission; Dose; Engineering; Environmental Hazards; Enzymes; Freezing; Goals; Growth; Gulf War; Human; Immunity; Immunization; Immunodeficient Mouse; Immunotherapy; In Vitro; Incidence; Individual; Infection; Inflammation; Inflammatory; Interleukin-2; Koreans; Ligands; Listeria; Listeria monocytogenes; Lymphocyte; Lymphoma; Malignant Neoplasms; Malignant neoplasm of lung; Malignant neoplasm of prostate; Membrane Proteins; Metabolism; Methods; Microbe; Military Personnel; Molecular; Monkeys; Natural Immunity; Normal Cell; Partial Remission; Pathology; Pathway interactions; Patients; Pattern recognition receptor; Persian Gulf; Play; Primates; Publishing; Renal carcinoma; Risk; Rodent; Role; Salmonella; Salmonella Vaccines; Salmonella enterica; Salmonella typhi; Salmonella typhimurium; Site; Solid Neoplasm; Stable Disease; T cell response; T cell therapy; T-Cell Receptor; T-Lymphocyte; Techniques; Testing; Tissues; Toxic effect; United States; Vaccines; Veterans; Vietnam; Work; World War II; Zoledronate; adaptive immunity; analog; anergy; bacterial metabolism; cancer immunotherapy; cancer type; cell transformation; chemical synthesis; chemokine; chimeric antigen receptor; cytokine; drug synthesis; experience; field study; high risk; humanized mouse; improved; in vivo; in vivo Model; inorganic phosphate; isopentenyl pyrophosphate; isoprenoid; killings; malignant breast neoplasm; melanoma; men; metabolic engineering; mevalonate; microbial; mortality; neoplastic cell; prenyl; public health relevance; receptor; response; success; tumor

 DESCRIPTION (provided by applicant): 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; yet therapies for lung cancer and other solid tumors are still limited. Recent successes in T cell cancer immunotherapy point to a potential breakthrough in treatment. T cells expressing chimeric antigen receptors or tumor-reactive αβ TCRs have cured patients with advanced metastatic disease. Intrinsic T cell immunity against tumors can be released using mAbs to remove inhibition by "checkpoint" CTLA-4 and PD-1 receptors which has resulted in a number of cures in melanoma and lung cancer. Yet, significant limitations exist for these therapies. Therapy is limited to certain cancers, not all patients respond to therapy, and there is significant toxicity. Although showing great promise, additional approaches to cancer immunotherapy are needed. Treatment with γδ T cells expressing Vγ2Vδ2 TCRs is one such therapy that shows promise. In contrast to αβ T cells, the antigen responses of γδ T cells expressing Vγ2Vδ2 TCRs are not MHC restricted. The major subset of human γδ T cells use their Vγ2Vδ2 T cell receptors to recognize the foreign-microbial isoprenoid metabolite, HMBPP, and the self-metabolite, IPP. Normal cells and tumor cells from a wide variety of tissues can stimulate Vγ2Vδ2 cells. Vγ2Vδ2 T cells expand to very high numbers during many infections (up to 1 in 2 circulating T cells) and can kill tumor cells and infected cells as well as secrete inflammatory cytokines, chemokines, and growth factors. Two approaches are being used to treat cancer with Vγ2Vδ2 T cells. The first is to immunize with stimulators such as the bromohydrin analog of HMBPP or the aminobisphosphonate, zoledronate, with low-dose IL-2. The second is to adoptively transfer Vγ2Vδ2 T cells grown ex vivo. This approach has cured a patient with metastatic kidney cancer, induced remission in another with breast cancer, and induced partial remissions or stable disease in other patients but needs to be made more effective. Metabolic engineering of bacteria is a new field of study that has focused on altering bacteria for drug or chemical synthesis. Changes in bacterial metabolism are made by modifying biochemical pathways or by introducing new ones. We have now provided proof-of-principle for this approach by metabolic engineering Salmonella to overproduce HMBPP and demonstrating responses in monkeys. We now propose to improve our Salmonella vaccine and to test a new Listeria vaccine. Both species have been used for cancer vaccines but differ significantly because Salmonella is given orally whereas Listeria is given intravenously. We will use the bacterial vaccines to target and activate adoptively transferred Vγ2Vδ 2 T cell in tumors because they preferentially localize to tumor cells. To accomplish our goals, we will: metabolically engineer bacteria to overproduce HMBPP, test engineered bacteria in vitro and in vivo in monkeys, and assess the ability of metabolically engineered bacteria to target and activate adoptively transferred Vγ2Vδ2 T cells to control tumors. 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. A proof-of-principle Salmonella vaccine has been derived and the results recently published. The molecular methods to create more vaccines are well developed. In conclusion, immunotherapy using metabolically engineered bacterial vaccines with Vγ2Vδ2 T cells has the potential to be broadly applicable for the treatment of many different tumors both by direct activation and through adoptive transfer.

 描述(由申请人提供)： 在美国和全世界，发达国家和发展中国家的癌症发病率和死亡率都大幅上升。癌症约占人类死亡的13%，2007年有760万人死于癌症。在美国，死于肺癌的人数超过了乳腺癌、结肠癌、肾癌和前列腺癌的总和。最近的研究表明，退伍军人患肺癌的可能性比没有在军队服役的人高25%到75%；然而，肺癌和其他实体肿瘤的治疗方法仍然有限。最近的成功案例 在T细胞癌方面，免疫治疗指出了治疗方面的潜在突破。表达嵌合抗原受体或肿瘤反应性αβTCR的T细胞已经治愈了晚期转移疾病的患者。抗肿瘤的内在T细胞免疫可以用mAbs释放，以消除“检查点”CTLA-4和PD-1受体的抑制，后者已导致许多黑色素瘤和肺癌的治愈。然而，这些疗法存在很大的局限性。治疗仅限于某些癌症，并不是所有的患者对治疗都有反应，而且有 毒性很大。尽管显示出巨大的希望，但还需要更多的癌症免疫治疗方法。用γδT细胞表达V-γ-2V-δ-2TCR是一种有希望的治疗方法。与αβT细胞相比，表达V-γδ-2V-γ-2TCR的δ-T细胞的抗原反应不受MHC的限制。人类γδT细胞的主要亚群使用它们的Vγ2Vδ2T细胞受体来识别外来微生物的异戊二烯代谢产物hMBPP和自身代谢产物ipp。来自多种组织的正常细胞和肿瘤细胞都能刺激Vγ2Vδ2细胞。Vγ2Vδ2T细胞在许多感染过程中扩增到非常高的数量(高达2个循环T细胞中的1个)，并能杀死肿瘤细胞和感染细胞，以及分泌炎性细胞因子、趋化因子和生长因子。使用Vγ2Vδ2T细胞治疗癌症有两种方法。第一种是用刺激物免疫，如HMBPP的溴代醇类似物或带有低剂量IL-2的氨基双膦酸唑来膦酸盐。二是过继转移体外培养的V-γ-2V-δ-2T细胞。这种方法已经治愈了一名转移性肾癌患者，诱导了另一名乳腺癌患者的缓解，并诱导了其他患者的部分缓解或稳定疾病，但需要更有效。 细菌代谢工程是一个新的研究领域，它专注于改变细菌以 药物或化学合成。细菌新陈代谢的变化是通过改变生化途径或引入新途径来实现的。我们现在已经通过代谢工程沙门氏菌来过量生产HMBPP并在猴子身上展示了反应，为这一方法提供了原则上的证据。我们现在提议改进我们的沙门氏菌疫苗，并测试一种新的李斯特菌疫苗。这两种细菌都被用于癌症疫苗，但有很大的不同，因为沙门氏菌是口服的，而李斯特菌是静脉注射的。我们将使用细菌疫苗在肿瘤中靶向和激活过继转移的Vγ2Vδ2T细胞，因为它们优先定位于肿瘤细胞。为了实现我们的目标，我们将：代谢工程细菌过度生产HMBPP，在猴子体内和体外测试工程菌，并评估代谢工程菌靶向和激活过继转移的Vγ2Vδ2T细胞以控制肿瘤的能力。我们有一支出色的团队，有着出色的记录，并拥有丰富的γδT细胞和类异戊二烯代谢工作经验。我们已经建立了体内模型和技术。一种原理验证的沙门氏菌疫苗已经衍生出来，并于最近公布了结果。制造更多疫苗的分子方法已经开发得很好了。综上所述，使用Vγ2Vδ2T细胞代谢工程细菌疫苗进行免疫治疗，无论是通过直接激活还是通过过继转移，都有可能广泛应用于多种不同肿瘤的治疗。