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: 9206071
• 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/9206071
• 关键词: Adoptive Immunotherapy; Adoptive Transfer; Age; Alpha Cell; Anabolism; 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 Factor; Gulf War; Human; Immunity; Immunization; Immunize; Immunodeficient Mouse; Immunotherapy; In Vitro; Incidence; Individual; Infection; Inflammation; Inflammatory; Interleukin-2; Intravenous; 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; Oral; Partial Remission; Pathology; Pathway interactions; Patients; Pattern recognition receptor; Persian Gulf; Play; Primates; Publishing; Renal carcinoma; Risk; 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; World War II; Zoledronate; adaptive immunity; alpha-beta T-Cell Receptor; 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; immune checkpoint blockade; 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; γδ T cells

 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，癌症的发病率和死亡率在发达国家和发展中国家都急剧增加。2007年，癌症导致约13%的人类死亡，760万人死于癌症。在美国，死于肺癌的人数比乳腺癌、结肠癌、肾癌和前列腺癌的总和还要多。最近的研究表明，退伍军人患肺癌的可能性比未在军队服役的人高25%至75%;然而，肺癌和其他实体瘤的治疗仍然有限。 最近取得的成功 在T细胞癌中，免疫疗法指出了治疗上潜在突破。表达嵌合抗原受体或肿瘤反应性αβ TCR的T细胞治愈了晚期转移性疾病患者。可以使用mAb释放针对肿瘤的固有T细胞免疫以消除“检查点”CTLA-4和PD-1受体的抑制，这已经导致黑色素瘤和肺癌的许多治愈。然而，这些疗法存在明显的局限性。治疗仅限于某些癌症，并非所有患者都对治疗有反应， 毒性显著。虽然显示出巨大的希望，但还需要更多的癌症免疫治疗方法。 用表达Vγ 2 VS 2 TCR的γδ T细胞治疗是一种显示出前景的此类疗法。与αβ T细胞相反，表达Vγ 2 V δ2 TCR的γδ T细胞的抗原反应不受MHC限制。人γδ T细胞的主要亚群使用其Vγ 2 V δ2 T细胞受体来识别外来微生物类异戊二烯代谢物HMBPP和自身代谢物IPP。来自多种组织的正常细胞和肿瘤细胞可以刺激Vγ 2 VS 2细胞。Vγ 2 VS 2 T细胞在许多感染期间扩增至非常高的数量（高达1/2的循环T细胞），并且可以杀死肿瘤细胞和感染的细胞以及分泌炎性细胞因子、趋化因子和生长因子。目前有两种方法用于用Vγ 2 V δ2 T细胞治疗癌症。第一种是用刺激剂如HMBPP的溴代醇类似物或氨基二膦酸盐、唑来膦酸盐与低剂量IL-2免疫。第二种是过继转移离体生长的Vγ 2 VS 2 T细胞。这种方法治愈了一名转移性肾癌患者，在另一名乳腺癌患者中诱导缓解，并在其他患者中诱导部分缓解或稳定疾病，但需要更有效。 细菌的代谢工程是一个新的研究领域，其重点是改变细菌， 药物或化学合成。细菌代谢的变化是通过改变生物化学途径或引入新的途径来实现的。我们现在已经通过代谢工程沙门氏菌过量生产HMBPP并在猴子中证明反应来证明这种方法的原理。我们现在建议改进沙门氏菌疫苗，并测试一种新的李斯特菌疫苗。这两个物种都被用于癌症疫苗，但差异显着，因为沙门氏菌是口服，而李斯特菌是静脉注射。我们将使用细菌疫苗靶向并激活肿瘤中过继转移的Vγ 2 V δ 2 T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：代谢工程化细菌以过量生产HMBPP，在猴子中体外和体内测试工程化细菌，并评估代谢工程化细菌靶向和激活过继转移的Vγ 2 VS 2 T细胞以控制肿瘤的能力。我们拥有一支优秀的团队，拥有出色的业绩记录，并在γδ T细胞和类异戊二烯代谢方面拥有丰富的经验。我们建立了体内模型和技术。一种沙门氏菌疫苗的原理证明已经被推导出来，结果最近发表。制造更多疫苗的分子方法已经发展得很好。总之，使用具有Vγ 2 VS 2 T细胞的代谢工程化细菌疫苗的免疫疗法具有通过直接活化和通过过继转移广泛适用于治疗许多不同肿瘤的潜力。