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）： 在美国乃至全世界，在发达国家和发展中国家中，癌症的发病率和死亡率都大大增加。癌症导致约13％的人死亡，2007年有760万人死于癌症。美国死于肺癌的人多于乳腺癌，结肠，肾脏和前列腺癌的癌症。最近的研究表明，与未在军队中服役的人相比，退伍军人患肺癌的可能性高25％至75％。然而，肺癌和其他实体瘤的疗法仍然有限。最近的成功 固有的T细胞免疫疗法表明治疗的潜在突破。表达嵌合抗原受体或肿瘤反应性αβTCR的T细胞已治愈患有晚期转移性疾病的患者。可以使用MABS释放固有的T细胞免疫疗法器，以通过“检查点” CTLA-4和PD-1受体去除抑制作用，从而导致黑色素瘤和肺癌的多种治疗方法。然而，这些疗法存在重大局限性。治疗仅限于某些癌症，并非所有患者都对治疗做出反应，并且 尽管表现出巨大的希望，但仍需要对癌症免疫疗法的其他方法。用表达Vγ2VΔ2TCR的γδT细胞治疗就是一种这种疗法，显示出有望的疗法。与αβT细胞相反，表达Vγ2VΔ2TCR的γδT细胞的抗原反应不受MHC的限制。人γδT细胞的主要子集使用其Vγ2VΔ2T细胞接收器来识别外毛发异源性代谢产物HMBPP和自由状化代谢物IPP。来自多种组织的正常细胞和肿瘤细胞可以刺激Vγ2Vδ2细胞。在许多感染期间（多达2个循环T细胞中有1个），Vγ2VΔ2T细胞扩展到很高的数量，可以杀死肿瘤细胞和感染细胞以及秘密炎症细胞因子，趋化因子和生长因子。正在使用两种方法用Vγ2V​​δ2T细胞治疗癌症。首先是用刺激剂（例如HMBPP的溴氢化蛋白类似物）或低剂量IL-2的氨基膦酸盐酸氨酸酯类似物进行免疫。第二个是适当地转移离体生长的Vγ2VΔ2T细胞。这种方法已经治愈了患有转移性肾癌的患者，在另一种患有乳腺癌的另一种患者中诱发了缓解，并诱发了其他患者的部分缓解或稳定的疾病，但需要更有效。 细菌的代谢工程是一个新的研究领域，重点是改变细菌 药物或化学合成。细菌代谢的变化是通过修改生化途径或引入新的变化。现在，我们已经通过代谢工程沙门氏菌为这种方法提供了原则证明，以过多生产HMBPP并证明猴子的反应。现在，我们建议改善沙门氏菌疫苗并测试一种新的李斯特菌疫苗。这两种物种都被用于癌症疫苗，但由于口服沙门氏菌，而李斯特氏菌静脉注射给予了明显不同。我们将使用细菌疫苗靶向和激活肿瘤中适当转移的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：代谢工程的细菌过量产生HMBPP，在猴子中测试了工程细菌和体内，并评估了代谢工程细菌靶向并激活适当地转移的Vγ2Vδ2T细胞控制肿瘤的能力。我们拥有一支出色的团队，具有出色的往绩，并且具有与γδT细胞和类异丙代谢合作的丰富经验。我们已经建立了体内模型和技术。原则证明沙门氏菌疫苗已得出，结果最近发表了。产生更多疫苗的分子方法是良好的。总之，使用具有Vγ2VΔ2T细胞的代谢工程细菌阴道的免疫疗法具有广泛适用于通过直接激活和通过自适应转移来治疗许多不同肿瘤的可能性。