Remotely controlled listerial bactodrones for cancer immunotherapy

用于癌症免疫治疗的远程控制李斯特菌杆菌

• 批准号: 10318673
• 负责人: Jason Gigley
• 金额: $ 15.94万
• 依托单位: UNIVERSITY OF WYOMING
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-01-01 至 2023-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10318673
• 关键词: 4T1; Actins; Agonist; Antigen-Presenting Cells; Attenuated; Bacteria; Benign; Blood; Breast Cancer Cell; Breast Cancer Model; Breast Cancer cell line; CCL2 gene; CD8-Positive T-Lymphocytes; Cancer Biology; Cancer Model; Cause of Death; Cell Death; Cells; Cervical; Chemicals; Clinical Trials; Cytosol; Dinucleoside Phosphates; Disseminated Malignant Neoplasm; Engineering; Ensure; Enzymes; Genes; Genetic; Genetic Engineering; Goals; Human; Immune; Immune checkpoint inhibitor; Immunooncology; Immunotherapy; In Vitro; Inflammatory Response; Injections; Innate Immune System; Interferon Type I; Interferon-beta; Interferons; Interleukin-12; Interleukin-2; Intravenous; Lead; Listeria monocytogenes; Malignant Neoplasms; Mediating; Metastatic breast cancer; Modality; Mus; Myeloid-derived suppressor cells; Natural Immunity; Neoplasm Metastasis; Non-Small-Cell Lung Carcinoma; Nucleotides; Pathway interactions; Performance; Periodicity; Plasma; Plasmids; Pre-Clinical Model; Primary Neoplasm; Process; Production; Prostate; Protocols documentation; Risk; STING agonists; Signal Transduction; Site; System; T-Lymphocyte; TNF gene; Technology; Testing; Therapeutic; Tissues; Toxic effect; Treatment Cost; Tumor Antigens; anti-cancer; base; cancer immunotherapy; cell motility; checkpoint therapy; chemical synthesis; cytokine; delivery vehicle; design; diguanylate cyclase; dimer; efficacy testing; genetic payload; immunogenic; improved; in vivo; innovation; intravenous injection; malignant breast neoplasm; neoplastic cell; protocol development; recruit; response; synthetic biology; tumor; tumor growth; tumor microenvironment; tumor-immune system interactions

PROJECT SUMMARY Attenuated and avirulent strains of Listeria monocytogenes (Lm), that are delivered via intravenous injections, accumulate and propagate in primary tumors and metastases while being quickly cleared from healthy tissues. We intend to use these strains as remotely controlled, tumor-specific anticancer payload delivery vehicles, bactodrones. In this project we will engineer Lm to synthesize and secrete cyclic dinucleotides (c-di-NMPs) as potent innate immune system stimulators inside tumor microenvironments. On-site accumulation of c-di-NMPs will induce production of type I interferon via the STING innate immunity pathway. This will improve the capacity of Lm to induce immunogenic tumor cell death and lead to the release of tumor-associated antigens, which will facilitate recruitment of tumor-specific CD8 T cells. The sustained tumor-localized c-di-NMP production will keep T cells and other anticancer immune cells activated. To assess feasibility and efficacy of delivering intratumoral c-di-NMP via genetically engineered Lm, we will pursue two aims. In aim 1, we will engineer Lm to secrete enzymes for c-di-NMP synthesis in immune and tumor cells. In aim 2, the engineered Lm will deliver plasmids encoding a c-di-NMP synthases, via a process known as bactofection. Both approaches are expected to turn infected cells in the tumor microenvironment into c-di-NMP producing factories and ensure durable STING activation. Importantly, Lm-mediated c-di-NMP delivery systems will be made inducible with a benign chemical inducer, which will enable temporal control of STING activation and limit toxicity associated with systemic c-di-NMP exposure. Following optimization of the Lm bactodrones in vitro, and in breast cancer cell line, we will test efficacy of periodic bactodrone injections in a mouse metastatic breast cancer model. We anticipate that Lm bactodrones will become efficient vehicles for tumor-localized, temporally controlled and inexpensive delivery of genetic payloads for various antitumor activities.

项目摘要 李斯特氏菌菌株（LM）的衰减和无毒的菌株，这些菌株是通过静脉注射的， 在原发性肿瘤和转移中积聚并传播，同时迅速从健康组织中清除。 我们打算将这些菌株用作远程控制的，肿瘤特异性的抗癌有效载荷递送车辆， Bactodrones。在这个项目中，我们将设计LM以合成和分泌环状二核苷酸（C-DI-NMP）为 肿瘤微环境内有效的先天免疫系统刺激剂。 C-DI-NMP的现场积累 将通过刺痛的先天免疫途径诱导I型干扰素的产生。这将改善 LM诱导免疫原性肿瘤细胞死亡的能力并导致肿瘤相关抗原的释放， 这将促进肿瘤特异性CD8 T细胞的募集。持续肿瘤定位的C-DI-NMP 生产将使T细胞和其他抗癌免疫细胞激活。评估可行性和功效 通过基因工程LM传递肿瘤内C-DI-NMP，我们将追求两个目标。在AIM 1中，我们将 LM工程师分泌酶以在免疫细胞和肿瘤细胞中分泌C-DI-NMP合成。在AIM 2中，工程 LM将通过称为Bactofection的过程传递编码C-DI-NMP合酶的质粒。两个都 预计方法将在肿瘤微环境中将受感染的细胞转化为产生C-DI-NMP 工厂并确保持久的刺激激活。重要的是，LM介导的C-DI-NMP输送系统将是 用良性化学诱导剂诱导，这将使刺激激活的时间控制和 限制与全身性C-DI-NMP暴露有关的毒性。在优化LM Bactodrones之后 体外和乳腺癌细胞系中，我们将测试小鼠转移性注射周期性双子酮的功效 乳腺癌模型。我们预计LM Bactodrones将成为有效的肿瘤定位的车辆， 各种抗肿瘤活动的遗传有效载荷的时间控制和廉价地交付。