Novel immunotherapeutic regimen combining the dendritic cell expansion power of Albumin-Flt3L and inflammatory cues of Salmonella

新型免疫治疗方案结合了白蛋白-Flt3L 的树突状细胞扩增能力和沙门氏菌的炎症信号

• 批准号: 10041196
• 负责人: Chien-Fu Hung
• 金额: $ 20.47万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-07-01 至 2022-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10041196
• 关键词: Adjuvant; Affect; Albumins; Animal Model; Antigens; Attenuated; Bacteria; Biological; Blood Vessels; Bone Marrow; CD4 Positive T Lymphocytes; CD44 gene; CD8-Positive T-Lymphocytes; Cells; Chimeric Proteins; Clonality; Cross Presentation; Cross-Priming; Cues; Cytotoxic T-Lymphocytes; Data; Dendritic Cells; Development; Drug Kinetics; Enterobacteriaceae; Exhibits; FLT3 ligand; Future; Generations; Genetic; HPV E7; Half-Life; Head and Neck Cancer; Human Papillomavirus; Hypoxia; Image; Immune; Immunity; Immunotherapeutic agent; In Situ; In Vitro; Inflammation; Inflammatory; Injections; Interferon Type II; Ligands; Literature; Lymphocyte; Malignant Neoplasms; Mediating; Medical; Modeling; Molecular; Monitor; Morbidity - disease rate; Mus; Mutation; Names; Neoplasm Transplantation; Outcome; Papillomavirus Transforming Protein E6; Pattern; Perfusion; Phenotype; Population; Positron-Emission Tomography; Pre-Clinical Model; Process; Property; Public Health; Recycling; Regimen; Resolution; Role; Salmonella; Serum Proteins; Stains; System; T cell response; T-Cell Proliferation; T-Lymphocyte; T-Lymphocyte Epitopes; TNF gene; Therapeutic; Tissues; Tracer; Transplantation; Treatment Efficacy; Treatment Protocols; Tumor Burden; Tumor Immunity; Tumor-Derived; Vaccination; Viral; antigen-specific T cells; antitumor effect; cancer therapy; cancer type; chemoradiation; cytokine; cytotoxic CD8 T cells; design; effective therapy; experimental study; fetal liver kinase-2; imaging agent; improved; in vivo; in vivo imaging system; innovation; lymph nodes; lymphoid neoplasm; mouse model; neonatal Fc receptor; neoplastic cell; novel; oral HPV-positive head and neck cancers; pathogen; pre-clinical; response; secondary lymphoid organ; source localization; trafficking; transplant model; treatment strategy; tumor; tumor progression; uptake

Project Summary/Abstract The overwhelming public health burden of HPV-associated head and neck squamous cell carcinoma has created great demand for novel, broadly effective therapies with reduced treatment morbidity and improved long-term survival. The generation of tumor-specific CD8+ T cell immunity requires potent antigen cross-presentation by dendritic cells (DCs) since tumor cells do not efficiently present relevant CD8+ T cell epitopes. cDC1s are known for their ability to process exogenous antigen and potently cross-present to CD8+ T cells. Because of this, innovative strategies to enhance cDC1s could robustly induce HPV-specific immunity and have great therapeutic potential in the treatment of HPV-associated HNC. FMS-like tyrosine kinase 3 ligand (Flt3L) is a cytokine that expands and differentiates DC precursors to murine cDC1s, but therapeutic potential of Flt3L is limited because of its short half-life and global distribution in vivo. We have overcome the described issues of Flt3L by generating a genetic fusion of Albumin (Alb) to Flt3L, named Albumin-Flt3L (Alb-Flt3L). Alb has a long half-life due to FcRn- mediated transcytolic recycling and exhibits selective trafficking to the LN and TM. Once cDC1s expand, they require a strong, tissue localized source of inflammation for activation, or they will present T cell epitopes without adequate costimulation, causing suppression. Enteric bacteria such as Salmonella serve as an ideal agent for tumor specific cDC1 activation as they can provide numerous pathogen-associated molecular patterns for cDC1 activation, and have been described to colonize the tumor efficiently likely due to hypoxia. One caveat of bacteria therapy in cancer is that consideration must be made regarding preclinical model selection, as colonization varies greatly between spontaneous and transplantable tumor models. Our preliminary data shows that Alb-Flt3L fusion protein exhibits increased half-life and selective accumulation in LN and tumor compared to native Flt3L. Alb- Flt3L is able to expand cross-presenting DC populations in vivo following a single injection and leads to enhanced HPV-specific T cells in a vaccination model and tumor control in a proof-of-principle experiment. The Salmonella strain used in our experiments successfully colonizes the tumor and can efficiently activate Alb-Flt3L-derived DCs. In this proposal, the ability of Alb-Flt3L + Salmonella to promote HPV-specific cytotoxic T cell antitumor immunity through the expansion and activation of cross-presenting DCs and subsequent tumor control in our spontaneous and transplantable HPV-associated HNC models will be investigated. HPV-specific CD8+ T cell responses will be evaluated using tetramer and TCR sequencing approaches. The mechanism by which Alb- Flt3L mediates its immunostimulatory function will also be interrogated using appropriate deficient mouse models. Studies regarding colonization of Salmonella in spontaneous vs transplantable tumor models will be investigated using our high-resolution nanoPET/CT system and appropriate PET tracers. Successful completion of this proposal will generate valuable preclinical and mechanistic data regarding the therapeutic potential of Alb- Flt3L + Salmonella as a universal strategy to treat HPV-associated HNC as well as many other types of cancer.

项目摘要/摘要 与HPV相关的头部和颈部鳞状细胞癌的压倒性公共卫生负担已经产生 对新颖的，广泛有效的疗法的巨大需求，以减少治疗的发病率并改善了长期 生存。肿瘤特异性CD8+ T细胞免疫的产生需要通过 树突状细胞（DC）由于肿瘤细胞没有有效地呈现相关的CD8+ T细胞表位。 CDC1是已知的 因为它们能够处理外源性抗原并有效地与CD8+ T细胞交叉。因为这， 增强CDC1的创新策略可以牢固地诱导HPV特异性免疫力并具有出色的治疗性 治疗与HPV相关的HNC的潜力。 FMS样酪氨酸激酶3配体（FLT3L）是一种细胞因子 扩展并区分DC前体与鼠CDC1，但FLT3L的治疗潜力有限，因为 其在体内的短期半衰期和全球分布。我们通过产生了FLT3L的描述问题 白蛋白（ALB）与FLT3L的遗传融合，称为白蛋白 - FLT3L（ALB-FLT3L）。由于fcrn- 介导的跨环循环利用并表现出对LN和TM的选择性贩运。一旦CDC1扩展，他们 需要强大的组织局部炎症来源以进行激活，否则它们会呈现T细胞表位而无需 足够的共刺激，导致抑制。肠道细菌（例如沙门氏菌）是理想的药物 肿瘤特异性CDC1激活，因为它们可以为CDC1提供许多与病原体相关的分子模式 激活，并已描述为由于缺氧而有效地定居肿瘤。一个警告 癌症的治疗是必须考虑临床前模型的选择，因为定植变化 在自发性和可移植肿瘤模型之间进行了极大的影响。我们的初步数据表明Alb-Flt3L融合 与天然FLT3L相比，蛋白质在LN和肿瘤中表现出增加的半衰期和选择性积累。 alb 单一注射后，FLT3L能够在体内扩展交叉呈递的直流种群，并导致增强 疫苗接种模型中的HPV特异性T细胞和原则证明实验中的肿瘤控制。沙门氏菌 我们的实验中使用的菌株成功地殖民了肿瘤，并可以有效激活Alb-Flt3L衍生 DCS。在此提案中，Alb-Flt3L +沙门氏菌促进HPV特异性细胞毒性T细胞抗肿瘤的能力 通过扩展和激活交叉呈递DC的免疫力以及我们在我们的随后的肿瘤控制 将研究自发和可移植HPV相关的HNC模型。 HPV特异性CD8+ T细胞 将使用四聚体和TCR测序方法评估响应。 Alb-的机制 FLT3L介导其免疫刺激功能也将使用适当的缺陷小鼠询问 型号。关于自发性与移植肿瘤模型中沙门氏菌定殖的研究将是 使用我们的高分辨率纳米特/CT系统和适当的宠物示踪剂进行了研究。成功完成 该提案将产生有关Alb-的治疗潜力的有价值的临床前和机械数据 FLT3L +沙门氏菌是治疗与HPV相关的HNC以及许多其他类型的癌症的普遍策略。