Defining a cross-primed anti-tumor T cell signature to guide immunotherapy development

定义交叉引发的抗肿瘤 T 细胞特征来指导免疫疗法的开发

• 批准号: 10677596
• 负责人: Gabrielle Lubitz
• 金额: $ 4.61万
• 依托单位: ICAHN SCHOOL OF MEDICINE AT MOUNT SINAI
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-07-11 至 2025-07-10
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10677596
• 关键词: Address; Adoptive Transfer; Allogenic; Animal Model; Antigen Presentation; Antigen-Presenting Cells; Antigens; Antitumor Response; Autoantigens; Automobile Driving; CD8-Positive T-Lymphocytes; CXCL9 gene; Cell Separation; Cell physiology; Cross Presentation; Cross-Priming; Defect; Dendritic Cells; Development; Exhibits; FLT3 ligand; Failure; Flow Cytometry; Immune response; Immunologic Monitoring; Immunotherapy; Inbred BALB C Mice; Interferon Type II; Interleukin-12; Interleukin-2; Intrinsic factor; Knock-out; Luciferases; Lymphoma; Measures; Mediating; Methods; Modeling; Monitor; Mus; Mutation; Outcome; Outcome Study; PD-1 blockade; Patients; Phenotype; Radiation therapy; Resistance; Role; Sorting; T cell response; T cell therapy; T-Cell Activation; T-Lymphocyte; Techniques; Tissues; Transfection; Tumor Antigens; Vaccination; anti-PD-1; antigen-specific T cells; antitumor effect; cancer infiltrating T cells; cancer therapy; chemokine; cytotoxicity; design; effective therapy; immune checkpoint blockade; improved; in situ vaccination; in situ vaccine; in vivo; malignant breast neoplasm; mouse model; neoantigens; neoplastic cell; new therapeutic target; novel; patient subsets; pre-clinical; predicting response; programs; receptor; recruit; response; success; targeted treatment; therapy design; tool; transcriptome sequencing; transcriptomics; tumor; tumor growth

PROJECT SUMMARY/ABSTRACT Immunotherapies such as checkpoint blockade have revolutionized cancer therapy, but responses are seen only in a subset of patients. Though tumor-intrinsic factors such as tumor mutational burden (TMB) or IFNγ “inflamed” signature partially predict sensitivity to checkpoint blockade, these correlations are limited—most patients with “inflamed” tumors or high TMB still fail to respond. A critical step for efficacy of T cell-mediated immunotherapies, including checkpoint blockade, is dendritic cell cross-presentation of tumor antigen (Ag) to CD8+ T cells. Cross- presentation in vivo requires Batf3-expressing type 1 dendritic cells (cDC1), though these DC have additional functions, including secretion of T cell-recruiting chemokines, driving tumor-reactive T cell (TRT) responses. Because patients with cDC1-enriched tumors have improved responses to anti-PD1, we developed an in situ vaccination (ISV) combining FLT3L, radiotherapy (XRT), and TLR agonism to enhance cDC1 cross-priming of TRT and observed that ISV potentiated anti-tumor effects of PD1 blockade and induced systemic tumor regressions in treated patients. Additionally, we have shown that adoptive transfer of tumor-specific T cells into syngeneic RAG-/- mice clears tumors, while transfer into allogeneic RAG-/- mice fails to control tumor growth, highlighting that APC cross-priming of CD8+ T cells is required for efficacy of antitumor T cells. Despite the critical role of cDC1 cross-priming of CD8+ T cells for effective therapy, there is no established method for measuring T cell cross- or direct-priming in vivo. Consequently, there is a critical need for methods to directly measure CD8+ T cell cross-priming for identifying novel therapeutic targets to enhance cross-priming, and to understand mechanisms of therapy resistance. We hypothesize that cross- and direct-primed T cells, and ISV-primed vs untreated T cells, will harbor distinct signatures, mirroring their differential antitumor efficacy. In Aim 1, we will develop mouse models of cross- and direct-priming using H2Kd knockout, H2Kb-transfected, GFP/OVA- expressing lymphoma and breast cancers and transfer of Ag-specific T cells into syngeneic and allogeneic RAG-/- mice. We will sort tumor-reactive T cells and perform bulk RNA-seq and spectral flow cytometry to identify a cross-primed CD8+ T cell signature. In Aim 2, we will use bulk RNA-seq and spectral flow cytometry to characterize the T cell response to ISV and checkpoint blockade across tumors (lymphoma, breast cancer), model antigens (GFP, OVA, luciferase), and endogenous tumor antigens. The outcome of this study will be elucidation of a cross-primed CD8+ T cell phenotype and a novel immune monitoring technique that allows targeted design of novel immunotherapies by targeting novel checkpoints or costimulators to increase cross- priming of tumor-reactive T cells.

项目概要/摘要 检查点封锁等免疫疗法彻底改变了癌症治疗，但仅能​​看到反应 尽管肿瘤内在因素如肿瘤突变负荷（TMB）或 IFNγ“发炎”。 签名部分预测对检查点封锁的敏感性，这些相关性是有限的——大多数患者 “发炎”肿瘤或高 TMB 仍然无法做出反应，这是 T 细胞介导的免疫疗法疗效的关键一步。 包括检查点阻断，是树突状细胞将肿瘤抗原 (Ag) 交叉呈递给 CD8+ T 细胞。 体内表达需要表达 Batf3 的 1 型树突状细胞 (cDC1)，尽管这些 DC 具有额外的 功能，包括分泌 T 细胞募集趋化因子，驱动肿瘤反应性 T 细胞 (TRT) 反应。 由于富含 cDC1 的肿瘤患者对抗 PD1 的反应有所改善，我们开发了一种原位 疫苗接种 (ISV) 结合 FLT3L、放射治疗 (XRT) 和 TLR 激动，以增强 cDC1 交叉启动 TRT 并观察到 ​​ISV 增强 PD1 阻断的抗肿瘤作用并诱发全身性肿瘤 此外，我们还证明了肿瘤特异性 T 细胞的过继转移。 同基因 RAG-/- 小鼠可以清除肿瘤，而转移到同种异体 RAG-/- 小鼠中无法控制肿瘤生长， 强调 CD8+ T 细胞的 APC 交叉引发对于抗肿瘤 T 细胞的功效至关重要，尽管这一点至关重要。 cDC1 交叉启动 CD8+ T 细胞对于有效治疗的作用，目前尚无确定的测量 T 的方法 由于体内细胞交叉或直接引发测试，迫切需要直接测量 CD8+ 的方法。 T 细胞交叉启动用于识别新的治疗靶点以增强交叉启动并了解 我们勇敢地面对交叉和直接引发的 T 细胞，以及 ISV 引发的 T 细胞和 ISV 引发的 T 细胞。 未经处理的 T 细胞将具有独特的特征，反映其不同的抗肿瘤功效。 使用 H2Kd 敲除、H2Kb 转染、GFP/OVA 开发交叉和直接引发的小鼠模型 表达淋巴瘤和乳腺癌，并将 Ag 特异性 T 细胞转移到同基因和同种异体中 我们将对 RAG-/- 小鼠进行分类，并进行批量 RNA 测序和光谱流式细胞术来识别。 在目标 2 中，我们将使用批量 RNA 测序和光谱流式细胞术来检测交叉引发的 CD8+ T 细胞特征。 表征 T 细胞对 ISV 的反应和跨肿瘤（淋巴瘤、乳腺癌）的检查点阻断， 模型抗原（GFP、OVA、荧光素酶）和内源性肿瘤抗原将是本研究的结果。 阐明交叉引发的 CD8+ T 细胞表型和一种新颖的免疫监测技术，该技术允许 通过针对新的检查点或共刺激物来增加交叉免疫疗法的靶向设计 肿瘤反应性 T 细胞的启动。

项目成果

Cancer vaccines: the next immunotherapy frontier.

癌症疫苗：下一个免疫治疗前沿。

• 发表时间: 2022-08
• 影响因子: 22.7
• 作者: Lin, Matthew J;Svensson;Lubitz, Gabrielle S;Marabelle, Aurélien;Melero, Ignacio;Brown, Brian D;Brody, Joshua D
• 通讯作者: Brody, Joshua D