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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

基本信息

批准号：
10537849
负责人：
Gabrielle Lubitz
金额：
$ 4.52万
依托单位：
ICAHN SCHOOL OF MEDICINE AT MOUNT SINAI
依托单位国家：
美国
项目类别：
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/10537849
关键词：
Address Adoptive Transfer Agonist Allogenic Animal Model Antigen Presentation Antigen-Presenting Cells Antigens Autoantigens Automobile Driving Breast Cancer Model CD8-Positive T-Lymphocytes CXCL9 gene Cross Presentation Cross-Priming 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 T cell response T cell therapy T-Cell Activation T-Lymphocyte Techniques Tissues Tumor Antigens Tumor-infiltrating immune cells Vaccination anti-PD-1 antigen-specific T cells antitumor effect base 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 treatment response tumor tumor growth

项目摘要

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.

项目总结/摘要免疫疗法，如检查点阻断，已经彻底改变了癌症治疗，但反应只出现在在一部分患者中。虽然肿瘤内在因素如肿瘤突变负荷（TMB）或IFNγ“发炎”，特征部分预测了对检查点阻滞的敏感性，这些相关性是有限的-大多数患者 “发炎”肿瘤或高TMB仍然不能响应。T细胞介导的免疫疗法的功效的关键步骤，包括检查点阻断在内的免疫抑制是树突细胞将肿瘤抗原（Ag）交叉呈递给CD 8 + T细胞。交叉- 体内呈递需要表达Batf 3的1型树突状细胞（cDC 1），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cDC 1富集肿瘤患者对抗PD 1的反应有所改善，我们开发了原位免疫荧光法。联合FLT 3L、放疗（XRT）和TLR激动的疫苗接种（ISV），以增强CDC 1交叉致敏，观察到ISV增强PD 1阻断的抗肿瘤作用并诱导全身性肿瘤在治疗的患者中回归。此外，我们已经表明，过继转移肿瘤特异性T细胞进入同基因RAG-/-小鼠清除肿瘤，而转移到异基因RAG-/-小鼠中不能控制肿瘤生长，这突出了CD 8 + T细胞的APC交叉致敏对于抗肿瘤T细胞的功效是必需的。尽管批评 CD 8 + T细胞的cDC 1交叉致敏对于有效治疗的作用，目前还没有建立测量T细胞的方法。体内细胞交叉或直接引发。因此，迫切需要直接测量CD 8+的方法。 T细胞交叉致敏，用于鉴定新的治疗靶点以增强交叉致敏，并了解治疗抵抗的机制。我们假设交叉和直接致敏的T细胞，以及ISV致敏的T细胞与未经处理的T细胞将具有不同的特征，反映出它们的差异抗肿瘤功效。在目标1中，我们使用H2Kd敲除、H2Kb转染、GFP/OVA- 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异基因的 RAG-/-小鼠我们将对肿瘤反应性T细胞进行分类，并进行批量RNA-seq和光谱流式细胞术，交叉致敏的CD 8 + T细胞特征。在目标2中，我们将使用批量RNA-seq和光谱流式细胞术，表征T细胞对ISV的应答和肿瘤（淋巴瘤、乳腺癌）中的检查点阻断，模型抗原（GFP、OVA、荧光素酶）和内源性肿瘤抗原。这项研究的结果将是阐明交叉致敏的CD 8 + T细胞表型和一种新的免疫监测技术，通过靶向新的检查点或共刺激因子来靶向设计新的免疫疗法，以增加交叉免疫应答。引发肿瘤反应性T细胞。