Detection of PD-1 inhibitory signaling and its molecular relays in T cells: Implications for cancer immunotherapy

T 细胞中 PD-1 抑制信号传导及其分子中继的检测：对癌症免疫治疗的影响

基本信息

批准号：
10605878
负责人：
VASSILIKI A BOUSSIOTIS
金额：
$ 72.26万
依托单位：
BETH ISRAEL DEACONESS MEDICAL CENTER
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-01-01 至 2027-12-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10605878
关键词：
Ablation Affect Antibodies Antigen-Presenting Cells Antitumor Response B-Lymphocytes Binding Biochemical Biopsy CD8-Positive T-Lymphocytes CD80 gene CD8B1 gene Cancer Patient Cells Clinical Trials Cytoplasmic Tail Detection Development Genes Human Immune Immune system Immunologics Immunosuppression Immunotherapy Individual Infiltration Knowledge Ligands Macrophage Malignant Neoplasms Mediating Memory Modeling Molecular Molecular Profiling Mus Neurotransmitters Outcome PD-1 blockade Pathway interactions Patients Pattern Phosphotyrosine Physiological Production Property Proteins Regulatory T-Lymphocyte Shapes Signal Induction Signal Transduction T cell response T memory cell T-Cell Activation T-Lymphocyte T-Lymphocyte Subsets Tumor Immunity Tumor-Infiltrating Lymphocytes Tyrosine Work anti-cancer therapeutic cancer cell cancer immunotherapy cancer infiltrating T cells cancer therapy cell type checkpoint therapy exhaustion fatty acid oxidation immune activation improved outcome inhibitor lipid metabolism monocyte novel outcome prediction peripheral blood prognostic significance programmed cell death ligand 1 programmed cell death protein 1 receptor response success transcriptomics tumor tumor growth tumor microenvironment

项目摘要

PD-1 blocking agents have achieved significant success as anti-cancer therapeutics. The mechanism(s) of how PD-1 compromises anti-tumor function remain poorly understood. Although in trans engagement of PD-1 expressed in T cells by its ligands expressed on APC or cancer cells inhibits T cell activation, evolving discoveries provide evidence that PD-L1:B7-1(CD80) in cis and PD-1:PD-L1 in cis non-canonical interactions occur when these molecules are co-expressed on APC, and disrupt the canonical interaction between PD-1 and PD-L1 in trans and T cell inhibitory signaling. Thus, expression of PD-L1 in the tumor microenvironment (TME) is not synonymous with PD-1-mediated T cell inhibition. We generated an antibody that recognizes PD-1pY248 (pPD-1), which is required for PD-1 inhibitory signaling, and detected pPD-1 in mouse and human. We found pPD-1+ T cells in cultures, in mouse tumor models and patient biopsies, but also in CD8+ T central memory (TCM) cells in the peripheral blood of healthy individuals. In tumor bearing mice, pPD-1 was expressed in tumor infiltrating CD8+ T lymphocytes but mostly in Treg. We generated mice with conditional targeting of Pdcd1 gene (PD-1f/f) and selectively eliminated PD-1 in Treg (Pdcd1f/fFoxP3cre). In tumor-bearing Pdcd1f/fFoxP3cre mice, Treg displayed enrichment in pathways regulating lipid metabolism, fatty acid oxidation, and production of monocyte/macrophage chemotactic protein-1 (MCP-1) and GABAergic neurotransmitter with known immunosuppressive function. These features correlated with a significant increase of B cells and M2-like macrophages, diminished activation of tumor infiltrating T cells, and increased tumor growth. Our results reveal a previously unappreciated network by which Treg-selective blockade of PD-1 signaling reshapes the immunological landscape and suggest that abrogation of PD-1 signaling in distinct cell types differentially impacts the TME. Our findings indicate that pPD-1 is a powerful marker to identify T cells subjected to PD-1 inhibitory signaling and support the novel hypothesis that cell-specific detection of PD-1 signaling by pPD-1 might predict the outcome of checkpoint immunotherapy. To investigate these, we will pursue the following specific aims: 1. To identify the immunological and biochemical properties of pPD-1+ T cells in the context of cancer. We will characterize pPD-1+ CD8+ TIL and Treg by single cell immunoprofiling, and investigate how T cell subset- specific PD-1 signaling reshapes the TME by using our Pdcd1f/fCD8cre and Pdcd1f/fFoxP3cre mice. 2. To identify the molecular and functional properties of pPD-1+ cells in healthy individuals. We will examine how PD-1 signaling shapes the properties of T cells in healthy individuals and in cancer patients and uncover why only cancer-mediated PD-1 signaling induces TEX. 3. To determine expression, function and prognostic significance of pPD-1+ TIL in cancer patients. We will examine cell-specific PD-1 expression and signaling in patient biopsies, co-expression of PD-1/pPD-1, PD- L1 and CD80, and determine their prognostic significance.

作为抗癌治疗剂，PD-1阻断剂已取得了巨大的成功。如何 PD-1损害抗肿瘤功能的理解仍然很少。虽然在PD-1的反式参与中在T细胞中通过其在APC或癌细胞中表达的配体表达的配体抑制T细胞激活，不断发展。提供证据表明顺式和PD-1中的PD-L1：B7-1（CD80）在顺式非传统相互作用中发生的PD-1：PD-L1。这些分子在APC上共表达，并破坏PD-1和PD-L1之间的规范相互作用反式和T细胞抑制信号传导。因此，PD-L1在肿瘤微环境（TME）中的表达不是 PD-1介导的T细胞抑制的代名词。我们生成了一种识别PD-1PY248（PPD-1）的抗体，这是PD-1抑制信号传导所需的，并且在小鼠和人类中检测到PPD-1。我们发现PPD-1+ T 培养物中的细胞，小鼠肿瘤模型和患者活检，以及CD8+ T中心记忆（TCM）细胞中的细胞健康个体的外周血。在肿瘤小鼠中，PPD-1在肿瘤浸润CD8+中表达 T淋巴细胞，但主要在Treg中。我们以有条件地靶向PDCD1基因（PD-1F/F）和在Treg（PDCD1F/FFOXP3CRE）中有选择地消除PD-1。在承受肿瘤的PDCD1F/FFOXP3CRE小鼠中，Treg显示了在调节脂质代谢，脂肪酸氧化和产生的途径中富集单核细胞/巨噬细胞趋化蛋白-1（MCP-1）和GABA能神经递质已知免疫抑制功能。这些特征与B细胞和M2样显着增加有关巨噬细胞，肿瘤浸润的T细胞的激活减少，并增加了肿瘤的生长。我们的结果揭示了先前未欣赏的网络，通过该网络，PD-1信号传导的Treg选择性阻断可重塑免疫学局势，并建议在不同的细胞类型中废除PD-1信号传导差异影响 TME。我们的发现表明，PPD-1是识别受PD-1抑制作用的T细胞的强大标记信号传导和支持新的假设，即PPD-1对PD-1信号的特异性检测可能会预测检查点免疫疗法的结果。为了调查这些，我们将追求以下具体目标： 1。在癌症的背景下确定PPD-1+ T细胞的免疫学和生化特性。我们将通过单细胞免疫局体来表征PPD-1+ CD8+ TIL和Treg，并研究T细胞子群特定的PD-1信号传导通过使用我们的PDCD1F/FCD8CRE和PDCD1F/FFOXP3CRE小鼠来重塑TME。 2。确定健康个体中PPD-1+细胞的分子和功能特性。我们将检查PD-1信号传导如何塑造健康个体和癌症患者中T细胞的性质以及发现为什么只有癌症介导的PD-1信号传导会引起Tex。 3。确定PPD-1+ TIL在癌症患者中的表达，功能和预后意义。我们将检查患者活检中的细胞特异性PD-1表达和信号传导，PD-1/PPD-1的共表达，PD- L1和CD80，并确定其预后意义。