Department of Medical Technology, Faculty of Associated Medical Sciences, Chiang Mai University, Chiang Mai, Thailand.
Laboratory of Molecular Pharmacology, Department of Pharmacology, School of Pharmacy, Southwest Medical University, Luzhou, China.
Sci Rep. 2022 Jul 6;12(1):11444. doi: 10.1038/s41598-022-15020-0.
As immune checkpoint inhibitors (ICIs) continue to advance, more evidence has emerged that anti-PD-1/PD-L1 immunotherapy is an effective treatment against cancers. Known as the programmed death ligand-1 (PD-L1), this co-inhibitory ligand contributes to T cell exhaustion by interacting with programmed death-1 (PD-1) receptor. However, cancer-intrinsic signaling pathways of the PD-L1 molecule are not well elucidated. Therefore, the present study aimed to evaluate the regulatory network of PD-L1 and lay the basis of successful use of anti-PD-L1 immunotherapy in acute myeloid leukemia (AML). Data for AML patients were extracted from TCGA and GTEx databases. The downstream signaling pathways of PD-L1 were identified via Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis. The key PD-L1 related genes were selected by weighted gene co-expression network analysis (WGCNA), MCC algorithm and Molecular Complex Detection (MCODE). The CCK-8 assay was used to assess cell proliferation. Flow cytometry was used to determine cell apoptosis and cell cycle. Western blotting was used to identify the expression of the PI3K-AKT signaling pathway. PD-L1 was shown to be elevated in AML patients when compared with the control group, and high PD-L1 expression was associated with poor overall survival rate. The ECM-receptor interaction, as well as the PI3K-AKT signaling pathway, were important PD-L1 downstream pathways. All three analyses found eight genes (ITGA2B, ITGB3, COL6A5, COL6A6, PF4, NMU, AGTR1, F2RL3) to be significantly associated with PD-L1. Knockdown of PD-L1 inhibited AML cell proliferation, induced cell apoptosis and G2/M cell cycle arrest. Importantly, PD-L1 knockdown reduced the expression of PI3K and p-AKT, but PD-L1 overexpression increased their expression. The current study elucidates the main regulatory network and downstream targets of PD-L1 in AML, assisting in the understanding of the underlying mechanism of anti-PD-1/PD-L1 immunotherapy and paving the way for clinical application of ICIs in AML.
随着免疫检查点抑制剂(ICIs)的不断发展,越来越多的证据表明抗 PD-1/PD-L1 免疫疗法是治疗癌症的有效方法。这种共抑制配体被称为程序性死亡配体 1(PD-L1),通过与程序性死亡-1(PD-1)受体相互作用,导致 T 细胞衰竭。然而,PD-L1 分子的癌症内在信号通路尚未得到很好的阐明。因此,本研究旨在评估 PD-L1 的调控网络,为抗 PD-L1 免疫疗法在急性髓细胞白血病(AML)中的成功应用奠定基础。从 TCGA 和 GTEx 数据库中提取 AML 患者的数据。通过基因本体论(GO)和京都基因与基因组百科全书(KEGG)富集分析确定 PD-L1 的下游信号通路。通过加权基因共表达网络分析(WGCNA)、MCC 算法和分子复合物检测(MCODE)选择关键的 PD-L1 相关基因。CCK-8 测定法用于评估细胞增殖。流式细胞术用于确定细胞凋亡和细胞周期。Western blot 用于鉴定 PI3K-AKT 信号通路的表达。与对照组相比,AML 患者的 PD-L1 表达升高,高 PD-L1 表达与总生存率降低相关。ECM-受体相互作用以及 PI3K-AKT 信号通路是 PD-L1 的重要下游途径。所有三种分析均发现与 PD-L1 显著相关的 8 个基因(ITGA2B、ITGB3、COL6A5、COL6A6、PF4、NMU、AGTR1、F2RL3)。PD-L1 的敲低抑制了 AML 细胞的增殖,诱导了细胞凋亡和 G2/M 细胞周期停滞。重要的是,PD-L1 敲低降低了 PI3K 和 p-AKT 的表达,但 PD-L1 过表达增加了它们的表达。本研究阐明了 PD-L1 在 AML 中的主要调控网络和下游靶标,有助于理解抗 PD-1/PD-L1 免疫疗法的潜在机制,并为 AML 中 ICIs 的临床应用铺平道路。