Qingfei Jiedu decoction inhibits PD-L1 expression in lung adenocarcinoma based on network pharmacology analysis, molecular docking and experimental verification

Abstract

Objective: We aim at investigating the molecular mechanisms through which the Qingfei Jiedu decoction (QFJDD) regulates PD-L1 expression in lung adenocarcinoma (LUAD).Methods: Bioactive compounds and targets of QFJDD were screened from TCMSP, BATMAN-TCM, and literature. Then, GeneCard, OMIM, PharmGKB, Therapeutic Target, and DrugBank databases were used to identify LUAD-related genes. The protein-protein interaction (PPI) network was constructed using overlapping targets of bioactive compounds in LUAD with the Cytoscape software and STRING database. The potential functions and pathways in which the hub genes were enriched by GO, KEGG, and DAVID pathway analyses. Molecular docking of bioactive compounds and key genes was executed via AutoDock Vina. Qualitative and quantitative analyses of QFJDD were performed using UPLC-Q-TOF-MS and UPLC. Expressions of key genes were determined by qRT-PCR, immunoreactivity score (IRS) of PD-L1 was assessed by immunohistochemistry (IHC), while the CD8+PD-1+T% derived from spleen tissues of Lewis lung cancer (LLC) bearing-mice was calculated using flow cytometry (FCM).Results: A total of 53 bioactive compounds and 288 targets of QFJDD as well as 8151 LUAD associated genes were obtained. Further, six bioactive compounds, including quercetin, luteolin, kaempferol, wogonin, baicalein, and acacetin, and 22 hub genes were identified. The GO analysis showed that the hub genes were mainly enriched in DNA or RNA transcription. KEGG and DAVID pathway analyses revealed that 20 hub genes were primarily enriched in virus, cancer, immune, endocrine, and cardiovascular pathways. The EGFR, JUN, RELA, HIF1A, NFKBIA, AKT1, MAPK1, and MAPK14 hub genes were identified as key genes in PD-L1 expression and PD-1 checkpoint pathway. Moreover, ideal affinity and regions were identified between core compounds and key genes. Notably, QFJDD downregulated EGFR, JUN, RELA, HIF1A, NFKBIA, and CD274 expressions (p < 0.05), while it upregulated AKT1 and MAPK1 (p < 0.05) levels in A549 cells. The PD-L1 IRS of LLC tissue in the QFJDD high dose (Hd) group was lower than model group (p < 0.01). CD8+PD-1+T% was higher in the QFJDD Hd group than in normal and model groups (p < 0.05).Conclusion: QFJDD downregulates PD-L1 expression and increases CD8+PD-1+T% via regulating HIF-1, EGFR, JUN and NFκB signaling pathways. Therefore, QFJDD is a potential treatment option for LUAD.

Computational Approaches Drive Developments in Immune-Oncology Therapies for PD-1/PD-L1 Immune Checkpoint Inhibitors

“…Several small molecules targeting the PD-1/PD-L1 interaction have been identified in the last years, some of them comprising moieties such as urea (e.g., CA-170 ( 1 )), guanidine (e.g., abemaciclib ( 4 ), ciforadenant ( 6 ), 6-thio-2′-deoxyguanosine ( 10 ), SF1126 ( 15 )), sulfone (e.g., navtemadlin ( 3 ), vismodegib ( 11 )), biphenyl (e.g., INCB086550 ( 5 )) and N -heterocycle (e.g., tomivosertib ( 2 ), pexidantinib ( 9 ), carbozanitinib ( 12 ), cediranib and olaparib ( 13 ), itacitinib ( 14 ), sitravatinib ( 16 ), talazopanib ( 17 ), apatinib ( 18 )), which are already in the clinical phases ( Table 1 and Figure 3 ). However, the clinical results of targeting multiple immune-related targets still need to be elucidated; therefore, some works have recently been reported to identify small molecule targets using computational approaches in this regard [ 101 , 102 , 103 ]. Zhang et al [ 101 ] reported using the PharmMapper database [ 104 ] to identify the targets of the small molecule lomustine (a nitrosourea derivative, 50 ), which is investigated for the treatment of primary glioblastoma (see Figure 13 ).…”

“…Zhang et al [ 101 ] reported using the PharmMapper database [ 104 ] to identify the targets of the small molecule lomustine (a nitrosourea derivative, 50 ), which is investigated for the treatment of primary glioblastoma (see Figure 13 ). Pan et al [ 102 ] used the BATMAN-TCM web tool [ 105 ] to identify 288 targets and 53 bioactive compounds from Qingfei Jiedu decoction (QFJDD, an empirical Chinese prescription prepared from several herbals) that regulates PD-L1 expression in lung adenocarcinoma (LUAD). In the end, six flavonoid derivatives, including quercetin ( 51 ), luteolin ( 52 ), kaempferol ( 53 ), wogonin ( 54 ), baicalein ( 55 ) and acacetin ( 56 ), and 22 hub genes were identified ( Figure 13 ).…”

“…PPI, gene enrichment analysis, gene difference analysis, molecular docking and MD simulation have been used by the authors to screen out three effective targets of lomustine ( 50 ): HMOX1 (heme oxygenase 1), AKT1(AKT serine/threonine kinase 1) and EGFR (epidermal growth factor receptor) [ 101 ]. The same target, EGFR, was proposed by a different approach for QFJDD bioactive compounds ( 51 – 56 ) through a network construction and analysis comprising PPI, GO, KEGG and DAVID pathway analyses and molecular docking [ 102 ]. Several selected targets were associated with numerous human cancers, e.g., AKT1 [ 101 ], EGFR [ 101 , 102 ], HIF-1 (hypoxia inducible factor 1 subunit alpha) [ 102 ], MYC [ 103 ] and CXCL8 [ 103 ].…”

“…The same target, EGFR, was proposed by a different approach for QFJDD bioactive compounds ( 51 – 56 ) through a network construction and analysis comprising PPI, GO, KEGG and DAVID pathway analyses and molecular docking [ 102 ]. Several selected targets were associated with numerous human cancers, e.g., AKT1 [ 101 ], EGFR [ 101 , 102 ], HIF-1 (hypoxia inducible factor 1 subunit alpha) [ 102 ], MYC [ 103 ] and CXCL8 [ 103 ]. Furthermore, some of the selected targets seem to be related to an inflammatory response (CXCL8 [ 103 ]) or responses to viral infection (JUN [ 102 ], NFκB [ 102 ], and CD80 [ 57 ]).…”

“…Several selected targets were associated with numerous human cancers, e.g., AKT1 [ 101 ], EGFR [ 101 , 102 ], HIF-1 (hypoxia inducible factor 1 subunit alpha) [ 102 ], MYC [ 103 ] and CXCL8 [ 103 ]. Furthermore, some of the selected targets seem to be related to an inflammatory response (CXCL8 [ 103 ]) or responses to viral infection (JUN [ 102 ], NFκB [ 102 ], and CD80 [ 57 ]).…”