Combination of betulinic acid and EGFR-TKIs exerts synergistic anti-tumor effects against wild-type EGFR NSCLC by inducing autophagy-related cell death via EGFR signaling pathway
Abstract Background Epidermal growth factor receptor (EGFR) tyrosine kinase inhibitors (TKIs) have revolutionized the treatment of lung cancer patients with mutated EGFR. However, the efficacy of EGFR-TKIs in wild-type EGFR tumors has been shown to be marginal. Methods that can sensitize EGFR-TKIs t...
Saved in:
| Main Authors: | , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
BMC
2024-05-01
|
| Series: | Respiratory Research |
| Subjects: | |
| Online Access: | https://doi.org/10.1186/s12931-024-02844-9 |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
| _version_ | 1849699761542660096 |
|---|---|
| author | Han Wang Xiaohui Du Wenwen Liu Congcong Zhang Ying Li Jingwen Hou Yi Yu Guiru Li Qi Wang |
| author_facet | Han Wang Xiaohui Du Wenwen Liu Congcong Zhang Ying Li Jingwen Hou Yi Yu Guiru Li Qi Wang |
| author_sort | Han Wang |
| collection | DOAJ |
| description | Abstract Background Epidermal growth factor receptor (EGFR) tyrosine kinase inhibitors (TKIs) have revolutionized the treatment of lung cancer patients with mutated EGFR. However, the efficacy of EGFR-TKIs in wild-type EGFR tumors has been shown to be marginal. Methods that can sensitize EGFR-TKIs to EGFR wild-type NSCLC remain rare. Hence, we determined whether combination treatment can maximize the therapeutic efficacy of EGFR-TKIs. Methods We established a focused drug screening system to investigate candidates for overcoming the intrinsic resistance of wild-type EGFR NSCLC to EGFR-TKIs. Molecular docking assays and western blotting were used to identify the binding mode and blocking effect of the candidate compounds. Proliferation assays, analyses of drug interactions, colony formation assays, flow cytometry and nude mice xenograft models were used to determine the effects and investigate the molecular mechanism of the combination treatment. Results Betulinic acid (BA) is effective at targeting EGFR and synergizes with EGFR-TKIs (gefitinib and osimertinib) preferentially against wild-type EGFR. BA showed inhibitory activity due to its interaction with the ATP-binding pocket of EGFR and dramatically enhanced the suppressive effects of EGFR-TKIs by blocking EGFR and modulating the EGFR-ATK-mTOR axis. Mechanistic studies revealed that the combination strategy activated EGFR-induced autophagic cell death and that the EGFR-AKT-mTOR signaling pathway was essential for completing autophagy and cell cycle arrest. Activation of the mTOR pathway or blockade of autophagy by specific chemical agents markedly attenuated the effect of cell cycle arrest. In vivo administration of the combination treatment caused marked tumor regression in the A549 xenografts. Conclusions BA is a potential wild-type EGFR inhibitor that plays a critical role in sensitizing EGFR-TKI activity. BA combined with an EGFR-TKI effectively suppressed the proliferation and survival of intrinsically resistant lung cancer cells via the inhibition of EGFR as well as the induction of autophagy-related cell death, indicating that BA combined with an EGFR-TKI may be a potential therapeutic strategy for overcoming the primary resistance of wild-type EGFR-positive lung cancers. Graphical abstract |
| format | Article |
| id | doaj-art-23613b7dab6b42a0af5b4bbc54e255b7 |
| institution | DOAJ |
| issn | 1465-993X |
| language | English |
| publishDate | 2024-05-01 |
| publisher | BMC |
| record_format | Article |
| series | Respiratory Research |
| spelling | doaj-art-23613b7dab6b42a0af5b4bbc54e255b72025-08-20T03:18:28ZengBMCRespiratory Research1465-993X2024-05-0125111610.1186/s12931-024-02844-9Combination of betulinic acid and EGFR-TKIs exerts synergistic anti-tumor effects against wild-type EGFR NSCLC by inducing autophagy-related cell death via EGFR signaling pathwayHan Wang0Xiaohui Du1Wenwen Liu2Congcong Zhang3Ying Li4Jingwen Hou5Yi Yu6Guiru Li7Qi Wang8The Second Hospital of Dalian Medical UniversityThe Second Hospital of Dalian Medical UniversityThe Second Hospital of Dalian Medical UniversityThe Second Hospital of Dalian Medical UniversityThe Second Hospital of Dalian Medical UniversityThe Second Hospital of Dalian Medical UniversityThe Second Hospital of Dalian Medical UniversityThe Second Hospital of Dalian Medical UniversityThe Second Hospital of Dalian Medical UniversityAbstract Background Epidermal growth factor receptor (EGFR) tyrosine kinase inhibitors (TKIs) have revolutionized the treatment of lung cancer patients with mutated EGFR. However, the efficacy of EGFR-TKIs in wild-type EGFR tumors has been shown to be marginal. Methods that can sensitize EGFR-TKIs to EGFR wild-type NSCLC remain rare. Hence, we determined whether combination treatment can maximize the therapeutic efficacy of EGFR-TKIs. Methods We established a focused drug screening system to investigate candidates for overcoming the intrinsic resistance of wild-type EGFR NSCLC to EGFR-TKIs. Molecular docking assays and western blotting were used to identify the binding mode and blocking effect of the candidate compounds. Proliferation assays, analyses of drug interactions, colony formation assays, flow cytometry and nude mice xenograft models were used to determine the effects and investigate the molecular mechanism of the combination treatment. Results Betulinic acid (BA) is effective at targeting EGFR and synergizes with EGFR-TKIs (gefitinib and osimertinib) preferentially against wild-type EGFR. BA showed inhibitory activity due to its interaction with the ATP-binding pocket of EGFR and dramatically enhanced the suppressive effects of EGFR-TKIs by blocking EGFR and modulating the EGFR-ATK-mTOR axis. Mechanistic studies revealed that the combination strategy activated EGFR-induced autophagic cell death and that the EGFR-AKT-mTOR signaling pathway was essential for completing autophagy and cell cycle arrest. Activation of the mTOR pathway or blockade of autophagy by specific chemical agents markedly attenuated the effect of cell cycle arrest. In vivo administration of the combination treatment caused marked tumor regression in the A549 xenografts. Conclusions BA is a potential wild-type EGFR inhibitor that plays a critical role in sensitizing EGFR-TKI activity. BA combined with an EGFR-TKI effectively suppressed the proliferation and survival of intrinsically resistant lung cancer cells via the inhibition of EGFR as well as the induction of autophagy-related cell death, indicating that BA combined with an EGFR-TKI may be a potential therapeutic strategy for overcoming the primary resistance of wild-type EGFR-positive lung cancers. Graphical abstracthttps://doi.org/10.1186/s12931-024-02844-9EGFR-TKIsBetulinic acidWild-type EGFRAutophagic cell deathCell cycle arrestNon-small cell lung cancer |
| spellingShingle | Han Wang Xiaohui Du Wenwen Liu Congcong Zhang Ying Li Jingwen Hou Yi Yu Guiru Li Qi Wang Combination of betulinic acid and EGFR-TKIs exerts synergistic anti-tumor effects against wild-type EGFR NSCLC by inducing autophagy-related cell death via EGFR signaling pathway Respiratory Research EGFR-TKIs Betulinic acid Wild-type EGFR Autophagic cell death Cell cycle arrest Non-small cell lung cancer |
| title | Combination of betulinic acid and EGFR-TKIs exerts synergistic anti-tumor effects against wild-type EGFR NSCLC by inducing autophagy-related cell death via EGFR signaling pathway |
| title_full | Combination of betulinic acid and EGFR-TKIs exerts synergistic anti-tumor effects against wild-type EGFR NSCLC by inducing autophagy-related cell death via EGFR signaling pathway |
| title_fullStr | Combination of betulinic acid and EGFR-TKIs exerts synergistic anti-tumor effects against wild-type EGFR NSCLC by inducing autophagy-related cell death via EGFR signaling pathway |
| title_full_unstemmed | Combination of betulinic acid and EGFR-TKIs exerts synergistic anti-tumor effects against wild-type EGFR NSCLC by inducing autophagy-related cell death via EGFR signaling pathway |
| title_short | Combination of betulinic acid and EGFR-TKIs exerts synergistic anti-tumor effects against wild-type EGFR NSCLC by inducing autophagy-related cell death via EGFR signaling pathway |
| title_sort | combination of betulinic acid and egfr tkis exerts synergistic anti tumor effects against wild type egfr nsclc by inducing autophagy related cell death via egfr signaling pathway |
| topic | EGFR-TKIs Betulinic acid Wild-type EGFR Autophagic cell death Cell cycle arrest Non-small cell lung cancer |
| url | https://doi.org/10.1186/s12931-024-02844-9 |
| work_keys_str_mv | AT hanwang combinationofbetulinicacidandegfrtkisexertssynergisticantitumoreffectsagainstwildtypeegfrnsclcbyinducingautophagyrelatedcelldeathviaegfrsignalingpathway AT xiaohuidu combinationofbetulinicacidandegfrtkisexertssynergisticantitumoreffectsagainstwildtypeegfrnsclcbyinducingautophagyrelatedcelldeathviaegfrsignalingpathway AT wenwenliu combinationofbetulinicacidandegfrtkisexertssynergisticantitumoreffectsagainstwildtypeegfrnsclcbyinducingautophagyrelatedcelldeathviaegfrsignalingpathway AT congcongzhang combinationofbetulinicacidandegfrtkisexertssynergisticantitumoreffectsagainstwildtypeegfrnsclcbyinducingautophagyrelatedcelldeathviaegfrsignalingpathway AT yingli combinationofbetulinicacidandegfrtkisexertssynergisticantitumoreffectsagainstwildtypeegfrnsclcbyinducingautophagyrelatedcelldeathviaegfrsignalingpathway AT jingwenhou combinationofbetulinicacidandegfrtkisexertssynergisticantitumoreffectsagainstwildtypeegfrnsclcbyinducingautophagyrelatedcelldeathviaegfrsignalingpathway AT yiyu combinationofbetulinicacidandegfrtkisexertssynergisticantitumoreffectsagainstwildtypeegfrnsclcbyinducingautophagyrelatedcelldeathviaegfrsignalingpathway AT guiruli combinationofbetulinicacidandegfrtkisexertssynergisticantitumoreffectsagainstwildtypeegfrnsclcbyinducingautophagyrelatedcelldeathviaegfrsignalingpathway AT qiwang combinationofbetulinicacidandegfrtkisexertssynergisticantitumoreffectsagainstwildtypeegfrnsclcbyinducingautophagyrelatedcelldeathviaegfrsignalingpathway |