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Cellular and Molecular Biology

Dual targeting of BCL-2 and MCL-1 in the presence of BAX breaks venetoclax resistance in human small cell lung cancer

Abstract

Background

No targeted drugs are currently available against small cell lung cancer (SCLC). BCL-2 family members are involved in apoptosis regulation and represent therapeutic targets in many malignancies.

Methods

Expression of BCL-2 family members in 27 SCLC cell lines representing all known four SCLC molecular subtypes was assessed by qPCR, Western blot and mass spectrometry-based proteomics. BCL-2 and MCL-1 inhibition (venetoclax and S63845, respectively) was assessed by MTT assay and flow cytometry and in mice bearing human SCLC tumours. Drug interactions were calculated using the Combenefit software. Ectopic BAX overexpression was achieved by expression plasmids.

Results

The highest BCL-2 expression levels were detected in ASCL1- and POU2F3-driven SCLC cells. Although sensitivity to venetoclax was reflected by BCL-2 levels, not all cell lines responded consistently despite their high BCL-2 expression. MCL-1 overexpression and low BAX levels were both characteristic for venetoclax resistance in SCLC, whereas the expression of other BCL-2 family members did not affect therapeutic efficacy. Combination of venetoclax and S63845 resulted in significant, synergistic in vitro and in vivo anti-tumour activity and apoptosis induction in double-resistant cells; however, this was seen only in a subset with detectable BAX. In non-responding cells, ectopic BAX overexpression sensitised to venetoclax and S63845 and, furthermore, induced synergistic drug interaction.

Conclusions

The current study reveals the subtype specificity of BCL-2 expression and sheds light on the mechanism of venetoclax resistance in SCLC. Additionally, we provide preclinical evidence that combined BCL-2 and MCL-1 targeting is an effective approach to overcome venetoclax resistance in high BCL-2-expressing SCLCs with intact BAX.

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Fig. 1: BCL-2 expression pattern poorly correlates with venetoclax resistance in SCLC cell lines.
Fig. 2: Venetoclax resistance is characterised by increased MCL-1 and decreased BAX levels.
Fig. 3: Venetoclax and S63845 synergise in a subset of SCLC cell lines in vitro and in vivo.
Fig. 4: Venetoclax and S63845 induce apoptosis in sensitive cells.
Fig. 5: BAX is required for synergistic interactions between venetoclax and S63845.

Data availability

Data were generated by the authors and are available on request.

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Acknowledgements

We thank Barbara Dekan, Barbara Peter-Vörösmarty and Erzsebet Nagy for technical assistance and Nicole Woldmar and Beáta Szeitz for their help with the proteomic analyses.

Funding

KS was supported the Austrian Science Fund (FWF No. T 1062-B33) and the City of Vienna Fund for Innovative Interdisciplinary Cancer Research. BD, ZM and AR acknowledge funding from the Hungarian National Research, Development and Innovation Office (KH130356 to BD; 2020-1.1.6-JÖVŐ and TKP2021-EGA-33 to BD and ZM; FK-143751 to BD, ZM and AH-R). BD was also supported by the Austrian Science Fund (FWF I3522, FWF I3977 and I4677). ZM was supported by the UNKP-20-3 and UNKP-21-3 New National Excellence Program of the Ministry for Innovation and Technology of Hungary and by the Hungarian Respiratory Society (MPA #2020). ZM is recipient of the IASLC/ILCF Young Investigator Grant 2023. VL is a recipient of the Bolyai Research Scholarship of the Hungarian Academy of Sciences and the UNKP-19-4 New National Excellence Program of the Ministry for Innovation and Technology. MG was supported by the City of Vienna Fund for Innovative Interdisciplinary Cancer Research and the Berndorf Private Foundation. MR acknowledges funding from the Mrs. Berta Kamprad´s Cancer Foundation (FBKS-2020-22-(291)).

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Contributions

ZV was responsible for conceptualisation, data curation, formal analysis, investigation, methodology, software, visualisation, writing the original draft, review and editing. ZM was responsible for conceptualisation, data curation, formal analysis, investigation, methodology, software, resources, writing the original draft, review and editing. AS, NB, BF, AH-R, IK, and ES were responsible for data curation, writing—review and editing. CL and VP were responsible for formal analysis, writing—review and editing. SP was responsible for investigation, resources, writing—review and editing. KB was responsible for investigation, writing—review and editing. MR was responsible for data curation, software, visualisation, resources, writing—review and editing. GM-V was responsible for resources, writing—review and editing. FR-V and KH were responsible for project administration, resources, writing—review and editing. MAH, TK, and MG were responsible for project administration, writing—review and editing. VL was responsible for conceptualisation, methodology, supervision, writing—review and editing. BD was responsible for conceptualisation, investigation, methodology, project administration, resources, supervision, writing—original draft and writing—review and editing. KS was responsible for conceptualisation, data curation, investigation, methodology, project administration, software, visualisation, validation, resources, supervision, writing—original draft and writing—review and editing.

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Correspondence to Balazs Dome or Karin Schelch.

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Valko, Z., Megyesfalvi, Z., Schwendenwein, A. et al. Dual targeting of BCL-2 and MCL-1 in the presence of BAX breaks venetoclax resistance in human small cell lung cancer. Br J Cancer (2023). https://doi.org/10.1038/s41416-023-02219-9

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