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Divergent effect of Birinapant, and BV6 SMAC mimetic on TNFα induced NF-κB signaling and cell viability in activated hepatic stellate cells

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Abstract

Background

Tumor necrosis factor-α (TNFα) is a pleiotropic cytokine involved in nuclear factor kappa B (NF-κB) mediated cell survival as well as cell death. High serum TNFα levels correlate with liver fibrosis and enhance hepatic stellate cell (HSC) viability. However, the regulatory role of cellular inhibitor of apoptosis-1/2 (cIAP1/2) during TNFα induced NF-κB signaling in activated HSCs is largely unknown.

Method and Results

Activated HSCs were treated with cIAP1/2 inhbitiors i.e., SMAC mimetic BV6, and Birinapant in the presence of TNFα and macrophage conditioned media. TNFα cytokine increased cIAP2 expression and enhanced cell viability through the canonical NF-κB signaling in activated HSCs. cIAP2 inhibition via BV6 decreased the TNFα induced canonical NF-κB signaling, and reduced cell viability in activated HSCs. SMAC mimetic, Birinapant alone did not affect the cell viability but treatment of TNFα sensitized HSCs with Birinapant induced cell death. While BV6 mediated cIAP2 ablation was able to decrease the TNFα induced canonical NF-κB signaling, this effect was not observed with Birinapant treatment. Secreted TNFα from M1 polarized macrophages sensitized activated HSCs to BV6 or Birinapant mediated cell death. However, M2 polarized macrophage conditioned medium rescued the activated HSCs from BV6 mediated cytotoxicity.

Conclusion

In this study, we describe the regulatory role of cIAP2 in TNFα induced NF-κB signaling in activated HSCs. Targeting cIAP2 may be a promising approach for liver fibrosis treatment via modulating NF-κB signaling in activated HSCs.

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Data availability

The data acquired and analyzed in the current study are available from the corresponding author on reasonable request.

Code availability

Not applicable.

Abbreviations

TNFα :

Tumor necrosis factor-α

cIAP1/2 :

Cellular inhibitor of apoptosis-1/2

NF-κB :

Nuclear factor kappa B

HSC :

Hepatic stellate cell

αSMA :

α-Smooth muscle actin

qHSC :

Quiescent HSC

SMAC :

Second mitochondria-derived activator of caspase

NASH :

Non-alcholic steatohepatitis

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Acknowledgements

The authors would would like appreciate laboratory members, Bornika Roy, Basundhra Das, Prachi Sharma, and Umesh Gautam for their helpful and constructive discussion. The authors are grateful to Dr. W. Selvamurthy and Prof B.C. Das for encouraging and supporting our research at the Amity Institute of Molecular Medicine and Stem Cell Research (AIMMSCR), Amity University, Uttar Pradesh.

Funding

This work was supported by fellowships and grant supports from the Council of Scientific & Industrial Research (CSIR-09/915(006)/2017-EMRI to S. Sharma), University Grants Commission (UGC Ref no-581/CSIR-UGC-NET June 2017 to S.M. Ghufran), Department of Biotechnology (DBT-102/IFD/SAN/3003/2017–2018 to S. Biswas) and Science and Engineering Research Board (SERB) (CRG/2018/003918 to S. Biswas and SERB- YSS/2015/000092 to S. Ghose) Indian Council of Medical Research (ICMR-2019–1306/SCR/ADHOC-BMS to S. Biswas), India.

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Author notes

  1. Sachin Sharma

    Present address: Department of Medicine, University of California, San Francisco, San Francisco, CA, USA

  2. Shaikh Maryam Ghufran and Sachin Sharma have equally contributed to this work.

Authors and Affiliations

  1. Amity Institute of Molecular Medicine and Stem Cell Research (AIMMSCR), Translational Cancer & Stem Cell Research Laboratory, Amity University Uttar Pradesh (AUUP), Amity University, Research Laboratory 101, J3 Block Sector 125, Noida, 201313, Uttar Pradesh, India

    Shaikh Maryam Ghufran, Sachin Sharma & Subhrajit Biswas

  2. Department of Medical Oncology, All India Institute of Medical Sciences, New Delhi, India

    Sampa Ghose

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Contributions

SMG and SS together designed the experiments, performed most of the biochemical and cellular experiments, acquired, analyzed, and interpreted the data with statistical validation. SG carried out ELISA, and critical revision of the manuscript. SB was involved in study concept and design, acquisition, analysis, and interpretation of data; drafting of the manuscript, critical revision of the manuscript for intellectual content.

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Correspondence to Subhrajit Biswas.

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Ghufran, S.M., Sharma, S., Ghose, S. et al. Divergent effect of Birinapant, and BV6 SMAC mimetic on TNFα induced NF-κB signaling and cell viability in activated hepatic stellate cells. Mol Biol Rep 50, 2107–2117 (2023). https://doi.org/10.1007/s11033-022-08210-6

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