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Secretome of senescent hepatoma cells modulate immune cell fate by macrophage polarization and neutrophil extracellular traps formation

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Abstract

Presence of dysfunctional senescent hepatocytes is a hallmark feature of liver cirrhosis which finally culminates in liver cancer. We now report the presence of senescent hepatocytes (p21 and p53 positive) in the vicinity of infiltrated immune cells in hepatocellular carcinoma tissue specimens by immunohistochemistry. Hence, we evaluated in vitro, the relevance of senescent hepatoma cells in altering the fate of monocytes and neutrophils by assaying for macrophage polarization and extracellular trap (NETs) formation, respectively. Premature senescence was induced in hepatoma cells (HepG2 and Huh7 cells) by treating cells with doxorubicin. Senescent hepatoma cells showed strong inflammatory phenotype with induced expression of cytokines (IL1β, IL6, IL8 and IL13) as evaluated by flow cytometry. The senescent secretome from hepatoma cells when incubated with healthy monocytes caused it to differentiate predominantly towards M2 fate (CD80low CD86low CD163high CD206high) when analysed by flow cytometry. This was corroborated by the finding in clinical samples where human hepatocellular carcinoma harbouring senescent hepatocytes showed presence of M2 macrophages, while M1 macrophages were predominant in non-tumorous region. Additionally, the senescent secretome from Huh7 cells enhanced the NETs formation, while HepG2 secretome had an inhibitory effect. In conclusion, the “pro-inflammatory” senescent secretome drives non-inflammatory type M2 macrophage polarization and modulated neutrophil traps which in turn can influence the tumor microenvironment.

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Acknowledgements

BS is a recipient of UGC senior research fellowship. We thank DST-FIST for infrastructural support. We are thankful to Dr. Ritesh Kumar Tiwari (CU-BD, CoE, CRNN) for helping with flow cytometry analysis. We thank the anonymous reviewers for giving expert inputs which helped in improvement of the manuscript.

Funding

The authors thank DST-FIST  (Dept Science and Technology, India) for providing infrastructural grant and UGC for providing fellowship to BS.

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Author notes

  1. Bijoya Sen

    Present address: Buck Institute for Research on Aging, Novato, CA, USA

Authors and Affiliations

  1. Department of Molecular and Cellular Medicine, Institute of Liver and Biliary Sciences (ILBS), D1 Block, Vasant Kunj, New Delhi, 110070, India

    Bijoya Sen, Savera Aggarwal, Rhisita Nath, Rashi Sehgal, Ravinder Singh, Khushboo Agrawal, Nirupma Trehanpati & Gayatri Ramakrishna

  2. Department of Pathology, Institute of Liver and Biliary Sciences, New Delhi, India

    Ashwini Nagaraghatta Shashidhara & Archana Rastogi

  3. Department of Transfusion Medicine, Institute of Liver and Biliary Sciences, New Delhi, India

    Meenu Bajpai

  4. Department of Hepatopancreatobiliary Surgery, Institute of Liver and Biliary Sciences, New Delhi, India

    Viniyendra Pamecha

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Contributions

BS: Performed all experiments, compiled and analysed data, and inputs in manuscript writing, GR: Conceived and designed study, analysed data and wrote manuscript, SA, RN, RS, KA, RS, AN: helped in experimental protocols; NT: provided valuable research inputs, helped with flow cytometry analysis and flowantibodies; AR: Read pathology slides and gave critical clinical inputs on interpretation of IHC results, MB: provided blood samples of healthy volunteers, VP: valuable clinical inputs on study design and provided resected specimens.

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Correspondence to Gayatri Ramakrishna.

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The study was approved by the Ethics Committee of Institute of Liver and Biliary Sciences, Delhi, India.

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Informed consent of healthy volunteers who consented to give their blood were included in this study.

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Sen, B., Aggarwal, S., Nath, R. et al. Secretome of senescent hepatoma cells modulate immune cell fate by macrophage polarization and neutrophil extracellular traps formation. Med Oncol 39, 134 (2022). https://doi.org/10.1007/s12032-022-01732-w

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