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ORIGINAL RESEARCH article

Front. Oncol.
Sec. Head and Neck Cancer
Volume 14 - 2024 | doi: 10.3389/fonc.2024.1484319

Mechanistic Insights into HPV-Positivity in Non-Smokers and HPV-Negativity in Smokers with Head and Neck Cancer

Provisionally accepted
Markus Hoffmann Markus Hoffmann 1,2*Susanne Hille Susanne Hille 3Asita Fazel Asita Fazel 2Martin Laudien Martin Laudien 2Susanne Wiegand Susanne Wiegand 2Martin Mueller Martin Mueller 4Oliver Mueller Oliver Mueller 3Elgar Susanne Quabius Elgar Susanne Quabius 2
  • 1 University of Kiel, Kiel, Germany
  • 2 Dept. of Otorhinolaryngology, Head and Neck Surgery, UKSH, Campus Kiel, Kiel, Germany
  • 3 Markus Hoffmann, Kiel, Germany
  • 4 Research Program Infection and Cancer, German Cancer Research Center, Heidelberg, Germany

The final, formatted version of the article will be published soon.

    Introduction Several aspects of the involvement of HPV in the pathogenesis of HPV-associated diseases remain poorly understood including mechanistic aspects of infection and the question of why the majority of HPV-positive HNSCC-patients are non-smokers, whereas HPV-negatives are smokers.Our previous research, based on 1,100 patient samples, hypothesized an explanation for this phenomenon: Smoking induces upregulation of a mucosal protective protein (SLPI), which competes with HPV for binding to Annexin A2 (AnxA2), pivotal for HPV cell entry. Here we investigate the mechanistic aspects of our hypothesis using transfection assays. Methods: HaCaT and HeLa cell lines were used to investigate the effects of shRNA transfection and nicotine exposure on HPV16-PsVuptake. Cells were treated with Lipofectamine™ RNAiMAX for 48 or 72 hours with specific shRNAconcentrations, while nicotine was added to the cell medium at the indicated concentrations. Protein isolation, SLPI-and AnxA2-quantification, LDH cytotoxicity assessment, HPV16-PsV-uptake measurement, mRNA-isolation, cDNA-synthesis and RT-qPCR were performed. Results: In vitro transfection experiments with HPV16 pseudovirions (PsVs) showed that PsVs entered cells significantly better when SLPI was downregulated and significantly less when AnxA2 was downregulated. Nicotine exposure increased SLPI levels and reduced PsV uptake. Conclusions: The overexpression of SLPI caused by tobacco-smoking can hinder HPV cell entry by binding to AnxA2 and thus prevent successful HPV infection. Conversely, non-smokers have lower SLPI-levels, associated with an excess of unbound AnxA2, favoring HPV cell-entry. These findings support our hypothesis, suggesting a paradigm shift in understanding virus-related pathogenesis, particularly in the head and neck region, and the nature of HPV infection.

    Keywords: HPV, Virus infection, viral cell entry, Smoking, SLPI, Annexin A2, HNSCC

    Received: 22 Aug 2024; Accepted: 16 Oct 2024.

    Copyright: © 2024 Hoffmann, Hille, Fazel, Laudien, Wiegand, Mueller, Mueller and Quabius. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

    * Correspondence: Markus Hoffmann, University of Kiel, Kiel, Germany

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