Abstract
A recent hypothesis suggesting that the pharmacological target TRPV1 (transient receptor potential vanilloid subfamily, member 1) may function as a tumour suppressor, which potentially impacts the development of TRPV1 antagonist therapeutics for a range of conditions. However, little is known about the long-term physiologic effects of TRPV1 blockade in the skin. In vitro and in vivo studies suggested that the potent TRPV1 competitive antagonist AMG-9810 promoted proliferation in N/TERT1 cells (telomerase-immortalised primary human keratinocytes 1) and tumour development in mouse skin that was mediated through EGFR/Akt/mTOR signalling. We attempted to reproduce the reported in vitro and in vivo findings to further explore this hypothesis to understand the underlying mechanism and the risk associated with TRPV1 antagonism in the skin. In vitro proliferation studies using multiple methods and topical application with AMG-9810 and structurally similar TRPV1 antagonists such as SB-705498 and PAC-14028 were performed. Although we confirmed expression of TRPV1 in primary human epidermal keratinocytes (HEKn) and spontaneously immortalised human keratinocytes (HaCaT), we were unable to demonstrate cell proliferation in either cell type or any clear evidence of increased expression of proteins in the EGFR/Akt/mTOR signalling pathway with these molecules. We were also unable to demonstrate skin tumour promotion or underlying molecular mechanisms involved in the EGFR/Akt/mTOR signalling pathway in a single-dose and two-stage carcinogenesis mouse study treated with TRPV1 antagonists. In conclusion, our data suggest that inhibiting the pharmacological function of TRPV1 in skin by specific antagonists has not been considered to be indicative of skin tumour development.
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Abbreviations
- Akt:
-
Family of serine/threonine protein kinases
- EGFR:
-
Epidermal growth factor receptor
- HaCaT:
-
Spontaneously immortalised human keratinocytes
- HEKn:
-
Primary human epidermal keratinocytes
- ICC:
-
Immunocytochemistry
- mTOR:
-
Mammalian target of rapamycin
- TRPV1:
-
Transient receptor potential vanilloid subfamily, member 1
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Acknowledgements
We kindly acknowledge Tracy Walker, Chloe Taylor, Paul Daly and Rhiannon Lowe for their technical assistance with studies carried out at GlaxoSmithKline R&D and Rebekah Austin-Sparks, Mark Lennon and Mike Aylott for the statistical analyses relating to studies carried out at GlaxoSmithKline R&D. We also kindly acknowledge KiWha Lee and Byoungyoung Woo for compound synthesis relating to studies carried out at AmorePacific R&D. HaCaT cells were purchased by Prof. Ok-Nam Bae (College of Pharmacy, Hanyang University Ansan, Gyeonggido) from CLS Cell Line Service, Germany, and kindly provided as original cryopreserved cells for these studies. The authors alone are responsible for the content and writing of this article. Part of this study was supported by a grant of the Korean Health Technology R&D Project, Ministry of Health and Welfare, Republic of Korea (HI13C2304).
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Park, M., Naidoo, A.A., Burns, A. et al. Do TRPV1 antagonists increase the risk for skin tumourigenesis? A collaborative in vitro and in vivo assessment. Cell Biol Toxicol 34, 143–162 (2018). https://doi.org/10.1007/s10565-017-9407-8
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DOI: https://doi.org/10.1007/s10565-017-9407-8