Oncostatin M sensitizes keratinocytes to UVB-induced inflammation via GSDME-mediated pyroptosis.

Abstract

BACKGROUND:

Oncostatin M (OSM), an interleukin-6 (IL-6) family proinflammatory cytokine, plays a critical role in inflammatory skin diseases, but its mechanism of action is not well understood.

OBJECTIVE:

To demonstrate the mechanism of OSM induced pyropotosis in normal human epidermal keratinocytes (NHEKs) and immortalized human keratinocytes (HaCaT cells).

METHODS:

NHEKs and HaCaT cells were treated with OSM. Knockout of OSM receptor (OSMR) with CRISPR/Cas9 system, knockdown of GSDME with small interfering RNA and primary keratinocytes from Osmr-/- and Gsdme-/- mice were used to study the effect of OSMR and GSDME. After treatment of OSM, NHEKs and HaCaT cells were irradiated with UVB. The mRNA was analyzed by quantitative real-time polymerase chain reaction (qRT-PCR) and RNA sequencing, protein level was detected by Western Blotting, Elisa and immunofluorescence. Cell death was examined by lactate dehydrogenase (LDH) releasing.

RESULTS:

Here we found that OSM induced pyropotosis in NHEKs and HaCaT cells, but knockout of OSMR abolished pyropotosis. RNA sequencing revealed an upregulation of several key genes involved in NLRP3 inflammasome activation following OSM treatment, among which NLRP3, GSDME, and IL-1β were confirmed by qRT-PCR and Western Blotting. Knockdown of GSDME alleviated OSM-induced pyropotosis. Pretreatment of OSM boosted UVB-induced pyroptosis and inflammation in NHEKs and HaCaT cells, and this priming function was lost in keratinocytes of Osmr-/- and Gsdme-/- mice. Similar results were obtained in a 3-dimensional culture of human epidermis.

CONCLUSION:

OSM functions as a priming cytokine to enhance UVB-induced inflammation in keratinocytes, providing insight into the pathogenesis of inflammatory skin diseases.

• Authors: Jun Liu, Yadan Zhong, Huiting Liu, Huan Yang, Ping Lu, Yanqiang Shi, Xuan Wang, Wen Zheng, Xiaoling Yu, Yingping Xu, Bin Yang
• Journal: Journal of dermatological science (J Dermatol Sci) Vol. 104 Issue 2 Pg. 95-103 (Nov 2021) ISSN: 1873-569X [Electronic] Netherlands
• PMID: 34674925 (Publication Type: Journal Article)
• Copyright: Copyright © 2021 Japanese Society for Investigative Dermatology. Published by Elsevier B.V. All rights reserved.

Chemical References

• GSDME protein, human
• Gsdme protein, mouse
• IL1B protein, human
• Interleukin-1beta
• NLR Family, Pyrin Domain-Containing 3 Protein
• NLRP3 protein, human
• OSMR protein, human
• Oncostatin M Receptor beta Subunit
• Osmr protein, mouse
• Pore Forming Cytotoxic Proteins
• RNA, Small Interfering
• Oncostatin M

Topics

• Animals
• Epidermis (metabolism)
• Gene Knockout Techniques
• HaCaT Cells
• Humans
• Interleukin-1beta (genetics, metabolism)
• Keratinocytes (radiation effects)
• Mice, Knockout
• NLR Family, Pyrin Domain-Containing 3 Protein (genetics, metabolism)
• Oncostatin M (pharmacology)
• Oncostatin M Receptor beta Subunit (genetics)
• Pore Forming Cytotoxic Proteins (genetics, metabolism)
• Pyroptosis (drug effects, genetics, radiation effects)
• RNA, Small Interfering
• Ultraviolet Rays
• Up-Regulation
• Mice