Downregulation of SHANK-associated RH domain-interacting protein elevates interleukin-33 expression by stimulating the Janus kinase 2/signal transducer and activator of transcription signalling pathway in HaCaT cells.

Publisher: Oxford University Press Country of Publication: England NLM ID: 7606847 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1365-2230 (Electronic) Linking ISSN: 03076938 NLM ISO Abbreviation: Clin Exp Dermatol Subsets: MEDLINE

Publication: 2023- : Oxford : Oxford University Press
Original Publication: Oxford : Blackwell Scientific Publications.

Dermatitis, Atopic/*drug therapy ; Dermatitis, Atopic/*metabolism ; Interleukin-33/*metabolism ; STAT2 Transcription Factor/*metabolism ; Ubiquitins/*pharmacology; Adolescent ; Adult ; Case-Control Studies ; Child ; Chromatin Immunoprecipitation/methods ; Dermatitis, Atopic/immunology ; Down-Regulation ; Female ; Fluorescent Antibody Technique/methods ; HaCaT Cells/metabolism ; HaCaT Cells/pathology ; Humans ; Immunohistochemistry/methods ; Inflammation/metabolism ; Janus Kinase 2/chemistry ; Janus Kinase 2/metabolism ; Male ; Middle Aged ; Nitriles/therapeutic use ; Pyrazoles/therapeutic use ; Pyrimidines/therapeutic use ; STAT3 Transcription Factor/chemistry ; STAT3 Transcription Factor/metabolism ; Signal Transduction ; Th2 Cells/immunology ; Th2 Cells/metabolism ; Ubiquitins/metabolism ; Young Adult

Background: Atopic dermatitis (AD) is a chronic inflammatory skin disease in which T-helper type 2 (Th2) immune responses are dominant. SH3 and multiple ankyrin repeat domains (SHANK)-associated RH domain-interacting protein (SHARPIN) is expressed at low levels in AD, resulting in the upregulation of the signal transducer and activator of transcription (STAT)3 protein and the Th2 cytokine, interleukin (IL)-33. However, the roles of SHARPIN in AD are not yet fully elucidated.
Aim: To evaluate the signalling interactions of SHARPIN and IL-33 in order to improve understanding of AD pathogenesis.
Methods: Western blotting was used to detect the Janus kinase (JAK)/STAT signalling proteins and IL-33 protein in HaCaT cells to determine the key proteins mediating the interaction between SHARPIN and IL-33. The findings were validated by immunofluorescence and immunohistochemical staining. Chromatin immunoprecipitation assays were used to evaluate the activity of STAT3 at the IL-33 promoter.
Results: We found that phosphorylated (p)JAK2 and pSTAT3 were upregulated in SHARPIN-knockdown HaCaT cells. Subsequent chromatin immunoprecipitation assays revealed that STAT3 binds to the IL-33 promoter to mediate IL-33 expression. Moreover, SHARPIN-mediated expression of IL-33 was reduced after treatment of HaCaT cells with the JAK/STAT inhibitor ruxolitinib. STAT3 and IL-33 expression levels were higher in AD skin lesion tissues than in normal skin tissues.
Conclusion: These findings suggest that SHARPIN modulates inflammation in HaCaT cells by inhibiting JAK/STAT signalling, supporting the application of SHARPIN as a potential therapeutic target for AD.
(© 2021 British Association of Dermatologists.)

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JCYJ20160429161334124 Science and Technology Project of Shenzhen; NSFC81703108 National Natural Science Foundation of China

0 (IL33 protein, human)
0 (Interleukin-33)
0 (Nitriles)
0 (Pyrazoles)
0 (Pyrimidines)
0 (SHARPIN protein, human)
0 (STAT2 Transcription Factor)
0 (STAT3 Transcription Factor)
0 (Ubiquitins)
82S8X8XX8H (ruxolitinib)
EC 2.7.10.2 (JAK2 protein, human)
EC 2.7.10.2 (Janus Kinase 2)

Date Created: 20210206 Date Completed: 20211129 Latest Revision: 20211129

20221213

10.1111/ced.14591

33548083