Human β-defensin-3 attenuates atopic dermatitis–like inflammation through autophagy activation and the aryl hydrocarbon receptor signaling pathway
Ge Peng, … , Shigaku Ikeda, François Niyonsaba
Ge Peng, … , Shigaku Ikeda, François Niyonsaba
Published July 14, 2022
Citation Information: J Clin Invest. 2022;132(17):e156501. https://doi.org/10.1172/JCI156501.
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Research Article Dermatology Inflammation

Human β-defensin-3 attenuates atopic dermatitis–like inflammation through autophagy activation and the aryl hydrocarbon receptor signaling pathway

Abstract

Human β-defensin-3 (hBD-3) exhibits antimicrobial and immunomodulatory activities; however, its contribution to autophagy regulation remains unclear, and the role of autophagy in the regulation of the epidermal barrier in atopic dermatitis (AD) is poorly understood. Here, keratinocyte autophagy was restrained in the skin lesions of patients with AD and murine models of AD. Interestingly, hBD-3 alleviated the IL-4– and IL-13–mediated impairment of the tight junction (TJ) barrier through keratinocyte autophagy activation, which involved aryl hydrocarbon receptor (AhR) signaling. While autophagy deficiency impaired the epidermal barrier and exacerbated inflammation, hBD-3 attenuated skin inflammation and enhanced the TJ barrier in AD. Importantly, hBD-3–mediated improvement of the TJ barrier was abolished in autophagy-deficient AD mice and in AhR-suppressed AD mice, suggesting a role for hBD-3–mediated autophagy in the regulation of the epidermal barrier and inflammation in AD. Thus, autophagy contributes to the pathogenesis of AD, and hBD-3 could be used for therapeutic purposes.

Authors

Ge Peng, Saya Tsukamoto, Risa Ikutama, Hai Le Thanh Nguyen, Yoshie Umehara, Juan V. Trujillo-Paez, Hainan Yue, Miho Takahashi, Takasuke Ogawa, Ryoma Kishi, Mitsutoshi Tominaga, Kenji Takamori, Jiro Kitaura, Shun Kageyama, Masaaki Komatsu, Ko Okumura, Hideoki Ogawa, Shigaku Ikeda, François Niyonsaba

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Figure 4

hBD-3–induced autophagy improves TJ barrier function in keratinocytes through AhR signaling.

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hBD-3–induced autophagy improves TJ barrier function in keratinocytes th...
(A and B) Keratinocytes were treated with 10 μg/mL hBD-3 or vehicle control (–) for 9 hours. (A) Representative immunofluorescence images (left) and quantification of LC3 puncta (right); n = 4 per group. Scale bars: 5 μm. (B) Representative electron microscopic images (left) and quantification of autophagic areas (right); n = 10 per group. Scale bars: 10 μm. The yellow arrowheads indicate autophagic vacuoles. (C) Keratinocytes were transfected with adenoviruses carrying Atg3 or mutant Atg3C264S for 48 hours and then treated with 10 μg/mL hBD-3 for 9 hours. Representative immunofluorescence images (left) and quantification of claudin-1 and ZO-1 (right); n = 3 per group. Scale bars: 5 μm. (D) Keratinocyte layers grown on Transwell inserts were transfected with adenoviruses carrying Atg3 or mutant Atg3C264S for 48 hours and then treated with 10 μg/mL hBD-3 for 48 hours, and transepithelial electrical resistance (TER) was assessed by CellZscope. (E) Cells were pretreated with CH-223191 (CH) for 2 hours and then treated with 10 μg/mL hBD-3 for 9 hours. Representative immunoblots of the indicated proteins are shown. Quantification of the band intensities is shown in the right panel; n = 3 per group. (F and G) Keratinocytes were treated with 10 μg/mL hBD-3 or 0.01% acetic acid as a vehicle control for 9 hours. (F) Representative proximity ligation assay images (left) and quantification (right) of the AhR-LC3 and AhR-p62 interactions in keratinocytes with or without AhR siRNA transfection; n = 3 per group. Scale bars: 20 μm. (G) Representative Western blot images (left) and quantification of the band intensities (right) of AhR ubiquitination; n = 3 per group. Mean ± SD. *P < 0.05, **P < 0.01, #P < 0.05, ##P < 0.01, ###P < 0.01. Statistical significance was determined by 2-tailed Student’s t test (A, B, F, and G) and 1-way ANOVA with Tukey’s multiple-comparison test (C–E). All of the data are representative of 3 independent experiments.