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. 2024 Jan 2;12(1):2203841.
doi: 10.1080/21688370.2023.2203841. Epub 2023 Apr 20.

House dust mite and Th2 cytokine-mediated epithelial barrier dysfunction attenuation by KL001 in 16-HBE cells

Santhosh Kumar Duraisamy  1 , Ashokkumar Srinivasan  1 , Isaac Kirubakaran Sundar  1
Affiliations

House dust mite and Th2 cytokine-mediated epithelial barrier dysfunction attenuation by KL001 in 16-HBE cells

Santhosh Kumar Duraisamy et al. Tissue Barriers. .

Abstract

House dust mite (HDM) is a common aeroallergen that can disrupt the airway epithelial barrier leading to dysregulated immune response, resulting in allergic lung diseases such as asthma. Cryptochrome (CRY), a circadian clock gene, plays an important role in the regulation of metabolism, and immune response. It remains unclear whether stabilizing CRY using KL001 can attenuate HDM/Th2 cytokine-induced epithelial barrier dysfunction in 16-HBE cells. We evaluate the effect of KL001 (20 µM) pre-treatment (4 hrs) in HDM/Th2 cytokine (IL-4 or IL-13)-mediated change in epithelial barrier function. HDM and Th2 cytokine-induced changes in transepithelial electrical resistance (TEER) were determined by an xCELLigence real-time cell analyzer and delocalization of adherens junction complex (AJC: E-cadherin and β-catenin) and tight junction proteins (TJP: Occludin and Zonula occludens-1) by immunostaining and confocal microscopy. Finally, quantitative real-time PCR (qRT-PCR) and Western blotting were used to measure altered gene expression and protein abundance of the epithelial barrier function and core clock genes, respectively. HDM and Th2 cytokine treatment significantly decreased TEER associated with altered gene expression and protein abundance of the selected epithelial barrier function and circadian clock genes. However, pre-treatment with KL001 attenuated HDM and Th2 cytokine-induced epithelial barrier dysfunction as early as 12-24 hrs. KL001 pre-treatment showed attenuation of HDM and Th2 cytokine-induced alteration in the localization and gene expression of AJP and TJP (Cdh1, Ocln, and Zo1) and core clock genes (Clock, Arntl/Bmal1, Cry1/2, Per1/2, Nr1d1/Rev-erbα, and Nfil3). We demonstrate, for the first time, the protective role of KL001 in HDM and Th2 cytokine-mediated epithelial barrier dysfunction.

Keywords: KL001; allergic asthma; circadian clock; epithelial barrier dysfunction; house dust mite; transepithelial electrical resistance.

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Conflict of interest statement

No potential conflict of interest was reported by the authors.

Figures

None
Graphical abstract
Figure 1.
Figure 1.
HDM and Th2 cytokine (IL-4 and IL-13)-induced epithelial barrier dysfunction attenuation by KL001 in 16-HBE cells.
Figure 2.
Figure 2.
KL001 treatment restored HDM and Th2 cytokine (IL-4 and IL-13)-induced disruption of adherens junction complex proteins in 16-HBE cells.
Figure 3.
Figure 3.
KL001 treatment restored HDM and Th2 cytokine (IL-4 and IL-13)-induced disruption of tight junction proteins in 16-HBE cells.
Figure 4.
Figure 4.
KL001 treatment attenuates HDM/Th2 cytokine (IL-4 and IL-13)-induced change in mRNA expression of adherens junction complex and tight junction protein genes in 16-HBE cells.
Figure 5.
Figure 5.
KL001 treatment attenuates HDM/Th2 cytokine (IL-4 and IL-13)-induced change in mRNA expression of circadian clock genes in 16-HBE cells.
See this image and copyright information in PMC

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