Myosin light chain kinase-mediated epithelial barrier dysfunction as a potential pathogenic mechanism of afatinib-induced diarrheas: A study in human colonoid model
Issued Date
2025-01-15
Resource Type
ISSN
00142999
eISSN
18790712
Scopus ID
2-s2.0-85211055206
Pubmed ID
39637932
Journal Title
European Journal of Pharmacology
Volume
987
Rights Holder(s)
SCOPUS
Bibliographic Citation
European Journal of Pharmacology Vol.987 (2025)
Suggested Citation
Worakajit N., Satitsri S., Kitiyakara T., Muanprasat C. Myosin light chain kinase-mediated epithelial barrier dysfunction as a potential pathogenic mechanism of afatinib-induced diarrheas: A study in human colonoid model. European Journal of Pharmacology Vol.987 (2025). doi:10.1016/j.ejphar.2024.177174 Retrieved from: https://repository.li.mahidol.ac.th/handle/20.500.14594/102937
Title
Myosin light chain kinase-mediated epithelial barrier dysfunction as a potential pathogenic mechanism of afatinib-induced diarrheas: A study in human colonoid model
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Abstract
Diarrheas are an important adverse effect of afatinib, a tyrosine kinase inhibitor (TKI) anti-cancer drug, leading to mortality and morbidity in cancer patients with their pathophysiological mechanisms related to intestinal barrier dysfunctions being poorly understood. This study aimed to investigate the effect of afatinib on intestinal epithelial barrier integrity using a human colon-derived organoid model (colonoids). Afatinib (0.5 μM) significantly decreased the transepithelial electrical resistance (TEER) by ∼60% and increased apical-to-basolateral dextran flux by > 20 folds without causing apparent cytotoxicity in human colonoids. The delocalization of zonula occludens-1 (ZO-1) and a decrease in mRNA and protein expression of claudin-4 and ZO-1 were also observed in the afatinib-treated human colonoids. Afatinib induced nuclear translocation of nuclear factor kappa B (NF-κB) as well as mRNA and protein expression of NF-κB targets including tumor necrosis factor (TNF)-alpha, interleukin-8 (IL-8), and inducible nitric oxide synthase (iNOS) indicating the initiation of the NF-κB-mediated epithelial inflammatory responses. Interestingly, afatinib induced mRNA and protein expression of myosin light chain (MLC) kinase (MLCK) and MLC phosphorylation, a known inducer of intestinal epithelial barrier disruption. Treatment with iNOS inhibitor (1400W) or MLCK inhibitor (ML-7) reversed the effect of afatinib on mRNA expressions of ZO-1 and claudin-4, and TEER. Collectively, our results indicate that afatinib induces intestinal epithelial barrier dysfunction via mechanisms involving NF-κB-iNOS-MLCK pathways. This finding may pave the way for developing therapeutic strategies to reduce adverse effects and enhance efficacy of TKI in cancer patients.