Abstract
Chronic non-healing wounds including diabetic, venous, and decubitus skin ulcers are currently lacking effective therapies. Non-healing diabetic ulcers can lead to amputations as progress into a highly chronic state before detection and existing treatments for these wounds often fail. Granzyme B (GzmB) is a serine protease that was, until recently, believed to function exclusively in cytotoxic lymphocyte-mediated apoptosis. However, during excessive or chronic inflammation, GzmB can accumulate in the extracellular milieu, retain its activity, and cleave a number of important extracellular proteins. Epidermal growth factor receptor (EGFR) is a transmembrane receptor involved in cellular processes such as proliferation and migration. EGFR signaling is integral to the wound healing process. The present study investigated the effects of GzmB on keratinocyte cell migration using HaCaT cell line. Using electric cell-substrate impedance sensing and scratch assays, the present study demonstrates that GzmB inhibits keratinocyte migration by interfering with the EGFR pathway. GzmB limited cell transition into a migratory morphology and was found to reduce ligand-induced EGFR phosphorylation. Inhibition of GzmB reversed the aforementioned effects. In summary, data from the present study suggest key role for GzmB in the pathogenesis of impaired wound healing through the impairment of EGFR signaling and cell migration.
Funding source: Canadian Institutes of Health Research
Award Identifier / Grant number: 332654
Funding statement: We thank Dr. Honglin Luo, Furquan Shaheen, Alana Jackson, Steve Kalloger for their technical assistance; Dr. Kevin Bennewith (BC Cancer Research Center, Vancouver, BC, Canada) for providing MCF-7 cells; This study was funded by a grants-in-aid from the Canadian Diabetes Association (D.J.G.), Rick Hansen Institute (D.J.G.) and Canadian Institutes of Health Research (CIHR Grant/Award Number: ‘332654’) (D.J.G., R.C.B.): CIHR IMPACT post-doctoral fellowship (S.S.); Frederick Banting and Charles Best Canada Graduate Scholarship (Y.M. and M.S.); MITACS Accelerate Post-doctoral fellowship (L.G.P.).
Acknowledgments
We thank Dr. Honglin Luo, Furquan Shaheen, Alana Jackson, Steve Kalloger for their technical assistance; Dr. Kevin Bennewith (BC Cancer Research Center, Vancouver, BC, Canada) for providing MCF-7 cells; This study was funded by a grants-in-aid from the Canadian Diabetes Association (D.J.G.), Rick Hansen Institute (D.J.G.) and Canadian Institutes of Health Research (CIHR Grant/Award Number: ‘332654’) (D.J.G., R.C.B.): CIHR IMPACT post-doctoral fellowship (S.S.); Frederick Banting and Charles Best Canada Graduate Scholarship (Y.M. and M.S.); MITACS Accelerate Post-doctoral fellowship (L.G.P.).
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