CCN4/WISP1 controls cutaneous wound healing by modulating proliferation, migration and ECM expression in dermal fibroblasts via α5β1 and TNF\α

Understanding the mechanisms that control cutaneous wound healing is crucial to successfully manage repair of damaged skin. The goal of the current study was to uncover novel extracellular matrix (ECM) components that control the wound healing process. Full thickness skin defects were created in mice and used to show CCN4 up-regulation during wound-healing as early as 1 day after surgery, suggesting a role in inflammation and subsequent dermal migration and proliferation. To determine how CCN4 could regulate wound healing we used Ccn4-KO mice and showed they had delayed wound closure accompanied by reduced expression of Col1a1 and Fn mRNA. Boyden chamber assays using Ccn4-deficient dermal fibroblasts showed they have reduced migration and proliferation compared to WT counterparts. To confirm CCN4 has a role in proliferation and migration of dermal cells, siRNA knockdown and transduction of CCN4 adenoviral transduction were used and resulted in reduced or enhanced migration of human adult dermal fibroblast (hADF) cells respectively. The induced migration of the dermal fibroblasts by CCN4 appears to work via alpha 5 beta 1 integrin receptors that further stimulates down-stream ERK/JNK signaling. The regulation of CCN4 by TNF-alpha prompted us look further at their potential relationship. Treatment of hADFs with CCN4 and TNF-alpha alone or together showed CCN4 counteracted the inhibition of TNF-alpha on COL1A1 and FN mRNA expression and the stimulation of TNF-alpha on MMP-1 and MMP3 mRNA expression. CCN4 appeared to counterbalance the effects of TNF-alpha by inhibiting downstream NF-kappa B/p-65 signaling. Taken together we show CCN4 stimulates dermal fibroblast cell migration, proliferation and inhibits TNF-alpha stimulation, all of which could regulate wound healing. Published by Elsevier B.V.