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

Abstract

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 α5β1 integrin receptors that further stimulates down-stream ERK/JNK signaling. The regulation of CCN4 by TNF-α prompted us look further at their potential relationship. Treatment of hADFs with CCN4 and TNF-α alone or together showed CCN4 counteracted the inhibition of TNF-α on COL1A1 and FN mRNA expression and the stimulation of TNF-α on MMP-1 and MMP3 mRNA expression. CCN4 appeared to counterbalance the effects of TNF-α by inhibiting downstream NF-κB/p-65 signaling. Taken together we show CCN4 stimulates dermal fibroblast cell migration, proliferation and inhibits TNF-α stimulation, all of which could regulate wound healing.

The tumor cell‐secreted matricellular protein WISP 1 drives pro‐metastatic collagen linearization

“…Therefore, although WISP1 may regulate signaling pathways and induce gene expression changes in other contexts (Konigshoff et al, 2009;Gurbuz & Chiquet-Ehrismann, 2015;Maeda et al, 2015;Zhang et al, 2015;Ono et al, 2018), our findings indicate that WISP1 can regulate cancer cell behavior via its capacity to directly interact with type I collagen and to promote fibril linearization, which in turn facilitates tumor cell invasion. Specifically, no significant changes in gene expression were induced by Wisp1 overexpression in tumor cells cultivated on Col I lattices, in the exact same conditions in which Wisp1 overexpression enhances cell motility.…”

“…Among these factors, WISP1 (CCN4) is highly upregulated by TGFb1 at both the mRNA and protein levels in 4T1 cells as well as in MDA-MB-231 cells and other tumor cell lines (Figs 2A-C and Appendix Fig S1A and B). Interestingly, WISP1 is a member of the CCN family of secreted ECM proteins, which contribute to several developmental and disease processes characterized by profound ECM remodeling, including wound healing, bone turnover, and lung fibrosis (Chen & Lau, 2009;Konigshoff et al, 2009;Maeda et al, 2015;Ono et al, 2018). WISP1 gene expression in breast cancers and other carcinomas is higher than in normal tissues and promotes cancer cell proliferation and invasion in vitro (Chiang et al, 2015;Gurbuz & Chiquet-Ehrismann, 2015).…”

“…Surprisingly, Wisp1 overexpression did not trigger any major gene expression changes in cells plated on either plastic or Col I, in the same conditions in which WISP1 enhances invasion ( Fig 3K and L). Furthermore, the phosphorylation status of members of 8 key pathways implicated in tumor cell motility and invasion, or proposed to be modulated by WISP1 in other cell types (Konigshoff et al, 2009;Gurbuz & Chiquet-Ehrismann, 2015;Zhang et al, 2015;Ono et al, 2018), remained substantially unaffected by Wisp1 overexpression (Fig EV4H). Thus, although WISP1 may regulate signaling pathways and induce gene expression changes in other contexts (Konigshoff et al, 2009;Gurbuz & Chiquet-Ehrismann, 2015;Maeda et al, 2015;Zhang et al, 2015;Ono et al, 2018), the data presented here indicate that it can promote tumor cell invasion by binding to type I collagen and altering its architecture rather than by inducing major gene expression changes.…”

Pseudomonas aeruginosa infection alters the macrophage phenotype switching process during wound healing in diabetic mice

“…The complete medium was supplemented with 20 ng/mL rmM-CSF (Invitrogen) and cultured in a humidified incubator at 37 C and 5% CO 2 . Skin fibroblasts were isolated as previously reported with modification (Ono et al, 2018). Briefly, skin tissue was collected from db/db mouse, after removed the hypodermis layer and associated blood vessels, the left tissue was cut into 2-3 mm pieces and placed in a culture dish and incubated for 3 min to allow attachment, then the culture medium DMEM plus 10% FBS, 50 IU/mL penicillin and 50 mg/mL streptomycin was gently added into dish.…”

CCN proteins as potential actionable targets in scleroderma

“…CCN4 has been identified in skin fibroblasts and seems to play role in a paracrine manner, by binding to decorin and biglycan . Moreover, CCN4 has been shown to regulate wound healing by modulating proliferation, migration and ECM expression in dermal fibroblasts via α5β1 integrin and TNFα …”