title: Decorin-expressing adenovirus decreases collagen synthesis and upregulates MMP expression in keloid fibroblasts and keloid spheroids.
abstract: Decorin is a natural transforming growth factor-β1 (TGF-β1) antagonist. Reduced decorin synthesis is associated with dermal scarring, and increased decorin expression appears to reduce scar tissue formation. To investigate the therapeutic potential of decorin for keloids, human dermal fibroblasts (HDFs) and keloid-derived fibroblasts (KFs) were transduced with decorin-expressing adenovirus (dE1-RGD/GFP/DCN), and we examined the therapeutic potential of decorin-expressing Ad for treating pathologic skin fibrosis. Decorin expression was examined by immunofluorescence assay on keloid tissues. HDFs and KFs were transduced with dE1-RGD/GFP/DCN or control virus, and protein levels of decorin, epidermal growth factor receptor (EGFR) and secreted TGF-β1 were assessed by Western blotting and ELISA. And type I and III collagen, and matrix metalloproteinase-1 (MMP-1) and matrix metalloproteinase-3 (MMP-3) mRNA levels were measured by real-time RT-PCR. Additionally, we immunohistochemically investigated the expression levels of the major extracellular matrix (ECM) proteins in keloid spheroids transduced with dE1-RGD/GFP/DCN. Lower decorin expression was observed in the keloid region compared to adjacent normal tissues. After treatment with dE1-RGD/GFP/DCN, secreted TGF-β1 and EGFR protein expressions were decreased in TGF-β1-treated HDFs and KFs. Also, type I and III collagen mRNA levels were decreased, and the expression of MMP-1 and MMP-3 mRNA was strongly upregulated. In addition, the expression of type I and III collagen, fibronectin and elastin was significantly reduced in dE1-RGD/GFP/DCN-transduced keloid spheroids. These results support the utility of decorin-expressing adenovirus to reduce collagen synthesis in KFs and keloid spheroid, which may be highly beneficial in treating keloids.
authors: Department of Plastic and Reconstructive Surgery, College of Medicine, Institute for Human Tissue Restoration, Yonsei University, Seoul, Korea.
authors: Won Jai
authors: WJ
authors: Lee
authors: Department of Bioengineering, College of Engineering, Hanyang University, Seoul, Korea.
authors: Hyo Min
authors: HM
authors: Ahn
authors: Department of Plastic and Reconstructive Surgery, College of Medicine, Institute for Human Tissue Restoration, Yonsei University, Seoul, Korea.
authors: Hyun
authors: H
authors: Roh
authors: Department of Bioengineering, College of Engineering, Hanyang University, Seoul, Korea.
authors: Youjin
authors: Y
authors: Na
authors: Department of Bioengineering, College of Engineering, Hanyang University, Seoul, Korea.
authors: Il-Kyu
authors: IK
authors: Choi
authors: Department of Dermatology and Cutaneous Biology Research Institute, College of Medicine, Yonsei University, Seoul, Korea.
authors: Ju Hee
authors: JH
authors: Lee
authors: Department of Plastic and Reconstructive Surgery, College of Medicine, Institute for Human Tissue Restoration, Yonsei University, Seoul, Korea.
authors: Yong Oock
authors: YO
authors: Kim
authors: Department of Plastic and Reconstructive Surgery, College of Medicine, Institute for Human Tissue Restoration, Yonsei University, Seoul, Korea.
authors: Dae Hyun
authors: DH
authors: Lew
authors: Department of Bioengineering, College of Engineering, Hanyang University, Seoul, Korea.
authors: Chae-Ok
authors: CO
authors: Yun
mesh_terms: D000256:Adenoviridae / Q000235:genetics*; D002478:Cells, Cultured; D024042:Collagen Type I / Q000096:biosynthesis / Q000235:genetics; D024061:Collagen Type III / Q000096:biosynthesis / Q000235:genetics; D058575:Decorin / Q000235:genetics / Q000627:therapeutic use*; D016326:Extracellular Matrix Proteins / Q000096:biosynthesis* / Q000235:genetics; D005347:Fibroblasts / Q000378:metabolism* / Q000473:pathology; D005786:Gene Expression Regulation; D015316:Genetic Therapy; D005822:Genetic Vectors / Q000235:genetics / Q000494:pharmacology*; D006801:Humans; D007627:Keloid / Q000378:metabolism / Q000473:pathology*; D020781:Matrix Metalloproteinase 1 / Q000096:biosynthesis* / Q000235:genetics; D019278:Matrix Metalloproteinase 3 / Q000096:biosynthesis* / Q000235:genetics; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics; D060888:Real-Time Polymerase Chain Reaction; D012867:Skin / Q000473:pathology; D018874:Spheroids, Cellular
chemical_list: D024042:Collagen Type I; D024061:Collagen Type III; C546878:DCN protein, human; D058575:Decorin; D016326:Extracellular Matrix Proteins; D012333:RNA, Messenger; C511768:MMP3 protein, human; D019278:Matrix Metalloproteinase 3; C522360:MMP1 protein, human; D020781:Matrix Metalloproteinase 1
keywords: MMP; collagen; decorin-expressing adenovirus; gene therapy; keloid; keloid spheroid