Treatment With Antisense Oligonucleotide Reduces the Expression of Type I Collagen in a Human-Skin Organ-Wound Model

Nath, Rahul K.; Xiong, W.; Humphries, Andrea D.; Beri, Rajinder

doi:10.1097/sap.0b013e31803bf66c

Cited by 7 publications

(4 citation statements)

References 44 publications

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“…Recently, the use of human skin explants for evaluating the effect of potential therapeutic agents has been reported. In an attempt to interfere with the fibrotic process, Nath et al recently reported using antisense oligonucleotides to reduce Collagen Type I expression ex vivo in a skin organ culture model that was maintained for 7 days [ 17 ]. To extend the application of human skin explants to in vivo gene therapy, Lippin et al obtained skin explants from patients with chronic renal failure and anemia, injected them with an adenovirus expressing human erythropoietin, and reimplanted them into the patients [ 18 ].…”

Section: Discussionmentioning

confidence: 99%

Human Skin Culture as an Ex Vivo Model for Assessing the Fibrotic Effects of Insulin-Like Growth Factor Binding Proteins

Yasuoka¹,

Larregina²,

Yamaguchi³

et al. 2008

TORJ

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Systemic sclerosis (SSc) is a connective tissue disease of unknown etiology. A hallmark of SSc is fibrosis of the skin and internal organs. We recently demonstrated increased expression of IGFBP-3 and IGFBP-5 in primary cultures of fibroblasts from the skin of patients with SSc. In vitro, IGFBP-3 and IGFBP-5 induced a fibrotic phenotype and IGFBP-5 triggered dermal fibrosis in mice. To assess the ability of IGFBPs to trigger fibrosis, we used an ex vivo human skin organ culture model. Our findings demonstrate that IGFBP-3 and IGFBP-5, but not IGFBP-4, increase dermal and collagen bundle thickness in human skin explants, resulting in substantial dermal fibrosis and thickening. These fibrotic effects were sustained for at least two weeks. Our findings demonstrate that human skin ex vivo is an appropriate model to assess the effects of fibrosis-inducing factors such as IGFBPs, and for evaluating the efficacy of inhibitors/therapies to halt the progression of fibrosis and potentially reverse it.

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Section: Discussionmentioning

confidence: 99%

Human Skin Culture as an Ex Vivo Model for Assessing the Fibrotic Effects of Insulin-Like Growth Factor Binding Proteins

Yasuoka¹,

Larregina²,

Yamaguchi³

et al. 2008

TORJ

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“…Human HS tissues that satisfied the following requirements were used in the study: 20–50 years; hypertrophic scars had not faded within a year; the patients did not administer any drugs before surgery; confirmation of clinical diagnosis according to the appearance (thickened, raised (>2 mm), red, hard, itchy, and the relevant pathological diagnosis) ( n = 6). Cultured HS tissues ex vivo were performed as described previously [ 24 – 26 ]. Subcutaneous fat tissues were removed, and HS tissues were cut into 10 × 10 mm sections and cultured with DMEM/80 % ADSC-CM in the presence of a p38 inhibitor (SB203580; 10 μM) and activator (anisomycin; 25 μg/ml).…”

Section: Methodsmentioning

confidence: 99%

Adipose tissue-derived stem cells suppress hypertrophic scar fibrosis via the p38/MAPK signaling pathway

Zhang

Gao

et al. 2016

Stem Cell Res Ther

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BackgroundHypertrophic scars (HS) generally occur after injury to the deep layers of the dermis, resulting in functional deficiency for patients. Growing evidence has been identified that the supernatant of adipose tissue-derived stem cells (ADSCs) significantly ameliorates fibrosis of different tissues, but limited attention has been paid to its efficacy on attenuating skin fibrosis. In this study, we explored the effect and possible mechanism of ADSC-conditioned medium (ADSC-CM) on HS.MethodReal-time quantitative polymerase chain reaction (qRT-PCR) and Western blotting were used to detect the expression of collagen I (Col1), collagen III (Col3), and α-smooth muscle actin (α-SMA) after fibroblasts and cultured HS tissues were stimulated with ADSC-CM and p38 inhibitor/activator. Immunofluorescence staining was performed to test the expression of α-SMA. Masson’s trichrome staining, hematoxylin and eosin (H&E) staining, and immunohistochemistry staining were carried out to assess the histological and pathological change of collagen in the BALB/c mouse excisional model. All data were analyzed by using SPSS17.0 software. Statistical analysis was performed by Student’s t tests.ResultsThe in vitro and ex vivo study revealed ADSC-CM decreased the expression of Col1, Col3, and α-SMA. Together, thinner and orderly arranged collagen was manifested in HS tissues cultured with ADSC-CM. Dramatically, the assessed morphology showed an accelerated healing rate, less collagen deposition, and col1- and col3-positive cells in the ADSC-CM treated group. Importantly, the protein level of p-p38 was downregulated in a concentration-dependent manner in HS-derived fibroblasts with ADSC-CM treatment, which further decreased the expression of p-p38 after the application of its inhibitor, SB203580. SB203580 led to an obvious decline in the expression of Col1, Col3, and α-SMA in fibroblasts and cultured HS tissues and presented more ordered arrangement and thinner collagen fibers in BALB/c mice. Lastly, anisomycin, an agonist of p38, upregulated the expression of fibrotic proteins and revealed more disordered structure and denser collagen fibers.ConclusionThis study demonstrated that ADSC-CM could decrease collagen deposition and scar formation in in vitro, ex vivo and in vivo experiments. The regulation of the p38/MAPK signaling pathway played an important role in the process. The application of ADSC-CM may provide a novel therapeutic strategy for HS treatment, and the anti-scarring effect can be achieved by inhibition of the p38/MAPK signaling pathway.Electronic supplementary materialThe online version of this article (doi:10.1186/s13287-016-0356-6) contains supplementary material, which is available to authorized users.

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“…Lowering collagen expression in these tissues may have implications in improving their mechanical properties during the healing process of ligaments and tendons. Our own study demonstrated the efficacy of directly targeting the COL1A1 collagen gene using antisense to type 1 collagen, reducing collagen expression in a human-skin organ model [20].…”

Section: Discussionmentioning

confidence: 79%

“…Hence there is a critical need for effective and safe therapies. We have previously demonstrated that the type 1 collagen antisense oligodeoxynucleotide, AS60-ODN (a human analog of mouse AS61-ODN) reduced the mRNA and protein expression of type 1 collagen in a novel ex vivo human wound injury model [20] .…”

Section: Introductionmentioning

confidence: 99%

Protective Effect of Type I Collagen Antisense Oligonucleotides on Bleomycin Induced Pulmonary Fibrosis

Nath¹,

Somasundaram²,

Xiong³

et al. 2010

TOPROCJ

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Background: Pulmonary fibrosis is a chronic and usually untreatable fatal lung disease. It can result from many endogenous or exogenous causes, such as infection, chronic inflammatory diseases, environmental toxins, chemical poisoning, radiation and chemotherapy. The accumulation of collagen, largely type I collagen rich extracellular matrix (ECM) is the hallmark of lung fibrosis. Current therapies for pulmonary fibrosis utilizing corticosteroids and immunosupressants have demonstrated only marginal effectiveness. Methods: In the present study, we tested the efficacy of the mouse type 1 collagen antisense oligodeoxynucleotides (AS61-ODN, a 20 mer) in bleomycin induced pulmonary fibrosis in mice. Bleomycin was instilled transtracheally by direct tracheal cut down in mice, and oligonucleotides (ODN) were injected through the tail vein. Hematoxylin and eosin (H & E) and Masson's trichrome staining were done to determine the cellular architecture of the lungs after bleomycin and oligonucleotide treatment. Type I procollagen gene COL1A1 expression was determined by Northern blot analysis. Type I collagen protein expression was determined by Western blot analysis and immunofluorescence. Results: There was much less disruption of the lung architecture as indicated by significant decrease in the Ashcroft score, when mice were injected with AS61-ODN following bleomycin instillation. AS61-ODN significantly reduced the mRNA and protein expression of type 1 collagen by 45.2 % (p < 0.015) and 35.0 % (p < 0.004) respectively 21 days after treatment in bleomycin induced fibrotic lung tissue. Immunofluorescence of bleomycin induced mouse lung tissue treated with AS61-ODN showed a specific reduction in collagen expression and fibrosis. Conclusion: Human type I collagen antisense oligodeoxynucleotides (AS60-ODN, a 20 mer), which is 99 percent homologous with mouse AS61-ODN, might be useful as a potential therapeutic agent to treat patients with pulmonary fibrosis.

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Treatment With Antisense Oligonucleotide Reduces the Expression of Type I Collagen in a Human-Skin Organ-Wound Model

Cited by 7 publications

References 44 publications

Human Skin Culture as an Ex Vivo Model for Assessing the Fibrotic Effects of Insulin-Like Growth Factor Binding Proteins

Human Skin Culture as an Ex Vivo Model for Assessing the Fibrotic Effects of Insulin-Like Growth Factor Binding Proteins

Adipose tissue-derived stem cells suppress hypertrophic scar fibrosis via the p38/MAPK signaling pathway

Protective Effect of Type I Collagen Antisense Oligonucleotides on Bleomycin Induced Pulmonary Fibrosis

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