IL-13 mediates collagen deposition via STAT6 and microRNA-135b: a role for epigenetics.
Systemic sclerosis is an autoimmune connective tissue disease in which T cells play a prominent role. We and others have previously demonstrated a role for T cell-derived IL-13 in mediating the induction of collagen in dermal fibroblasts and that blockade with IL-13 antibodies attenuates this increase. In this study we want to probe the signalling that underpins IL-13 mediated matrix deposition. Isolated dermal fibroblasts were incubated with recombinant IL-13 and gene expression by qRT-PCR was performed for collagen1A1 and TGF-β1. Small interfering RNA (siRNA) was used to knock down STAT6 and a small molecule inhibitor was also used to block this pathway. MiR-135b was transfected into fibroblasts plus and minus IL-13 to see if this miR plays a role. miR-135b was measured in systemic sclerosis fibroblasts isolated from patients and also in serum. Results showed that IL-13 increased collagen expression and that this is independent from TGF-β1. This is dependent on STAT6 as targeting this blocked induction. MiR-135b reduces collagen induction in fibroblasts and scleroderma fibroblasts have lower constitutive levels of the miR. We further demonstrate that miR135b is repressed by methylation and may include MeCP2. In conclusion we show that STAT6 and miR-135b regulate IL-13-mediated collagen production by fibroblasts.
Faculty of Health and Life Sciences, Northumbria University, Newcastle Upon Tyne, NE1 8ST, United Kingdom.
Steven
S
O'Reilly
Institute of Cellular Medicine Faculty of Medical Sciences, MRG 4th Floor Cookson Building Framlington Place, Newcastle Upon Tyne, NE2 4HH.|National Institute of geriatrics rheumatology and rehabilitation, Warsaw, Poland.|Institute of Immunology and Experimental Therapy, Polish Academy of Science, Wroclaw, Poland.
Marzena
M
Ciechomska
School of Biological and Biomedical Sciences, Durham University, Durham United Kingdom.
Nicola
N
Fullard
School of Biological and Biomedical Sciences, Durham University, Durham United Kingdom.
Stefan
S
Przyborski
Department of Rheumatology and Clinical Immunology University Medical Centre Utrecht, Utrecht, The Netherlands.
Jacob M
JM
van Laar
D000368:Aged; D024042:Collagen Type I / Q000235:genetics*; D000090602:Collagen Type I, alpha 1 Chain; D019175:DNA Methylation; D044127:Epigenesis, Genetic; D005260:Female; D005347:Fibroblasts / Q000166:cytology / Q000378:metabolism; D005786:Gene Expression Regulation; D006801:Humans; D018793:Interleukin-13 / Q000378:metabolism*; D035683:MicroRNAs / Q000235:genetics*; D008875:Middle Aged; D050800:STAT6 Transcription Factor / Q000235:genetics* / Q000378:metabolism; D012595:Scleroderma, Systemic / Q000235:genetics*; D015398:Signal Transduction; D053773:Transforming Growth Factor beta1 / Q000235:genetics
D024042:Collagen Type I; D000090602:Collagen Type I, alpha 1 Chain; C578095:IL13 protein, human; D018793:Interleukin-13; C544965:MIRN135 microRNA, human; D035683:MicroRNAs; D050800:STAT6 Transcription Factor; C494101:STAT6 protein, human; C508960:TGFB1 protein, human; D053773:Transforming Growth Factor beta1