TGFBR2 but not SPP1 genotype modulates osteopontin expression in Duchenne muscular dystrophy muscle

A polymorphism (rs28357094) in the promoter region of the SPP1 gene coding for osteopontin (OPN) is a strong determinant of disease severity in Duchenne muscular dystrophy (DMD). The rare G allele of rs28357094 alters gene promoter function and reduces mRNA expression in transfected HeLa cells. To dissect the molecular mechanisms of increased disease severity associated with the G allele, we characterized SPP1 mRNA and protein in DMD muscle biopsies of patients with defined rs28357094 genotype. We did not find significant differences in osteopontin mRNA or protein expression between patients carrying the T (ancestral allele) or TG/GG genotypes at rs28357094. The G allele was significantly associated with reduced CD4+ and CD68+ cells on patient muscle biopsy. We also quantified transforming growth factor‐β (TGFB) and TGFB receptor‐2 (TGFBR2) mRNA in DMD muscle biopsies, given the ability of TGFB and TGFBR2 to activate SPP1 promoter region and their role in DMD pathogenesis. The amount of TGFB and TGFBR2 mRNA did not predict the amount of SPP1 mRNA or protein, while a polymorphism in the TGFBR2 gene (rs4522809) was found to be a strong predictor of SPP1 mRNA level. Our findings suggest that OPN mediates inflammatory changes in DMD and that TGFB signalling has a role in the complex regulation of osteopontin expression. Copyright © 2012 Pathological Society of Great Britain and Ireland. Published by John Wiley & Sons, Ltd.

[1]  G. Livshits,et al.  Quantitative genetic study of the circulating osteopontin in community-selected families , 2011, Osteoporosis International.

[2]  A. Chambers,et al.  Pre- and post-translational regulation of osteopontin in cancer , 2011, Journal of Cell Communication and Signaling.

[3]  G. Lanfranchi,et al.  SPP1 genotype is a determinant of disease severity in Duchenne muscular dystrophy , 2010, Neurology.

[4]  G. Kundu,et al.  Transcriptional regulation of human osteopontin promoter by histone deacetylase inhibitor, trichostatin A in cervical cancer cells , 2010, Molecular Cancer.

[5]  M. Colombo,et al.  Matricellular proteins: from homeostasis to inflammation, cancer, and metastasis , 2010, Cancer and Metastasis Reviews.

[6]  Y. Matsui,et al.  Osteopontin; as a target molecule for the treatment of inflammatory diseases. , 2010, Current drug targets.

[7]  M. Mora,et al.  Altered production of extra-cellular matrix components by muscle-derived Duchenne muscular dystrophy fibroblasts before and after TGF-β1 treatment , 2010, Cell and Tissue Research.

[8]  J. Golledge,et al.  A genetic polymorphism in transforming growth factor beta receptor‐2 is associated with serum osteopontin , 2009, International journal of immunogenetics.

[9]  E. Hoffman,et al.  Osteopontin promotes fibrosis in dystrophic mouse muscle by modulating immune cell subsets and intramuscular TGF-beta. , 2009, The Journal of clinical investigation.

[10]  J. Aubin,et al.  Estrogen receptor-related receptor alpha (ERRalpha) regulates osteopontin expression through a non-canonical ERRalpha response element in a cell context-dependent manner. , 2008, Journal of molecular endocrinology.

[11]  C. Angelini,et al.  Decorin and biglycan expression is differentially altered in several muscular dystrophies. , 2005, Brain : a journal of neurology.

[12]  Y. W. Chen,et al.  Early onset of inflammation and later involvement of TGFβ in Duchenne muscular dystrophy , 2005, Neurology.

[13]  M. Mastrogiacomo,et al.  Bone and cartilage formation by skeletal muscle derived cells , 2005, Journal of cellular physiology.

[14]  R. Ravazzolo,et al.  Polymorphisms in the osteopontin promoter affect its transcriptional activity. , 2004, Physiological genomics.

[15]  Lin Li,et al.  Gene regulation by Sp1 and Sp3. , 2004, Biochemistry and cell biology = Biochimie et biologie cellulaire.

[16]  M. Grounds,et al.  Anti‐TNFα (Remicade®) therapy protects dystrophic skeletal muscle from necrosis , 2004 .

[17]  G. Vita,et al.  Activation of nuclear factor-κB in inflammatory myopathies and Duchenne muscular dystrophy , 2003, Neurology.

[18]  J. D. Porter,et al.  A chronic inflammatory response dominates the skeletal muscle molecular signature in dystrophin-deficient mdx mice. , 2002, Human molecular genetics.

[19]  Thomas D. Schmittgen,et al.  Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method. , 2001, Methods.

[20]  W. Winckler,et al.  Altered pathological progression of diaphragm and quadriceps muscle in TNF-deficient, dystrophin-deficient mice , 2000, Neuromuscular Disorders.

[21]  L. Morandi,et al.  Expression of transforming growth factor-beta 1 in dystrophic patient muscles correlates with fibrosis. Pathogenetic role of a fibrogenic cytokine. , 1995, The Journal of clinical investigation.

[22]  P. Weissberg,et al.  Transforming growth factor beta decreases the rate of proliferation of rat vascular smooth muscle cells by extending the G2 phase of the cell cycle and delays the rise in cyclic AMP before entry into M phase. , 1994, The Biochemical journal.

[23]  K. Campbell,et al.  Dystrophin–glycoprotein complex: Its role in the molecular pathogenesis of muscular dystrophies , 1994, Muscle & nerve.

[24]  G. Proetzel,et al.  Targeted disruption of the mouse transforming growth factor-β1 gene results in multifocal inflammatory disease , 1992, Nature.

[25]  Eric P. Hoffman,et al.  Dystrophin: The protein product of the duchenne muscular dystrophy locus , 1987, Cell.

[26]  L. Boxhorn,et al.  Inhibition of skeletal muscle satellite cell differentiation by transforming growth factor‐beta , 1987, Journal of cellular physiology.

[27]  K. Uaesoontrachoon,et al.  Osteopontin and skeletal muscle myoblasts: association with muscle regeneration and regulation of myoblast function in vitro. , 2008, The international journal of biochemistry & cell biology.

[28]  A. C. Stumbo,et al.  Osteopontin expression in coculture of differentiating rat fetal skeletal fibroblasts and myoblasts , 2006, In Vitro Cellular & Developmental Biology - Animal.

[29]  P. Goldschmidt-Clermont,et al.  Transforming growth factor-beta-induced inhibition of myogenesis is mediated through Smad pathway and is modulated by microtubule dynamic stability. , 2004, Circulation research.

[30]  J. Huard,et al.  Transforming growth factor-beta1 induces the differentiation of myogenic cells into fibrotic cells in injured skeletal muscle: a key event in muscle fibrogenesis. , 2004, The American journal of pathology.

[31]  M. Somerman,et al.  TGFbeta and BMP-2 activation of the OPN promoter: roles of smad- and hox-binding elements. , 2001, Experimental cell research.

[32]  Thomas D. Schmittgen,et al.  Analysis of Relative Gene Expression Data Using Real-Time Quantitative PCR and the 2 2 DD C T Method , 2022 .