Gene expression profiling highlights defective myogenesis in DMD patients and a possible role for bone morphogenetic protein 4

Duchenne Muscular Dystrophy (DMD) is characterized by progressive muscle weakness and wasting. Despite the sustained presence of satellite cells in their skeletal muscles, muscle regeneration in DMD patients seems inefficient and unable to compensate for the continuous muscle fiber loss. To find a molecular explanation, we compared the gene expression profiles of myoblasts from healthy individuals and DMD patients during activation and differentiation in culture. DMD cultures showed significant gene expression changes, even before dystrophin is expressed. We found a higher expression level of bone morphogenetic protein 4 (BMP4) in DMD cultures, which we demonstrate to inhibit differentiation into myotubes. In the later stages of differentiation, we observed a significant decline in expression of sarcomeric genes in the absence of dystrophin, probably contributing to sarcomeric instability. These results support the hypothesis that inefficient muscle regeneration is caused by impaired myoblast differentiation and impaired maintenance of the myotubes.

[1]  Seumas McCroskery,et al.  Myostatin negatively regulates satellite cell activation and self-renewal , 2003, The Journal of cell biology.

[2]  R. Fässler,et al.  Absence of integrin alpha 7 causes a novel form of muscular dystrophy. , 1997, Nature genetics.

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

[4]  B. Benabdallah,et al.  Improved Success of Myoblast Transplantation in mdx Mice by Blocking the Myostatin Signal , 2005, Transplantation.

[5]  C. Auffray,et al.  Gene expression profiling of human satellite cells during muscular aging using cDNA arrays. , 2003, Gene.

[6]  M. Fardeau,et al.  Comparison between the growth pattern of cell cultures from normal and Duchenne dystrophy muscle , 1984, Journal of the Neurological Sciences.

[7]  A S Verkman,et al.  Structure and function of aquaporin water channels. , 2000, American journal of physiology. Renal physiology.

[8]  R. Seger,et al.  Fibroblast Growth Factor Promotes Recruitment of Skeletal Muscle Satellite Cells in Young and Old Rats , 1999, The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society.

[9]  Johan T den Dunnen,et al.  A common reference for cDNA microarray hybridizations. , 2002, Nucleic acids research.

[10]  F. Zara,et al.  Mutations in the caveolin-3 gene cause autosomal dominant limb-girdle muscular dystrophy , 1998, Nature Genetics.

[11]  K. Campbell,et al.  Association of dystrophin and an integral membrane glycoprotein , 1989, Nature.

[12]  J. Bouchard,et al.  Dystrophin expression in myofibers of Duchenne muscular dystrophy patients following intramuscular injections of normal myogenic cells. , 2004, Molecular therapy : the journal of the American Society of Gene Therapy.

[13]  M. Ohta,et al.  A quantitative study of the muscle satellite cells in various neuromuscular disorders , 1983, Journal of the Neurological Sciences.

[14]  Johan T. den Dunnen,et al.  Large-scale gene expression analysis of human skeletal myoblast differentiation , 2004, Neuromuscular Disorders.

[15]  S. Winokur,et al.  Facioscapulohumeral muscular dystrophy (FSHD) myoblasts demonstrate increased susceptibility to oxidative stress , 2003, Neuromuscular Disorders.

[16]  J. D. den Dunnen,et al.  Characterization and cell type distribution of a novel, major transcript of the Duchenne muscular dystrophy gene. , 1992, Differentiation; research in biological diversity.

[17]  J. Lefaucheur,et al.  Basic fibroblast growth factor promotes in vivo muscle regeneration in murine muscular dystrophy , 1995, Neuroscience Letters.

[18]  Laminin-alpha2 but not -alpha1-mediated adhesion of human (Duchenne) and murine (mdx) dystrophic myotubes is seriously defective. , 1997, FEBS letters.

[19]  H. Blau,et al.  Primary mouse myoblast purification, characterization, and transplantation for cell-mediated gene therapy , 1994, The Journal of cell biology.

[20]  D. Zwijnenburg,et al.  Relative quantification of 40 nucleic acid sequences by multiplex ligation-dependent probe amplification. , 2002, Nucleic acids research.

[21]  H. Eppenberger,et al.  Proliferation and differentiation of chick skeletal muscle cells cultured in a chemically defined medium. , 1981, Experimental cell research.

[22]  G. V. Ommen,et al.  Targeted exon skipping as a potential gene correction therapy for Duchenne muscular dystrophy , 2002, Neuromuscular Disorders.

[23]  Juan Li,et al.  Adeno-associated virus vector carrying human minidystrophin genes effectively ameliorates muscular dystrophy in mdx mouse model. , 2000, Proceedings of the National Academy of Sciences of the United States of America.

[24]  T. Rando The dystrophin–glycoprotein complex, cellular signaling, and the regulation of cell survival in the muscular dystrophies , 2001, Muscle & nerve.

[25]  Michael G. Walker,et al.  Cell adhesion and matrix remodeling genes identified by co‐expression analysis , 2002 .

[26]  R. Ahima,et al.  Functional improvement of dystrophic muscle by myostatin blockade , 2002, Nature.

[27]  D. Srivastava,et al.  Members of the HRT family of basic helix-loop-helix proteins act as transcriptional repressors downstream of Notch signaling. , 2000, Proceedings of the National Academy of Sciences of the United States of America.

[28]  K. Wagner,et al.  Loss of myostatin attenuates severity of muscular dystrophy in mdx mice , 2002, Annals of neurology.

[29]  L. Chandler,et al.  Delivery of FGF genes to wound repair cells enhances arteriogenesis and myogenesis in skeletal muscle. , 2002, Molecular therapy : the journal of the American Society of Gene Therapy.

[30]  T. Rando,et al.  The regulation of Notch signaling controls satellite cell activation and cell fate determination in postnatal myogenesis. , 2002, Developmental cell.

[31]  E. Metter,et al.  Skeletal muscle satellite cell populations in healthy young and older men and women , 2000, The Anatomical record.

[32]  M. Koenig,et al.  Complete cloning of the duchenne muscular dystrophy (DMD) cDNA and preliminary genomic organization of the DMD gene in normal and affected individuals , 1987, Cell.

[33]  G. Butler-Browne,et al.  Skeletal muscle regeneration and the mitotic clock , 2000, Experimental Gerontology.

[34]  U. Lendahl,et al.  Functional Notch signaling is required for BMP4-induced inhibition of myogenic differentiation , 2003, Development.

[35]  K. Oexle,et al.  Cause of progression in Duchenne muscular dystrophy: impaired differentiation more probable than replicative aging. , 2001, Neuropediatrics.

[36]  C. Minetti,et al.  Altered aquaporin‐4 expression in human muscular dystrophies: a common feature? , 2002, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[37]  J. Schouten,et al.  Expression profiling via novel multiplex assay allows rapid assessment of gene regulation in defined signalling pathways. , 2003, Nucleic acids research.

[38]  Gayle M. Smythe,et al.  Notch-Mediated Restoration of Regenerative Potential to Aged Muscle , 2003, Science.

[39]  James M. Allen,et al.  Systemic delivery of genes to striated muscles using adeno-associated viral vectors , 2004, Nature Medicine.

[40]  Larry Kedes,et al.  HES and HERP families: Multiple effectors of the notch signaling pathway , 2003, Journal of cellular physiology.

[41]  J. E. Anderson,et al.  Deflazacort Increases Laminin Expression and Myogenic Repair, and Induces Early Persistent Functional Gain in mdx Mouse Muscular Dystrophy , 2000, Cell transplantation.

[42]  Eric P. Hoffman,et al.  Expression Profiling in the Muscular Dystrophies Identification of Novel Aspects of Molecular Pathophysiology , 2000 .

[43]  Luis Garcia,et al.  Rescue of Dystrophic Muscle Through U7 snRNA-Mediated Exon Skipping , 2004, Science.

[44]  Alex E. Lash,et al.  Gene Expression Omnibus: NCBI gene expression and hybridization array data repository , 2002, Nucleic Acids Res..

[45]  F. Baas,et al.  Therapeutic antisense-induced exon skipping in cultured muscle cells from six different DMD patients. , 2003, Human molecular genetics.

[46]  H. Blau,et al.  Defective myoblasts identified in Duchenne muscular dystrophy. , 1983, Proceedings of the National Academy of Sciences of the United States of America.

[47]  A. Monaco,et al.  An explanation for the phenotypic differences between patients bearing partial deletions of the DMD locus. , 1988, Genomics.

[48]  M. Fardeau,et al.  Changes in surface morphology and basal lamina of cultured muscle cells from Duchenne muscular dystrophy patients , 1990, Journal of the Neurological Sciences.

[49]  Tatsushi Toda,et al.  cDNA microarray analysis of individual Duchenne muscular dystrophy patients. , 2003, Human molecular genetics.

[50]  C. Mann,et al.  Functional amounts of dystrophin produced by skipping the mutated exon in the mdx dystrophic mouse , 2003, Nature Medicine.

[51]  L. McIntosh,et al.  Deflazacort but not prednisone improves both muscle repair and fiber growth in diaphragm and limb muscle in vivo in the mdx dystrophic mouse , 1996, Muscle & nerve.

[52]  L. Kunkel,et al.  Gene expression comparison of biopsies from Duchenne muscular dystrophy (DMD) and normal skeletal muscle , 2002, Proceedings of the National Academy of Sciences of the United States of America.

[53]  E. Wanke,et al.  Laminin‐α2 but not ‐α1‐mediated adhesion of human (Duchenne) and murine (mdx) dystrophic myotubes is seriously defective , 1997 .

[54]  H. Blau,et al.  Differentiation properties of pure populations of human dystrophic muscle cells. , 1983, Experimental cell research.

[55]  M. Gessler,et al.  Comparative analysis of the human and mouse Hey1 promoter: Hey genes are new Notch target genes. , 2000, Biochemical and biophysical research communications.

[56]  B. Nico,et al.  Aquaporins in skeletal muscle: reassessment of the functional role of aquaporin‐4 , 2004, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[57]  A. Verkman More than just water channels: unexpected cellular roles of aquaporins , 2005, Journal of Cell Science.

[58]  Bing Zhang,et al.  GOTree Machine (GOTM): a web-based platform for interpreting sets of interesting genes using Gene Ontology hierarchies , 2004, BMC Bioinformatics.

[59]  K. Campbell,et al.  Muscular dystrophies and the dystrophin-glycoprotein complex. , 1997, Current opinion in neurology.

[60]  B. Hogan Bone morphogenetic proteins in development. , 1996, Current opinion in genetics & development.

[61]  J. Schouten,et al.  Two‐color multiplex ligation‐dependent probe amplification: Detecting genomic rearrangements in hereditary multiple exostoses , 2004, Human mutation.

[62]  Ross Ihaka,et al.  Gentleman R: R: A language for data analysis and graphics , 1996 .

[63]  Z. Yablonka-Reuveni,et al.  Proliferative Dynamics and the Role of FGF2 During Myogenesis of Rat Satellite Cells on Isolated Fibers. , 1997, Basic and applied myology : BAM.