Tumour necrosis factor alpha-stimulated gene-6 inhibits osteoblastic differentiation of human mesenchymal stem cells induced by osteogenic differentiation medium and BMP-2.

To better understand the molecular pathogenesis of OPLL (ossification of the posterior longitudinal ligament) of the spine, an ectopic bone formation disease, we performed cDNA microarray analysis on cultured ligament cells from OPLL patients. We found that TSG-6 (tumour necrosis factor alpha-stimulated gene-6) is down-regulated during osteoblastic differentiation. Adenovirus vector-mediated overexpression of TSG-6 inhibited osteoblastic differentiation of human mesenchymal stem cells induced by BMP (bone morphogenetic protein)-2 or OS (osteogenic differentiation medium). TSG-6 suppressed phosphorylation and nuclear accumulation of Smad 1/5 induced by BMP-2, probably by inhibiting binding of the ligand to the receptor, since interaction between TSG-6 and BMP-2 was observed in vitro. TSG-6 has two functional domains, a Link domain (a hyaluronan binding domain) and a CUB domain implicated in protein interaction. The inhibitory effect on osteoblastic differentiation was completely lost with exogenously added Link domain-truncated TSG-6, while partial inhibition was retained by the CUB domain-truncated protein. In addition, the inhibitory action of TSG-6 and the in vitro interaction of TSG-6 with BMP-2 were abolished by the addition of hyaluronan. Thus, TSG-6, identified as a down-regulated gene during osteoblastic differentiation, suppresses osteoblastic differentiation induced by both BMP-2 and OS and is a plausible target for therapeutic intervention in OPLL.

[1]  Kenichi Yoshida,et al.  The Promyelotic Leukemia Zinc Finger Promotes Osteoblastic Differentiation of Human Mesenchymal Stem Cells as an Upstream Regulator of CBFA1*[boxs] , 2005, Journal of Biological Chemistry.

[2]  James D. Kang,et al.  Ossification of the posterior longitudinal ligament. Results of anterior cervical decompression and arthrodesis in sixty-one North American patients. , 2005, The Journal of bone and joint surgery. American volume.

[3]  Huan-Cheng Chang,et al.  Measuring masses of single bacterial whole cells with a quadrupole ion trap. , 2004, Journal of the American Chemical Society.

[4]  M. Nanes.  Tumor necrosis factor-alpha: molecular and cellular mechanisms in skeletal pathology. , 2003, Gene.

[5]  M. Zaidi,et al.  TSH Is a Negative Regulator of Skeletal Remodeling , 2003, Cell.

[6]  Toshihiro Tanaka,et al.  Genomewide linkage and linkage disequilibrium analyses identify COL6A1, on chromosome 21, as the locus for ossification of the posterior longitudinal ligament of the spine. , 2003, American journal of human genetics.

[7]  M. Khokha,et al.  Gremlin is the BMP antagonist required for maintenance of Shh and Fgf signals during limb patterning , 2003, Nature Genetics.

[8]  C. Milner,et al.  TSG-6: a multifunctional protein associated with inflammation , 2003, Journal of Cell Science.

[9]  T. Glant,et al.  Impaired cumulus mucification and female sterility in tumor necrosis factor-induced protein-6 deficient mice , 2003, Development.

[10]  M. Longaker,et al.  The BMP antagonist noggin regulates cranial suture fusion , 2003, Nature.

[11]  J. Clohisy,et al.  Tumor necrosis factor-α inhibits pre-osteoblast differentiation through its type-1 receptor , 2003 .

[12]  Markus Affolter,et al.  Structural basis of BMP signalling inhibition by the cystine knot protein Noggin , 2002, Nature.

[13]  Masaki Noda,et al.  Negative Regulation of Bone Morphogenetic Protein/Smad Signaling by Cas-interacting Zinc Finger Protein in Osteoblasts* , 2002, The Journal of Biological Chemistry.

[14]  P. Roughley,et al.  Cartilage-specific constitutive expression of TSG-6 protein (product of tumor necrosis factor alpha-stimulated gene 6) provides a chondroprotective, but not antiinflammatory, effect in antigen-induced arthritis. , 2002, Arthritis and rheumatism.

[15]  Marilyn S. Rugg,et al.  A Novel Allelic Variant of the Human TSG-6 Gene Encoding an Amino Acid Difference in the CUB Module , 2002, The Journal of Biological Chemistry.

[16]  A. J. Day,et al.  Hyaluronan-binding Proteins: Tying Up the Giant* , 2002, The Journal of Biological Chemistry.

[17]  J. Puzas,et al.  A Role for the BMP Antagonist Chordin in Endochondral Ossification , 2002, Journal of bone and mineral research : the official journal of the American Society for Bone and Mineral Research.

[18]  K. Miyazono,et al.  Divergence and convergence of TGF‐β/BMP signaling , 2001, Journal of cellular physiology.

[19]  K. Ikari,et al.  Functional Impact of Human Collagen α2(XI) Gene Polymorphism in Pathogenesis of Ossification of the Posterior Longitudinal Ligament of the Spine , 2001, Journal of bone and mineral research : the official journal of the American Society for Bone and Mineral Research.

[20]  R. Harland,et al.  Mutation and Analysis of Dan, the Founding Member of the Dan Family of Transforming Growth Factor β Antagonists , 2001, Molecular and Cellular Biology.

[21]  Sakae Tanaka,et al.  Negative Regulation of BMP/Smad Signaling by Tob in Osteoblasts , 2000, Cell.

[22]  J. Sinsheimer,et al.  Linkage analysis of candidate genes as susceptibility loci for osteoarthritis-suggestive linkage of COL9A1 to female hip osteoarthritis. , 2000, Rheumatology.

[23]  L. Cardon,et al.  Osteoarthritis-susceptibility locus on chromosome 11q, detected by linkage. , 1999, American Journal of Human Genetics.

[24]  T. Bouwmeester,et al.  The head inducer Cerberus is a multifunctional antagonist of Nodal, BMP and Wnt signals , 1999, Nature.

[25]  M. Leppert,et al.  Genetic mapping of ossification of the posterior longitudinal ligament of the spine. , 1998, American journal of human genetics.

[26]  Kohei Miyazono,et al.  TGF-β signalling from cell membrane to nucleus through SMAD proteins , 1997, Nature.

[27]  K. Miyazono,et al.  Cloning and characterization of a human type II receptor for bone morphogenetic proteins. , 1995, Proceedings of the National Academy of Sciences of the United States of America.

[28]  L. Klampfer,et al.  TSG-6: a TNF-, IL-1-, and LPS-inducible secreted glycoprotein associated with arthritis. , 1993, Journal of immunology.

[29]  P. Bork,et al.  The CUB domain. A widespread module in developmentally regulated proteins. , 1993, Journal of molecular biology.

[30]  T. Shows,et al.  Transcriptional regulation of TSG6, a tumor necrosis factor- and interleukin-1-inducible primary response gene coding for a secreted hyaluronan-binding protein. , 1993, The Journal of biological chemistry.

[31]  Tae H. Lee,et al.  A novel secretory tumor necrosis factor-inducible protein (TSG-6) is a member of the family of hyaluronate binding proteins, closely related to the adhesion receptor CD44 , 1992, The Journal of cell biology.

[32]  J. Vilček,et al.  Tumor necrosis factor. New insights into the molecular mechanisms of its multiple actions. , 1991, The Journal of biological chemistry.

[33]  J. Vilček,et al.  Isolation and characterization of eight tumor necrosis factor-induced gene sequences from human fibroblasts , 1990, Molecular and cellular biology.

[34]  K. Terayama.  Genetic Studies on Ossification of the Posterior Longitudinal Ligament of the Spine , 1989, Spine.

[35]  K. Wada.  [Genetic studies on the ossification of the posterior longitudinal ligament]. , 1987, Nihon Seikeigeka Gakkai zasshi.

[36]  L. Old,et al.  Tumor necrosis factor (TNF). , 1985, Science.

[37]  F. Barry,et al.  Up-regulation and differential expression of the hyaluronan-binding protein TSG-6 in cartilage and synovium in rheumatoid arthritis and osteoarthritis. , 2001, Osteoarthritis and cartilage.

[38]  T. Martin,et al.  Tumor Necrosis Factor (cid:97) Stimulates Osteoclast Differentiation by a Mechanism Independent of the ODF/RANKL–RANK Interaction , 2022 .

[39]  S. Teitelbaum,et al.  Tumor Necrosis Factor-α Mediates Orthopedic Implant Osteolysis , 1999 .

[40]  Printed in U.S.A. Copyright © 2000 by The Endocrine Society Inhibition of Osteoblast Differentiation by Tumor Necrosis Factor-�* , 2022 .