Impaired bone remodeling in children with osteogenesis imperfecta treated and untreated with bisphosphonates: the role of DKK1, RANKL, and TNF-α

SummaryIn this study, we investigated the bone cell activity in patients with osteogenesis imperfecta (OI) treated and untreated with neridronate. We demonstrated the key role of Dickkopf-1 (DKK1), receptor activator of nuclear factor-κB ligand (RANKL), and tumor necrosis factor alpha (TNF-α) in regulating bone cell of untreated and treated OI subjects. These cytokines could represent new pharmacological targets for OI.IntroductionBisphosphonates are widely used in the treatment of children with osteogenesis imperfecta (OI) with the objective of reducing the risk of fractures. Although bisphosphonates increase bone mineral density in OI subjects, the effects on fracture incidence are conflicting. The aim of this study was to investigate the mechanisms underlying bone cell activity in subjects with mild untreated forms of OI and in a group of subjects with severe OI treated with cycles of intravenous neridronate.MethodsSclerostin, DKK1, TNF-α, RANKL, osteoprotegerin (OPG), and bone turnover markers were quantified in serum of 18 OI patients (12 females, mean age 8.86 ± 3.90), 8 of which were receiving cyclic intravenous neridronate, and 21 sex- and age-matched controls. The effects on osteoblastogenesis and OPG expression of media conditioned by the serum of OI patients and anti-DKK1 neutralizing antibody were evaluated. Osteoclastogenesis was assessed in cultures from patients and controls.ResultsDKK1 and RANKL levels were significantly increased both in untreated and in treated OI subjects with respect to controls. The serum from patients with high DKK1 levels inhibited both osteoblast differentiation and OPG expression in vitro. High RANKL and low OPG messenger RNA (mRNA) levels were found in lymphomonocytes from patients. High amounts of TNF-α were expressed by monocytes, and an elevated percentage of circulating CD11b-CD51/CD61+ osteoclast precursors was observed in patients.ConclusionsOur study demonstrated the key role of DKK1, RANKL, and TNF-α in regulating bone cell activity of subjects with OI untreated and treated with bisphosphonates. These cytokines could represent new pharmacological targets for OI patients.

[1]  Christof Niehrs,et al.  Kremen proteins are Dickkopf receptors that regulate Wnt/beta-catenin signalling. , 2002, Nature.

[2]  M. Dimopoulos,et al.  Bortezomib reduces serum dickkopf‐1 and receptor activator of nuclear factor‐κB ligand concentrations and normalises indices of bone remodelling in patients with relapsed multiple myeloma , 2006, British journal of haematology.

[3]  A. Oberbauer,et al.  Alendronate affects long bone length and growth plate morphology in the oim mouse model for Osteogenesis Imperfecta. , 2003, Bone.

[4]  C. Giunta,et al.  Recessive osteogenesis imperfecta: Clinical, radiological, and molecular findings , 2012, American journal of medical genetics. Part C, Seminars in medical genetics.

[5]  Xu Yang,et al.  The effects of RANKL inhibition on fracture healing and bone strength in a mouse model of osteogenesis imperfecta , 2008, Journal of orthopaedic research : official publication of the Orthopaedic Research Society.

[6]  Georg Schett,et al.  Dickkopf-1 is a master regulator of joint remodeling , 2007, Nature Medicine.

[7]  G. Eghbali-Fatourechi.  Bisphosphonate therapy in pediatric patients , 2014, Journal of Diabetes & Metabolic Disorders.

[8]  A. Boskey,et al.  Comparable outcomes in fracture reduction and bone properties with RANKL inhibition and alendronate treatment in a mouse model of osteogenesis imperfecta , 2012, Osteoporosis International.

[9]  A. Boskey,et al.  RANKL Inhibition Improves Bone Properties in a Mouse Model of Osteogenesis Imperfecta , 2010, Connective tissue research.

[10]  Haitao Li,et al.  Immature osteoblast lineage cells increase osteoclastogenesis in osteogenesis imperfecta murine. , 2010, The American journal of pathology.

[11]  W. Taylor,et al.  Circulating mediators of bone remodeling in psoriatic arthritis: implications for disordered osteoclastogenesis and bone erosion , 2010, Arthritis research & therapy.

[12]  R. Kutner,et al.  A potential role for Dkk-1 in the pathogenesis of osteosarcoma predicts novel diagnostic and treatment strategies , 2007, British Journal of Cancer.

[13]  L. Tatò,et al.  Intravenous Neridronate in Children With Osteogenesis Imperfecta: A Randomized Controlled Study , 2004, Journal of bone and mineral research : the official journal of the American Society for Bone and Mineral Research.

[14]  I. Lazzareschi,et al.  High serum sclerostin levels in children with haemophilia A , 2016, British Journal of Haematology.

[15]  Peng Liu,et al.  Sclerostin Binds to LRP5/6 and Antagonizes Canonical Wnt Signaling* , 2005, Journal of Biological Chemistry.

[16]  Joan C. Marini,et al.  New perspectives on osteogenesis imperfecta , 2011, Nature Reviews Endocrinology.

[17]  A. Boskey,et al.  Alendronate Treatment for Infants with Osteogenesis Imperfecta: Demonstration of Efficacy in a Mouse Model , 2002, Pediatric Research.

[18]  Z. Rumboldt,et al.  Osteoblastic response to the defective matrix in the osteogenesis imperfecta murine (oim) mouse. , 2002, Endocrinology.

[19]  F. Glorieux,et al.  Osteogenesis Imperfecta: Update on presentation and management , 2008, Reviews in Endocrine and Metabolic Disorders.

[20]  D. Dempster,et al.  Increased resorptive activity and accompanying morphological alterations in osteoclasts derived from the oim/oim mouse model of osteogenesis imperfecta , 2007, Journal of cellular biochemistry.

[21]  A. Boskey,et al.  High- and low-dose OPG–Fc cause osteopetrosis-like changes in infant mice , 2012, Pediatric Research.

[22]  D. Sillence,et al.  Genetic heterogeneity in osteogenesis imperfecta. , 1979, Journal of medical genetics.

[23]  Christof Niehrs,et al.  Kremen proteins are Dickkopf receptors that regulate Wnt/β-catenin signalling , 2002, Nature.

[24]  A. Forlino,et al.  Cellular Mechanism of Decreased Bone in Brtl Mouse Model of OI: Imbalance of Decreased Osteoblast Function and Increased Osteoclasts and Their Precursors , 2008, Journal of bone and mineral research : the official journal of the American Society for Bone and Mineral Research.

[25]  Lester R Curtin,et al.  Centers for Disease Control and Prevention 2000 growth charts for the United States: improvements to the 1977 National Center for Health Statistics version. , 2002, Pediatrics.

[26]  L. Idolazzi,et al.  Bisphosphonate treatment of postmenopausal osteoporosis is associated with a dose dependent increase in serum sclerostin. , 2012, Bone.

[27]  K. Dwan,et al.  Bisphosphonate therapy for osteogenesis imperfecta. , 2016, The Cochrane database of systematic reviews.

[28]  D. Janerich,et al.  Shifting genetic patterns in anencephaly and spina bifida. , 1978, Journal of medical genetics.

[29]  Roland Baron,et al.  WNT signaling in bone homeostasis and disease: from human mutations to treatments , 2013, Nature Medicine.

[30]  R. Baron,et al.  Deletion of a Single Allele of the Dkk1 Gene Leads to an Increase in Bone Formation and Bone Mass , 2006, Journal of bone and mineral research : the official journal of the American Society for Bone and Mineral Research.

[31]  F. Glorieux,et al.  Static and dynamic bone histomorphometry in children with osteogenesis imperfecta. , 2000, Bone.

[32]  S. Colucci,et al.  Osteogenic Differentiation of Dental Follicle Stem Cells , 2012, International journal of medical sciences.

[33]  O. Stephens,et al.  Myeloma-derived Dickkopf-1 disrupts Wnt-regulated osteoprotegerin and RANKL production by osteoblasts: a potential mechanism underlying osteolytic bone lesions in multiple myeloma. , 2008, Blood.

[34]  S. Janz,et al.  Attenuation of WNT signaling by DKK-1 and -2 regulates BMP2-induced osteoblast differentiation and expression of OPG, RANKL and M-CSF , 2007, Molecular Cancer.

[35]  D. Vignali.  Multiplexed particle-based flow cytometric assays. , 2000, Journal of immunological methods.

[36]  L. Idolazzi,et al.  Dickkopf-1 and Sclerostin Serum Levels in Patients with Systemic Mastocytosis , 2015, Calcified Tissue International.

[37]  L. Root,et al.  A Controlled Study of the Effects of Alendronate in a Growing Mouse Model of Osteogenesis Imperfecta , 2001, Calcified Tissue International.

[38]  H. Ryoo,et al.  BMP2 Protein Regulates Osteocalcin Expression via Runx2-mediated Atf6 Gene Transcription* , 2011, The Journal of Biological Chemistry.

[39]  L. Gesualdo,et al.  High dickkopf-1 levels in sera and leukocytes from children with 21-hydroxylase deficiency on chronic glucocorticoid treatment. , 2013, American journal of physiology. Endocrinology and metabolism.

[40]  M. Tetti,et al.  High Levels of Serum Prostaglandin E2 in Children with Osteogenesis Imperfecta Are Reduced by Neridronate Treatment , 2008, Pediatric Research.

[41]  S. Adami,et al.  Bone turnover markers in patients with osteogenesis imperfecta. , 2004, Bone.

[42]  F. Glorieux,et al.  Cyclical Intravenous Pamidronate Treatment Affects Metaphyseal Modeling in Growing Patients With Osteogenesis Imperfecta , 2005, Journal of bone and mineral research : the official journal of the American Society for Bone and Mineral Research.

[43]  Roland Baron,et al.  Increased Bone Turnover with Decreased Bone Formation by Osteoblasts in Children with Osteogenesis Imperfecta Tarda , 1983, Pediatric Research.

[44]  L. Idolazzi,et al.  In patients with rheumatoid arthritis, Dickkopf-1 serum levels are correlated with parathyroid hormone, bone erosions and bone mineral density. , 2015, Clinical and experimental rheumatology.

[45]  F. Glorieux,et al.  Modern approach to children with osteogenesis imperfecta , 2003, Journal of pediatric orthopedics. Part B.

[46]  C. Heeschen,et al.  The Wnt Antagonist Dickkopf-1 Mobilizes Vasculogenic Progenitor Cells via Activation of the Bone Marrow Endosteal Stem Cell Niche , 2008, Circulation research.