Abstract
Neurofibromatosis type 1 (NF1) is a pandemic genetic disorder characterized by malignant and nonmalignant manifestations, including skeletal abnormalities, such as osteoporosis, scoliosis, short stature, and pseudarthrosis. Recent studies in genetically inbred mice and from human patients with NF1 have identified multiple gains in osteoclast (OCL) functions both in vitro and in vivo. Given that osteoblasts secrete cytokines that promote OCL maturation/activation, we sought to identify whether haploinsufficiency of Nf1 (Nf1+/−) osteoblasts and their precursors secrete cytokines that have a central role in this process. Osteoblast conditioned media (OBCM) from Nf1+/− osteoblasts promoted OCL migration and bone resorption compared with WT OBCM. Osteopontin (OPN), a matrix protein found in mineralized tissues and pivotal in modulating OCL functions, was present in increased concentrations in Nf1+/− osteoblasts. Addition of OPN neutralizing antibody to Nf1+/− OBCM diminished the gain in bioactivity on OCL functions, including OCL migration and bone resorption. Our study identifies an important paracrine loop whereby elevated secretion of OPN by osteoblasts activate Nf1+/− OCLs that already have an intrinsic propensity for bone resorption leading to osteopenia and osteoporosis.
Similar content being viewed by others
Log in or create a free account to read this content
Gain free access to this article, as well as selected content from this journal and more on nature.com
or
Abbreviations
- BMMNCs:
-
bone marrow mononuclear cells
- Erk:
-
extracellular signal-regulated kinase
- GRD:
-
GAP related domain
- NF1:
-
neurofibromatosis type 1
- Nf1+/− :
-
haploinsufficiency of Nf1
- OBCM:
-
osteoblast conditional media
- OCL:
-
osteoclast
- OPN:
-
osteopontin
- TRAP:
-
tartrate resistant acid phosphatase
References
Bollag G, Clapp DW, Shih S, Adler F, Zhang YY, Thompson P, Lange BJ, Freedman MH, McCormick F, Jacks T, Shannon K 1996 Loss of NF1 results in activation of the Ras signaling pathway and leads to aberrant growth in haematopoietic cells. Nat Genet 12: 144–148
Largaespada DA, Brannan CI, Jenkins NA, Copeland NG 1996 Nf1 deficiency causes Ras-mediated granulocyte/macrophage colony stimulating factor hypersensitivity and chronic myeloid leukaemia. Nat Genet 12: 137–143
Yu X, Chen S, Potter OL, Murthy SM, Li J, Pulcini JM, Ohashi N, Winata T, Everett ET, Ingram D, Clapp WD, Hock JM 2005 Neurofibromin and its inactivation of Ras are prerequisites for osteoblast functioning. Bone 36: 793–802
Crawford AH Jr, Bagamery N 1986 Osseous manifestations of neurofibromatosis in childhood. J Pediatr Orthop 6: 72–88
Kuorilehto T, Ekholm E, Nissinen M, Hietaniemi K, Hiltunen A, Paavolainen P, Penttinen R, Peltonen J 2006 NF1 gene expression in mouse fracture healing and in experimental rat pseudarthrosis. J Histochem Cytochem 54: 363–370
Stevenson DA, Moyer-Mileur LJ, Murray M, Slater H, Sheng X, Carey JC, Dube B, Viskochil DH 2007 Bone mineral density in children and adolescents with neurofibromatosis type 1. J Pediatr 150: 83–88
Illes T, Halmai V, de Jonge T, Dubousset J 2001 Decreased bone mineral density in neurofibromatosis-1 patients with spinal deformities. Osteoporos Int 12: 823–827
Stevenson DA, Zhou H, Ashrafi S, Messiaen LM, Carey JC, D'Astous JL, Santora SD, Viskochil DH 2006 Double inactivation of NF1 in tibial pseudarthrosis. Am J Hum Genet 79: 143–148
Teitelbaum SL 2000 Osteoclasts, integrins, and osteoporosis. J Bone Miner Metab 18: 344–349
Yang FC, Chen S, Robling AG, Yu X, Nebesio TD, Yan J, Morgan T, Li X, Yuan J, Hock J, Ingram DA, Clapp DW 2006 Hyperactivation of p21 and PI3K cooperate to alter murine and human neurofibromatosis type 1-haploinsufficient osteoclast functions. J Clin Invest 116: 2880–2891
Weber GF, Cantor H 1996 The immunology of Eta-1/osteopontin. Cytokine Growth Factor Rev 7: 241–248
Liaw L, Birk DE, Ballas CB, Whitsitt JS, Davidson JM, Hogan BL 1998 Altered wound healing in mice lacking a functional osteopontin gene (spp1). J Clin Invest 101: 1468–1478
Chellaiah MA, Hruska KA 2003 The integrin alpha (v) beta(3) and CD44 regulate the actions of osteopontin on osteoclast motility. Calcif Tissue Int 72: 197–205
Shapses SA, Cifuentes M, Spevak L, Chowdhury H, Brittingham J, Boskey AL, Denhardt DT 2003 Osteopontin facilitates bone resorption, decreasing bone mineral crystallinity and content during calcium deficiency. Calcif Tissue Int 73: 86–92
Standal T, Borset M, Sundan A 2004 Role of osteopontin in adhesion, migration, cell survival and bone remodeling. Exp Oncol 26: 179–184
Chellaiah MA, Kizer N, Biswas R, Alvarez U, Strauss-Schoenberger J, Rifas L, Rittling SR, Denhardt DT, Hruska KA 2003 Osteopontin deficiency produces osteoclast dysfunction due to reduced CD44 surface expression. Mol Biol Cell 14: 173–189
Yoshitake H, Rittling SR, Denhardt DT, Noda M 1999 Osteopontin-deficient mice are resistant to ovariectomy-induced bone resorption. Proc Natl Acad Sci USA 96: 8156–8160
Wu X, Estwick SA, Chen S, Yu M, Ming W, Nebesio TD, Li Y, Yuan J, Kapur R, Ingram D, Yoder MC, Yang FC 2006 Neurofibromin plays a critical role in modulating osteoblast differentiation of mesenchymal stem/progenitor cells. Hum Mol Genet 15: 2837–2845
Hiatt KK, Ingram DA, Zhang Y, Bollag G, Clapp DW 2001 Neurofibromin GTPase-activating protein-related domains restore normal growth in Nf1−/− cells. J Biol Chem 276: 7240–7245
Cao JJ, Wronski TJ, Iwaniec U, Phleger L, Kurimoto P, Boudignon B, Halloran BP 2005 Aging increases stromal/osteoblastic cell-induced osteoclastogenesis and alters the osteoclast precursor pool in the mouse. J Bone Miner Res 20: 1659–1668
Kolanczyk M, Kossler N, Kuhnisch J, Lavitas L, Stricker S, Wilkening U, Manjubala I, Fratzl P, Sporle R, Herrmann BG, Parada LF, Kornak U, Mundlos S 2007 Multiple roles for neurofibromin in skeletal development and growth. Hum Mol Genet 16: 874–886
Ishii T, Ohshima S, Ishida T, Mima T, Tabunoki Y, Kobayashi H, Maeda M, Uede T, Liaw L, Kinoshita N, Kawase I, Saeki Y 2004 Osteopontin as a positive regulator in the osteoclastogenesis of arthritis. Biochem Biophys Res Commun 316: 809–815
Sugatani T, Alvarez UM, Hruska KA 2003 Activin A stimulates IkappaB-alpha/NFkappaB and RANK expression for osteoclast differentiation, but not AKT survival pathway in osteoclast precursors. J Cell Biochem 90: 59–67
Rangaswami H, Bulbule A, Kundu GC 2005 JNK1 differentially regulates osteopontin-induced nuclear factor-inducing kinase/MEKK1-dependent activating protein-1-mediated promatrix metalloproteinase-9 activation. J Biol Chem 280: 19381–19392
Scherpereel A, Lee YC 2007 Biomarkers for mesothelioma. Curr Opin Pulm Med 13: 339–443
El-Tanani MK, Campbell FC, Kurisetty V, Jin D, McCann M, Rudland PS 2006 The regulation and role of osteopontin in malignant transformation and cancer. Cytokine Growth Factor Rev 17: 463–474
Gutmann DH, Wu YL, Hedrick NM, Zhu Y, Guha A, Parada LF 2001 Heterozygosity for the neurofibromatosis 1 (NF1) tumor suppressor results in abnormalities in cell attachment, spreading and motility in astrocytes. Hum Mol Genet 10: 3009–3016
Gingery A, Bradley E, Shaw A, Oursler MJ 2003 Phosphatidylinositol 3-kinase coordinately activates the MEK/ERK and AKT/NFkappaB pathways to maintain osteoclast survival. J Cell Biochem 89: 165–179
Abbas S, Clohisy JC, Abu-Amer Y 2003 Mitogen-activated protein (MAP) kinases mediate PMMA-induction of osteoclasts. J Orthop Res 21: 1041–1048
Digilio MC, Sarkozy A, Capolino R, Chiarini Testa MB, Esposito G, de Zorzi A, Cutrera R, Marino B, Dallapiccola B 2008 Costello syndrome: clinical diagnosis in the first year of life. Eur J Pediatr 167: 621–628
Hou JW 2008 Rapidly progressive scoliosis after successful treatment for osteopenia in Costello syndrome. Am J Med Genet A 146: 393–396
Acknowledgements
We thank Marilyn Wales for administrative support.
Author information
Authors and Affiliations
Additional information
Supported by March of Dimes Foundation #6-FY08-246 (F.C.Y.).
Rights and permissions
About this article
Cite this article
Li, H., Liu, Y., Zhang, Q. et al. Ras Dependent Paracrine Secretion of Osteopontin by Nf1+/− Osteoblasts Promote Osteoclast Activation in a Neurofibromatosis Type I Murine Model. Pediatr Res 65, 613–618 (2009). https://doi.org/10.1203/PDR.0b013e3181a1c607
Received:
Accepted:
Issue date:
DOI: https://doi.org/10.1203/PDR.0b013e3181a1c607
This article is cited by
-
Progressive bone impairment with age and pubertal development in neurofibromatosis type I
Archives of Osteoporosis (2018)
-
Aberrant Myeloid Differentiation Contributes to the Development of Osteoporosis in Neurofibromatosis Type 1
Current Osteoporosis Reports (2016)
-
The effect of five proteins on stem cells used for osteoblast differentiation and proliferation: a current review of the literature
Cellular and Molecular Life Sciences (2014)
-
The ecology of brain tumors: lessons learned from neurofibromatosis-1
Oncogene (2011)
