The incidence of craniosynostosis is one in every 1,800–2500 births. The gene-environment model proposes that if a genetic predisposition is coupled with environmental exposures, the effects can be multiplicative resulting in severely abnormal phenotypes. At present, very little is known about the role of gene-environment interactions in modulating craniosynostosis phenotypes, but prior evidence suggests a role for endocrine factors. Here we provide a report of the effects of thyroid hormone exposure on murine calvaria cells. Murine derived calvaria cells were exposed to critical doses of pharmaceutical thyroxine and analyzed after 3 and 7 days of treatment. Endpoint assays were designed to determine the effects of the hormone exposure on markers of osteogenesis and included, proliferation assay, quantitative ALP activity assay, targeted qPCR for mRNA expression of Runx2, Alp, Ocn, and Twist1, genechip array for 28,853 targets, and targeted osteogenic microarray with qPCR confirmations. Exposure to thyroxine stimulated the cells to express ALP in a dose dependent manner. There were no patterns of difference observed for proliferation. Targeted RNA expression data confirmed expression increases for Alp and Ocn at 7 days in culture. The genechip array suggests substantive expression differences for 46 gene targets and the targeted osteogenesis microarray indicated 23 targets with substantive differences. 11 gene targets were chosen for qPCR confirmation because of their known association with bone or craniosynostosis (Col2a1, Dmp1, Fgf1, 2, Igf1, Mmp9, Phex, Tnf, Htra1, Por, and Dcn). We confirmed substantive increases in mRNA for Phex, FGF1, 2, Tnf, Dmp1, Htra1, Por, Igf1 and Mmp9, and substantive decreases for Dcn. It appears thyroid hormone may exert its effects through increasing osteogenesis. Targets isolated suggest a possible interaction for those gene products associated with calvarial suture growth and homeostasis as well as craniosynostosis.
References
[1]
Radetti G, Zavallone A, Gentili L, Beck-Peccoz P, Bona G (2002) Foetal and neonatal thyroid disorders. Minerva pediatrica 54: 383–400.
[2]
Menking M, Wiebel J, Schmid WU, Schmidt WT, Ebel KD, et al. (1972) Premature craniosynostosis associated with hyperthyroidism in 4 children with reference to 5 further cases in the literature. Monatsschrift fur Kinderheilkunde 120: 106–110.
[3]
Rasmussen SA, Yazdy MM, Carmichael SL, Jamieson DJ, Canfield MA, et al. (2007) Maternal thyroid disease as a risk factor for craniosynostosis. Obstetrics and gynecology 110: 369–377.
[4]
Robinson DC, Hall R, Munro DS (1969) Grave'S's disease, an unusual complication: raised intracranial pressure due to premature fusion of skull sutures. Archives of disease in childhood 44: 252–257.
[5]
Riggs W Jr, Wilroy RS Jr, Etteldorf JN (1972) Neonatal hyperthyroidism with accelerated skeletal maturation, craniosynostosis, and brachydactyly. Radiology 105: 621–625.
[6]
Camfield PR, Camfield CS, Cohen MM, Jr. (2000) Neurosurgical aspects of craniosynostosis. In: Cohen MM, Jr, MacLean RE, editors. Craniosynostosis: Diagnosis, Evaluation, and Management. New York: Oxford University Press. 177–183.
[7]
Cohen MM Jr (2005) Editorial: perspectives on craniosynostosis. Am J Med Genet A 136A: 313–326.
[8]
Jabs EW (2002) Genetic Etiologies of Craniosynostosis. In: Mooney MP, Siegel MI, editors. Understanding Craniofacial Anomalies: The Etiopathogenesis of Craniosynostoses and Facial Clefting. New York: John W. Wiley and Sons. 125–146.
[9]
Boyadjiev SA (2007) Genetic analysis of non-syndromic craniosynostosis. Orthod Craniofac Res 10: 129–137.
[10]
Richtsmeier JT (2002) Cranial vault dysmorphology and growth in craniosynostosis. In: Mooney MP, Siegel MI, editors. Understanding Craniofacial Anomalies: The Etiopathogenesis of Craniosynostoses and Facial Clefting. New York: John W. Wiley and Sons. 321–342.
[11]
Gault DT, Renier D, Marchac D, Jones BM (1992) Intracranial pressure and intracranial volume in children with craniosynostosis. Plast Reconstr Surg 90: 377–381.
[12]
Chadduck WM, Chadduck JB, Boop FA (1992) The subarachnoid spaces in craniosynostosis. Neurosurgery 30: 867–871.
[13]
Miller MT (2000) Ocular findings in craniosynostosis. In: Cohen MM, Jr, MacLean RE, editors. Craniosynostosis: Diagnosis, Evaluation, and Management. New York: Oxford University Press. 184–196.
[14]
Kapp-Simon KA, Speltz ML, Cunningham ML, Patel PK, Tomita T (2007) Neurodevelopment of children with single suture craniosynostosis: a review. Child's nervous system: ChNS: official journal of the International Society for Pediatric Neurosurgery 23: 269–281.
[15]
Esparza J, Hinojosa J, Garcia-Recuero I, Romance A, Pascual B, et al. (2008) Surgical treatment of isolated and syndromic craniosynostosis. Results and complications in 283 consecutive cases. Neurocirugia (Astur) 19: 509–529.
[16]
Persing JA, Jane JA (2000) Neurosurgical treatment of craniosynostosis. In: Cohen MM, Jr, MacLean RE, editors. Craniosynostosis: Diagnosis, Evaluation, and Management. New York: Oxford University Press. 209–227.
[17]
Bassett JH, Williams AJ, Murphy E, Boyde A, Howell PG, et al. (2008) A lack of thyroid hormones rather than excess thyrotropin causes abnormal skeletal development in hypothyroidism. Molecular endocrinology 22: 501–512.
[18]
Brent GA (2000) Tissue-specific actions of thyroid hormone: insights from animal models. Reviews in endocrine & metabolic disorders 1: 27–33.
[19]
Harvey CB, Stevens DA, Williams AJ, Jackson DJ, O'Shea P, et al. (2003) Analysis of thyroid hormone responsive gene expression in osteoblastic cells. Molecular and cellular endocrinology 213: 87–97.
[20]
Williams GR (2009) Actions of thyroid hormones in bone. Endokrynologia Polska 60: 380–388.
[21]
Hashmi SS, Canfield MA, Marengo L, Moffitt KB, Belmont JW, et al.. (2012) The association between neonatal thyroxine and craniosynostosis, Texas, 2004–2007. Birth defects research Part A, Clinical and molecular teratology.
[22]
Penfold JL, Simpson DA (1975) Premature craniosynostosis-a complication of thyroid replacement therapy. The Journal of pediatrics 86: 360–363.
[23]
Segni M, Leonardi E, Mazzoncini B, Pucarelli I, Pasquino AM (1999) Special features of Graves' disease in early childhood. Thyroid: official journal of the American Thyroid Association 9: 871–877.
[24]
Akita S, Hirano A, Fujii T (1996) Identification of IGF-I in the calvarial suture of young rats: histochemical analysis of the cranial sagittal sutures in a hyperthyroid rat model. Plastic and reconstructive surgery 97: 1–12.
[25]
Stamper BD, Park SS, Beyer RP, Bammler TK, Farin FM, et al. (2011) Differential expression of extracellular matrix-mediated pathways in single-suture craniosynostosis. PloS one 6: e26557.
[26]
Xiong Y, Ibhazehiebo K, Iwasaki T, Koibuchi N (2012) An in vitro method to study the effects of thyroid hormone-disrupting chemicals on neuronal development. Neurotoxicology 33: 753–757.
[27]
Yao C, Zhang J, Wang L, Guo Y, Tian Z (2007) Inhibitory effects of thyroxine on cytokine production by T cells in mice. International immunopharmacology 7: 1747–1754.
Kasono K, Sato K, Han DC, Fujii Y, Tsushima T, et al. (1988) Stimulation of alkaline phosphatase activity by thyroid hormone in mouse osteoblast-like cells (MC3T3-E1): a possible mechanism of hyperalkaline phosphatasia in hyperthyroidism. Bone and mineral 4: 355–363.
[30]
Miraoui H, Marie PJ (2010) Pivotal role of Twist in skeletal biology and pathology. Gene 468: 1–7.
[31]
Johnson D, Iseki S, Wilkie AO, Morriss–Kay GM (2000) Expression patterns of Twist and Fgfr1, ？2 and ？3 in the developing mouse coronal suture suggest a key role for twist in suture initiation and biogenesis. Mechanisms of development 91: 341–345.
[32]
Zumbrunn J, Trueb B (1996) Primary structure of a putative serine protease specific for IGF-binding proteins. FEBS letters 398: 187–192.
[33]
Ramajayam G, Vignesh RC, Karthikeyan S, Kumar KS, Karthikeyan GD, et al. (2012) Regulation of insulin-like growth factors and their binding proteins by thyroid stimulating hormone in human osteoblast-like (SaOS2) cells. Molecular and cellular biochemistry 368: 77–88.
[34]
Huang BK, Golden LA, Tarjan G, Madison LD, Stern PH (2000) Insulin-like growth factor I production is essential for anabolic effects of thyroid hormone in osteoblasts. Journal of bone and mineral research: the official journal of the American Society for Bone and Mineral Research 15: 188–197.
[35]
Shackleton CH (2012) Role of a disordered steroid metabolome in the elucidation of sterol and steroid biosynthesis. Lipids 47: 1–12.
[36]
Arlt W (2007) P450 oxidoreductase deficiency and Antley-Bixler syndrome. Reviews in endocrine & metabolic disorders 8: 301–307.
[37]
von Marschall Z, Fisher LW (2010) Decorin is processed by three isoforms of bone morphogenetic protein-1 (BMP1). Biochemical and biophysical research communications 391: 1374–1378.
[38]
Gomez D, Al Haj Zen A, Borges LF, Philippe M, Gutierrez PS, et al. (2009) Syndromic and non-syndromic aneurysms of the human ascending aorta share activation of the Smad2 pathway. The Journal of pathology 218: 131–142.
[39]
Wadhwa S, Bi Y, Ortiz AT, Embree MC, Kilts T, et al. (2007) Impaired posterior frontal sutural fusion in the biglycan/decorin double deficient mice. Bone 40: 861–866.
[40]
Bodo M, Baroni T, Carinci F, Becchetti E, Bellucci C, et al. (1999) A regulatory role of fibroblast growth factor in the expression of decorin, biglycan, betaglycan and syndecan in osteoblasts from patients with Crouzon's syndrome. European journal of cell biology 78: 323–330.
[41]
Morey JS, Ryan JC, Van Dolah FM (2006) Microarray validation: factors influencing correlation between oligonucleotide microarrays and real-time PCR. Biological procedures online 8: 175–193.
[42]
Kohara H, Kitaura H, Fujimura Y, Yoshimatsu M, Morita Y, et al. (2011) IFN-gamma directly inhibits TNF-alpha-induced osteoclastogenesis in vitro and in vivo and induces apoptosis mediated by Fas/Fas ligand interactions. Immunology letters 137: 53–61.
[43]
Li CW, Xia W, Huo L, Lim SO, Wu Y, et al. (2012) Epithelial-mesenchymal transition induced by TNF-alpha requires NF-kappaB-mediated transcriptional upregulation of Twist1. Cancer research 72: 1290–1300.
[44]
Oh JH, Park SY, de Crombrugghe B, Kim JE (2012) Chondrocyte-specific ablation of Osterix leads to impaired endochondral ossification. Biochemical and biophysical research communications 418: 634–640.
[45]
Weiss S, Hennig T, Bock R, Steck E, Richter W (2010) Impact of growth factors and PTHrP on early and late chondrogenic differentiation of human mesenchymal stem cells. Journal of cellular physiology 223: 84–93.
[46]
Owen C, Chen F, Flenniken AM, Osborne LR, Ichikawa S, et al. (2012) A novel Phex mutation in a new mouse model of hypophosphatemic rickets. Journal of cellular biochemistry 113: 2432–2441.
[47]
Sheng MH, Wergedal JE, Mohan S, Amoui M, Baylink DJ, et al. (2012) Targeted overexpression of osteoactivin in cells of osteoclastic lineage promotes osteoclastic resorption and bone loss in mice. PloS one 7: e35280.
[48]
Feng JQ, Ward LM, Liu S, Lu Y, Xie Y, et al. (2006) Loss of DMP1 causes rickets and osteomalacia and identifies a role for osteocytes in mineral metabolism. Nature genetics 38: 1310–1315.
[49]
Coussens AK, Hughes IP, Wilkinson CR, Morris CP, Anderson PJ, et al. (2008) Identification of genes differentially expressed by prematurely fused human sutures using a novel in vivo – in vitro approach. Differentiation; research in biological diversity 76: 531–545.
[50]
Connerney J, Andreeva V, Leshem Y, Mercado MA, Dowell K, et al. (2008) Twist1 homodimers enhance FGF responsiveness of the cranial sutures and promote suture closure. Developmental biology 318: 323–334.
[51]
Rice R, Rice DP, Thesleff I (2005) Foxc1 integrates Fgf and Bmp signalling independently of twist or noggin during calvarial bone development. Developmental dynamics: an official publication of the American Association of Anatomists 233: 847–852.
[52]
Ang BU, Spivak RM, Nah HD, Kirschner RE (2010) Dura in the pathogenesis of syndromic craniosynostosis: fibroblast growth factor receptor 2 mutations in dural cells promote osteogenic proliferation and differentiation of osteoblasts. The Journal of craniofacial surgery 21: 462–467.
[53]
Greenwald JA, Mehrara BJ, Spector JA, Warren SM, Fagenholz PJ, et al. (2001) In vivo modulation of FGF biological activity alters cranial suture fate. The American journal of pathology 158: 441–452.
[54]
Kim HJ, Rice DP, Kettunen PJ, Thesleff I (1998) FGF-, BMP- and Shh-mediated signalling pathways in the regulation of cranial suture morphogenesis and calvarial bone development. Development 125: 1241–1251.
[55]
Morriss-Kay GM, Wilkie AO (2005) Growth of the normal skull vault and its alteration in craniosynostosis: insights from human genetics and experimental studies. J Anat 207: 637–653.
[56]
Marie PJ, Kaabeche K, Guenou H (2008) Roles of FGFR2 and twist in human craniosynostosis: insights from genetic mutations in cranial osteoblasts. Frontiers of oral biology 12: 144–159.
[57]
Rice DP, Aberg T, Chan Y, Tang Z, Kettunen PJ, et al. (2000) Integration of FGF and TWIST in calvarial bone and suture development. Development 127: 1845–1855.
[58]
Lee KM, Santos-Ruiz L, Ferretti P (2010) A single-point mutation in FGFR2 affects cell cycle and Tgfbeta signalling in osteoblasts. Biochimica et biophysica acta 1802: 347–355.
[59]
Mansukhani A, Bellosta P, Sahni M, Basilico C (2000) Signaling by fibroblast growth factors (FGF) and fibroblast growth factor receptor 2 (FGFR2)-activating mutations blocks mineralization and induces apoptosis in osteoblasts. The Journal of cell biology 149: 1297–1308.
