Deficiency for the ER-stress transducer OASIS causes severe recessive osteogenesis imperfecta in humans
© Symoens et al.; licensee BioMed Central Ltd. 2013
Received: 27 May 2013
Accepted: 15 September 2013
Published: 30 September 2013
Osteogenesis imperfecta (OI) is a clinically and genetically heterogeneous brittle bone disorder. Whereas dominant OI is mostly due to heterozygous mutations in either COL1A1 or COL1A2, encoding type I procollagen, recessive OI is caused by biallelic mutations in genes encoding proteins involved in type I procollagen processing or chaperoning. Hitherto, some OI cases remain molecularly unexplained. We detected a homozygous genomic deletion of CREB3L1 in a family with severe OI. CREB3L1 encodes OASIS, an endoplasmic reticulum-stress transducer that regulates type I procollagen expression during murine bone formation. This is the first report linking CREB3L1 to human recessive OI, thereby expanding the OI gene spectrum.
Osteogenesis imperfecta (OI) is a genetically heterogeneous brittle bone disorder with varying degrees of clinical severity, ranging from perinatal lethality to generalized osteopenia. The predominant autosomal dominant forms display mutations in either COL1A1 or COL1A2, encoding the α1- and α2-chains of type I procollagen, while rarer autosomal recessive forms mostly result from defective endoplasmic reticulum (ER)-resident proteins involved in post-translational processing or chaperoning of these α(I)-chains[1, 2]. Processing defects prevent normal collagen fibrillogenesis and on biochemical analysis often show perturbed modification of the collagen α-chain. Known defects include biallelic mutations in LEPRE1[3–5], CRTAP[5, 6], PPIB[7, 8], BMP1[9, 10], and PLOD2. Mutations in chaperones (including Hsp47 (SERPINH1) and FKBP10) impair intracellular collagen trafficking with intracellular retention or aggregation of collagen molecules and show dilation of the ER on electron microscopy, resulting in OI or related phenotypes[12–14]. Finally, rare other defects linked to distinct mechanisms involve the transcription factor osterix (SP7), pigment epithelium derived factor (SERPINF1) and transmembrane protein 38B (TMEM38B)[17, 18]. A recurrent mutation in a gene encoding the Interferon-inducible transmembrane protein 5 (IFITM5), which is involved in bone growth during prenatal murine development, was recently shown to cause autosomal (AD) dominant OI[19–21]. Recently, heterozygous and homozygous mutations in WNT1 (WNT1), which is a key signalling molecule in osteoblast function and bone development, were shown to underlie certain forms of AD early-onset osteoporosis and AR OI, which was in some patients associated with severe intellectual disability[22–26]. However, a small proportion of OI patients remain molecularly unexplained.
The parents have a healthy daughter (III:1) and have had one miscarriage (III:2, cause unknown). The adolescent daughter has blue sclerae but had not experienced any fractures. The mother (II:5) at 38 years of age and the father (II:6) at 47 years have blue sclerae, a soft and velvety skin and normal teeth. While the mother has small joint hypermobility, the father has conductive hearing loss.
Subsequently, all known OI genes (COL1A1, COL1A2, BMP1, LEPRE1, CRTAP, PPIB, PLOD2, SERPINH1, FKBP10, SP7, SERPINF1, TMEM38B, IFITM5 and WNT1) were sequenced by direct Sanger sequencing (ABI3730XL automated sequencer, Applied Biosystems), but no causal mutation(s) were detected.
We selected the CREB3L1 gene [GenBank:NM_052854.2], encoding the ER-stress transducer OASIS (Old Astrocyte Specifically Induced Substance), as an excellent candidate gene based on the observation that OASIS-/- mice were born with severe osteopenia and spontaneous fractures, reminiscent of severe human OI. In those mice, OASIS was shown to be crucial for bone formation through activating col1a1 transcription and facilitating the secretion of matrix proteins. Treatment of murine osteoblasts with BMP-2 (bone morphogenic protein 2) causes mild ER-stress and is associated with accelerated RIP (regulated intramembrane proteolysis) of OASIS. The N-terminal part of OASIS is subsequently translocated to the nucleus, where it binds to the osteoblast-specific UPRE (unfolded protein response element) regulatory region in the murine Col1a1 promoter thereby causing high levels of type I procollagen expression. While the amount of type I procollagen is normal in the murine OASIS-/- skin, reduced amounts of type I procollagen were detected in OASIS-/- calvaria and tibia, which suggested tissue-specific decrease of type I procollagen in the bone matrix but also failure of the OASIS-/- osteoblasts to produce high levels of type I procollagen. OASIS further functions as a tissue-specific ER-stress transducer that alters transcription of target genes involved in developmental processes, differentiation, or maturation upon mild ER-stress. PCR amplification of all exons and flanking introns of CREB3L1 failed in foetus III:4, suggesting a homozygous whole gene deletion. ArrayCGH analysis (1M SurePrint G3 Human CGH Microarray, Agilent Technologies) and copy number profiling (arrayCGHbase) confirmed this genomic deletion, which encompasses CREB3L1 and the first exon of DGKZ (arr11p11.2(46268141–46359490)×0, Figure 2B)[30, 31]. Whereas the arr11p11.2(46268141–46359490)×0 homozygous deletion was not reported before, heterozygous deletions or gains of this genomic region are described in the Decipher database and the Database of Genomic Variants but encompassing large genomic regions comprising multiple genes (6 to 86 genes and/or multiple chromosomal abnormalities) which, in some cases, are associated with intellectual disability. Both parents and the healthy sister were heterozygous for the deletion (data not shown). DGKZ encodes diacylglycerol kinase zeta, an ubiquitously expressed enzyme that is most abundantly present in the brain, thymus and skeletal muscle and which has a regulatory role in T-cell receptor signalling and T-cell activation. Two different isoforms (DGKζ1 in immune cells and DGKζ2 in other cells) are known, in which exon 1 is either present or absent and which have a tissue- and developmental stage-specific expression. Hitherto, no known function in bone formation has been ascribed to DGKζ and thus a possible contributing role to (the severity of) the bone phenotype of patient III:3 and foetus III:4 cannot completely be excluded. Expression analysis by real time-quantitative PCR (RT-qPCR) on total RNA isolated from dermal fibroblasts of foetus III:4 confirmed complete absence of the CREB3L1 transcript. In order to investigate the expression of the two DGKZ isoforms (DGKζ1 and DGKζ2), two different primer pairs were designed, of which one was specific for exon 1 that is only present in the DGKζ1 isoform. RT-qPCR experiments revealed no amplification for the primer pair specific for exon 1 in cultured dermal fibroblasts, suggesting that the DGKζ1 isoform is not expressed in these cells. For the second primer pair normal DGKZ expression was observed, which implies normal expression of the DGKζ2 isoform in cultured human dermal fibroblasts (Figure 2C). RT-qPCR analysis of the ER-stress markers BiP, CHOP and the spliced form of XBP1 showed levels comparable to controls, even after stimulation of confluent fibroblasts for 4 hours with the ER-stress inducers Tunicamycin (Tu, 10 μg/ml, Sigma-Aldrich) and Thapsigargin (Th, 1 μM, Sigma-Aldrich) (Figure 2C). This is in accordance to the observations in OASIS-/- mice. The expression level of CREB3L1 was unchanged in control fibroblasts after treatment with Tu and Th (Figure 2C), suggesting that OASIS does not play a major role in the ER-stress pathways previously linked to disease pathogenesis. Additionally, our finding that type I (pro)collagen production is normal in human dermal fibroblasts (Figure 2A) confirms that OASIS has a tissue-specific effect on type I (pro)collagen production.
In conclusion, the identification of CREB3L1 (encoding the ER-stress transducer OASIS) as a novel gene for autosomal recessive OI expands the spectrum of genes linked to OI and reinforces the role of ER-stress in the pathophysiology of OI.
We wish to thank J. Weytens, P. Van Acker, S. Baute, L. Demuynck and P. Vermassen for excellent technical assistance. We thank Dr. K. Vleminckx for critical review of the manuscript. FM and BC are post-doctoral fellows of the Fund for Scientific Research-Flanders. Contract grant sponsor: FWO grant number G.0171.05 and Methusalem grant number 08/01M01108.
- Forlino A, Cabral WA, Barnes AM, Marini JC: New perspectives on osteogenesis imperfecta. Nat Rev Endocrinol. 2011, 7: 540-557. 10.1038/nrendo.2011.81.PubMedView ArticleGoogle Scholar
- Marini JC, Forlino A, Cabral WA, Barnes AM, San Antonio JD, Milgrom S, Hyland JC, Korkko J, Prockop DJ, De Paepe A, et al: Consortium for osteogenesis imperfecta mutations in the helical domain of type I collagen: regions rich in lethal mutations align with collagen binding sites for integrins and proteoglycans. Hum Mutat. 2007, 28: 209-221. 10.1002/humu.20429.PubMedView ArticleGoogle Scholar
- Cabral WA, Chang W, Barnes AM, Weis M, Scott MA, Leikin S, Makareeva E, Kuznetsova NV, Rosenbaum KN, Tifft CJ, et al: Prolyl 3-hydroxylase 1 deficiency causes a recessive metabolic bone disorder resembling lethal/severe osteogenesis imperfecta. Nat Genet. 2007, 39: 359-365. 10.1038/ng1968.PubMedView ArticleGoogle Scholar
- Van Dijk F, Nikkels PG, den Hollander NS, Nesbitt IM, van Rijn RR, Cobben JM, Pals G: Lethal/ severe osteogenesis imperfecta in a large family: a novel homozygous LEPRE1 mutation and bone histological findings. Pediatr Dev Pathol. 2010, 14 (3): 228-234.PubMedView ArticleGoogle Scholar
- Baldridge D, Schwarze U, Morello R, Lennington J, Bertin TK, Pace JM, Pepin MG, Weis M, Eyre DR, Walsh J, et al: CRTAP and LEPRE1 mutations in recessive osteogenesis imperfecta. Hum Mutat. 2008, 29: 1435-1442. 10.1002/humu.20799.PubMedView ArticleGoogle Scholar
- Barnes AM, Chang W, Morello R, Cabral WA, Weis M, Eyre DR, Leikin S, Makareeva E, Kuznetsova N, Uveges TE, et al: Deficiency of cartilage-associated protein in recessive lethal osteogenesis imperfecta. N Engl J Med. 2006, 355: 2757-2764. 10.1056/NEJMoa063804.PubMedView ArticleGoogle Scholar
- Barnes AM, Carter EM, Cabral WA, Weis M, Chang W, Makareeva E, Leikin S, Rotimi CN, Eyre DR, Raggio CL, Marini JC: Lack of cyclophilin B in osteogenesis imperfecta with normal collagen folding. N Engl J Med. 2010, 362: 521-528. 10.1056/NEJMoa0907705.PubMedView ArticleGoogle Scholar
- van Dijk FS, Nesbitt IM, Zwikstra EH, Nikkels PG, Piersma SR, Fratantoni SA, Jimenez CR, Huizer M, Morsman AC, Cobben JM, et al: PPIB mutations cause severe osteogenesis imperfecta. Am J Hum Genet. 2009, 85: 521-527. 10.1016/j.ajhg.2009.09.001.PubMedView ArticleGoogle Scholar
- Martinez-Glez V, Valencia M, Caparros-Martin JA, Aglan M, Temtamy S, Tenorio J, Pulido V, Lindert U, Rohrbach M, Eyre D, et al: Identification of a mutation causing deficient BMP1/mTLD proteolytic activity in autosomal recessive osteogenesis imperfecta. Hum Mutat. 2012, 33: 343-350. 10.1002/humu.21647.PubMedView ArticleGoogle Scholar
- Asharani PV, Keupp K, Semler O, Wang W, Li Y, Thiele H, Yigit G, Pohl E, Becker J, Frommolt P, et al: Attenuated BMP1 function compromises osteogenesis, leading to bone fragility in humans and zebrafish. Am J Hum Genet. 2012, 90: 661-674. 10.1016/j.ajhg.2012.02.026.PubMedView ArticleGoogle Scholar
- Bank RA, Robins SP, Wijmenga C, Breslau-Siderius LJ, Bardoel AF, van der Sluijs HA, Pruijs HE, TeKoppele JM: Defective collagen crosslinking in bone, but not in ligament or cartilage, in Bruck syndrome: indications for a bone-specific telopeptide lysyl hydroxylase on chromosome 17. Proc Natl Acad Sci U S A. 1999, 96: 1054-1058. 10.1073/pnas.96.3.1054.PubMedView ArticleGoogle Scholar
- Alanay Y, Avaygan H, Camacho N, Utine GE, Boduroglu K, Aktas D, Alikasifoglu M, Tuncbilek E, Orhan D, Bakar FT, et al: Mutations in the gene encoding the RER protein FKBP65 cause autosomal-recessive osteogenesis imperfecta. Am J Hum Genet. 2010, 87: 572-573. 10.1016/j.ajhg.2010.09.002.View ArticleGoogle Scholar
- Kelley BP, Malfait F, Bonafe L, Baldridge D, Homan E, Symoens S, Willaert A, Elcioglu N, Van Maldergem L, Verellen-Dumoulin C, et al: Mutations in FKBP10 cause recessive osteogenesis imperfecta and Bruck syndrome. J Bone Miner Res. 2011, 26: 666-672. 10.1002/jbmr.250.PubMedView ArticleGoogle Scholar
- Christiansen HE, Schwarze U, Pyott SM, AlSwaid A, Al Balwi M, Alrasheed S, Pepin MG, Weis MA, Eyre DR, Byers PH: Homozygosity for a missense mutation in SERPINH1, which encodes the collagen chaperone protein HSP47, results in severe recessive osteogenesis imperfecta. Am J Hum Genet. 2010, 86: 389-398. 10.1016/j.ajhg.2010.01.034.PubMedView ArticleGoogle Scholar
- Lapunzina P, Aglan M, Temtamy S, Caparros-Martin JA, Valencia M, Leton R, Martinez-Glez V, Elhossini R, Amr K, Vilaboa N, Ruiz-Perez VL: Identification of a frameshift mutation in Osterix in a patient with recessive osteogenesis imperfecta. Am J Hum Genet. 2010, 87: 110-114. 10.1016/j.ajhg.2010.05.016.PubMedView ArticleGoogle Scholar
- Becker J, Semler O, Gilissen C, Li Y, Bolz HJ, Giunta C, Bergmann C, Rohrbach M, Koerber F, Zimmermann K, et al: Exome sequencing identifies truncating mutations in human SERPINF1 in autosomal-recessive osteogenesis imperfecta. Am J Hum Genet. 2011, 88: 362-371. 10.1016/j.ajhg.2011.01.015.PubMedView ArticleGoogle Scholar
- Shaheen R, Alazami AM, Alshammari MJ, Faqeih E, Alhashmi N, Mousa N, Alsinani A, Ansari S, Alzahrani F, Al-Owain M, et al: Study of autosomal recessive osteogenesis imperfecta in Arabia reveals a novel locus defined by TMEM38B mutation. J Med Genet. 2012, 49: 630-635. 10.1136/jmedgenet-2012-101142.PubMedView ArticleGoogle Scholar
- Volodarsky M, Markus B, Cohen I, Staretz-Chacham O, Flusser H, Landau D, Shelef I, Langer Y, Birk OS: A deletion mutation in TMEM38B associated with autosomal recessive osteogenesis imperfecta. Hum Mutat. 2013, 34: 582-586.PubMedGoogle Scholar
- Cho TJ, Lee KE, Lee SK, Song SJ, Kim KJ, Jeon D, Lee G, Kim HN, Lee HR, Eom HH, et al: A single recurrent mutation in the 5'-UTR of IFITM5 causes osteogenesis imperfecta type V. Am J Hum Genet. 2012, 91: 343-348. 10.1016/j.ajhg.2012.06.005.PubMedView ArticleGoogle Scholar
- Semler O, Garbes L, Keupp K, Swan D, Zimmermann K, Becker J, Iden S, Wirth B, Eysel P, Koerber F, et al: A mutation in the 5'-UTR of IFITM5 creates an in-frame start codon and causes autosomal-dominant osteogenesis imperfecta type V with hyperplastic callus. Am J Hum Genet. 2012, 91: 349-357. 10.1016/j.ajhg.2012.06.011.PubMedView ArticleGoogle Scholar
- Hanagata N, Li X, Morita H, Takemura T, Li J, Minowa T: Characterization of the osteoblast-specific transmembrane protein IFITM5 and analysis of IFITM5-deficient mice. J Bone Miner Metab. 2011, 29: 279-290. 10.1007/s00774-010-0221-0.PubMedView ArticleGoogle Scholar
- Fahiminiya S, Majewski J, Mort J, Moffatt P, Glorieux FH, Rauch F: Mutations in WNT1 are a cause of osteogenesis imperfecta. J Med Genet. 2013, 50: 345-348. 10.1136/jmedgenet-2013-101567.PubMedView ArticleGoogle Scholar
- Keupp K, Beleggia F, Kayserili H, Barnes AM, Steiner M, Semler O, Fischer B, Yigit G, Janda CY, Becker J, et al: Mutations in WNT1 cause different forms of bone fragility. Am J Hum Genet. 2013, 92: 565-574. 10.1016/j.ajhg.2013.02.010.PubMedView ArticleGoogle Scholar
- Pyott SM, Tran TT, Leistritz DF, Pepin MG, Mendelsohn NJ, Temme RT, Fernandez BA, Elsayed SM, Elsobky E, Verma I, et al: WNT1 mutations in families affected by moderately severe and progressive recessive osteogenesis imperfecta. Am J Hum Genet. 2013, 92: 590-597. 10.1016/j.ajhg.2013.02.009.PubMedView ArticleGoogle Scholar
- Laine CM, Joeng KS, Campeau PM, Kiviranta R, Tarkkonen K, Grover M, Lu JT, Pekkinen M, Wessman M, Heino TJ, et al: WNT1 mutations in early-onset osteoporosis and osteogenesis imperfecta. N Engl J Med. 2013, 368: 1809-1816. 10.1056/NEJMoa1215458.PubMedView ArticleGoogle Scholar
- Faqeih E, Shaheen R, Alkuraya FS: WNT1 mutation with recessive osteogenesis imperfecta and profound neurological phenotype. J Med Genet. 2013, 50: 491-492. 10.1136/jmedgenet-2013-101750.PubMedView ArticleGoogle Scholar
- Hellemans J, Mortier G, De Paepe A, Speleman F, Vandesompele J: qBase relative quantification framework and software for management and automated analysis of real-time quantitative PCR data. Genome Biol. 2007, 8: R19. 10.1186/gb-2007-8-2-r19.PubMedView ArticleGoogle Scholar
- Ding L, Bunting M, Topham MK, McIntyre TM, Zimmerman GA, Prescott SM: Alternative splicing of the human diacylglycerol kinase zeta gene in muscle. Proc Natl Acad Sci U S A. 1997, 94: 5519-5524. 10.1073/pnas.94.11.5519.PubMedView ArticleGoogle Scholar
- Murakami T, Saito A, Hino S, Kondo S, Kanemoto S, Chihara K, Sekiya H, Tsumagari K, Ochiai K, Yoshinaga K, et al: Signalling mediated by the endoplasmic reticulum stress transducer OASIS is involved in bone formation. Nat Cell Biol. 2009, 11: 1205-1211. 10.1038/ncb1963.PubMedView ArticleGoogle Scholar
- Buysse K, Delle Chiaie B, Van Coster R, Loeys B, De Paepe A, Mortier G, Speleman F, Menten B: Challenges for CNV interpretation in clinical molecular karyotyping: lessons learned from a 1001 sample experience. Eur J Med Genet. 2009, 52: 398-403. 10.1016/j.ejmg.2009.09.002.PubMedView ArticleGoogle Scholar
- Menten B, Pattyn F, De Preter K, Robbrecht P, Michels E, Buysse K, Mortier G, De Paepe A, van Vooren S, Vermeesch J, et al: arrayCGHbase: an analysis platform for comparative genomic hybridization microarrays. BMC Bioinforma. 2005, 6: 124. 10.1186/1471-2105-6-124.View ArticleGoogle Scholar
- Firth HV, Richards SM, Bevan AP, Clayton S, Corpas M, Rajan D, Van Vooren S, Moreau Y, Pettett RM, Carter NP: DECIPHER: database of chromosomal imbalance and phenotype in humans using ensembl resources. Am J Hum Genet. 2009, 84: 524-533. 10.1016/j.ajhg.2009.03.010.PubMedView ArticleGoogle Scholar
- Iafrate AJ, Feuk L, Rivera MN, Listewnik ML, Donahoe PK, Qi Y, Scherer SW, Lee C: Detection of large-scale variation in the human genome. Nat Genet. 2004, 36: 949-951. 10.1038/ng1416.PubMedView ArticleGoogle Scholar
- Rincon E, Gharbi SI, Santos-Mendoza T, Merida I: Diacylglycerol kinase zeta: at the crossroads of lipid signaling and protein complex organization. Prog Lipid Res. 2012, 51: 1-10. 10.1016/j.plipres.2011.10.001.PubMedView ArticleGoogle Scholar
- Kanoh H, Yamada K, Sakane F: Diacylglycerol kinase: a key modulator of signal transduction?. Trends Biochem Sci. 1990, 15: 47-50. 10.1016/0968-0004(90)90172-8.PubMedView ArticleGoogle Scholar
This article is published under license to BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.