Skip to main content
SpringerLink
Log in

Genetic Disruption of Anoctamin 5 in Mice Replicates Human Gnathodiaphyseal Dysplasia (GDD)

  • Original Research
  • Published:
Calcified Tissue International Aims and scope Submit manuscript

Abstract

Gnathodiaphyseal dysplasia (GDD; OMIM#166260) is a rare skeletal disorder which is mainly characterized by cemento-osseous lesions in mandibles, bone fragility, bowing and diaphyseal sclerosis of tubular bones. GDD is caused by point mutations in Anoctamin-5 (ANO5); however, the disease mechanisms remain unclear. Here we generated Ano5-knockout (KO) mice using a CRISPR/Cas 9 approach to study loss of function aspects of GDD mutations. Homozygous Ano5 knockout mice (Ano5−/−) replicate some typical traits of human GDD including massive jawbones, bowing tibia, sclerosis and cortical thickening of femoral and tibial diaphyses. Serum alkaline phosphatase (ALP) levels were elevated in Ano5−/− mice as in GDD patients. Calvaria-derived Ano5−/− osteoblast cultures show increased osteoblastogenesis, which is consistent with our previous in vitro observations. Bone matrix is hypermineralized, and the expression of bone formation-related factors is enhanced in Ano5−/− mice, suggesting that the osteogenic anomaly arises from a genetic disruption of Ano5. We believe this new mouse model will shed more light on the development of skeletal abnormalities in GDD on a cellular and molecular level.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Subscribe and save

Springer+ Basic
$34.99 /Month
  • Get 10 units per month
  • Download Article/Chapter or eBook
  • 1 Unit = 1 Article or 1 Chapter
  • Cancel anytime
Subscribe now

Buy Now

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6

Similar content being viewed by others

References

  1. Akasaka Y, Nakajima T, Koyama K et al (1969) Familial cases of a new systemic bone disease, hereditary gnathodiaphyseal sclerosis. Nippon Seikeigeka Gakkai Zasshi 43:381–394

    CAS  PubMed  Google Scholar 

  2. Riminucci M, Collins MA, Boyde A et al (2001) Gnathodiaphyseal dysplasia: a syndrome of fibro-osseous lesions of jawbones, bone fragility, and long bone bowing. J Bone Miner Res 16:1710–1718

    Article  CAS  PubMed  Google Scholar 

  3. Ahluwalia J, Ly JQ, Norman E et al (2001) Gnathodiaphyseal dysplasia. Clin Imaging 31:67–69

    Article  Google Scholar 

  4. Tsutsumi S, Kamata N, Maruoka Y et al (2003) Autosomal dominant gnathodiaphyseal dysplasia maps to chromosome 11p14.3–15.1. J Bone Miner Res 18:413–418

    Article  CAS  PubMed  Google Scholar 

  5. Jin L, Liu Y, Sun F et al (2017) Three novel ANO5 missense mutations in Caucasian and Chinese families and sporadic cases with gnathodiaphyseal dysplasia. Sci Rep 7:40935

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  6. Tsutsumi S, Kamata N, Vokes TJ et al (2004) The novel gene encoding a putative transmembrane protein is mutated in gnathodiaphyseal dysplasia (GDD). Am J Hum Genet 74:1255–1261

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  7. Marconi C, Brunamonti Binello P, Badiali G et al (2013) A novel missense mutation in ANO5/TMEM16E is causative for gnathodiaphyseal dysplasia in a large Italian pedigree. Eur J Hum Genet 21:613–619

    Article  CAS  PubMed  Google Scholar 

  8. Rolvien T, Koehne T, Kornak U et al (2017) A novel ANO5 mutation causing gnathodiaphyseal dysplasia with high bone turnover osteosclerosis. J Bone Miner Res 32:277–284

    Article  CAS  PubMed  Google Scholar 

  9. Vengoechea J, Carpenter L (2015) Gnathodiaphyseal dysplasia presenting as polyostotic fbrous dysplasia. Am J Med Genet A 167:1421–1422

    Article  PubMed  Google Scholar 

  10. Andreeva TV, Tyazhelova TV, Rykalina VN et al (2016) Whole exome sequencing links dental tumor to an autosomal-dominant mutation in ANO5 gene associated with gnathodiaphyseal dysplasia and muscle dystrophies. Sci Rep 6:26440

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  11. Duong HA, Le KT, Soulema AL et al (2016) Gnathodiaphyseal dysplasia: report of a family with a novel mutation of the ANO5 gene. Oral Surg Oral Med Oral Pathol Oral Radiol 121:123–128

    Article  Google Scholar 

  12. Hartzell HC, Yu K, Xiao Q et al (2009) Anoctamin/TMEM16 family members are Ca2+-activated Cl– channels. J Physiol 587:2127–2139

    Article  CAS  PubMed  Google Scholar 

  13. Tran TT, Tobiume K, Hirono C et al (2014) TMEM16E (GDD1) exhibits protein instability and distinct characteristics in chloride channel/pore forming ability. J Cell Physiol 229:181–190

    Article  CAS  PubMed  Google Scholar 

  14. Duran C, Qu Z, Osunkoya AO et al (2012) ANOs 3–7 in the anoctamin/Tmem16 Cl– channel family are intracellular proteins. Am J Physiol Cell Physiol 302:C482–C493

    Article  CAS  PubMed  Google Scholar 

  15. Di Zanni E, Gradogna A, Scholz-Starke J et al (2018) Gain of function of TMEM16E/ANO5 scrambling activity caused by a mutation associated with gnathodiaphyseal dysplasia. Cell Mol Life Sci 75:1657–1670

    Article  CAS  PubMed  Google Scholar 

  16. Mizuta K, Tsutsumi S, Inoue H et al (2007) Molecular characterization of GDD1/TMEM16E, the gene product responsible for autosomal dominant gnathodiaphyseal dysplasia. Biochem Biophys Res Commun 357:126–132

    Article  CAS  PubMed  Google Scholar 

  17. Ehlen HW, Chinenkova M, Moser M et al (2013) Inactivation of anoctamin-6/Tmem16f, a regulator of phosphatidylserine scrambling in osteoblasts, leads to decreased mineral deposition in skeletal tissues. J Bone Miner Res 28:246–259

    Article  CAS  PubMed  Google Scholar 

  18. Ousingsawat J, Wanitchakool P, Schreiber R et al (2015) Anoctamin-6 controls bone mineralization by activating the calcium transporter NCX1. J Biol Chem 290:6270–6280

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  19. Zhao P, Torcaso A, Mariano A et al (2014) Anoctamin 6 regulates C2C12 myoblast proliferation. PLoS ONE 9:e92749

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  20. Hicks D, Sarkozy A, Muelas N et al (2011) A founder mutation in Anoctamin 5 is a major cause of limb-girdle muscular dystrophy. Brain 134:171–182

    Article  PubMed  PubMed Central  Google Scholar 

  21. Little AA, McKeever PE, Gruis KL (2013) Novel mutations in the Anoctamin 5 gene (ANO5) associated with limb-girdle muscular dystrophy 2L. Muscle Nerve 47:287–291

    Article  PubMed  Google Scholar 

  22. Magri F, Del Bo R, D’Angelo MG et al (2012) Frequency and characterisation of anoctamin 5 mutations in a cohort of Italian limb-girdle muscular dystrophy patients. Neuromuscul Disord 22:934–943

    Article  PubMed  PubMed Central  Google Scholar 

  23. Griffin DA, Johnson RW, Whitlock JM et al (2016) Defective membrane fusion and repair in Anoctamin5-deficient muscular dystrophy. Hum Mol Genet 25:1900–1911

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  24. Xu J, El Refaey M, Xu L et al (2015) Genetic disruption of Ano5 in mice does not recapitulate human ANO5-deficient musculardystrophy. Skelet Muscle 5:43

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  25. Gyobu S, Miyata H, Ikawa M et al (2015) A role of TMEM16E carrying a scrambling domain in sperm motility. Mol Cell Biol 36:645–659

    Article  CAS  PubMed  Google Scholar 

  26. Sui T, Xu L, Lau YS et al (2018) Development of muscular dystrophy in a CRISPR-engineered mutant rabbit model with frame-disrupting ANO5 mutations. Cell Death Dis 9:609

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  27. Otaify GA, Whyte MP, Gottesman GS et al (2018) Gnathodiaphyseal dysplasia: Severe atypical presentation with novel heterozygous mutation of the anoctamin gene (ANO5). Bone 107:167–171

    Article  CAS  Google Scholar 

  28. Schessl J, Kress W, Schoser B (2012) Novel ANO5 mutations causing hyper-CK-emia, limb girdle muscular weakness and Miyoshi type of muscular dystrophy. Muscle Nerve 45:740–742

    Article  CAS  PubMed  Google Scholar 

  29. Bolduc V, Marlow G, Boycott KM et al (2010) Recessive mutations in the putative calcium-activated chloride channel Anoctamin 5 cause proximal LGMD2L and distal MMD3 muscular dystrophies. Am J Hum Genet 86:213–221

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  30. Witting N, Duno M, Petri H et al (2013) Anoctamin 5 muscular dystrophy in Denmark: prevalence, genotypes, phenotypes, cardiac findings, and muscle protein expression. J Neurol 260:2084–2093

    Article  CAS  PubMed  Google Scholar 

  31. Sarkozy A, Hicks D, Hudson J et al (2013) ANO5 gene analysis in a large cohort of patients with anoctaminopathy: confirmation of male prevalence and high occurrence of the common exon 5 gene mutation. Hum Mutat 34:1111–1118

    Article  CAS  PubMed  Google Scholar 

  32. Penisson-Besnier I, Saint-Andre JP, Hicks D et al (2012) Myopathy caused by anoctamin 5 mutations and necrotizing vasculitis. J Neurol 259:1988–1990

    Article  PubMed  Google Scholar 

  33. Liao BY, Zhang J (2008) Null mutations in human and mouse orthologs frequently result in different phenotypes. Proc Natl Acad Sci USA 105:6987–6992

    Article  PubMed  Google Scholar 

  34. Chipman SD, Sweet HO, McBride DJ Jr et al (1993) Defective pro alpha 2(I) collagen synthesis in a recessivemutation in mice: a model of human osteogenesis imperfecta. Proc Natl Acad Sci USA 90:1701–1705

    Article  CAS  PubMed  Google Scholar 

  35. Ueki Y, Lin CY, Senoo M et al (2007) Increased myeloid cell responses to M-CSF and RANKL cause bone loss and inflammation in SH3BP2 “cherubism” mice. Cell 128:71–83

    Article  CAS  PubMed  Google Scholar 

  36. Chen IP, Wang CJ, Strecker S (2009) Introduction of a Phe377del mutation in ANK creates a mouse model for craniometaphysealdysplasia. J Bone Miner Res 24:1206–1215

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  37. Mestas J, Hughes CCW (2004) Of mice and not men: differences between mouse and human immunology. J Immunol 172:2731–2738

    Article  CAS  Google Scholar 

  38. Terpstra AHM (2001) Differences between humans and mice in efficacy of the body fat lowering effect of conjugated linoleic acid: role of metabolic rate. J Nutr 131:2067–2068

    Article  CAS  PubMed  Google Scholar 

  39. Wahbi K, Behin A, Becane HM (2013) Dilated cardiomyopathy in patients with mutations in anoctamin 5. Int J Cardiol 16876–16879

  40. Campos-Xavier B, Saraiva JM, Savarirayan R et al (2001) Phenotypic variability at the TGF-beta1 locus in Camurati-Engelmann disease. Hum Genet 109:653–658

    Article  CAS  PubMed  Google Scholar 

  41. Sodek J, Ganss B, McKee MD (2000) Osteopontin. Crit Rev Oral Biol Med 11:279–303

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgements

This work was supported by the National Natural Science Foundation of China (Grant # 81570958); Beijing Natural Science Foundation (Grant #7162075); High-level Talents of Beijing Health System (Grant #2013-3-036); and Scientific Foundation for the Returned Overseas Chinese Scholars, State Education Ministry (Grant # 2015-1098).

Author information

Authors and Affiliations

  1. Beijing Stomatological Hospital, Capital Medical University, Beijing, People’s Republic of China

    Xiaoyu Wang, Xiu Liu, Rui Dong, Chao Liang & Ying Hu

  2. Department of Reconstructive Sciences, Center for Regenerative Medicine and Skeletal Development, University of Connecticut Health, Farmington, CT, USA

    Ernst J. Reichenberger

  3. Beijing Stomatological Hospital, Beijing Institute of Dental Research, Capital Medical University, No 4 Tiantanxili, Dongcheng District, Beijing, 100050, People’s Republic of China

    Ying Hu

Authors
  1. Xiaoyu Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Xiu Liu
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Rui Dong
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Chao Liang
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Ernst J. Reichenberger
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Ying Hu
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Ying Hu.

Ethics declarations

Conflict of interest

Xiaoyu Wang, Xiu Liu, Rui Dong, Chao Liang, Ernst J. Reichenberger and Ying Hu declare that they have no conflict of interest.

Human and Animal Rights and Informed Consent

The animal experimentation protocols were approved by the Institutional Animal Care and Use Committee of the Beijing Stomatological Hospital (the approval number: KQYY-201611-001) and were strictly undertaken in accordance with the ethical guidelines of the Caring for Laboratory Animals by the Ministry of Science and Technology of the People’s Republic of China.

Additional information

Publisher’s Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Electronic supplementary material

Below is the link to the electronic supplementary material.

223_2019_528_MOESM1_ESM.docx

Supplementary material 1. Fig. S1 Skeletal muscle changes in Ano5+/+ and Ano5-/- mice. (A) H&E staining of gastrocnemius muscle of 16-week-old mice. Scale bar = 50 μm. (B) qPCR analysis of Dysferlin and Dystrophin in gastrocnemius (GAS), quadriceps (QD) and biceps muscles of 16-week-old mice. aP < 0.05, bP < 0.01. (C) Elevated serum creatine kinase (CK) level was detected in 16-week-old Ano5-/- mice. (n = 5). bP < 0.01 (DOCX 1084 KB)

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Wang, X., Liu, X., Dong, R. et al. Genetic Disruption of Anoctamin 5 in Mice Replicates Human Gnathodiaphyseal Dysplasia (GDD). Calcif Tissue Int 104, 679–689 (2019). https://doi.org/10.1007/s00223-019-00528-x

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00223-019-00528-x

Keywords

Search

Navigation