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

  • Xiaoyu Wang
  • Xiu Liu
  • Rui Dong
  • Chao Liang
  • Ernst J. Reichenberger
  • Ying HuEmail author
Original Research


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.


Gnathodiaphyseal dysplasia Ano5 Genetic disorder Skeletal phenotype Osteoblastogenesis 



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).

Compliance with Ethical Standards

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.

Supplementary material

223_2019_528_MOESM1_ESM.docx (1.1 mb)
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)


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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  • Xiaoyu Wang
    • 1
  • Xiu Liu
    • 1
  • Rui Dong
    • 1
  • Chao Liang
    • 1
  • Ernst J. Reichenberger
    • 2
  • Ying Hu
    • 1
    • 3
    Email author
  1. 1.Beijing Stomatological HospitalCapital Medical UniversityBeijingPeople’s Republic of China
  2. 2.Department of Reconstructive Sciences, Center for Regenerative Medicine and Skeletal DevelopmentUniversity of Connecticut HealthFarmingtonUSA
  3. 3.Beijing Stomatological HospitalBeijing Institute of Dental Research, Capital Medical UniversityBeijingPeople’s Republic of China

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