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  • Basic Science Article
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Functional analysis of a novel pathogenic variant in CREBBP associated with bone development

Pediatric Research volume 96, pages 1626–1635 (2024)Cite this article

Abstract

Background

CREBBP has been extensively studied in syndromic diseases associated with skeletal dysplasia. However, there is limited research on the molecular mechanisms through which CREBBP may impact bone development. We identified a novel pathogenic CREBBP variant (c.C3862T/p.R1288W, which is orthologous to mouse c.3789 C > T/p.R1289W) in a patient with non-syndromic polydactyly.

Methods

We created a homozygous Crebbp p.R1289W mouse model and compared their skeletal phenotypes to wild-type (WT) animals. Bone marrow stem cells (BMSCs) were isolated and assessed for their proliferative capacity, proportion of apoptotic cells in culture, and differentiation to chondrocytes and osteocytes.

Results

We observed a significant decrease in body length in 8-week-old homozygous Crebbp p.R1289W mice. The relative length of cartilage of the digits of Crebbp p.R1289W mice was significantly increased compared to WT mice. BMSCs derived from Crebbp p.R1289W mice had significantly decreased cell proliferation and an elevated rate of apoptosis. Consistently, cell proliferative capacity was decreased and the proportion of apoptotic cells was increased in the distal femoral growth plate of Crebbp p.R1289W compared to WT mice. Chemical induction of BMSCs indicated that Crebbp p.R1289W may promote chondrocyte differentiation.

Conclusion

The Crebbp p.R1289W variant plays a pathogenic role in skeletal development in mice.

Impact

  • CREBBP has been extensively studied in syndromic diseases characterized by skeletal dysplasia.

  • There is limited research regarding the molecular mechanism through which CREBBP may affect bone development.

  • To our knowledge, we generated the first animal model of a novel Crebbp variant, which is predicted to be pathogenic for skeletal diseases.

  • Certain pathogenic variants, such as Crebbp p.R1289W, can independently lead to variant-specific non-syndromic skeletal dysplasia.

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Fig. 1: Identification of novel heterozygous CREBBP variants in patients with polydactyly.
Fig. 2: Relative cartilage length is significantly longer in digits of Crebbp p.R1289W compared to WT mice.
Fig. 3: Crebbp p.R1289W does not affect the characteristics of trabecular bone.
Fig. 4: Crebbp p.R1289W inhibits the proliferation of BMSCs derived from mice and promotes apoptosis.
Fig. 5: Crebbp p.R1289W promotes the differentiation of BMSCs derived from mice into cartilage.

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Data availability

Data that support the findings of this study are available from the corresponding author upon reasonable request.

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Acknowledgements

We express our gratitude to the patients and their families who willingly participated in this study.

Funding

The research was financially supported by grants from the National Natural Science Foundation of China (No. 82372328), the Medicine Guide Project (Chinese and Western medicine) of the Shanghai Science and Technology Committee (No. 18411961400), and the Science and Technology Commission Fund of Shanghai (No. 20ZR1434800). The authors declare no conflicts of interest.

Author information

Author notes

  1. These authors contributed equally: Qing Fang, Yunlan Xu, Jue Feng.

Authors and Affiliations

  1. Pediatric Translational Medicine Institute, Shanghai Children’s Medical Center, Shanghai Jiao Tong University School of Medicine, Shanghai, 200127, P. R. China

    Qing Fang, Xiaoqing Zhang, Bo Wang, Qihua Fu & Ying Xiang

  2. Department of Pediatric Orthopedic, Shanghai Children’s Medical Center, Shanghai Jiao Tong University School of Medicine, Shanghai, 200127, P. R. China

    Yunlan Xu & Jue Feng

Authors
  1. Qing Fang
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  2. Yunlan Xu
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  3. Jue Feng
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  5. Bo Wang
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Contributions

Conceptualization: Q.F., J.F. and Y.L.X. Formal analysis: B.W., Q.H.F., Y.X. Investigation: F.Q. and X.Q.Z. Writing-original draft preparation: Y.X. Writing-review and editing: Y.X., B.W., Y.L.X. and Q.H.F.

Corresponding authors

Correspondence to Bo Wang, Qihua Fu or Ying Xiang.

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Competing interests

The authors declare no competing interests.

Ethical approval and informed consent

The study protocol was approved by the Ethics Committee of Shanghai Children’s Medical Center, School of Medicine, Shanghai Jiao Tong University(no. SCMCIRB-K2017008),and written informed consent was obtained from all participants or their legal guardians. All animal procedures were approved by the animal ethics committee of Shanghai Children’s Medical Center (no. SCMC-LAWEC-2019-015).

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Fang, Q., Xu, Y., Feng, J. et al. Functional analysis of a novel pathogenic variant in CREBBP associated with bone development. Pediatr Res 96, 1626–1635 (2024). https://doi.org/10.1038/s41390-024-03490-z

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