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Expanding the genetic and phenotypic relevance of CLCN4 variants in neurodevelopmental condition: 13 new patients

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Abstract

Objectives

CLCN4 variations have recently been identified as a genetic cause of X-linked neurodevelopmental disorders. This study aims to broaden the phenotypic spectrum of CLCN4-related condition and correlate it with functional consequences of CLCN4 variants.

Methods

We described 13 individuals with CLCN4-related neurodevelopmental disorder. We analyzed the functional consequence of the unreported variants using heterologous expression, biochemistry, confocal fluorescent microscopy, patch-clamp electrophysiology, and minigene splicing assay.

Results

We identified five novel (p.R41W, p.L348V, p.G480R, p.R603W, c.1576 + 5G > A) and three known (p.T203I, p.V275M, p.A555V) pathogenic CLCN4 variants in 13 Chinese patients. The p.V275M variant is found at high frequency and seen in four unrelated individuals. All had global developmental delay (GDD)/intellectual disability (ID). Seizures were present in eight individuals, and 62.5% of them developed refractory epilepsy. Five individuals without seizures showed moderate to severe GDD/ID. Developmental delay precedes seizure onset in most patients. The variants p.R41W, p.L348V, and p.R603W compromise the anion/exchange function of ClC-4. p.R41W partially impairs ClC-3/ClC-4 association. p.G480R reduces ClC-4 expression levels and impairs the heterodimerization with ClC-3. The c.1576 + 5G > A variant causes 22 bp deletion of exon 10.

Conclusions

We further define and broaden the clinical and mutational spectrum of CLCN4-related neurodevelopmental conditions. The p.V275M variant may be a potential hotspot CLCN4 variant in Chinese patients. The five novel variants cause loss of function of ClC-4. Transport dysfunction, protein instability, intracellular trafficking defect, or failure of ClC-4 to oligomerize may contribute to the pathophysiological events leading to CLCN4-related neurodevelopmental disorder.

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Data availability

The anonymized data are available from the corresponding author upon reasonable request.

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Acknowledgements

We thank the patients and their parents for kindly giving us permission to publish this data. We also thank Tingting Yang and other staff at Beijing Chigene Translational Medicine Research Center for their expert technical assistance. We are grateful to Dr. Daniel Kortzak for generating the ClC-4 structure (Fig. 1a).

Funding

This work was funded by the National Natural Science Foundation of China (82071462, 81771409 to PJ and 82201316 to HH), the Natural Science Foundation of Hunan Province (2022JJ40785 to HH), the China Postdoctoral Science Foundation (2023M733950 to HH), and the Key R&D Program of Hunan Province (2022SK2036 to PJ), the German Research Foundation (DFG) (GU 2042/2-1 to REG).

Author information

Authors and Affiliations

  1. Department of Pediatrics, Xiangya Hospital, Central South University, Changsha, China

    Hailan He, Xinyi Li, XueQin Lin & Jing Peng

  2. Institute of Biological Information Processing (IBI-7), Structural Biochemistry, Jülich Research Center, Jülich, Germany

    G. A. Guzman

  3. Institute of Biological Information Processing (IBI-1), Molecular and Cell Physiology, Jülich Research Center, Jülich, Germany

    Stefanie Bungert-Plümke, Arne Franzen & Raul E. Guzman

  4. Department of Rehabilitation, Wuhan Children’s Hospital, Tongji Medical College, Huazhong University of Science & Technology, Wuhan, China

    Hongmin Zhu

  5. Department of Neurology, Xiamen Maternal and Child Health Care Hospital, Xiamen, China

    Guilan Peng & Zhongqin Huang

  6. Epilepsy Center, Children’s Hospital Affiliated to Shandong University, Jinan, China

    Hongwei Zhang

  7. Department of Rehabilitation, Children’s Hospital, Zhejiang University School of Medicine, Hangzhou, China

    Yonglin Yu

  8. Department of Pediatric Neurology, Hebei Children’s Hospital, Hebei Medical University, Shijiazhuang, China

    Suzhen Sun

  9. Department of Pediatrics, Guangdong Provincial People’s Hospital, Guangzhou, China

    Qiongxiang Zhai

  10. Department of Pediatrics, The First Affiliated Hospital of Zhengzhou University, Zhengzhou, China

    Zheng Chen

Authors
  1. Hailan He
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  2. Xinyi Li
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  3. G. A. Guzman
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  4. Stefanie Bungert-Plümke
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  5. Arne Franzen
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  6. XueQin Lin
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  7. Hongmin Zhu
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  8. Guilan Peng
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  9. Hongwei Zhang
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  10. Yonglin Yu
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  11. Suzhen Sun
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  13. Qiongxiang Zhai
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  14. Zheng Chen
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  15. Jing Peng
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  16. Raul E. Guzman
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Contributions

Conceptualization: Hailan He, Jing Peng, and Raul E. Guzman; investigation: Hailan He, Raul E. Guzman, Guzman G. A., Stefanie Bungert-Plümke, and Arne Franzen; resources: Hongmin Zhu, Hongwei Zhang, yonglin Yu, Suzhen Sun, Guilan Peng, Zheng Chen, Zhongqin Huang and Jing Peng; collecting and analyzing data: Hailan He, Raul E. Guzman, XueQin Lin, Guzman G. A., Stefanie Bungert-Plümke, and Arne Franzen; writing—original draft preparation: Hailan He; writing—review and editing: Raul E. Guzman, and Jing Peng.

Corresponding authors

Correspondence to Jing Peng or Raul E. Guzman.

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Conflicts of interest

The authors declare no conflicts of interest.

Ethics approval

This study was conducted following the Declaration of Helsinki and approved by the ethics committee of Xiangya Hospital of Central South University and relevant local ethics committee. Written informed consent was obtained from their parents.

Supplementary Information

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Supplementary file 1 (DOCX 789 kb)

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He, H., Li, X., Guzman, G.A. et al. Expanding the genetic and phenotypic relevance of CLCN4 variants in neurodevelopmental condition: 13 new patients. J Neurol (2024). https://doi.org/10.1007/s00415-024-12383-4

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  • DOI: https://doi.org/10.1007/s00415-024-12383-4

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