cdh23 affects congenital hearing loss through regulating purine metabolism

Shu Yang; Bing-Lin Xie; Xiao-Ping Dong; Ling-Xiang Wang; Gang-Hua Zhu; Tian Wang; Wei-Jing Wu; Ruo-Sha Lai; Rong Tao; Min-Xin Guan; Fang-Yi Chen; Dong-Hui Tan; Zhong Deng; Hua-Ping Xie; Yong Zeng; Zi-An Xiao; Ding-Hua Xie

doi:10.3389/fnmol.2023.1079529

cdh23 affects congenital hearing loss through regulating purine metabolism

Front Mol Neurosci. 2023 Jul 27:16:1079529. doi: 10.3389/fnmol.2023.1079529. eCollection 2023.

Authors

Shu Yang¹, Bing-Lin Xie², Xiao-Ping Dong³, Ling-Xiang Wang³, Gang-Hua Zhu¹, Tian Wang¹, Wei-Jing Wu¹, Ruo-Sha Lai¹, Rong Tao¹, Min-Xin Guan^{4

5}, Fang-Yi Chen⁶, Dong-Hui Tan⁷, Zhong Deng⁷, Hua-Ping Xie², Yong Zeng³, Zi-An Xiao¹, Ding-Hua Xie¹

Affiliations

¹ Department of Otorhinolaryngology-Head & Neck Surgery, The Second Xiangya Hospital of Central South University, Changsha, Hunan, China.
² Laboratory of Animal Nutrition and Human Health, Hunan International Joint Laboratory of Animal Intestinal Ecology and Health, College of Life Science, Hunan Normal University, Changsha, Hunan, China.
³ The National & Local Joint Engineering Laboratory of Animal Peptide Drug Development, College of Life Science, Hunan Normal University, Changsha, Hunan, China.
⁴ Institute of Genetics, Zhejiang University, Hangzhou, Zhejiang, China.
⁵ Department of Human Genetics, Zhejiang University School of Medicine, Zhejiang Provincial Key Laboratory of Genetic & Developmental Disorders, Hangzhou, Zhejiang, China.
⁶ Shenzhen Key Laboratory of Smart Healthcare Engineering, Department of Biomedical Engineering, Southern University of Science and Technology, Shenzhen, China.
⁷ Department of Otolaryngology-Head and Neck Surgery, The Affiliated Hospital of Xiang Nan College, Chenzhou, China.

Abstract

Introduction: The pathogenic gene CDH23 plays a pivotal role in tip links, which is indispensable for mechanoelectrical transduction in the hair cells. However, the underlying molecular mechanism and signal regulatory networks that influence deafness is still largely unknown.

Methods: In this study, a congenital deafness family, whole exome sequencing revealed a new mutation in the pathogenic gene CDH23, subsequently; the mutation has been validated using Sanger sequencing method. Then CRISPR/Cas9 technology was employed to knockout zebrafish cdh23 gene. Startle response experiment was used to compare with wide-type, the response to sound stimulation between wide-type and cdh23^-/-. To further illustrate the molecular mechanisms underlying congenital deafness, comparative transcriptomic profiling and multiple bioinformatics analyses were performed.

Results: The YO-PRO-1 assay result showed that in cdh23 deficient embryos, the YO-PRO-1 signal in inner ear and lateral line neuromast hair cells were completely lost. Startle response experiment showed that compared with wide-type, the response to sound stimulation decreased significantly in cdh23 mutant larvae. Comparative transcriptomic showed that the candidate genes such as atp1b2b and myof could affect hearing by regulating ATP production and purine metabolism in a synergetic way with cdh23. RT-qPCR results further confirmed the transcriptomics results. Further compensatory experiment showed that ATP treated cdh23^-/- embryos can partially recover the mutant phenotype.

Conclusion: In conclusion, our study may shed light on deciphering the principal mechanism and provide a potential therapeutic method for congenital hearing loss under the condition of CDH23 mutation.

Keywords: ATP; Cdh23; congenital hearing loss; gene knockout; metabolism; regulatory network.