PAX3 mutation suppress otic progenitors proliferation and induce apoptosis by inhibiting WNT1/β-catenin signaling pathway in WS1 patient iPSC-derived inner ear organoids

SiJun Li; Chufeng He; Lingyun Mei; Xuewen Wu; Yong Feng; Jian Song

doi:10.1016/j.bbrc.2024.149510

PAX3 mutation suppress otic progenitors proliferation and induce apoptosis by inhibiting WNT1/β-catenin signaling pathway in WS1 patient iPSC-derived inner ear organoids

Biochem Biophys Res Commun. 2024 Feb 26:698:149510. doi: 10.1016/j.bbrc.2024.149510. Epub 2024 Jan 13.

Authors

SiJun Li¹, Chufeng He², Lingyun Mei³, Xuewen Wu⁴, Yong Feng⁵, Jian Song⁶

Affiliations

¹ Department of Otolaryngology-Head and Neck Surgery, Xiangya Hospital, Central South University, Changsha, Hunan, China; Otolaryngology Major Disease Research Key Laboratory of Hunan Province, Changsha, Hunan, China; Clinical Research Center for Pharyngolaryngeal Diseases and Voice Disorders in Hunan Province, Changsha, Hunan, China; National Clinical Research Center for Geriatric Disorders, Department of Geriatrics, Xiangya Hospital, Central South University, Changsha, Hunan, China. Electronic address: 1006038473@qq.com.
² Department of Otolaryngology-Head and Neck Surgery, Xiangya Hospital, Central South University, Changsha, Hunan, China; Otolaryngology Major Disease Research Key Laboratory of Hunan Province, Changsha, Hunan, China; Clinical Research Center for Pharyngolaryngeal Diseases and Voice Disorders in Hunan Province, Changsha, Hunan, China; National Clinical Research Center for Geriatric Disorders, Department of Geriatrics, Xiangya Hospital, Central South University, Changsha, Hunan, China. Electronic address: hechufeng2013@163.com.
³ Department of Otolaryngology-Head and Neck Surgery, Xiangya Hospital, Central South University, Changsha, Hunan, China; Otolaryngology Major Disease Research Key Laboratory of Hunan Province, Changsha, Hunan, China; Clinical Research Center for Pharyngolaryngeal Diseases and Voice Disorders in Hunan Province, Changsha, Hunan, China; National Clinical Research Center for Geriatric Disorders, Department of Geriatrics, Xiangya Hospital, Central South University, Changsha, Hunan, China. Electronic address: entmly@csu.edu.cn.
⁴ Department of Otolaryngology-Head and Neck Surgery, Xiangya Hospital, Central South University, Changsha, Hunan, China; Otolaryngology Major Disease Research Key Laboratory of Hunan Province, Changsha, Hunan, China; Clinical Research Center for Pharyngolaryngeal Diseases and Voice Disorders in Hunan Province, Changsha, Hunan, China; National Clinical Research Center for Geriatric Disorders, Department of Geriatrics, Xiangya Hospital, Central South University, Changsha, Hunan, China. Electronic address: xwwu840903@hotmail.com.
⁵ Department of Otolaryngology-Head and Neck Surgery, Xiangya Hospital, Central South University, Changsha, Hunan, China; Department of Otorhinolaryngology, The Affiliated Maternal and Child Health Hospital of Hunan Province, Hengyang Medical School, University of South China, Changsha, Hunan, China; MOE Key Lab of Rare Pediatric Diseases & Institute of Otorhinolaryngology, Head and Neck Surgery, University of South China, Changsha, Hunan, China. Electronic address: fengyong_hn@hotmail.com.
⁶ Department of Otolaryngology-Head and Neck Surgery, Xiangya Hospital, Central South University, Changsha, Hunan, China; Otolaryngology Major Disease Research Key Laboratory of Hunan Province, Changsha, Hunan, China; Clinical Research Center for Pharyngolaryngeal Diseases and Voice Disorders in Hunan Province, Changsha, Hunan, China; National Clinical Research Center for Geriatric Disorders, Department of Geriatrics, Xiangya Hospital, Central South University, Changsha, Hunan, China. Electronic address: joe0426@126.com.

PMID: 38278051
DOI: 10.1016/j.bbrc.2024.149510

Abstract

Waardenburg syndrome type 1 (WS1) is a hereditary disease mainly characterized by sensorineural hearing loss, dystopia canthorum, and pigmentary defects. To elucidate molecular mechanisms underlying PAX3-associated hearing loss, we developed inner ear organoids model using induced pluripotent stem cells (iPSCs) derived from WS1 patient and healthy individual. Our results revealed a significant reduction in the size of inner ear organoids, accompanied by an increased level of apoptosis in organoids derived from WS1 patient-iPSCs carrying PAX3 c.214A > G. Transcriptome profiling analysis by RNA-seq indicated that inner ear organoids from WS1 patients were associated with suppression of inner ear development and WNT signaling pathway. Furthermore, the upregulation of the WNT1/β-catenin pathway which was achieved through the correction of PAX3 isogenic mutant iPSCs using CRISPR/Cas9, contributed to an increased size of inner ear organoids and a reduction in apoptosis. Together, our results provide insight into the underlying mechanisms of hearing loss in WS.

Keywords: Apoptosis; CRISPR/Cas9; Organoid; WNT1/β-catenin; Waardenburg syndrome 1; iPSCs.

MeSH terms

Apoptosis
Cell Proliferation
Deafness*
Ear, Inner*
Humans
Induced Pluripotent Stem Cells*
Mutation
Organoids
PAX3 Transcription Factor / genetics
Waardenburg Syndrome* / genetics
Wnt Signaling Pathway
beta Catenin / genetics

Substances

PAX3 Transcription Factor
beta Catenin
PAX3 protein, human