Impaired AIF-CHCHD4 interaction and mitochondrial calcium overload contribute to auditory neuropathy spectrum disorder in patient-iPSC-derived neurons with AIFM1 variant
Abstract Auditory neuropathy spectrum disorder (ANSD) is a hearing impairment caused by dysfunction of inner hair cells, ribbon synapses, spiral ganglion neurons and/or the auditory nerve itself. Approximately 1/7000 newborns have abnormal auditory nerve function, accounting for 10%-14% of cases of...
| Main Authors: | , , , , , , , , , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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Nature Publishing Group
2023-06-01
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| Series: | Cell Death and Disease |
| Online Access: | https://doi.org/10.1038/s41419-023-05899-6 |
| _version_ | 1797789586454216704 |
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| author | Yue Qiu Hongyang Wang Mingjie Fan Huaye Pan Jing Guan Yangwei Jiang Zexiao Jia Kaiwen Wu Hui Zhou Qianqian Zhuang Zhaoying Lei Xue Ding Huajian Cai Yufei Dong Lei Yan Aifu Lin Yong Fu Dong Zhang Qingfeng Yan Qiuju Wang |
| author_facet | Yue Qiu Hongyang Wang Mingjie Fan Huaye Pan Jing Guan Yangwei Jiang Zexiao Jia Kaiwen Wu Hui Zhou Qianqian Zhuang Zhaoying Lei Xue Ding Huajian Cai Yufei Dong Lei Yan Aifu Lin Yong Fu Dong Zhang Qingfeng Yan Qiuju Wang |
| author_sort | Yue Qiu |
| collection | DOAJ |
| description | Abstract Auditory neuropathy spectrum disorder (ANSD) is a hearing impairment caused by dysfunction of inner hair cells, ribbon synapses, spiral ganglion neurons and/or the auditory nerve itself. Approximately 1/7000 newborns have abnormal auditory nerve function, accounting for 10%-14% of cases of permanent hearing loss in children. Although we previously identified the AIFM1 c.1265 G > A variant to be associated with ANSD, the mechanism by which ANSD is associated with AIFM1 is poorly understood. We generated induced pluripotent stem cells (iPSCs) from peripheral blood mononuclear cells (PBMCs) via nucleofection with episomal plasmids. The patient-specific iPSCs were edited via CRISPR/Cas9 technology to generate gene-corrected isogenic iPSCs. These iPSCs were further differentiated into neurons via neural stem cells (NSCs). The pathogenic mechanism was explored in these neurons. In patient cells (PBMCs, iPSCs, and neurons), the AIFM1 c.1265 G > A variant caused a novel splicing variant (c.1267-1305del), resulting in AIF p.R422Q and p.423-435del proteins, which impaired AIF dimerization. Such impaired AIF dimerization then weakened the interaction between AIF and coiled-coil-helix-coiled-coil-helix domain-containing protein 4 (CHCHD4). On the one hand, the mitochondrial import of ETC complex subunits was inhibited, subsequently leading to an increased ADP/ATP ratio and elevated ROS levels. On the other hand, MICU1-MICU2 heterodimerization was impaired, leading to mCa2+ overload. Calpain was activated by mCa2+ and subsequently cleaved AIF for its translocation into the nucleus, ultimately resulting in caspase-independent apoptosis. Interestingly, correction of the AIFM1 variant significantly restored the structure and function of AIF, further improving the physiological state of patient-specific iPSC-derived neurons. This study demonstrates that the AIFM1 variant is one of the molecular bases of ANSD. Mitochondrial dysfunction, especially mCa2+ overload, plays a prominent role in ANSD associated with AIFM1. Our findings help elucidate the mechanism of ANSD and may lead to the provision of novel therapies. |
| first_indexed | 2024-03-13T01:51:53Z |
| format | Article |
| id | doaj.art-b6d5ad11b2a84e4fb3d54c6e65e8be61 |
| institution | Directory Open Access Journal |
| issn | 2041-4889 |
| language | English |
| last_indexed | 2024-03-13T01:51:53Z |
| publishDate | 2023-06-01 |
| publisher | Nature Publishing Group |
| record_format | Article |
| series | Cell Death and Disease |
| spelling | doaj.art-b6d5ad11b2a84e4fb3d54c6e65e8be612023-07-02T11:28:10ZengNature Publishing GroupCell Death and Disease2041-48892023-06-0114611410.1038/s41419-023-05899-6Impaired AIF-CHCHD4 interaction and mitochondrial calcium overload contribute to auditory neuropathy spectrum disorder in patient-iPSC-derived neurons with AIFM1 variantYue Qiu0Hongyang Wang1Mingjie Fan2Huaye Pan3Jing Guan4Yangwei Jiang5Zexiao Jia6Kaiwen Wu7Hui Zhou8Qianqian Zhuang9Zhaoying Lei10Xue Ding11Huajian Cai12Yufei Dong13Lei Yan14Aifu Lin15Yong Fu16Dong Zhang17Qingfeng Yan18Qiuju Wang19College of Life Sciences, Zhejiang UniversitySenior Department of Otolaryngology, Head and Neck Surgery, Chinese PLA Institute of Otolaryngology, Chinese PLA General HospitalCollege of Life Sciences, Zhejiang UniversityCollege of Life Sciences, Zhejiang UniversitySenior Department of Otolaryngology, Head and Neck Surgery, Chinese PLA Institute of Otolaryngology, Chinese PLA General HospitalCollege of Life Sciences, Zhejiang UniversityCollege of Life Sciences, Zhejiang UniversitySenior Department of Otolaryngology, Head and Neck Surgery, Chinese PLA Institute of Otolaryngology, Chinese PLA General HospitalCollege of Life Sciences, Zhejiang UniversityCollege of Life Sciences, Zhejiang UniversityCollege of Life Sciences, Zhejiang UniversityCollege of Life Sciences, Zhejiang UniversityCollege of Life Sciences, Zhejiang UniversityCollege of Life Sciences, Zhejiang UniversityCollege of Life Sciences, Zhejiang UniversityCollege of Life Sciences, Zhejiang UniversityThe Children’s Hospital of Zhejiang University School of MedicineCollege of Life Sciences, Zhejiang UniversityCollege of Life Sciences, Zhejiang UniversitySenior Department of Otolaryngology, Head and Neck Surgery, Chinese PLA Institute of Otolaryngology, Chinese PLA General HospitalAbstract Auditory neuropathy spectrum disorder (ANSD) is a hearing impairment caused by dysfunction of inner hair cells, ribbon synapses, spiral ganglion neurons and/or the auditory nerve itself. Approximately 1/7000 newborns have abnormal auditory nerve function, accounting for 10%-14% of cases of permanent hearing loss in children. Although we previously identified the AIFM1 c.1265 G > A variant to be associated with ANSD, the mechanism by which ANSD is associated with AIFM1 is poorly understood. We generated induced pluripotent stem cells (iPSCs) from peripheral blood mononuclear cells (PBMCs) via nucleofection with episomal plasmids. The patient-specific iPSCs were edited via CRISPR/Cas9 technology to generate gene-corrected isogenic iPSCs. These iPSCs were further differentiated into neurons via neural stem cells (NSCs). The pathogenic mechanism was explored in these neurons. In patient cells (PBMCs, iPSCs, and neurons), the AIFM1 c.1265 G > A variant caused a novel splicing variant (c.1267-1305del), resulting in AIF p.R422Q and p.423-435del proteins, which impaired AIF dimerization. Such impaired AIF dimerization then weakened the interaction between AIF and coiled-coil-helix-coiled-coil-helix domain-containing protein 4 (CHCHD4). On the one hand, the mitochondrial import of ETC complex subunits was inhibited, subsequently leading to an increased ADP/ATP ratio and elevated ROS levels. On the other hand, MICU1-MICU2 heterodimerization was impaired, leading to mCa2+ overload. Calpain was activated by mCa2+ and subsequently cleaved AIF for its translocation into the nucleus, ultimately resulting in caspase-independent apoptosis. Interestingly, correction of the AIFM1 variant significantly restored the structure and function of AIF, further improving the physiological state of patient-specific iPSC-derived neurons. This study demonstrates that the AIFM1 variant is one of the molecular bases of ANSD. Mitochondrial dysfunction, especially mCa2+ overload, plays a prominent role in ANSD associated with AIFM1. Our findings help elucidate the mechanism of ANSD and may lead to the provision of novel therapies.https://doi.org/10.1038/s41419-023-05899-6 |
| spellingShingle | Yue Qiu Hongyang Wang Mingjie Fan Huaye Pan Jing Guan Yangwei Jiang Zexiao Jia Kaiwen Wu Hui Zhou Qianqian Zhuang Zhaoying Lei Xue Ding Huajian Cai Yufei Dong Lei Yan Aifu Lin Yong Fu Dong Zhang Qingfeng Yan Qiuju Wang Impaired AIF-CHCHD4 interaction and mitochondrial calcium overload contribute to auditory neuropathy spectrum disorder in patient-iPSC-derived neurons with AIFM1 variant Cell Death and Disease |
| title | Impaired AIF-CHCHD4 interaction and mitochondrial calcium overload contribute to auditory neuropathy spectrum disorder in patient-iPSC-derived neurons with AIFM1 variant |
| title_full | Impaired AIF-CHCHD4 interaction and mitochondrial calcium overload contribute to auditory neuropathy spectrum disorder in patient-iPSC-derived neurons with AIFM1 variant |
| title_fullStr | Impaired AIF-CHCHD4 interaction and mitochondrial calcium overload contribute to auditory neuropathy spectrum disorder in patient-iPSC-derived neurons with AIFM1 variant |
| title_full_unstemmed | Impaired AIF-CHCHD4 interaction and mitochondrial calcium overload contribute to auditory neuropathy spectrum disorder in patient-iPSC-derived neurons with AIFM1 variant |
| title_short | Impaired AIF-CHCHD4 interaction and mitochondrial calcium overload contribute to auditory neuropathy spectrum disorder in patient-iPSC-derived neurons with AIFM1 variant |
| title_sort | impaired aif chchd4 interaction and mitochondrial calcium overload contribute to auditory neuropathy spectrum disorder in patient ipsc derived neurons with aifm1 variant |
| url | https://doi.org/10.1038/s41419-023-05899-6 |
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