Anti-Excitotoxic Effects of N-Butylidenephthalide Revealed by Chemically Insulted Purkinje Progenitor Cells Derived from SCA3 iPSCs
Spinocerebellar ataxia type 3 (SCA3) is characterized by the over-repetitive CAG codon in the ataxin-3 gene (<i>ATXN3</i>), which encodes the mutant ATXN3 protein. The pathological defects of SCA3 such as the impaired aggresomes, autophagy, and the proteasome have been reported previousl...
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MDPI AG
2022-01-01
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| Series: | International Journal of Molecular Sciences |
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| Online Access: | https://www.mdpi.com/1422-0067/23/3/1391 |
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| author | Hsin-Han Yang I-Tsang Chiang Jen-Wei Liu Jeanne Hsieh Jui-Hao Lee Huai-En Lu Hwa-Sung Tso Yu-Chen Deng Jo-Chi Kao Jhen-Rong Wu Horng-Jyh Harn Tzyy-Wen Chiou |
| author_facet | Hsin-Han Yang I-Tsang Chiang Jen-Wei Liu Jeanne Hsieh Jui-Hao Lee Huai-En Lu Hwa-Sung Tso Yu-Chen Deng Jo-Chi Kao Jhen-Rong Wu Horng-Jyh Harn Tzyy-Wen Chiou |
| author_sort | Hsin-Han Yang |
| collection | DOAJ |
| description | Spinocerebellar ataxia type 3 (SCA3) is characterized by the over-repetitive CAG codon in the ataxin-3 gene (<i>ATXN3</i>), which encodes the mutant ATXN3 protein. The pathological defects of SCA3 such as the impaired aggresomes, autophagy, and the proteasome have been reported previously. To date, no effective treatment is available for SCA3 disease. This study aimed to study anti-excitotoxic effects of n-butylidenephthalide by chemically insulted Purkinje progenitor cells derived from SCA3 iPSCs. We successfully generated Purkinje progenitor cells (PPs) from SCA3 patient-derived iPSCs. The PPs, expressing both neural and Purkinje progenitor’s markers, were acquired after 35 days of differentiation. In comparison with the PPs derived from control iPSCs, SCA3 iPSCs-derived PPs were more sensitive to the excitotoxicity induced by quinolinic acid (QA). The observations of QA-treated SCA3 PPs showing neural degeneration including neurite shrinkage and cell number decrease could be used to quickly and efficiently identify drug candidates. Given that the QA-induced neural cell death of SCA3 PPs was established, the activity of calpain in SCA3 PPs was revealed. Furthermore, the expression of cleaved poly (ADP-ribose) polymerase 1 (PARP1), a marker of apoptotic pathway, and the accumulation of ATXN3 proteolytic fragments were observed. When SCA3 PPs were treated with n-butylidenephthalide (n-BP), upregulated expression of calpain 2 and concurrent decreased level of calpastatin could be reversed, and the overall calpain activity was accordingly suppressed. Such findings reveal that n-BP could not only inhibit the cleavage of ATXN3 but also protect the QA-induced excitotoxicity from the Purkinje progenitor loss. |
| first_indexed | 2024-03-09T23:47:55Z |
| format | Article |
| id | doaj.art-99c025697dde47e4bec9d38f6fd28839 |
| institution | Directory Open Access Journal |
| issn | 1661-6596 1422-0067 |
| language | English |
| last_indexed | 2024-03-09T23:47:55Z |
| publishDate | 2022-01-01 |
| publisher | MDPI AG |
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| series | International Journal of Molecular Sciences |
| spelling | doaj.art-99c025697dde47e4bec9d38f6fd288392023-11-23T16:39:40ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672022-01-01233139110.3390/ijms23031391Anti-Excitotoxic Effects of N-Butylidenephthalide Revealed by Chemically Insulted Purkinje Progenitor Cells Derived from SCA3 iPSCsHsin-Han Yang0I-Tsang Chiang1Jen-Wei Liu2Jeanne Hsieh3Jui-Hao Lee4Huai-En Lu5Hwa-Sung Tso6Yu-Chen Deng7Jo-Chi Kao8Jhen-Rong Wu9Horng-Jyh Harn10Tzyy-Wen Chiou11Department of Life Science and Graduate Institute of Biotechnology, National Dong Hwa University, Hualien 974, TaiwanDepartment of Life Science and Graduate Institute of Biotechnology, National Dong Hwa University, Hualien 974, TaiwanEverfront Biotech Inc., New Taipei City 221, TaiwanEverfront Biotech Inc., New Taipei City 221, TaiwanEverfront Biotech Inc., New Taipei City 221, TaiwanBioresource Collection and Research Center, Food Industry Research and Development Institute, Hsinchu 300, TaiwanDepartment of Life Science and Graduate Institute of Biotechnology, National Dong Hwa University, Hualien 974, TaiwanDepartment of Life Science and Graduate Institute of Biotechnology, National Dong Hwa University, Hualien 974, TaiwanDepartment of Life Science and Graduate Institute of Biotechnology, National Dong Hwa University, Hualien 974, TaiwanDepartment of Life Science and Graduate Institute of Biotechnology, National Dong Hwa University, Hualien 974, TaiwanBioinnovation Center, Buddhist Tzu Chi Foundation, Hualien 970, TaiwanDepartment of Life Science and Graduate Institute of Biotechnology, National Dong Hwa University, Hualien 974, TaiwanSpinocerebellar ataxia type 3 (SCA3) is characterized by the over-repetitive CAG codon in the ataxin-3 gene (<i>ATXN3</i>), which encodes the mutant ATXN3 protein. The pathological defects of SCA3 such as the impaired aggresomes, autophagy, and the proteasome have been reported previously. To date, no effective treatment is available for SCA3 disease. This study aimed to study anti-excitotoxic effects of n-butylidenephthalide by chemically insulted Purkinje progenitor cells derived from SCA3 iPSCs. We successfully generated Purkinje progenitor cells (PPs) from SCA3 patient-derived iPSCs. The PPs, expressing both neural and Purkinje progenitor’s markers, were acquired after 35 days of differentiation. In comparison with the PPs derived from control iPSCs, SCA3 iPSCs-derived PPs were more sensitive to the excitotoxicity induced by quinolinic acid (QA). The observations of QA-treated SCA3 PPs showing neural degeneration including neurite shrinkage and cell number decrease could be used to quickly and efficiently identify drug candidates. Given that the QA-induced neural cell death of SCA3 PPs was established, the activity of calpain in SCA3 PPs was revealed. Furthermore, the expression of cleaved poly (ADP-ribose) polymerase 1 (PARP1), a marker of apoptotic pathway, and the accumulation of ATXN3 proteolytic fragments were observed. When SCA3 PPs were treated with n-butylidenephthalide (n-BP), upregulated expression of calpain 2 and concurrent decreased level of calpastatin could be reversed, and the overall calpain activity was accordingly suppressed. Such findings reveal that n-BP could not only inhibit the cleavage of ATXN3 but also protect the QA-induced excitotoxicity from the Purkinje progenitor loss.https://www.mdpi.com/1422-0067/23/3/1391n-butylidenephthalideSCA3ATXN3iPSCsPurkinje progenitorquinolinic acid |
| spellingShingle | Hsin-Han Yang I-Tsang Chiang Jen-Wei Liu Jeanne Hsieh Jui-Hao Lee Huai-En Lu Hwa-Sung Tso Yu-Chen Deng Jo-Chi Kao Jhen-Rong Wu Horng-Jyh Harn Tzyy-Wen Chiou Anti-Excitotoxic Effects of N-Butylidenephthalide Revealed by Chemically Insulted Purkinje Progenitor Cells Derived from SCA3 iPSCs International Journal of Molecular Sciences n-butylidenephthalide SCA3 ATXN3 iPSCs Purkinje progenitor quinolinic acid |
| title | Anti-Excitotoxic Effects of N-Butylidenephthalide Revealed by Chemically Insulted Purkinje Progenitor Cells Derived from SCA3 iPSCs |
| title_full | Anti-Excitotoxic Effects of N-Butylidenephthalide Revealed by Chemically Insulted Purkinje Progenitor Cells Derived from SCA3 iPSCs |
| title_fullStr | Anti-Excitotoxic Effects of N-Butylidenephthalide Revealed by Chemically Insulted Purkinje Progenitor Cells Derived from SCA3 iPSCs |
| title_full_unstemmed | Anti-Excitotoxic Effects of N-Butylidenephthalide Revealed by Chemically Insulted Purkinje Progenitor Cells Derived from SCA3 iPSCs |
| title_short | Anti-Excitotoxic Effects of N-Butylidenephthalide Revealed by Chemically Insulted Purkinje Progenitor Cells Derived from SCA3 iPSCs |
| title_sort | anti excitotoxic effects of n butylidenephthalide revealed by chemically insulted purkinje progenitor cells derived from sca3 ipscs |
| topic | n-butylidenephthalide SCA3 ATXN3 iPSCs Purkinje progenitor quinolinic acid |
| url | https://www.mdpi.com/1422-0067/23/3/1391 |
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