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Updated: Oct 3, 2025
Chemogenetic Regulation in Reprogrammed Stem Cell-derived Precursor Cells in Treating Neurodegenerative Diseases
Published on: May 2, 2025
Anti-Excitotoxic Effects of N-Butylidenephthalide Revealed by Chemically Insulted Purkinje Progenitor Cells Derived
Hsin-Han Yang1, I-Tsang Chiang1, Jen-Wei Liu2
1Department of Life Science and Graduate Institute of Biotechnology, National Dong Hwa University, Hualien 974, Taiwan.
Abstract:
Spinocerebellar ataxia type 3 (SCA3) is characterized by the over-repetitive CAG codon in the ataxin-3 gene (ATXN3), 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.
Insights
N-butylidenephthalide (n-BP) shows promise in treating spinocerebellar ataxia type 3 (SCA3). This study found n-BP protects against quinolinic acid-induced excitotoxicity in SCA3 patient-derived cells, offering a potential therapeutic avenue.
Area of Science:
- Neuroscience
- Genetics
- Pharmacology
Background:
- Spinocerebellar ataxia type 3 (SCA3) is a neurodegenerative disorder caused by CAG repeat expansion in the ATXN3 gene.
- Existing research highlights pathological defects in SCA3, including impaired aggresomes, autophagy, and proteasome function.
- Currently, no effective treatments are available for SCA3 disease.
Purpose of the Study:
- To investigate the anti-excitotoxic effects of n-butylidenephthalide (n-BP) on Purkinje progenitor cells (PPs) derived from SCA3 patient-induced pluripotent stem cells (iPSCs).
- To establish a cellular model for identifying potential drug candidates for SCA3.
Main Methods:
- Generation of Purkinje progenitor cells (PPs) from SCA3 patient-derived iPSCs.
- Induction of excitotoxicity using quinolinic acid (QA) in SCA3 PPs.
- Assessment of cellular degeneration, calpain activity, ATXN3 proteolytic fragments, and apoptotic markers (PARP1).
- Treatment of QA-exposed SCA3 PPs with n-butylidenephthalide (n-BP).
Main Results:
- SCA3 PPs exhibited increased sensitivity to QA-induced excitotoxicity, characterized by neurite shrinkage and cell death.
- QA treatment led to increased calpain activity, cleaved PARP1 expression, and accumulation of ATXN3 fragments in SCA3 PPs.
- n-BP treatment reversed upregulated calpain 2 and downregulated calpastatin, suppressing calpain activity and inhibiting ATXN3 cleavage.
Conclusions:
- n-Butylidenephthalide (n-BP) demonstrates protective effects against quinolinic acid-induced excitotoxicity in SCA3 Purkinje progenitor cells.
- n-BP mitigates SCA3 pathology by inhibiting ATXN3 cleavage and suppressing calpain activity.
- This study establishes a valuable cellular model for SCA3 drug discovery and highlights n-BP as a potential therapeutic agent.
