Phenylbutyric Acid Modulates Apoptosis and ER Stress‐Related Gene Expression in Glycogen Storage Disease Type Ib In Vitro Model
ABSTRACT Introduction Chronic endoplasmic reticulum (ER) stress and increased apoptosis are involved in the pathogenesis of glycogen storage disease Ib (GSD Ib), whereas small molecule phenylbutyrate (4‐PBA) showed the capability of reducing ER stress‐induced apoptosis. The objective was to generate...
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Wiley
2025-01-01
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| Series: | Molecular Genetics & Genomic Medicine |
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| Online Access: | https://doi.org/10.1002/mgg3.70054 |
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| author | Marina Parezanovic Nina Stevanovic Marina Andjelkovic Milena Ugrin Sonja Pavlovic Maja Stojiljkovic Anita Skakic |
| author_facet | Marina Parezanovic Nina Stevanovic Marina Andjelkovic Milena Ugrin Sonja Pavlovic Maja Stojiljkovic Anita Skakic |
| author_sort | Marina Parezanovic |
| collection | DOAJ |
| description | ABSTRACT Introduction Chronic endoplasmic reticulum (ER) stress and increased apoptosis are involved in the pathogenesis of glycogen storage disease Ib (GSD Ib), whereas small molecule phenylbutyrate (4‐PBA) showed the capability of reducing ER stress‐induced apoptosis. The objective was to generate an in vitro system in which capability of small molecules (SMs) to influence ER stress and apoptosis could be screened at the expression level. Methods G6PT‐deficient FlpInHEK293 cell line was created and validated using the CRISPR/Cas9 knockout method. Molecular markers of unfolded protein response (ATF4, DDIT3, HSPA5, XBP1s), and apoptosis (BCL2/BAX, CASP3, CASP7) in G6PT‐deficient cells were analyzed using RT‐qPCR method before and upon the treatment with 4‐PBA. Results Treatment with the most effective dose of 1 mM 4‐PBA reduced the expression of UPR markers and executioner caspases, while increased BCL2/BAX ratio in G6PT‐deficient cells. Our results proved the concept that 4‐PBA could alleviate markers of ER stress detected in the GSD Ib in vitro model system and prevent cell death. Conclusion This cost‐effective in vitro model screens the therapeutic potential of SMs affecting ER stress and apoptosis in G6PT‐deficient kidney cells, offering a first‐line screening assay for promising compounds. 4‐PBA's potential repurposing for GSD Ib patients opens new research directions. |
| format | Article |
| id | doaj-art-8b6c494a2e344f8b87a717093e5530da |
| institution | Kabale University |
| issn | 2324-9269 |
| language | English |
| publishDate | 2025-01-01 |
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| series | Molecular Genetics & Genomic Medicine |
| spelling | doaj-art-8b6c494a2e344f8b87a717093e5530da2025-01-24T08:16:42ZengWileyMolecular Genetics & Genomic Medicine2324-92692025-01-01131n/an/a10.1002/mgg3.70054Phenylbutyric Acid Modulates Apoptosis and ER Stress‐Related Gene Expression in Glycogen Storage Disease Type Ib In Vitro ModelMarina Parezanovic0Nina Stevanovic1Marina Andjelkovic2Milena Ugrin3Sonja Pavlovic4Maja Stojiljkovic5Anita Skakic6Group for Rare Disease Research and Therapeutics Development, Institute of Molecular Genetics and Genetic Engineering University of Belgrade Belgrade Republic of SerbiaGroup for Rare Disease Research and Therapeutics Development, Institute of Molecular Genetics and Genetic Engineering University of Belgrade Belgrade Republic of SerbiaGroup for Rare Disease Research and Therapeutics Development, Institute of Molecular Genetics and Genetic Engineering University of Belgrade Belgrade Republic of SerbiaGroup for Rare Disease Research and Therapeutics Development, Institute of Molecular Genetics and Genetic Engineering University of Belgrade Belgrade Republic of SerbiaGroup for Rare Disease Research and Therapeutics Development, Institute of Molecular Genetics and Genetic Engineering University of Belgrade Belgrade Republic of SerbiaGroup for Rare Disease Research and Therapeutics Development, Institute of Molecular Genetics and Genetic Engineering University of Belgrade Belgrade Republic of SerbiaGroup for Rare Disease Research and Therapeutics Development, Institute of Molecular Genetics and Genetic Engineering University of Belgrade Belgrade Republic of SerbiaABSTRACT Introduction Chronic endoplasmic reticulum (ER) stress and increased apoptosis are involved in the pathogenesis of glycogen storage disease Ib (GSD Ib), whereas small molecule phenylbutyrate (4‐PBA) showed the capability of reducing ER stress‐induced apoptosis. The objective was to generate an in vitro system in which capability of small molecules (SMs) to influence ER stress and apoptosis could be screened at the expression level. Methods G6PT‐deficient FlpInHEK293 cell line was created and validated using the CRISPR/Cas9 knockout method. Molecular markers of unfolded protein response (ATF4, DDIT3, HSPA5, XBP1s), and apoptosis (BCL2/BAX, CASP3, CASP7) in G6PT‐deficient cells were analyzed using RT‐qPCR method before and upon the treatment with 4‐PBA. Results Treatment with the most effective dose of 1 mM 4‐PBA reduced the expression of UPR markers and executioner caspases, while increased BCL2/BAX ratio in G6PT‐deficient cells. Our results proved the concept that 4‐PBA could alleviate markers of ER stress detected in the GSD Ib in vitro model system and prevent cell death. Conclusion This cost‐effective in vitro model screens the therapeutic potential of SMs affecting ER stress and apoptosis in G6PT‐deficient kidney cells, offering a first‐line screening assay for promising compounds. 4‐PBA's potential repurposing for GSD Ib patients opens new research directions.https://doi.org/10.1002/mgg3.700544‐PBAapoptosisCRISPR/Cas9ER stressGSD Ib in vitro model system |
| spellingShingle | Marina Parezanovic Nina Stevanovic Marina Andjelkovic Milena Ugrin Sonja Pavlovic Maja Stojiljkovic Anita Skakic Phenylbutyric Acid Modulates Apoptosis and ER Stress‐Related Gene Expression in Glycogen Storage Disease Type Ib In Vitro Model Molecular Genetics & Genomic Medicine 4‐PBA apoptosis CRISPR/Cas9 ER stress GSD Ib in vitro model system |
| title | Phenylbutyric Acid Modulates Apoptosis and ER Stress‐Related Gene Expression in Glycogen Storage Disease Type Ib In Vitro Model |
| title_full | Phenylbutyric Acid Modulates Apoptosis and ER Stress‐Related Gene Expression in Glycogen Storage Disease Type Ib In Vitro Model |
| title_fullStr | Phenylbutyric Acid Modulates Apoptosis and ER Stress‐Related Gene Expression in Glycogen Storage Disease Type Ib In Vitro Model |
| title_full_unstemmed | Phenylbutyric Acid Modulates Apoptosis and ER Stress‐Related Gene Expression in Glycogen Storage Disease Type Ib In Vitro Model |
| title_short | Phenylbutyric Acid Modulates Apoptosis and ER Stress‐Related Gene Expression in Glycogen Storage Disease Type Ib In Vitro Model |
| title_sort | phenylbutyric acid modulates apoptosis and er stress related gene expression in glycogen storage disease type ib in vitro model |
| topic | 4‐PBA apoptosis CRISPR/Cas9 ER stress GSD Ib in vitro model system |
| url | https://doi.org/10.1002/mgg3.70054 |
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