Engineered tRNAs efficiently suppress CDKL5 premature termination codons
Abstract The CDKL5 deficiency disorder (CDD) is a severe neurodevelopmental disorder characterized by early-onset epilepsy, intellectual disability, motor and visual dysfunctions. The causative gene is CDKL5, which codes for a kinase required for brain development. There is no cure for CDD patients;...
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Nature Portfolio
2024-12-01
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| Series: | Scientific Reports |
| Online Access: | https://doi.org/10.1038/s41598-024-82766-0 |
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| author | Stefano Pezzini Aurora Mustaccia Pierre Aboa Giorgia Faustini Alessio Branchini Mirko Pinotti Angelisa Frasca Joseph J. Porter John D. Lueck Nicoletta Landsberger |
| author_facet | Stefano Pezzini Aurora Mustaccia Pierre Aboa Giorgia Faustini Alessio Branchini Mirko Pinotti Angelisa Frasca Joseph J. Porter John D. Lueck Nicoletta Landsberger |
| author_sort | Stefano Pezzini |
| collection | DOAJ |
| description | Abstract The CDKL5 deficiency disorder (CDD) is a severe neurodevelopmental disorder characterized by early-onset epilepsy, intellectual disability, motor and visual dysfunctions. The causative gene is CDKL5, which codes for a kinase required for brain development. There is no cure for CDD patients; treatments are symptomatic and focus mainly on seizure control. Several pathogenic variants are loss-of-function, but recent studies suggest that the CDD phenotype is sensitive to the CDKL5 gene dosage. Therefore, mRNA-targeted correction strategies that respect the physiological regulation of CDKL5 could be a valid alternative to augmentative gene therapy. Nonsense mutations cause ~ 11% of CDD cases, and these patients might benefit from readthrough therapies. We proved that drug-mediated readthrough efficiently suppresses premature CDKL5 nonsense codons, but the recoded kinase remained highly hypomorphic, curtailing the translational value of this pharmacological approach. In this study we explored if the recently developed Anticodon-edited tRNAs (ACE-tRNAs) offer an alternative readthrough strategy for CDD. Transfecting cells expressing different CDKL5 nonsense variants, we demonstrated that ACE-tRNAs efficiently restore full-length kinase synthesis. The recoded CDKL5 is correctly localized and catalytically active, thereby bringing tRNA-based therapy back into the spotlight for future investigations to assess the efficacy of this approach in correcting the pathological phenotype of CDD. |
| format | Article |
| id | doaj-art-2d15c2d856304fa9a38438f55a5c29d5 |
| institution | Kabale University |
| issn | 2045-2322 |
| language | English |
| publishDate | 2024-12-01 |
| publisher | Nature Portfolio |
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| series | Scientific Reports |
| spelling | doaj-art-2d15c2d856304fa9a38438f55a5c29d52025-01-05T12:28:34ZengNature PortfolioScientific Reports2045-23222024-12-0114111110.1038/s41598-024-82766-0Engineered tRNAs efficiently suppress CDKL5 premature termination codonsStefano Pezzini0Aurora Mustaccia1Pierre Aboa2Giorgia Faustini3Alessio Branchini4Mirko Pinotti5Angelisa Frasca6Joseph J. Porter7John D. Lueck8Nicoletta Landsberger9Department of Medical Biotechnology and Translational Medicine, University of MilanSan Raffaele Rett Research Unit, Neuroscience Division, IRCCS San Raffaele Scientific InstituteDepartment of Medical Biotechnology and Translational Medicine, University of MilanDepartment of Medical Biotechnology and Translational Medicine, University of MilanDepartment of Life Sciences and Biotechnology, University of FerraraDepartment of Life Sciences and Biotechnology, University of FerraraDepartment of Medical Biotechnology and Translational Medicine, University of MilanDepartments of Pharmacology and Physiology and Neurology, School of Medicine and Dentistry, University of RochesterDepartments of Pharmacology and Physiology and Neurology, School of Medicine and Dentistry, University of RochesterDepartment of Medical Biotechnology and Translational Medicine, University of MilanAbstract The CDKL5 deficiency disorder (CDD) is a severe neurodevelopmental disorder characterized by early-onset epilepsy, intellectual disability, motor and visual dysfunctions. The causative gene is CDKL5, which codes for a kinase required for brain development. There is no cure for CDD patients; treatments are symptomatic and focus mainly on seizure control. Several pathogenic variants are loss-of-function, but recent studies suggest that the CDD phenotype is sensitive to the CDKL5 gene dosage. Therefore, mRNA-targeted correction strategies that respect the physiological regulation of CDKL5 could be a valid alternative to augmentative gene therapy. Nonsense mutations cause ~ 11% of CDD cases, and these patients might benefit from readthrough therapies. We proved that drug-mediated readthrough efficiently suppresses premature CDKL5 nonsense codons, but the recoded kinase remained highly hypomorphic, curtailing the translational value of this pharmacological approach. In this study we explored if the recently developed Anticodon-edited tRNAs (ACE-tRNAs) offer an alternative readthrough strategy for CDD. Transfecting cells expressing different CDKL5 nonsense variants, we demonstrated that ACE-tRNAs efficiently restore full-length kinase synthesis. The recoded CDKL5 is correctly localized and catalytically active, thereby bringing tRNA-based therapy back into the spotlight for future investigations to assess the efficacy of this approach in correcting the pathological phenotype of CDD.https://doi.org/10.1038/s41598-024-82766-0 |
| spellingShingle | Stefano Pezzini Aurora Mustaccia Pierre Aboa Giorgia Faustini Alessio Branchini Mirko Pinotti Angelisa Frasca Joseph J. Porter John D. Lueck Nicoletta Landsberger Engineered tRNAs efficiently suppress CDKL5 premature termination codons Scientific Reports |
| title | Engineered tRNAs efficiently suppress CDKL5 premature termination codons |
| title_full | Engineered tRNAs efficiently suppress CDKL5 premature termination codons |
| title_fullStr | Engineered tRNAs efficiently suppress CDKL5 premature termination codons |
| title_full_unstemmed | Engineered tRNAs efficiently suppress CDKL5 premature termination codons |
| title_short | Engineered tRNAs efficiently suppress CDKL5 premature termination codons |
| title_sort | engineered trnas efficiently suppress cdkl5 premature termination codons |
| url | https://doi.org/10.1038/s41598-024-82766-0 |
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