Search

Your search keyword '"Miriam Centrulo"' showing total 4 results
Start Over Author "Miriam Centrulo"
4 results on '"Miriam Centrulo"'

1. Therapeutic homology-independent targeted integration in retina and liver

Author

Patrizia Tornabene, Rita Ferla, Manel Llado-Santaeularia, Miriam Centrulo, Margherita Dell’Anno, Federica Esposito, Elena Marrocco, Emanuela Pone, Renato Minopoli, Carolina Iodice, Edoardo Nusco, Settimio Rossi, Hristiana Lyubenova, Anna Manfredi, Lucio Di Filippo, Antonella Iuliano, Annalaura Torella, Giulio Piluso, Francesco Musacchia, Enrico Maria Surace, Davide Cacchiarelli, Vincenzo Nigro, and Alberto Auricchio

Subjects
Science
Abstract

Limits of AAV-mediated gene therapy include targeting dominant mutations and inducing long-term transgene expression. Here, the authors show that AAV-HITI results in efficient allele-independent integration of a donor DNA in both retina and liver providing therapeutic benefit in mouse models of either a genetic form of blindness or a lysosomal storage disease, respectively.

Published
2022
Full Text
View/download PDF

2. Inclusion of a degron reduces levelsof undesired inteins after AAV-mediated proteintrans-splicing in the retina

Author

Patrizia Tornabene, Ivana Trapani, Miriam Centrulo, Elena Marrocco, Renato Minopoli, Mariangela Lupo, Carolina Iodice, Carlo Gesualdo, Francesca Simonelli, Enrico M. Surace, and Alberto Auricchio

Subjects
protein trans-splicing, split-inteins, intein degradation, ecDHFR, gene therapy, AAV, Genetics, QH426-470, Cytology, QH573-671
Abstract

Split intein-mediated protein trans-splicing expands AAV transfer capacity, thus overcoming the limited AAV cargo. However, non-mammalian inteins persist as trans-splicing by-products, and this could raise safety concerns for AAV intein clinical applications. In this study, we tested the ability of several degrons to selectively decrease levels of inteins after protein trans-splicing and found that a version of E. coli dihydrofolate reductase, which we have shortened to better fit into the AAV vector, is the most effective. We show that subretinal administration of AAV intein armed with this short degron is both safe and effective in a mouse model of Stargardt disease (STGD1), which is the most common form of inherited macular degeneration in humans. This supports the use of optimized AAV intein for gene therapy of both STGD1 and other conditions that require transfer of large genes.

Published
2021
Full Text
View/download PDF

3. Inclusion of a degron reduces levelsof undesired inteins after AAV-mediated proteintrans-splicing in the retina

Author

Mariangela Lupo, Ivana Trapani, Enrico Maria Surace, Alberto Auricchio, Carlo Gesualdo, Carolina Iodice, Miriam Centrulo, Renato Minopoli, Patrizia Tornabene, Francesca Simonelli, Elena Marrocco, Tornabene, P., Trapani, I., Centrulo, M., Marrocco, E., Minopoli, R., Lupo, M., Iodice, C., Gesualdo, C., Simonelli, F., Surace, E. M., and Auricchio, A.

Subjects
Genetic enhancement, viruses, split-inteins, QH426-470, 03 medical and health sciences, 0302 clinical medicine, Dihydrofolate reductase, intein degradation, medicine, Genetics, Vector (molecular biology), Molecular Biology, Gene, 030304 developmental biology, 0303 health sciences, biology, QH573-671, AAV, Stargardt disease (STGD1), medicine.disease, gene therapy, Cell biology, Stargardt disease, ecDHFR, inherited retinal disease, RNA splicing, 030221 ophthalmology & optometry, biology.protein, Molecular Medicine, Original Article, split-intein, protein trans-splicing, Degron, Intein, Cytology
Abstract

Split intein-mediated protein trans-splicing expands AAV transfer capacity, thus overcoming the limited AAV cargo. However, non-mammalian inteins persist as trans-splicing by-products, and this could raise safety concerns for AAV intein clinical applications. In this study, we tested the ability of several degrons to selectively decrease levels of inteins after protein trans-splicing and found that a version of E. coli dihydrofolate reductase, which we have shortened to better fit into the AAV vector, is the most effective. We show that subretinal administration of AAV intein armed with this short degron is both safe and effective in a mouse model of Stargardt disease (STGD1), which is the most common form of inherited macular degeneration in humans. This supports the use of optimized AAV intein for gene therapy of both STGD1 and other conditions that require transfer of large genes., Graphical abstract, Intein-mediated protein trans-splicing (PTS) expands AAV gene transfer capacity. However, non-mammalian inteins persist as trans-splicing by-products. Here, we show that ecDHFR selectively degrades inteins after PTS and that AAV intein vectors armed with this degron are both safe and effective in the retina of a mouse model of genetic blindness.

Published
2021

4. Intein-mediated protein trans-splicing expands adeno-associated virus transfer capacity in the retina

Author

Elena Polishchuk, Carel B. Hoyng, Elena Marrocco, Alberto Auricchio, Ivana Trapani, Silvia Albert, Carlo Gesualdo, Carolina Iodice, Laura Giaquinto, Miriam Centrulo, Francesca Simonelli, Renato Minopoli, Enrico Maria Surace, Mariangela Lupo, Settimio Rossi, Maria Antonietta De Matteis, Frans P.M. Cremers, Paola Tiberi, Patrizia Tornabene, Antonella Iuliano, Roman S. Polishchuk, Sonia de Simone, Fabio Dell'Aquila, Tornabene, Patrizia, Trapani, I., Minopoli, R., Centrulo, M., Lupo, M., De Simone, S., Tiberi, Mario, Dell'Aquila, F., Marrocco, E., Iodice, C., Iuliano, A., Gesualdo, C., Rossi, S., Giaquinto, L., Albert, S., Hoyng, C. B., Polishchuk, E., Cremers, F. P. M., Surace, E. M., Simonelli, F., De Matteis, M. A., Polishchuk, R., Auricchio, A., Trapani, Ivana, Minopoli, Renato, Centrulo, Miriam, Lupo, Mariangela, de Simone, Sonia, Tiberi, Paola, Dell'Aquila, Fabio, Marrocco, Elena, Iodice, Carolina, Iuliano, Antonella, Gesualdo, Carlo, Rossi, Settimio, Giaquinto, Laura, Albert, Silvia, Hoyng, Carel B., Polishchuk, Elena, Cremers, Frans P. M., Surace, Enrico M., Simonelli, Francesca, De Matteis, Maria A., Polishchuk, Roman, and Auricchio, Alberto

Subjects
0301 basic medicine, Swine, Genetic enhancement, Genetic Vectors, Green Fluorescent Proteins, Induced Pluripotent Stem Cells, Protein reconstitution, Trans-splicing, Biology, medicine.disease_cause, Retina, Article, Sensory disorders Donders Center for Medical Neuroscience [Radboudumc 12], Inteins, Trans-Splicing, Mice, 03 medical and health sciences, chemistry.chemical_compound, 0302 clinical medicine, All institutes and research themes of the Radboud University Medical Center, medicine, Animals, Humans, Adeno-associated virus, Gene, Gene Transfer Techniques, Retinal, General Medicine, Dependovirus, 3. Good health, Cell biology, Organoids, Phenotype, 030104 developmental biology, medicine.anatomical_structure, chemistry, Intein, 030217 neurology & neurosurgery, Photoreceptor Cells, Vertebrate
Abstract

Retinal gene therapy with adeno-associated viral (AAV) vectors holds promises for treating inherited and noninherited diseases of the eye. Although clinical data suggest that retinal gene therapy is safe and effective, delivery of large genes is hindered by the limited AAV cargo capacity. Protein trans-splicing mediated by split inteins is used by single-cell organisms to reconstitute proteins. Here, we show that delivery of multiple AAV vectors each encoding one of the fragments of target proteins flanked by short split inteins results in protein trans-splicing and full-length protein reconstitution in the retina of mice and pigs and in human retinal organoids. The reconstitution of large therapeutic proteins using this approach improved the phenotype of two mouse models of inherited retinal diseases. Our data support the use of split intein–mediated protein trans-splicing in combination with AAV subretinal delivery for gene therapy of inherited blindness due to mutations in large genes.

Published
2019

Catalog

Books, media, physical & digital resources
See catalog results