المؤلفون: Carlo Gesualdo, Nicola de Prisco, Settimio Rossi, Salvatore Botta, Martina Sofia, Fabiola Curion, Enrico Maria Surace, Elena Marrocco, Cathal Wilson, Maria Laura Bacci, Domenico Ventrella, Mario Renda, Francesca Simonelli

المساهمون: Botta, Salvatore, Nicola de Prisco, Marrocco, Elena, Renda, Mario, Sofia, Martina, Curion, Fabiola, Maria Laura Bacci, Ventrella, Domenico, Wilson, Cathal, Gesualdo, Carlo, Rossi, Settimio, Simonelli, Francesca, Enrico Maria Surace, de Prisco, Nicola, Bacci, Maria Laura, Surace, Enrico Maria

مصطلحات موضوعية: 0301 basic medicine, Photoreceptors, Rhodopsin, Swine, Genetic Vectors, Kruppel-Like Transcription Factors, Mice, Transgenic, Biology, Ectopic Gene Expression, 03 medical and health sciences, Transcription (biology), Retinal Rod Photoreceptor Cells, Gene expression, Gene silencing, Animals, Gene Silencing, Neurodegeneration, Transcription factor, Gene, Nuclear Proteins, General Medicine, Genetic Therapy, Dependovirus, Phenotype, Cell biology, Ophthalmology, 030104 developmental biology, Gene Targeting, Mutation, biology.protein, Ectopic expression, Female, Therapeutic, Transcription, Retinitis Pigmentosa, Research Article

الوصف: The genome-wide activity of transcription factors (TFs) on multiple regulatory elements precludes their use as gene-specific regulators. Here we show that ectopic expression of a TF in a cell-specific context can be used to silence the expression of a specific gene as a therapeutic approach to regulate gene expression in human disease. We selected the TF Krüppel-like factor 15 (KLF15) based on its putative ability to recognize a specific DNA sequence motif present in the rhodopsin (RHO) promoter and its lack of expression in terminally differentiated rod photoreceptors (the RHO-expressing cells). Adeno-associated virus (AAV) vector-mediated ectopic expression of KLF15 in rod photoreceptors of pigs enables Rho silencing with limited genome-wide transcriptional perturbations. Suppression of a RHO mutant allele by KLF15 corrects the phenotype of a mouse model of retinitis pigmentosa with no observed toxicity. Cell-specific-context conditioning of TF activity may prove a novel mode for somatic gene-targeted manipulation.

وصف الملف: ELETTRONICO

الوصول الحر: https://explore.openaire.eu/search/publication?articleId=doi_dedup___::3dbebb9546ffd12a9a35ba4f0b2a400bTest
http://hdl.handle.net/11585/614148Test

المؤلفون: J. Fraser Wright, Alberto Auricchio, Katherine A. High, Paolo Melillo, Albert M. Maguire, Francesca Simonelli, Eric A. Pierce, Jennifer Wellman, Kathleen A. Marshall, Sandro Banfi, Francesco Testa, Settimio Rossi, Jean Bennett, Carmela Acerra, Junwei Sun, Enrico Maria Surace

المساهمون: Testa, F, Maguire, Am, Rossi, S, Pierce, Ea, Melillo, P, Marshall, K, Banfi, S, Surace, Enrico Maria, Sun, J, Acerra, C, Wright, Jf, Wellman, J, High, Ka, Auricchio, Alberto, Bennett, J, Simonelli, F., Testa, Francesco, Rossi, Settimio, Melillo, Paolo, Banfi, Sandro, Surace, Em, Wright, Fj, Auricchio, A, Simonelli, Francesca

المصدر: Ophthalmology. 120:1283-1291

مصطلحات موضوعية: Adult, Male, cis-trans-Isomerases, medicine.medical_specialty, Visual acuity, Adolescent, genetic structures, Genetic Vectors, Leber Congenital Amaurosis, Visual Acuity, Nystagmus, Gene mutation, Reflex, Pupillary, Transfection, Article, Young Adult, Ophthalmology, medicine, Humans, Pupillary light reflex, Child, business.industry, Genetic Therapy, Dependovirus, eye diseases, Visual field, Clinical trial, Cis-trans-Isomerases, Mutation, Cohort, Female, Injections, Intraocular, Visual Fields, medicine.symptom, business, Tomography, Optical Coherence, Follow-Up Studies

الوصف: OBJECTIVE: The aim of this study was to show the clinical data of long-term (3-year) follow-up of 5 patients affected by Leber congenital amaurosis type 2 (LCA2) treated with a single unilateral injection of adeno-associated virus AAV2-hRPE65v2. DESIGN: Clinical trial. PARTICIPANTS: Five LCA2 patients with RPE65 gene mutations. METHODS: After informed consent and confirmation of trial eligibility criteria, the eye with worse visual function was selected for subretinal delivery of adeno-associated virus (AAV2-hRPE65v2). Subjects were evaluated before and after surgery at designated follow-up visits (1, 2, 3, 14, 30, 60, 90, 180, 270, and 365 days, 1.5 years, and 3 years) by complete ophthalmic examination. Efficacy for each subject was monitored with best-corrected visual acuity, kinetic visual field, nystagmus testing, and pupillary light reflex. MAIN OUTCOME MEASURES: Best-corrected visual acuity, kinetic visual field, nystagmus testing, and pupillary light reflex. RESULTS: The data showed a statistically significant improvement of best-corrected visual acuity between baseline and 3 years after treatment in the treated eye (P

الوصول الحر: https://explore.openaire.eu/search/publication?articleId=doi_dedup___::05a60be709fc62c35116da6cbfb1fb0dTest
https://doi.org/10.1016/j.ophtha.2012.11.048Test

المؤلفون: Francesca Simonelli, Roman S. Polishchuk, Massimo Giunti, Elena V. Polishchuk, Maria Laura Bacci, Elisabetta Toriello, Andrea Sommella, Carolina Iodice, Sonia de Simone, Settimio Rossi, Alberto Auricchio, Linda Colecchi, Ivana Trapani, Pasqualina Colella

المساهمون: Trapani, Ivana, Toriello, Elisabetta, de Simone, Sonia, Colella, Pasqualina, Iodice, Carolina, Polishchuk, Elena V, Sommella, Andrea, Colecchi, Linda, Rossi, Settimio, Simonelli, Francesca, Giunti, Massimo, Bacci, Maria L, Polishchuk, Roman S, Auricchio, Alberto, Bacci, MARIA LAURA, De Simone, Sonia, Polishchuk, Elena V., Bacci, Maria L., Polishchuk, Roman S.

المصدر: Human Molecular Genetics

مصطلحات موضوعية: Swine, Genetic enhancement, viruses, Administration, Ophthalmic, Homology (biology), Macular Degeneration, Mice, 0302 clinical medicine, HEK293 Cell, Protein biosynthesis, Transgenes, Genetics (clinical), 0303 health sciences, Terminal Repeat Sequence, General Medicine, Articles, Dependovirus, Dependoviru, Stargardt disease, Female, Genetic Vector, AAV Vector, Human, ATP-Binding Cassette Transporter, Transgene, Genetic Vectors, Heterologous, Biology, Retina, Mouse model, 03 medical and health sciences, Gene therapy, Genetic, Genetics, medicine, Animals, Humans, Gene, Molecular Biology, 030304 developmental biology, Animal, HEK 293 cells, Terminal Repeat Sequences, Genetic Therapy, medicine.disease, Molecular biology, Disease Models, Animal, HEK293 Cells, ATP-Binding Cassette Transporters, 030217 neurology & neurosurgery

الوصف: Stargardt disease (STGD1) due to mutations in the large ABCA4 gene is the most common inherited macular degeneration in humans. We have shown that dual adeno-associated viral (AAV) vectors effectively transfer ABCA4 to the retina of Abca4-/- mice. However, they express both lower levels of transgene compared with a single AAV and truncated proteins. To increase productive dual AAV concatemerization, which would overcome these limitations, we have explored the use of either various regions of homology or heterologous inverted terminal repeats (ITR). In addition, we tested the ability of various degradation signals to decrease the expression of truncated proteins. We found the highest levels of transgene expression using regions of homology based on either alkaline phosphatase or the F1 phage (AK). The use of heterologous ITR does not decrease the levels of truncated proteins relative to full-length ABCA4 and impairs AAV vector production. Conversely, the inclusion of the CL1 degradation signal results in the selective degradation of truncated proteins from the 5'-half without affecting full-length protein production. Therefore, we developed dual AAV hybrid ABCA4 vectors including homologous ITR2, the photoreceptor-specific G protein-coupled receptor kinase 1 promoter, the AK region of homology and the CL1 degradation signal. We show that upon subretinal administration these vectors are both safe in pigs and effective in Abca4-/- mice. Our data support the use of improved dual AAV vectors for gene therapy of STGD1.

وصف الملف: STAMPA

الوصول الحر: https://explore.openaire.eu/search/publication?articleId=doi_dedup___::508edf1e50db352683d8fb83b8b86487Test
http://hdl.handle.net/11588/623722Test

المؤلفون: Jean Bennett, Jessica I. W. Morgan, Carmela Acerra, Kathleen A. Marshall, Katherine A. High, Daniel C. Chung, Xiaosong Zhu, Alberto Auricchio, Enrico Maria Surace, Bernd Hauck, Frauke Coppieters, Gui-Shuang Ying, Albert M. Maguire, Kenneth S. Shindler, Sandro Banfi, Jenni Fer Uvellman Mcdonnell, Nicholas J. Volpe, Arkady Lyubarsky, Ann Fulton, Leslie Raffini, Eric A. Pierce, Elfride De Baere, Olga Zelenaia, Jeannette L. Bennicelli, J. Fraser Wright, Francesco Testa, T. Michael Redmond, Junwei Sun, Edwin M. Stone, Federico Mingozzi, Francesca Simonelli, Bart P. Leroy, Settimio Rossi

المساهمون: Maguire, Am, High, Ka, Auricchio, A, Wright, Jf, Pierce, Ea, Testa, Francesco, Mingozzi, F, Bennicelli, Jl, Ying, G, Rossi, Settimio, Fulton, A, Marshall, Ka, Banfi, Sandro, Chung, Dc, Morgan, Ji, Hauck, B, Zelenaia, O, Zhu, X, Raffini, L, Coppieters, F, De Baere, E, Shindler, K, Volpe, Nj, Surace, Em, Acerra, C, Lyubarsky, A, Redmond, Tm, Stone, E, Sun, J, Mcdonnell, Jw, Leroy, Bp, Simonelli, Francesca, Bennett, J., Auricchio, Alberto, Testa, F, Rossi, S, Banfi, S, Surace, Enrico Maria, Simonelli, F

المصدر: Lancet (London, England). 374(9701)

مصطلحات موضوعية: Retinal degeneration, Male, Visual acuity, genetic structures, Visual Acuity, Blindness, chemistry.chemical_compound, Mice, Medicine, Child, media_common, Age Factors, General Medicine, Dependovirus, gene therapy, Leber’s congenital amaurosi, Treatment Outcome, Disease Progression, Leber's congenital amaurosis, Female, medicine.symptom, Safety, Retinopathy, Adult, cis-trans-Isomerases, medicine.medical_specialty, Adolescent, Genetic Vectors, Dark Adaptation, Optic Atrophy, Hereditary, Leber, Article, Injections, Young Adult, Nystagmus, Physiologic, Ophthalmology, Electroretinography, media_common.cataloged_instance, Animals, Humans, European union, Eye Proteins, Dose-Response Relationship, Drug, business.industry, Infant, Newborn, Retinal, Genetic Therapy, medicine.disease, eye diseases, Disease Models, Animal, RPE65 gene, RPE65, chemistry, Cis-trans-Isomerases, Mutation, business, Carrier Proteins

الوصف: Background Gene therapy has the potential to reverse disease or prevent further deterioration of vision in patients with incurable inherited retinal degeneration. We therefore did a phase 1 trial to assess the effect of gene therapy on retinal and visual function in children and adults with Leber's congenital amaurosis.Methods We assessed the retinal and visual function in 12 patients (aged 8-44 years) with RPE65-associated Leber's congenital amaurosis given one subretinal injection of adeno-associated virus (AAV) containing a gene encoding a protein needed for the isomerohydrolase activity of the retinal pigment epithelium (AAV2-hRPE65v2) in the worst eye at low (1.5x10(10) vector genomes), medium (4.8x10(10) vector genomes), or high dose (1.5x10(11) vector genomes) for up to 2 years.Findings AAV2-hRPE65v2 was well tolerated and all patients showed sustained improvement in subjective and objective measurements of vision (ie, dark adaptometry, pupillometry, electroretinography, nystagmus, and ambulatory behaviour). Patients had at least a 2 log unit increase in pupillary light responses, and an 8-year-old child had nearly the same level of light sensitivity as that in age-matched normal-sighted individuals. The greatest improvement was noted in children, all of whom gained ambulatory vision. The study is registered with ClinicalTrials.gov, number NCT00516477.Interpretation The safety, extent, and stability of improvement in vision in all patients support the use of AAV mediated gene therapy for treatment of inherited retinal diseases, with early intervention resulting in the best potential gain. Background: Gene therapy has the potential to reverse disease or prevent further deterioration of vision in patients with incurable inherited retinal degeneration. We therefore did a phase 1 trial to assess the effect of gene therapy on retinal and visual function in children and adults with Leber's congenital amaurosis. Methods: We assessed the retinal and visual function in 12 patients (aged 8-44 years) with RPE65-associated Leber's congenital amaurosis given one subretinal injection of adeno-associated virus (AAV) containing a gene encoding a protein needed for the isomerohydrolase activity of the retinal pigment epithelium (AAV2-hRPE65v2) in the worst eye at low (1·5×1010 vector genomes), medium (4·8×1010 vector genomes), or high dose (1·5×1011 vector genomes) for up to 2 years. Findings: AAV2-hRPE65v2 was well tolerated and all patients showed sustained improvement in subjective and objective measurements of vision (ie, dark adaptometry, pupillometry, electroretinography, nystagmus, and ambulatory behaviour). Patients had at least a 2 log unit increase in pupillary light responses, and an 8-year-old child had nearly the same level of light sensitivity as that in age-matched normal-sighted individuals. The greatest improvement was noted in children, all of whom gained ambulatory vision. The study is registered with ClinicalTrials.gov, number NCT00516477. Interpretation: The safety, extent, and stability of improvement in vision in all patients support the use of AAV-mediated gene therapy for treatment of inherited retinal diseases, with early intervention resulting in the best potential gain. Funding: Center for Cellular and Molecular Therapeutics at the Children's Hospital of Philadelphia, Foundation Fighting Blindness, Telethon, Research to Prevent Blindness, F M Kirby Foundation, Mackall Foundation Trust, Regione Campania Convenzione, European Union, Associazione Italiana Amaurosi Congenita di Leber, Fund for Scientific Research, Fund for Research in Ophthalmology, and National Center for Research Resources. © 2009 Elsevier Ltd. All rights reserved.

الوصول الحر: https://explore.openaire.eu/search/publication?articleId=doi_dedup___::83e3ef7f273825996ee1f6bcdf200da5Test
https://pubmed.ncbi.nlm.nih.gov/19854500Test