29 results on '"Covello G"'
Search Results
2. Characterization of a bidirectional promoter shared between two human genes related to aging: SIRT3 and PSMD13
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Bellizzi, D., Dato, S., Cavalcante, P., Covello, G., Di Cianni, F., Passarino, G., Rose, G., and De Benedictis, G.
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HEREDITY , *PROTEOLYTIC enzymes , *GENETIC transformation , *CANCER cells - Abstract
Abstract: The human SIRT3 gene contains an intronic VNTR enhancer whose variability is correlated with life span. The SIRT3 5′ flanking region encompasses the PSMD13 gene encoding the p40.5 regulator subunit of the 26S proteasome. Proteasome is a multicatalytic proteinase whose function declines with aging. SIRT3 and PSMD13 are linked in a head-to-head configuration (788-bp intergenic region). The molecular configuration of two genes that are both related to aging prompted us to search for shared regulatory mechanisms between them. Transfection experiments carried out in HeLa cells by deletion mutants of the PSMD13–SIRT3 intergenic region showed a complex pathway of coregulation acting in both directions. Furthermore, linkage disequilibrium (LD) analyses carried out in a sample of 710 subjects (18–108 years of age) screened for A21631G (marker of PSMD13), and for G477T and VNTRintron5 (markers of SIRT3), revealed high LD, with significantly different PSMD13–SIRT3 haplotype pools between samples of centenarians and younger people. [Copyright &y& Elsevier]
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- 2007
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3. Evaluation of Anatomical and Tomographic Biomarkers as Predictive Visual Acuity Factors in Eyes with Retinal Vein Occlusion Treated with Dexamethasone Implant.
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Covello G, Maglionico MN, Figus M, Busoni C, Sartini MS, Lupidi M, and Posarelli C
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Background: This prospective study evaluated the impact of anatomical and tomographic biomarkers on clinical outcomes of intravitreal dexamethasone implants in patients with macular edema secondary to retinal vein occlusion (RVO). Methods : The study included 46 patients (28 with branch RVO (BRVO) and 18 with central RVO (CRVO)). Best corrected visual acuity (BCVA) significantly improved from a mean baseline of 0.817 ± 0.220 logMAR to 0.663 ± 0.267 logMAR at six months and 0.639 ± 0.321 logMAR at twelve months ( p < 0.05). Central retinal thickness (CRT) showed a significant reduction from 666.2 ± 212.2 µm to 471.1 ± 215.6 µm at six months and 467 ± 175.7 µm at twelve months ( p < 0.05). No significant differences were found in OCT biomarkers between baseline and follow-ups. Results : The study analysed improvements in visual acuity relative to baseline biomarkers. At six months, ellipsoid zone disruption (EZD) was significant for all subgroups. Disorganization of retinal inner layers (DRIL), external limiting membrane (ELM) disruption, macular ischemia (MI), CRT, and BRVO showed significance for any improvement, while DRIL and ELM were significant for changes greater than 0.3 logMAR ( p < 0.05). At twelve months, EZD remained significant for all subgroups. ELM, MI, CRT, and BRVO were significant for any improvement, while MI and BRVO were significant for changes greater than 0.3 logMAR ( p < 0.05). Hyperreflective foci were not statistically significant at either time point ( p > 0.05). Conclusions : The regression model suggested that MI and CRVO could be negative predictive factors for visual outcomes, while ELM and EZD were associated with BCVA improvement one-year post-treatment.
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- 2024
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4. Is the Advent of New Surgical Procedures Changing the Baseline Features of Patients Undergoing First-Time Glaucoma Surgery?
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Palma A, Covello G, Posarelli C, Maglionico MN, Agnifili L, and Figus M
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Background : The aim of this study was to determine if the rise in new surgical procedures for glaucoma is changing the baseline features of patients. Methods : In this retrospective study, we reviewed the baseline features of patients undergoing their first glaucoma surgery in 2011 and 2021, collecting data regarding intraocular pressure (IOP), visual field (VF) parameters, stage of disease, and the type of surgery. Results : In the study, 455 patients were included in the analysis. From these, 230 eyes had glaucoma surgery performed in 2011 (Group A) and 225 eyes in 2021 (Group B). When considering the baseline features, Group A was older than Group B (72.7 ± 10.7 and 70 ± 12.4 years; p = 0.02, respectively), and showed a significantly more advanced VF mean defect (-16.4 ± 8.8 and -13.8 ± 8.7 dB; p < 0.01, respectively) and a higher IOP (25.9 ± 6.6 and 24.9 ± 7.8 mmHg; p = 0.02, respectively). Overall, severe VF damage at the time of surgery was more frequent in Group A (74.3%) than in Group B (60.8%) ( p < 0.01). The overall number of traditional glaucoma surgeries was 211 in 2011, reducing to 94 ten years later, with similar severe pre-operative VF defects. In 2021, minimally invasive bleb surgery (MIBS) represented 58% of all surgeries. Conclusions : In the last ten years, patients receiving glaucoma surgery for the first time were younger, had less severe disease, and a more contained IOP. The baseline feature modifications were probably related to the diffusion of new procedures, especially MIBS, which allowed for treating patients at an earlier stage, reserving traditional procedures for advanced cases.
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- 2024
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5. Effectiveness and safety of XEN45 implant over 12 months of follow-up: data from the XEN-Glaucoma Treatment Registry.
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Oddone F, Roberti G, Giammaria S, Posarelli C, Ghirelli G, Mastropasqua L, Agnifili L, Micelli Ferrari T, Pace V, Nucci P, Sacchi M, Monsellato G, Altafini R, Scuderi G, Perdicchi A, Uva M, Carnevale C, Covello G, Maglionico MN, Fea A, and Figus M
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- Humans, Prospective Studies, Stents, Intraocular Pressure, Treatment Outcome, Retrospective Studies, Glaucoma, Open-Angle drug therapy, Glaucoma, Open-Angle surgery, Glaucoma, Open-Angle complications, Glaucoma Drainage Implants, Glaucoma drug therapy, Glaucoma surgery, Glaucoma complications
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Objectives: To evaluate the 1-year effectiveness and safety of the XEN45, either alone or in combination with phacoemulsification, in glaucoma patients., Methods: This multicentre, prospective, observational study included consecutive eyes of glaucoma patients from the Italian XEN-Glaucoma Treatment Registry (XEN-GTR) who underwent XEN45 alone or in combination with phacoemulsification, with at least 1 year of follow-up. Surgical success was defined as intraocular pressure (IOP) < 18 mmHg and ≥20% reduction from preoperative IOP, over 1 year of follow-up., Results: Two hundred thirty-nine eyes (239 patients) were analyzed, 144 (60.2%) eyes in the XEN-solo and 95 (39.8%) eyes in the XEN+Phaco groups. One hundred-sixty-eight (70.3%) eyes achieved overall success, without statistically significant differences between study groups (p = 0.07). Preoperative IOP dropped from a median (IQR) of 23.0 (20.0-26.0) mmHg to 14.0 (12.0-16.0) mmHg at month 12 (p < 0.001), with overall 39.9 ± 18.3% IOP reduction. The mean number of preoperative ocular hypotensive medications (OHM) was significantly reduced from 2.7 ± 0.9 to 0.5 ± 0.9 at month 12 (p < 0.001). Preoperative IOP < 15 mmHg (HR: 6.63; 95%CI: 2.61-16.84, p < 0.001) and temporal position of the surgeon (HR: 4.25; 95%CI: 2.62-6.88, p < 0.001) were significantly associated with surgery failure. One hundred-forty-six (61.1%) eyes had no intraoperative complications, whereas 91 (38.1%) and 56 (23.4%) eyes experienced at least one complication, respectively early (< month 1) and late (≥ month 1), all self-limiting or successfully treated without sequelae. Needling occurred in 55 (23.0%) eyes at least once during follow-up., Conclusion: Over 1-year follow-up, XEN45 alone or in combination with phacoemulsification, had comparable success rates and effectively and safely lowered IOP and the need for OHM., (© 2023. The Author(s).)
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- 2024
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6. A machine learning approach to predict the glaucoma filtration surgery outcome.
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Agnifili L, Figus M, Porreca A, Brescia L, Sacchi M, Covello G, Posarelli C, Di Nicola M, Mastropasqua R, Nucci P, and Mastropasqua L
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- Humans, Meibomian Glands, Conjunctiva surgery, Fluorescein, Glaucoma surgery, Filtering Surgery
- Abstract
This study aimed at predicting the filtration surgery (FS) outcome using a machine learning (ML) approach. 102 glaucomatous patients undergoing FS were enrolled and underwent ocular surface clinical tests (OSCTs), determination of surgical site-related biometric parameters (SSPs) and conjunctival vascularization. Break-up-time, Schirmer test I, corneal fluorescein staining, Meibomian gland expressibility; conjunctival hyperemia, upper bulbar conjunctiva area of exposure, limbus to superior eyelid distance; and conjunctival epithelial and stromal (CET, CST) thickness and reflectivity (ECR, SCR) at AS-OCT were considered. Successful FS required a 30% baseline intraocular pressure reduction, with values ≤ 18 mmHg with or without medications. The classification tree (CT) was the ML algorithm used to analyze data. At the twelfth month, FS was successful in 60.8% of cases, whereas failed in 39.2%. At the variable importance ranking, CST and SCR were the predictors with the greater relative importance to the CART tree construction, followed by age. CET and ECR showed less relative importance, whereas OSCTs and SSPs were not important features. Within the CT, CST turned out the most important variable for discriminating success from failure, followed by SCR and age, with cut-off values of 75 µm, 169 on gray scale, and 62 years, respectively. The ROC curve for the classifier showed an AUC of 0.784 (0.692-0.860). In this ML approach, CT analysis found that conjunctival stroma thickness and reflectivity, along with age, can predict the FS outcome with good accuracy. A pre-operative thick and hyper-reflective stroma, and a younger age increase the risk of FS failure., (© 2023. Springer Nature Limited.)
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- 2023
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7. HCS-Splice: A High-Content Screening Method to Advance the Discovery of RNA Splicing-Modulating Therapeutics.
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Covello G, Siva K, Adami V, and Denti MA
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- Humans, Mutation genetics, RNA Splicing genetics, Alternative Splicing genetics
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Nucleic acid therapeutics have demonstrated an impressive acceleration in recent years. They work through multiple mechanisms of action, including the downregulation of gene expression and the modulation of RNA splicing. While several drugs based on the former mechanism have been approved, few target the latter, despite the promise of RNA splicing modulation. To improve our ability to discover novel RNA splicing-modulating therapies, we developed HCS-Splice, a robust cell-based High-Content Screening (HCS) assay. By implementing the use of a two-colour (GFP/RFP) fluorescent splicing reporter plasmid, we developed a versatile, effective, rapid, and robust high-throughput strategy for the identification of potent splicing-modulating molecules. The HCS-Splice strategy can also be used to functionally confirm splicing mutations in human genetic disorders or to screen drug candidates. As a proof-of-concept, we introduced a dementia-related splice-switching mutation in the Microtubule-Associated Protein Tau ( MAPT ) exon 10 splicing reporter. We applied HCS-Splice to the wild-type and mutant reporters and measured the functional change in exon 10 inclusion. To demonstrate the applicability of the method in cell-based drug discovery, HCS-Splice was used to evaluate the efficacy of an exon 10-targeting siRNA, which was able to restore the correct alternative splicing balance.
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- 2023
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8. Ultrasound Cyclo Plasty in Patients with Open Angle Glaucoma and High Myopia.
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Figus M, Palma A, Covello G, Agnifili L, and Posarelli C
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- Humans, Intraocular Pressure, Retrospective Studies, Tonometry, Ocular, Ciliary Body surgery, Treatment Outcome, Glaucoma, Open-Angle complications, Glaucoma, Open-Angle diagnosis, Glaucoma, Open-Angle surgery, Glaucoma surgery, Myopia complications, Myopia diagnosis, Myopia surgery
- Abstract
Prcis: Ultrasound cyclo plasty (UCP) using high-intensity focused ultrasound is an effective and safe procedure in lowering intraocular pressure (IOP) in patients with glaucoma, even in those with high myopia., Purpose: This study aimed to evaluate the efficacy and safety of UCP in glaucoma patients with high myopia., Materials and Methods: In this retrospective, single-center study, we enrolled 36 eyes divided into 2 groups based on axial length: group A (≥26.00 mm) and group B (<26.00 mm). We collected data about visual acuity, Goldmann applanation tonometry, biomicroscopy, and visual field before the procedure and at 1, 7, 30, 60, 90, 180, and 365 days after the procedure., Results: Mean IOP significantly decreased in both groups after treatment ( P <0.001). Mean IOP reduction from baseline to the last visit was 9.8±6.6 mmHg (38.7%) in group A and 9.6±6.3 mmHg (34.8%) in group B ( P <0.001). Mean IOP at the last visit was 15.8±4.1 mmHg in the myopic group and 18.1±5.6 mmHg in the non-myopic one. Regarding the number of IOP-lowering eyedrops being taken by our patients, statistically significant differences were found between groups A and B neither at baseline (2.8±0.9 and 2.6±1.0; P =0.568) nor 1 year after the procedure (2.5±1.1 and 2.6±1.1; P =0.762). No major complications occurred. All minor adverse events resolved within a few days., Conclusion: UCP seems to be an effective and well-tolerated strategy to lower IOP in glaucoma patients with high myopia., Competing Interests: Disclosure: The authors declare no conflict of interest., (Copyright © 2023 Wolters Kluwer Health, Inc. All rights reserved.)
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- 2023
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9. High-Throughput Microscopy Analysis of Mitochondrial Membrane Potential in 2D and 3D Models.
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Vianello C, Dal Bello F, Shin SH, Schiavon S, Bean C, Magalhães Rebelo AP, Knedlík T, Esfahani EN, Costiniti V, Lacruz RS, Covello G, Munari F, Scolaro T, Viola A, Rampazzo E, Persano L, Zumerle S, Scorrano L, Gianelle A, and Giacomello M
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- Humans, Membrane Potential, Mitochondrial, Mitochondria metabolism, Fibroblasts metabolism, Microscopy, Proteomics
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Recent proteomic, metabolomic, and transcriptomic studies have highlighted a connection between changes in mitochondria physiology and cellular pathophysiological mechanisms. Secondary assays to assess the function of these organelles appear fundamental to validate these -omics findings. Although mitochondrial membrane potential is widely recognized as an indicator of mitochondrial activity, high-content imaging-based approaches coupled to multiparametric to measure it have not been established yet. In this paper, we describe a methodology for the unbiased high-throughput quantification of mitochondrial membrane potential in vitro, which is suitable for 2D to 3D models. We successfully used our method to analyze mitochondrial membrane potential in monolayers of human fibroblasts, neural stem cells, spheroids, and isolated muscle fibers. Moreover, by combining automated image analysis and machine learning, we were able to discriminate melanoma cells from macrophages in co-culture and to analyze the subpopulations separately. Our data demonstrated that our method is a widely applicable strategy for large-scale profiling of mitochondrial activity.
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- 2023
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10. Validation of an Automated Artificial Intelligence Algorithm for the Quantification of Major OCT Parameters in Diabetic Macular Edema.
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Midena E, Toto L, Frizziero L, Covello G, Torresin T, Midena G, Danieli L, Pilotto E, Figus M, Mariotti C, and Lupidi M
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Artificial intelligence (AI) and deep learning (DL)-based systems have gained wide interest in macular disorders, including diabetic macular edema (DME). This paper aims to validate an AI algorithm for identifying and quantifying different major optical coherence tomography (OCT) biomarkers in DME eyes by comparing the algorithm to human expert manual examination. Intraretinal (IRF) and subretinal fluid (SRF) detection and volumes, external limiting-membrane (ELM) and ellipsoid zone (EZ) integrity, and hyperreflective retina foci (HRF) quantification were analyzed. Three-hundred three DME eyes were included. The mean central subfield thickness was 386.5 ± 130.2 µm. IRF was present in all eyes and confirmed by AI software. The agreement (kappa value) (95% confidence interval) for SRF presence and ELM and EZ interruption were 0.831 (0.738-0.924), 0.934 (0.886-0.982), and 0.936 (0.894-0.977), respectively. The accuracy of the automatic quantification of IRF, SRF, ELM, and EZ ranged between 94.7% and 95.7%, while accuracy of quality parameters ranged between 99.0% (OCT layer segmentation) and 100.0% (fovea centering). The Intraclass Correlation Coefficient between clinical and automated HRF count was excellent (0.97). This AI algorithm provides a reliable and reproducible assessment of the most relevant OCT biomarkers in DME. It may allow clinicians to routinely identify and quantify these parameters, offering an objective way of diagnosing and following DME eyes.
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- 2023
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11. Italian Candidates for the XEN Implant: An Overview from the Glaucoma Treatment Registry (XEN-GTR).
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Posarelli C, Figus M, Roberti G, Giammaria S, Ghirelli G, Quercioli P, Micelli Ferrari T, Pace V, Mastropasqua L, Agnifili L, Sacchi M, Scuderi G, Perdicchi A, Altafini R, Uva M, D'Andrea D, Covello G, Maglionico MN, Fea AM, Carnevale C, and Oddone F
- Abstract
Background The Italian XEN Glaucoma Treatment Registry (XEN-GTR) was created to acquire a comprehensive prospective dataset that includes the patient characteristics, intraoperative variables, and postoperative management of glaucoma patients undergoing the XEN gel stent implantation. Methods This was a prospective observational, longitudinal clinical study involving 10 centres throughout Italy. The baseline examination included a comprehensive evaluation of demographic parameters (age, sex, ethnicity, and systemic condition), specific ophthalmological parameters, and quality of life questionnaire score collection. Results The baseline data of 273 patients were analysed. The median (IQR) age was 72 (65.0 to 78.0) years. Of the 273 patients, 123 (45%) were female and 150 (55%) were male. A total of 86% of the patients had open-angle glaucoma with a mean intraocular pressure of 24 ± 6 (range 12.0-60.0) mmHg. The mean number of medications was 2.7 ± 0.9 at baseline for the patients with a prevalence of prostaglandin analogues combined with a beta-blocker and anhydrase carbonic inhibitor (31.8%). The mean scores of the NEI-VFQ 25 and GSS questionnaires were 78 ± 18 (range 26.5-100) and 85 ± 14 (range 79-93), respectively. Combined XEN/cataract surgeries were scheduled in 73.7% of the patients. The preferred place for the XEN implant was the supero-nasal quadrant (91.6%). Conclusions Observing the baseline characteristics of the typical Italian candidates for the XEN gel implant shows that they are patients affected by POAG and cataracts, with moderate to severe glaucoma damage, all of which has an impact on their quality of life.
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- 2022
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12. Choroidal Vascularity Index in Central and Branch Retinal Vein Occlusion.
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Loiudice P, Covello G, Figus M, Posarelli C, Sartini MS, and Casini G
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(1) Background: we aimed to evaluate choroidal vascularity change in eyes with central and branch retinal vein occlusion (RVO). (2) Methods: in this retrospective cross-sectional study, we reviewed the records of 47 patients with recent-onset, naïve, unilateral retinal vein occlusion. Enhanced-depth imaging optical coherence tomography scans were binarized using the ImageJ software; luminal area (LA) and total choroidal area (TCA) were measured. The choroidal vascularity index (CVI) was calculated as the proportion of LA to TCA. Depending on the pattern of macular oedema, eyes were classified as having no macular oedema (nME), cystoid macular oedema (CME), cystoid macular oedema with serous retinal detachment (mixed). (3) Results: CVI, TCA and LA were greater in eyes with RVO than in fellow, unaffected eyes. No difference was found between central and branch RVO except for central macular thickness (CMT). When compared with controls, eyes with CME presented a significant increase in subfoveal choroidal thickness, CMT, TCA, LA and CVI; eyes with mixed macular oedema had greater CMT and CVI than contralateral eyes; no significant differences in any of the considered parameters were observed in eyes with nME. (4) Conclusions: The results suggest that RVO alters the vascularity of the choroid that varies according to the type of macular oedema.
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- 2022
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13. Exon Skipping Through Chimeric Antisense U1 snRNAs to Correct Retinitis Pigmentosa GTPase-Regulator ( RPGR ) Splice Defect.
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Covello G, Ibrahim GH, Bacchi N, Casarosa S, and Denti MA
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- Exons genetics, GTP Phosphohydrolases genetics, Humans, Mutation, RNA, Small Nuclear genetics, Eye Proteins genetics, Eye Proteins metabolism, Retinitis Pigmentosa genetics, Retinitis Pigmentosa therapy
- Abstract
Inherited retinal dystrophies are caused by mutations in more than 250 genes, each of them carrying several types of mutations that can lead to different clinical phenotypes. Mutations in Retinitis Pigmentosa GTPase-Regulator ( RPGR ) cause X-linked Retinitis pigmentosa (RP). A nucleotide substitution in intron 9 of RPGR causes the increase of an alternatively spliced isoform of the mature mRNA, bearing exon 9a (E9a). This introduces a stop codon, leading to truncation of the protein. Aiming at restoring impaired gene expression, we developed an antisense RNA-based therapeutic approach for the skipping of RPGR E9a. We designed a set of specific U1 antisense snRNAs (U1_asRNAs) and tested their efficacy in vitro , upon transient cotransfection with RPGR minigene reporter systems in HEK-293T, 661W, and PC-12 cell lines. We thus identified three chimeric U1_asRNAs that efficiently mediate E9a skipping, correcting the genetic defect. Unexpectedly, the U1-5'antisense construct, which exhibited the highest exon-skipping efficiency in PC-12 cells, induced E9a inclusion in HEK-293T and 661W cells, indicating caution in the choice of preclinical model systems when testing RNA splicing-correcting therapies. Our data provide a proof of principle for the application of U1_snRNA exon skipping-based approach to correct splicing defects in RPGR .
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- 2022
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14. Choroidal vascularity index in thyroid-associated ophthalmopathy: a cross-sectional study.
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Loiudice P, Pellegrini M, Marinò M, Mazzi B, Ionni I, Covello G, Figus M, Nardi M, and Casini G
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Background: Hemodynamic changes have been observed in patients with Graves' disease. The aim of our study was to evaluate choroidal vascular change using the choroidal vascularity index (CVI) in patients with thyroid-associated ophthalmopathy (TAO)., Methods: In this cross-sectional observational study, 40 patients affected by TAO were recruited. Forty healthy individuals, matched for age and sex, served as controls. Foveal enhanced-depth imaging optical coherence tomography scans were obtained from all participants. Images were binarized using the ImageJ software and luminal area (LA) and total choroidal area (TCA) were measured. CVI was calculated as the proportion of LA to TCA. The relation between CVI or subfoveal choroidal thickness (SFCT) and clinical activity score, exophthalmometric value, diplopia status, gender, and age was evaluated., Results: CVI was significantly higher in patients with TAO (P = 0.004). No significant difference was observed in SFCT (P = 0.200) and TCA (P = 0.153) comparing TAO patients and healthy controls. LA was significantly higher in TAO group (P = 0.045). On multiple regression analysis, CVI was associated with TCA (P = 0.043). No association was found between SFCT or CVI and TCA, clinical activity score, exophthalmometric value, Inami value, diplopia status, gender or age (P > 0.05)., Conclusions: This is the first study that has demonstrated an increase in CVI in eyes with TAO compared with healthy controls and has assessed its association with clinical features.
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- 2021
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15. Delivery of oligonucleotide-based therapeutics: challenges and opportunities.
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Hammond SM, Aartsma-Rus A, Alves S, Borgos SE, Buijsen RAM, Collin RWJ, Covello G, Denti MA, Desviat LR, Echevarría L, Foged C, Gaina G, Garanto A, Goyenvalle AT, Guzowska M, Holodnuka I, Jones DR, Krause S, Lehto T, Montolio M, Van Roon-Mom W, and Arechavala-Gomeza V
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- Gene Expression, Oligonucleotides, Antisense, RNA, Small Interfering, Nanoparticles, Oligonucleotides
- Abstract
Nucleic acid-based therapeutics that regulate gene expression have been developed towards clinical use at a steady pace for several decades, but in recent years the field has been accelerating. To date, there are 11 marketed products based on antisense oligonucleotides, aptamers and small interfering RNAs, and many others are in the pipeline for both academia and industry. A major technology trigger for this development has been progress in oligonucleotide chemistry to improve the drug properties and reduce cost of goods, but the main hurdle for the application to a wider range of disorders is delivery to target tissues. The adoption of delivery technologies, such as conjugates or nanoparticles, has been a game changer for many therapeutic indications, but many others are still awaiting their eureka moment. Here, we cover the variety of methods developed to deliver nucleic acid-based therapeutics across biological barriers and the model systems used to test them. We discuss important safety considerations and regulatory requirements for synthetic oligonucleotide chemistries and the hurdles for translating laboratory breakthroughs to the clinic. Recent advances in the delivery of nucleic acid-based therapeutics and in the development of model systems, as well as safety considerations and regulatory requirements for synthetic oligonucleotide chemistries are discussed in this review on oligonucleotide-based therapeutics., (© 2021 The Authors. Published under the terms of the CC BY 4.0 license.)
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- 2021
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16. Combined Phacoemulsification and Ex-PRESS Implant with Everting Suture in Primary Angle-Closure Glaucoma: Survival Analysis and Predictive Factors.
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Covello G, Loiudice P, Maglionico MN, Nardi M, Figus M, and Posarelli C
- Abstract
The purpose of this study was to evaluate the efficacy and safety of combined phacoemulsification and Ex-PRESS implant with everting suture in primary angle-closure glaucoma (PACG) and to examine predictive factors of failure. Twenty-three eyes of 18 patients were enrolled. Data about time of removal of releasable sutures and traction of the everting suture and about changes in intraocular pressure (IOP) were collected, as well as comprehensive ophthalmic examinations. Success was defined by the following criteria: IOP ≤ 18 mmHg (criterion 1); IOP ≤ 15 mmHg (criterion 2); and IOP ≤ 12 mmHg (criterion 3). Success was categorized as complete or qualified, depending on whether it was reached without or with drugs, respectively. Success rate was assessed with Kaplan-Meier survival analysis with a Cox proportional hazard model to adjust for potential confounders. The lowering of IOP and the reduction of medications were statistically significant at every follow-up visit compared with the baseline ( p < 0.05). The complete success rates were 87%, 70%, and 17% accordingly to criterion 1, 2, and 3; the qualified success rates were 93%, 70%, and 20%, respectively. Most of the complications resolved spontaneously and conservatively. In conclusion, combined phacoemulsification and Ex-PRESS Minishunt implant with everting suture is a safe and effective surgery, even in PACG, lowering IOP and number of medications.
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- 2021
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17. Ophthalmological emergencies and the SARS-CoV-2 outbreak.
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Posarelli C, Maglionico MN, Covello G, Loiudice P, Cipriano A, Santini M, and Figus M
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- Adolescent, Adult, Aged, Betacoronavirus, COVID-19, Child, Child, Preschool, Female, Humans, Infant, Infant, Newborn, International Classification of Diseases, Italy epidemiology, Male, Middle Aged, Pandemics, Retrospective Studies, SARS-CoV-2, Young Adult, Coronavirus Infections epidemiology, Emergency Service, Hospital statistics & numerical data, Ophthalmology statistics & numerical data, Pneumonia, Viral epidemiology, Tertiary Care Centers statistics & numerical data
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Since the end of 2019, an outbreak of the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), originating in the Chinese city of Wuhan has spread rapidly worldwide causing thousands of deaths. Coronavirus disease (COVID-19) is supported by SARS-CoV-2 and represents the causative agent of a potentially fatal disease that is of great global public health concern. Italy has been the first European country recording an elevated number of infected forcing the Italian Government to call for total lockdown. The lockdown had the aim to limit the spread of infection through social distancing. The purpose of this study is to analyze how the pandemic has affected the patient's accesses to the Ophthalmological Emergency Department of a tertiary referral center in central-northern Italy, during the lockdown period. The charts of all patients that came to the Emergency Department during the lockdown period (March 10 -May 4, 2020) have been retrospectively collected and compared with those in the same period of 2019 and the period from 15 January- 9 March 2020. A significant reduction of visits during the lockdown has been observed, compared with those of pre-lockdown period (reduction of 65.4%) and with those of the same period of 2019 (reduction of 74.3%). Particularly, during the lockdown, minor and not urgency visits decreased whereas the undeferrable urgency ones increased. These pieces of evidence could be explained by the fear of patients to be infected; but also revealed patients misuse of emergency services., Competing Interests: The authors have declared that no competing interests exist.
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- 2020
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18. Behçet's syndrome and ocular involvement: changes over time.
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Posarelli C, Maglionico MN, Talarico R, Covello G, and Figus M
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- Humans, Behcet Syndrome diagnosis, Behcet Syndrome drug therapy
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Ocular involvement in Behçet's syndrome still represents a challenge for both rheumatologists and ophthalmologists; over the past 20 years the availability of new diagnostic tools and the concomitant introduction of biologic drugs led to a significant improvement in the management of these patients. The lack of uniform definitions and the diversity of the outcome measures still represent an obstacle for the prompt and correct management of ocular manifestations. The aim of the present review is to summarise the current evidences related to correct diagnosis and proper management of patients with Behçet's syndrome and ocular involvement.
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- 2020
19. Retinal, choroidal and optic disc analysis in patients with Graves' disease with or without orbitopathy.
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Casini G, Marinò M, Rubino M, Licari S, Covello G, Mazzi B, Ionni I, Rocchi R, Sframeli AT, Figus M, and Loiudice P
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- Cross-Sectional Studies, Humans, Retina, Tomography, Optical Coherence, Graves Ophthalmopathy complications, Graves Ophthalmopathy diagnosis, Optic Disk
- Abstract
Purpose: To compare optic disc, retinal and choroidal measurements in patients with Graves' disease with or without orbitopathy, and healthy controls., Methods: Optical coherence tomography and Heidelberg retinal tomography were performed in 40 patients with Graves' orbitopathy (GO), 40 subjects with Graves's disease (GD) with no sign of orbitopathy and 40 healthy controls. Degree of exophthalmos, ocular alignment, clinical activity score (CAS), choroidal thickness, retinal thickness, ganglion cell layer (GCL) thickness, disc area, cup area, rim area, cup/disc area ratio, linear cup/disc ratio and mean peripapillary retinal nerve fibre layer thickness were analysed., Results: GO patients and healthy controls significantly differ regarding mean central retinal thickness (275 ± 19 µm and 285 ± 20 µm, P = 0.017); mean central GCL thickness (14.87 ± 3.0 µm and 17.92 ± 5.02 µm, P = 0.001); mean disc area (2.00 ± 0.44 mm
2 and 1.72 ± 0.37 mm2 , P = 0.003); mean cup area (0.53 ± 0.52 mm2 and 0.31 ± 0.20 mm2 , P = 0.003); cup/disc area ratio (0.22 ± 0.10 and 0.17 ± 0.08, P = 0.010); and linear cup/disc ratio (0.47 ± 0.15 and 0.40 ± 0.13, respectively, P = 0.011). No difference was found between patients without orbitopathy and healthy controls. No significant difference was found regarding the choroidal thickness between the three groups. There was no statistically significant relationship between retinal thickness, ganglion cell layer thickness, mean disc area, mean cup area, cup/disc area ratio, linear cup/disc ratio, CAS, exophthalmometric value and ocular alignment., Conclusion: GO patients showed significant changes in foveal and GCL thickness, and optic nerve head morphology suggesting a possible influence of the orbital inflammatory process.- Published
- 2020
- Full Text
- View/download PDF
20. Transcriptome analysis of the zebrafish atoh7-/- Mutant, lakritz , highlights Atoh7-dependent genetic networks with potential implications for human eye diseases.
- Author
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Covello G, Rossello FJ, Filosi M, Gajardo F, Duchemin AL, Tremonti BF, Eichenlaub M, Polo JM, Powell D, Ngai J, Allende ML, Domenici E, Ramialison M, and Poggi L
- Abstract
Expression of the bHLH transcription protein Atoh7 is a crucial factor conferring competence to retinal progenitor cells for the development of retinal ganglion cells. Several studies have emerged establishing ATOH7 as a retinal disease gene. Remarkably, such studies uncovered ATOH7 variants associated with global eye defects including optic nerve hypoplasia, microphthalmia, retinal vascular disorders, and glaucoma. The complex genetic networks and cellular decisions arising downstream of atoh7 expression, and how their dysregulation cause development of such disease traits remains unknown. To begin to understand such Atoh7-dependent events in vivo, we performed transcriptome analysis of wild-type and atoh7 mutant ( lakritz ) zebrafish embryos at the onset of retinal ganglion cell differentiation. We investigated in silico interplays of atoh7 and other disease-related genes and pathways. By network reconstruction analysis of differentially expressed genes, we identified gene clusters enriched in retinal development, cell cycle, chromatin remodeling, stress response, and Wnt pathways. By weighted gene coexpression network, we identified coexpression modules affected by the mutation and enriched in retina development genes tightly connected to atoh7 . We established the groundwork whereby Atoh7-linked cellular and molecular processes can be investigated in the dynamic multi-tissue environment of the developing normal and diseased vertebrate eye., (© 2020 The Authors.)
- Published
- 2020
- Full Text
- View/download PDF
21. New device for ciliary sulcus suture fixation of intraocular lens.
- Author
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Nardi M, Covello G, Figus M, and Posarelli C
- Subjects
- Sutures, Lenses, Intraocular
- Published
- 2019
- Full Text
- View/download PDF
22. High-intensity focused ultrasound procedure: The rise of a new noninvasive glaucoma procedure and its possible future applications.
- Author
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Posarelli C, Covello G, Bendinelli A, Fogagnolo P, Nardi M, and Figus M
- Subjects
- Aqueous Humor metabolism, Glaucoma metabolism, Glaucoma physiopathology, High-Intensity Focused Ultrasound Ablation trends, Humans, Intraocular Pressure physiology, Glaucoma surgery, High-Intensity Focused Ultrasound Ablation methods
- Abstract
Glaucoma, one of the leading causes of blindness, is usually first managed medically, with incisional surgery as a second step. Noninvasive glaucoma procedures attempt to fill the gap between medical and surgical treatments and may work synergistically with them. High-intensity focused ultrasound induces a selective and controlled thermal ablation of the distal part of the ciliary body, and this effect is independent from the degree of tissue pigmentation with limited damage to adjacent structures. This selective and innovative treatment decreases intraocular pressure by reducing aqueous humor production and by increasing uveoscleral outflow. We review the current literature on the use of high-intensity focused ultrasound in glaucoma, exploring present use, safety, efficacy, and future clinical applications., (Copyright © 2019 Elsevier Inc. All rights reserved.)
- Published
- 2019
- Full Text
- View/download PDF
23. The short-term impact of probiotic consumption on the oral cavity microbiome.
- Author
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Dassi E, Ferretti P, Covello G, Bertorelli R, Denti MA, De Sanctis V, Tett A, and Segata N
- Subjects
- Biodiversity, Female, Healthy Volunteers, Humans, Longitudinal Studies, Male, Probiotics administration & dosage, RNA, Ribosomal, 16S genetics, Time Factors, Young Adult, Microbiota drug effects, Mouth microbiology, Probiotics pharmacology, Saliva microbiology
- Abstract
The dysbiosis of the oral microbiome is associated with both localized and systemic diseases. Modulating the resident microbial communities by the dietary consumption of probiotics has become an appealing means to promote host health by either restoring host-microbe balance or preventing dysbiosis. Most probiotics strategies target the intestinal microbiome, but little is known about their impact on the oral microbiome. We analyzed here the saliva microbiome from 21 volunteers, longitudinally collected before, during, and after consumption of a commercial probiotic and a standard yoghurt using 16S amplicon sequencing. The alpha diversity of the saliva microbiome had a statistically significant increase (P-value = 0.0011) in one of the groups that consumed the probiotic. The overall structure of the microbiome was however not significantly impacted by the probiotic, although oligotyping analysis revealed that both Streptococci and Lactobacilli present in the probiotic product persisted in the saliva microbiome. In contrast, non-probiotic yoghurt consumption had a lesser impact on the overall diversity and Lactobacillus and Streptococcus persistence. Our results suggest that consumption of commercial probiotics in healthy subjects increase the overall diversity of the oral cavity microbiome in the short term, but such dietary interventions are not able to substantially modify the structure of the microbiome.
- Published
- 2018
- Full Text
- View/download PDF
24. Double-target Antisense U1snRNAs Correct Mis-splicing Due to c.639+861C>T and c.639+919G>A GLA Deep Intronic Mutations.
- Author
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Ferri L, Covello G, Caciotti A, Guerrini R, Denti MA, and Morrone A
- Abstract
Fabry disease is a rare X-linked lysosomal storage disorder caused by deficiency of the α-galactosidase A (α-Gal A) enzyme, which is encoded by the GLA gene. GLA transcription in humans produces a major mRNA encoding α-Gal A and a minor mRNA of unknown function, which retains a 57-nucleotide-long cryptic exon between exons 4 and 5, bearing a premature termination codon. NM_000169.2:c.639+861C>T and NM_000169.2:c.639+919G>A GLA deep intronic mutations have been described to cause Fabry disease by inducing overexpression of the alternatively spliced mRNA, along with a dramatic decrease in the major one. Here, we built a wild-type GLA minigene and two minigenes that carry mutations c.639+861C>T and c.639+919G>A. Once transfected into cells, the minigenes recapitulate the molecular patterns observed in patients, at the mRNA, protein, and enzymatic level. We constructed a set of specific double-target U1asRNAs to correct c.639+861C>T and c.639+919G>A GLA mutations. Efficacy of U1asRNAs in inducing the skipping of the cryptic exon was evaluated upon their transient co-transfection with the minigenes in COS-1 cells, by real-time polymerase chain reaction (PCR), western blot analysis, and α-Gal A enzyme assay. We identified a set of U1asRNAs that efficiently restored α-Gal A enzyme activity and the correct splicing pathways in reporter minigenes. We also identified a unique U1asRNA correcting both mutations as efficently as the mutation-specific U1asRNAs. Our study proves that an exon skipping-based approach recovering α-Gal A activity in the c.639+861C>T and c.639+919G>A GLA mutations is active.
- Published
- 2016
- Full Text
- View/download PDF
25. Enhanced microbial diversity in the saliva microbiome induced by short-term probiotic intake revealed by 16S rRNA sequencing on the IonTorrent PGM platform.
- Author
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Dassi E, Ballarini A, Covello G, Quattrone A, Jousson O, De Sanctis V, Bertorelli R, Denti MA, and Segata N
- Subjects
- Adult, Bacteria classification, Bacteria genetics, Female, Humans, Male, Sequence Analysis, DNA methods, DNA, Bacterial chemistry, High-Throughput Nucleotide Sequencing, Microbiota genetics, Probiotics administration & dosage, RNA, Ribosomal, 16S genetics, Saliva microbiology
- Abstract
Microbial communities populating several human body habitats are important determinants of human health. Cultivation-free community-wide approaches like bacterial 16S rRNA sequencing recently revolutionized the study of such human-associated microbial diversity, and the continuously decreasing cost/throughput ratio of current sequencing platforms is further enhancing the availability and effectiveness of microbiome research. The IonTorrent PGM platform is among the latest available commercial high-throughput sequencing tools, but it is just starting to be used for 16S rRNA surveys with only episodic assessments of its performance. We present here the first IonTorrent profiling of the human saliva microbiome collected from 12 healthy individuals. In this cohort, a subset of the volunteers was asked to assume a probiotic product, in order to investigate its impact on the composition and the structure of the saliva microbiome. Analysis of the generated dataset suggests the suitability of the IonTorrent platform for 16S rRNA surveys, even though some platform-specific choices are required to optimize the consistency of the obtained bacterial profiles. Interestingly, we found a marked and statistically significant increase of the overall bacterial diversity in the saliva of individuals who received the probiotic product compared to the control group, suggesting a short-term effect of probiotic product administration on oral microbiome composition., (Copyright © 2014 Elsevier B.V. All rights reserved.)
- Published
- 2014
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- View/download PDF
26. An electroporation protocol for efficient DNA transfection in PC12 cells.
- Author
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Covello G, Siva K, Adami V, and Denti MA
- Abstract
A wide variety of mammalian cell types is used in gene transfection studies. Establishing transfection methods that enable highly efficient DNA uptake has become increasingly important. PC12 is an established rat pheochromocytoma cell line, which responds to exposure to NGF with cessation of growth, expression of cytoplasmic processes, and differentiation into cells resembling sympathetic neurons. Although PC12 cells represent an important model system to study a variety of neuronal functions, they proved relatively difficult to transfect. We have compared the efficiency of three different chemical transfection reagents (Lipofectamine 2000, Lipofectamine LTX and TransIT-LT1) and of two electroporation systems (Neon and Gene Pulser Xcell) in transiently transfecting undifferentiated PC12 cells. By comparing efficiencies from replicate experiments we proved electroporation (in particular Neon) to be the method of choice. By optimizing different parameters (voltage, pulse width and number of pulses) we reached high efficiency of transfection (90 %) and viability (99 %). We also demonstrated that, upon electroporation, cells are not altered by the transfection and maintain their ability to differentiate.
- Published
- 2014
- Full Text
- View/download PDF
27. Exon-skipping antisense oligonucleotides to correct missplicing in neurogenetic diseases.
- Author
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Siva K, Covello G, and Denti MA
- Subjects
- Animals, Ataxia Telangiectasia genetics, Ataxia Telangiectasia therapy, Blood-Brain Barrier, Congenital Disorders of Glycosylation genetics, Congenital Disorders of Glycosylation therapy, Cysts genetics, Cysts therapy, Drug Delivery Systems, Exons, Frontotemporal Dementia genetics, Frontotemporal Dementia therapy, Hereditary Central Nervous System Demyelinating Diseases genetics, Hereditary Central Nervous System Demyelinating Diseases therapy, Humans, Muscular Dystrophy, Duchenne genetics, Muscular Dystrophy, Duchenne therapy, Mutation, Neurofibromatoses genetics, Neurofibromatoses therapy, Niemann-Pick Disease, Type C genetics, Niemann-Pick Disease, Type C therapy, Oligonucleotides, Antisense chemistry, Pelizaeus-Merzbacher Disease genetics, Pelizaeus-Merzbacher Disease therapy, Phosphotransferases (Phosphomutases) deficiency, Phosphotransferases (Phosphomutases) genetics, RNA Splicing, Genetic Diseases, Inborn genetics, Genetic Diseases, Inborn therapy, Nervous System Diseases genetics, Nervous System Diseases therapy, Oligonucleotides, Antisense genetics, Oligonucleotides, Antisense therapeutic use, Targeted Gene Repair methods
- Abstract
Alternative splicing is an important regulator of the transcriptome. However, mutations may cause alteration of splicing patterns, which in turn leads to disease. During the past 10 years, exon skipping has been looked upon as a powerful tool for correction of missplicing in disease and progress has been made towards clinical trials. In this review, we discuss the use of antisense oligonucleotides to correct splicing defects through exon skipping, with a special focus on diseases affecting the nervous system, and the latest stage achieved in its progress.
- Published
- 2014
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- View/download PDF
28. Identification of GATA2 and AP-1 Activator elements within the enhancer VNTR occurring in intron 5 of the human SIRT3 gene.
- Author
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Bellizzi D, Covello G, Di Cianni F, Tong Q, and De Benedictis G
- Subjects
- Binding Sites genetics, Blotting, Western, GATA2 Transcription Factor genetics, HeLa Cells, Humans, Introns genetics, Minisatellite Repeats genetics, Point Mutation, Protein Binding, Proto-Oncogene Proteins c-fos genetics, Proto-Oncogene Proteins c-jun genetics, Transcription Factor AP-1 genetics, Transcription Factor AP-1 metabolism, Transfection, Enhancer Elements, Genetic genetics, GATA2 Transcription Factor metabolism, Proto-Oncogene Proteins c-fos metabolism, Proto-Oncogene Proteins c-jun metabolism, Sirtuin 3 genetics
- Abstract
Human SIRT3 gene contains an intronic VNTR enhancer. A T > C transition occurring in the second repeat of each VNTR allele implies the presence/absence of a putative GATA binding motif. A partially overlapping AP-1 site, not affected by the transition, was also identified. Aims of the present study were: 1) to verify if GATA and AP-1 sites could bind GATA2 and c-Jun/c-Fos factors, respectively; 2) to investigate whether such sites modulate the enhancer activity of the SIRT3-VNTR alleles. DAPA assay proved that GATA2 and c-Jun/c-Fos factors are able to bind the corresponding sites. Moreover, co-transfection experiments showed that the over-expression of GATA2 and c-Jun/c-Fos factors boosts the VNTR enhancer activity in an allelic-specific way. Furthermore, we established that GATA2 and c-Jun/c-Fos act additively in modulating the SIRT3-VNTR enhancer function. Therefore, GATA2 and AP-1 are functional sites and the T S> C transition of the second VNTR repeat affects their activity.
- Published
- 2009
- Full Text
- View/download PDF
29. A novel VNTR enhancer within the SIRT3 gene, a human homologue of SIR2, is associated with survival at oldest ages.
- Author
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Bellizzi D, Rose G, Cavalcante P, Covello G, Dato S, De Rango F, Greco V, Maggiolini M, Feraco E, Mari V, Franceschi C, Passarino G, and De Benedictis G
- Subjects
- Adult, Aged, Aged, 80 and over, Base Sequence, Female, Humans, Male, Middle Aged, Molecular Sequence Data, Sirtuin 3, Sirtuins, Enhancer Elements, Genetic genetics, Histone Deacetylases genetics, Longevity genetics, Minisatellite Repeats, Mitochondrial Proteins genetics, Polymorphism, Genetic
- Abstract
SIR2 genes control life span in model organisms, playing a central role in evolutionarily conserved pathways of aging and longevity. We wanted to verify whether similar effects may act in humans too. First, we searched for variability in the human sirtuin 3 gene (SIRT3) and discovered a VNTR polymorphism (72-bp repeat core) in intron 5. The alleles differed both for the number of repeats and for presence/absence of potential regulatory sites. Second, by transient transfection experiments, we demonstrated that the VNTR region has an allele-specific enhancer activity. Third, by analyzing allele frequencies as a function of age in a sample of 945 individuals (20-106 years), we found that the allele completely lacking enhancer activity is virtually absent in males older than 90 years. Thus the underexpression of a human sirtuin gene seems to be detrimental for longevity as it occurs in model organisms.
- Published
- 2005
- Full Text
- View/download PDF
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