Estudio de la expresión de micrornas y su modulacion como potencial diana terapéutica en la retinosis pigmentaria

217 p.La retinosis pigmentaria (RP) es la principal causa genética de degeneración de la visión en todo el mundo con más de 1.5 millones de afectados, para la que aún no existe un tratamiento estandarizado y eficaz. Esta enfermedad se caracteriza por su gran heterogeneidad clínica y genética, con más de 70 genes relacionados en su desarrollo.Partimos de la hipótesis de que en la RP se ve alterado un conjunto de mecanismos moleculares comunes a distintas formas de la enfermedad, con independencia del gen mutado. Esta idea se apoya en distintos trabajos en los que se ha descrito un patrón común de expresión diferencial tanto de genes, como de microRNAs en distintos modelos animales de RP, causados por mutaciones en genes diferentes.El desarrollo de esta tesis se ha planteado alrededor de dos objetivos principales; por una parte el estudio de la expresión de microRNAs para la identificación de microRNAs diferencialmente expresados que sirvan como potencial diana terapéutica en la RP, y por otra parte la modulación in vivo de un grupo de microRNAs y el análisis del efecto de dicha modulación en un modelo animal de la enfermedad, el ratón rd10

Estudio de la expresión de micrornas y su modulacion como potencial diana terapéutica en la retinosis pigmentaria

217 p.La retinosis pigmentaria (RP) es la principal causa genética de degeneración de la visión en todo el mundo con más de 1.5 millones de afectados, para la que aún no existe un tratamiento estandarizado y eficaz. Esta enfermedad se caracteriza por su gran heterogeneidad clínica y genética, con más de 70 genes relacionados en su desarrollo.Partimos de la hipótesis de que en la RP se ve alterado un conjunto de mecanismos moleculares comunes a distintas formas de la enfermedad, con independencia del gen mutado. Esta idea se apoya en distintos trabajos en los que se ha descrito un patrón común de expresión diferencial tanto de genes, como de microRNAs en distintos modelos animales de RP, causados por mutaciones en genes diferentes.El desarrollo de esta tesis se ha planteado alrededor de dos objetivos principales; por una parte el estudio de la expresión de microRNAs para la identificación de microRNAs diferencialmente expresados que sirvan como potencial diana terapéutica en la RP, y por otra parte la modulación in vivo de un grupo de microRNAs y el análisis del efecto de dicha modulación en un modelo animal de la enfermedad, el ratón rd10

A new approach based on targeted pooled DNA sequencing identifies novel mutations in patients with Inherited Retinal Dystrophies

Inherited retinal diseases (IRD) are a heterogeneous group of diseases that mainly affect the retina; more than 250 genes have been linked to the disease and more than 20 different clinical phenotypes have been described. This heterogeneity both at the clinical and genetic levels complicates the identification of causative mutations. Therefore, a detailed genetic characterization is important for genetic counselling and decisions regarding treatment. In this study, we developed a method consisting on pooled targeted next generation sequencing (NGS) that we applied to 316 eye disease related genes, followed by High Resolution Melting and copy number variation analysis. DNA from 115 unrelated test samples was pooled and samples with known mutations were used as positive controls to assess the sensitivity of our approach. Causal mutations for IRDs were found in 36 patients achieving a detection rate of 31.3%. Overall, 49 likely causative mutations were identified in characterized patients, 14 of which were first described in this study (28.6%). Our study shows that this new approach is a cost-effective tool for detection of causative mutations in patients with inherited retinopathies.This work was supported by grants from the National Institute of Health Carlos III (Institute of Health Carlos III/ISCIII) (CP10/00572, PI13/02621 and RD16/0008/0027 to JRE, PI17/01413 to CI, and a Research Intensification Contract to ALdM); the Basque Government's Industry Department (SAIOTEK: SAIO11-PE11BN002; and SAIO12-PC12BN001 to JRE), a grant from the Mutua Madrilena Foundation and support from the Retinitis Pigmentosa Patients of Gipuzkoa Foundation (BEGISARE). JR-E is a Miguel Servet II Fellow, National Institute of Health Carlos III (ISCIII). MEI was supported by grants from the Basque Government's Department of Education (DEDUC14/309). OB is supported by funding from the Retinitis Pigmentosa Patients of Gipuzkoa Foundation (BEGISARE) and a grant from the Mutua Madrilena Foundation. AA was supported by grants from the Fundacion Jesus de Gangoiti Barrera and from the Basque Government's Departments of Industry and Education (SAIOTEK-11BN002/PC12BN001/DEPLC13/002). CI is partially supported by a Research Intensification Contract (INTBIO15/001). The authors are grateful to Xabier Elcoroaristizabal and Marta Fernandez-Mercado for their helpful advice on developing the base-calling setup. Maribel Gomez; Naiara Telletxea and Nahikari Pastoriza at the Basque Biobank for isolating DNA samples; and Dr. Carmen Ayuso for kindly providing control samples. We also give special thanks to all patients with IRD and their families involved in the study

Expression Profiling Analysis Reveals Key MicroRNA– mRNA Interactions in Early Retinal Degeneration in Retinitis Pigmentosa

PURPOSE. The aim of this study was to identify differentially expressed microRNAs (miRNAs) that might play an important role in the etiology of retinal degeneration in a genetic mouse model of retinitis pigmentosa (rd10 mice) at initial stages of the disease. Methods. miRNAs-mRNA interaction networks were generated for analysis of biological pathways involved in retinal degeneration. RESULTS. Of more than 1900 miRNAs analyzed, we selected 19 miRNAs on the basis of (1) a significant differential expression in rd10 retinas compared with control samples and (2) an inverse expression relationship with predicted mRNA targets involved in biological pathways relevant to retinal biology and/or degeneration. Seven of the selected miRNAs have been associated with retinal dystrophies, whereas, to our knowledge, nine have not been previously linked to any disease. CONCLUSIONS. This study contributes to our understanding of the etiology and progression of retinal degeneration.Supported by the Fundacion Jesus de Gangoiti Barrera and from the Basque Government's Department of Industry and Education Grants SAIOTEK-PE11BN002, PC12BN001, and DEPLC13/002 (AA, JRE); funds from Foundation of Patients of Retinitis Pigmentosa of Gipuzkoa (Retinosis Gipuzkoa Begisare) (OB); a grant from the Fundacion Mutua Madrilena (OB); Basque Government's Department of Education grants DEDUC14/309 (MEI), Diputacion Foral de Gipuzkoa DFG15/006 (MM-C), and ELKARTEK 16/014 (MM-C); National Institute of Health Carlos III (Instituto de Salud Carlos III) Grants ISCIII: CP10/00572 (JRE), PI13/02621 (JRE); an Intensificacion Contract (ALdM) from the Basque Government's Department of Industry; and a grant from the Foundation of Patients of Retinitis Pigmentosa of Gipuzkoa (Retinosis Gipuzkoa Begisare) (JRE). JR-E is a Miguel Servet II Fellow, National Institute of Health Carlos III (Instituto de Salud Carlos III), ISCIII: CPII16/00012

Inhibition of MicroRNA 6937 Delays Photoreceptor and Vision Loss in a Mouse Model of Retinitis Pigmentosa

Inherited retinal dystrophies (IRDs) are a group of rare retinal conditions, including retinitis pigmentosa (RP), caused by monogenic mutations in 1 out of more than 250 genes. Despite recent advancements in gene therapy, there is still a lack of an effective treatment for this group of retinal conditions. MicroRNAs (miRNAs) are a class of highly conserved small non-coding RNAs that inhibit gene expression. Control of miRNAs-mediated protein expression has been described as a widely used mechanism for post-transcriptional regulation in many physiological and pathological processes in different organs, including the retina. Our main purpose was to test the hypothesis that modulation of a group of miRNAs can protect photoreceptor cells from death in the rd10 mouse model of retinitis pigmentosa. For this, we incorporated modulators of three miRNAs in adeno-associated viruses (AAVs), which were administered through sub-retinal injections. The results obtained indicate that inhibition of the miR-6937-5p slows down the visual deterioration of rd10 mice, reflected by an increased electroretinogram (ERG) wave response under scotopic conditions and significant preservation of the outer nuclear layer thickness. This work contributes to broadening our knowledge on the molecular mechanisms underlying retinitis pigmentosa and supports the development of novel therapeutic approaches for RP based on miRNA modulation

Estudio de la expresión de micrornas y su modulacion como potencial diana terapéutica en la retinosis pigmentaria

217 p.La retinosis pigmentaria (RP) es la principal causa genética de degeneración de la visión en todo el mundo con más de 1.5 millones de afectados, para la que aún no existe un tratamiento estandarizado y eficaz. Esta enfermedad se caracteriza por su gran heterogeneidad clínica y genética, con más de 70 genes relacionados en su desarrollo.Partimos de la hipótesis de que en la RP se ve alterado un conjunto de mecanismos moleculares comunes a distintas formas de la enfermedad, con independencia del gen mutado. Esta idea se apoya en distintos trabajos en los que se ha descrito un patrón común de expresión diferencial tanto de genes, como de microRNAs en distintos modelos animales de RP, causados por mutaciones en genes diferentes.El desarrollo de esta tesis se ha planteado alrededor de dos objetivos principales; por una parte el estudio de la expresión de microRNAs para la identificación de microRNAs diferencialmente expresados que sirvan como potencial diana terapéutica en la RP, y por otra parte la modulación in vivo de un grupo de microRNAs y el análisis del efecto de dicha modulación en un modelo animal de la enfermedad, el ratón rd10

Inhibition of MicroRNA 6937 Delays Photoreceptor and Vision Loss in a Mouse Model of Retinitis Pigmentosa

Inherited retinal dystrophies (IRDs) are a group of rare retinal conditions, including retinitis pigmentosa (RP), caused by monogenic mutations in 1 out of more than 250 genes. Despite recent advancements in gene therapy, there is still a lack of an effective treatment for this group of retinal conditions. MicroRNAs (miRNAs) are a class of highly conserved small non-coding RNAs that inhibit gene expression. Control of miRNAs-mediated protein expression has been described as a widely used mechanism for post-transcriptional regulation in many physiological and pathological processes in different organs, including the retina. Our main purpose was to test the hypothesis that modulation of a group of miRNAs can protect photoreceptor cells from death in the rd10 mouse model of retinitis pigmentosa. For this, we incorporated modulators of three miRNAs in adeno-associated viruses (AAVs), which were administered through sub-retinal injections. The results obtained indicate that inhibition of the miR-6937-5p slows down the visual deterioration of rd10 mice, reflected by an increased electroretinogram (ERG) wave response under scotopic conditions and significant preservation of the outer nuclear layer thickness. This work contributes to broadening our knowledge on the molecular mechanisms underlying retinitis pigmentosa and supports the development of novel therapeutic approaches for RP based on miRNA modulation

Inhibition of MicroRNA 6937 Delays Photoreceptor and Vision Loss in a Mouse Model of Retinitis Pigmentosa

Inherited retinal dystrophies (IRDs) are a group of rare retinal conditions, including retinitis pigmentosa (RP), caused by monogenic mutations in 1 out of more than 250 genes. Despite recent advancements in gene therapy, there is still a lack of an effective treatment for this group of retinal conditions. MicroRNAs (miRNAs) are a class of highly conserved small non-coding RNAs that inhibit gene expression. Control of miRNAs-mediated protein expression has been described as a widely used mechanism for post-transcriptional regulation in many physiological and pathological processes in different organs, including the retina. Our main purpose was to test the hypothesis that modulation of a group of miRNAs can protect photoreceptor cells from death in the rd10 mouse model of retinitis pigmentosa. For this, we incorporated modulators of three miRNAs in adeno-associated viruses (AAVs), which were administered through sub-retinal injections. The results obtained indicate that inhibition of the miR-6937-5p slows down the visual deterioration of rd10 mice, reflected by an increased electroretinogram (ERG) wave response under scotopic conditions and significant preservation of the outer nuclear layer thickness. This work contributes to broadening our knowledge on the molecular mechanisms underlying retinitis pigmentosa and supports the development of novel therapeutic approaches for RP based on miRNA modulation