AAV-Mediated Clarin-1 Expression in the Mouse Retina: Implications for USH3A Gene Therapy
| Autor: | William W. Hauswirth, Qiuhong Li, Carolina Abrahan, Wen-Tao Deng, Elena Novelli, Ping Zhu, Uwe Wolfrum, W. Clay Smith, Frank M. Dyka, Kerstin Nagel-Wolfrum, Enrica Strettoi, Rachel M Stupay, Vince A. Chiodo, Seok-Hong Min, Sanford L. Boye, Astra Dinculescu |
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| Rok vydání: | 2016 |
| Předmět: |
Photoreceptors
0301 basic medicine Retinal degeneration Sensory Receptors Physiology Usher syndrome Cell Membranes lcsh:Medicine Social Sciences Nervous System Photoreceptor cell Mice chemistry.chemical_compound 0302 clinical medicine Animal Cells Medicine and Health Sciences Psychology lcsh:Science Neurons Regulation of gene expression Genetics Multidisciplinary Retinal Degeneration Animal Models Dependovirus Cell biology Electrophysiology medicine.anatomical_structure Sensory Perception Cellular Types Anatomy Cellular Structures and Organelles Usher Syndromes Research Article Signal Transduction Cell type Imaging Techniques Ocular Anatomy Neurophysiology Outer plexiform layer Mouse Models Biology Research and Analysis Methods Retina 03 medical and health sciences Model Organisms Ocular System Fluorescence Imaging medicine Animals Humans lcsh:R Membrane Proteins Biology and Life Sciences Afferent Neurons Retinal Genetic Therapy Cell Biology medicine.disease Disease Models Animal 030104 developmental biology Gene Expression Regulation chemistry Synapses Eyes lcsh:Q Head 030217 neurology & neurosurgery Neuroscience |
| Zdroj: | PLoS ONE PLoS ONE, Vol 11, Iss 2, p e0148874 (2016) |
| ISSN: | 1932-6203 |
| DOI: | 10.1371/journal.pone.0148874 |
| Popis: | Usher syndrome type III (USH3A) is an autosomal recessive disorder caused by mutations in clarin-1 (CLRN1) gene, leading to progressive retinal degeneration and sensorineural deafness. Efforts to develop therapies for preventing photoreceptor cell loss are hampered by the lack of a retinal phenotype in the existing USH3 mouse models and by conflicting reports regarding the endogenous retinal localization of clarin-1, a transmembrane protein of unknown function. In this study, we used an AAV-based approach to express CLRN1 in the mouse retina in order to determine the pattern of its subcellular localization in different cell types. We found that all major classes of retinal cells express AAV-delivered CLRN1 driven by the ubiquitous, constitutive small chicken β-actin promoter, which has important implications for the design of future USH3 gene therapy studies. Within photoreceptor cells, AAV-expressed CLRN1 is mainly localized at the inner segment region and outer plexiform layer, similar to the endogenous expression of other usher proteins. Subretinal delivery using a full strength viral titer led to significant loss of retinal function as evidenced by ERG analysis, suggesting that there is a critical limit for CLRN1 expression in photoreceptor cells. Taken together, these results suggest that CLRN1 expression is potentially supported by a variety of retinal cells, and the right combination of AAV vector dose, promoter, and delivery method needs to be selected to develop safe therapies for USH3 disorder. |
| Databáze: | OpenAIRE |
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