Journal article
Functional validation of a human CAPN5 exome variant by lentiviral transduction into mouse retina
Human molecular genetics, Vol.23(10), pp.2665-2677
05/15/2014
DOI: 10.1093/hmg/ddt661
PMCID: PMC3990166
PMID: 24381307
Abstract
Exome sequencing indicated that the gene encoding the calpain-5 protease, CAPN5, is the likely cause of retinal degeneration and autoimmune uveitis in human patients with autosomal dominant neovascular inflammatory vitreoretinopathy (ADNIV, OMIM #193235). To explore the mechanism of ADNIV, a human CAPN5 disease allele was expressed in mouse retinas with a lentiviral vector created to express either the wild-type human (h) CAPN5 or the ADNIV mutant hCAPN5-R243L allele under a rhodopsin promoter with tandem green fluorescent protein (GFP) expression. Vectors were injected into the subretinal space of perinatal mice. Mouse phenotypes were analyzed using electroretinography, histology and inflammatory gene expression profiling. Mouse calpain-5 showed high homology to its human ortholog with >98% sequence identity that includes the ADNIV mutant residue. Calpain-5 protein was expressed in the inner and outer segments of the photoreceptors and in the outer plexiform layer. Expression of the hCAPN5-R243L allele caused loss of the electroretinogram b-wave, photoreceptor degeneration and induction of immune cell infiltration and inflammatory genes in the retina, recapitulating major features of the ADNIV phenotype. Intraocular neovascularization and fibrosis were not observed during the study period. Our study shows that expression of the hCAPN5-R243L disease allele elicits an ADNIV-like disease in mice. It further suggests that ADNIV is due to CAPN5 gain-of-function rather than haploinsufficiency, and retinal expression may be sufficient to generate an autoimmune response. Genetic models of ADNIV in the mouse can be used to explore protease mechanisms in retinal degeneration and inflammation as well as preclinical therapeutic testing.
Details
- Title: Subtitle
- Functional validation of a human CAPN5 exome variant by lentiviral transduction into mouse retina
- Creators
- Katherine J Wert - 1 Bernard and Shirlee Brown Glaucoma Laboratory, Department of Pathology and Cell Biology, College of Physicians and SurgeonsJessica M Skeie - 4 Omics LaboratoryAlexander G Bassuk - 4 Omics LaboratoryAlicia K Olivier - 7 Department of Pathology, University of Iowa, Iowa City, IA, USAStephen H Tsang - 1 Bernard and Shirlee Brown Glaucoma Laboratory, Department of Pathology and Cell Biology, College of Physicians and SurgeonsVinit B Mahajan - 4 Omics Laboratory
- Resource Type
- Journal article
- Publication Details
- Human molecular genetics, Vol.23(10), pp.2665-2677
- Publisher
- Oxford University Press
- DOI
- 10.1093/hmg/ddt661
- PMID
- 24381307
- PMCID
- PMC3990166
- ISSN
- 0964-6906
- eISSN
- 1460-2083
- Language
- English
- Date published
- 05/15/2014
- Academic Unit
- Neurology; Stead Family Department of Pediatrics; Iowa Neuroscience Institute; Neurology (Pediatrics); Ophthalmology and Visual Sciences
- Record Identifier
- 9984020613902771
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