CAPN5 mutation in hereditary uveitis: the R243L mutation increases calpain catalytic activity and triggers intraocular inflammation in a mouse model

Katherine J Wert; Alexander G Bassuk; Wen-Hsuan Wu; Lokesh Gakhar; Diana Coglan; MaryAnn Mahajan; Shu Wu; Jing Yang; Chyuan-Sheng Lin; Stephen H Tsang; Vinit B Mahajan

doi:10.1093/hmg/ddv189

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CAPN5 mutation in hereditary uveitis: the R243L mutation increases calpain catalytic activity and triggers intraocular inflammation in a mouse model

Journal article

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CAPN5 mutation in hereditary uveitis: the R243L mutation increases calpain catalytic activity and triggers intraocular inflammation in a mouse model

Katherine J Wert, Alexander G Bassuk, Wen-Hsuan Wu, Lokesh Gakhar, Diana Coglan, MaryAnn Mahajan, Shu Wu, Jing Yang, Chyuan-Sheng Lin, Stephen H Tsang, …

Human molecular genetics, Vol.24(16), pp.4584-4598

08/15/2015

DOI: 10.1093/hmg/ddv189

PMCID: PMC4512628

PMID: 25994508

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Abstract

A single amino acid mutation near the active site of the CAPN5 protease was linked to the inherited blinding disorder, autosomal dominant neovascular inflammatory vitreoretinopathy (ADNIV, OMIM #193235). In homology modeling with other calpains, this R243L CAPN5 mutation was situated in a mobile loop that gates substrate access to the calcium-regulated active site. In in vitro activity assays, the mutation increased calpain protease activity and made it far more active at low concentrations of calcium. To test whether the disease allele could yield an animal model of ADNIV, we created transgenic mice expressing human (h) CAPN5(R243L) only in the retina. The resulting hCAPN5(R243L) transgenic mice developed a phenotype consistent with human uveitis and ADNIV, at the clinical, histological and molecular levels. The fundus of hCAPN5(R243L) mice showed enhanced autofluorescence (AF) and pigment changes indicative of reactive retinal pigment epithelial cells and photoreceptor degeneration. Electroretinography showed mutant mouse eyes had a selective loss of the b-wave indicating an inner-retina signaling defect. Histological analysis of mutant mouse eyes showed protein extravasation from dilated vessels into the anterior chamber and vitreous, vitreous inflammation, vitreous and retinal fibrosis and retinal degeneration. Analysis of gene expression changes in the hCAPN5(R243L) mouse retina showed upregulation of several markers, including members of the Toll-like receptor pathway, chemokines and cytokines, indicative of both an innate and adaptive immune response. Since many forms of uveitis share phenotypic characteristics of ADNIV, this mouse offers a model with therapeutic testing utility for ADNIV and uveitis patients.

Inflammation

Cell Line

Calpain - metabolism

Humans

Genetic Diseases, Inborn - genetics

Mice, Transgenic

Genetic Diseases, Inborn - pathology

Calpain - genetics

Animals

Genetic Diseases, Inborn - metabolism

Mice

Uveitis - genetics

Uveitis - pathology

Uveitis - metabolism

Details

Title: Subtitle: CAPN5 mutation in hereditary uveitis: the R243L mutation increases calpain catalytic activity and triggers intraocular inflammation in a mouse model
Creators: Katherine J Wert - Barbara and Donald Jonas Laboratory of Stem Cells and Regenerative Medicine and Bernard and Shirlee Brown Glaucoma Laboratory, Edward S. Harkness Eye Institute, Institute of Human Nutrition, College of Physicians and Surgeons
Alexander G Bassuk - Department of Pediatrics and Neurology
Wen-Hsuan Wu - Barbara and Donald Jonas Laboratory of Stem Cells and Regenerative Medicine and Bernard and Shirlee Brown Glaucoma Laboratory, Edward S. Harkness Eye Institute
Lokesh Gakhar - Department of Biochemistry, Protein Crystallography Facility
Diana Coglan - Omics Laboratory and Department of Ophthalmology and Visual Sciences, University of Iowa, Iowa City, IA, USA
MaryAnn Mahajan - Omics Laboratory and Department of Ophthalmology and Visual Sciences, University of Iowa, Iowa City, IA, USA
Shu Wu - Department of Pediatrics and Neurology
Jing Yang - Protein Crystallography Facility, Omics Laboratory and
Chyuan-Sheng Lin - Herbert Irving Comprehensive Cancer Center
Stephen H Tsang - Barbara and Donald Jonas Laboratory of Stem Cells and Regenerative Medicine and Bernard and Shirlee Brown Glaucoma Laboratory, Edward S. Harkness Eye Institute, Institute of Human Nutrition, College of Physicians and Surgeons, Department of Pathology and Cell Biology, College of Physicians and Surgeons, Columbia University, New York, NY, USA, mahajanlab@gmail.com gene.targeting@gmail.com
Vinit B Mahajan - Omics Laboratory and Department of Ophthalmology and Visual Sciences, University of Iowa, Iowa City, IA, USA mahajanlab@gmail.com gene.targeting@gmail.com
Resource Type: Journal article
Publication Details: Human molecular genetics, Vol.24(16), pp.4584-4598
Publisher: England
DOI: 10.1093/hmg/ddv189
PMID: 25994508
PMCID: PMC4512628
ISSN: 0964-6906
eISSN: 1460-2083
Grant note: T32 EY013933 / NEI NIH HHS 5T32EY013933 / NEI NIH HHS F32CA196065 / NCI NIH HHS R01EY018213 / NEI NIH HHS R01EY024665 / NEI NIH HHS 5P30CA013696 / NCI NIH HHS K08EY020530 / NEI NIH HHS 5T32DK007647-20 / NIDDK NIH HHS R01 EY024665 / NEI NIH HHS 5P30EY019007 / NEI NIH HHS R01EY025225 / NEI NIH HHS
Language: English
Date published: 08/15/2015
Academic Unit: Neurology; Stead Family Department of Pediatrics; Iowa Neuroscience Institute; Biochemistry and Molecular Biology; Neurology (Pediatrics); Medicine Administration; Internal Medicine
Record Identifier: 9984020718402771

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