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, Mary Ann Mahajan, Shu Wu, Jing Yang, Chyuan Sheng Lin, Stephen H. Tsang, Vinit B. Mahajan

Research output: Contribution to journalArticle

20 Scopus citations

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) CAPN5R243L only in the retina. The resulting hCAPN5R243L transgenic mice developed a phenotype consistent with human uveitis and ADNIV, at the clinical, histological and molecular levels. The fundus of hCAPN5R243L 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 hCAPN5R243L 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.

Original languageEnglish (US)
Pages (from-to)4584-4598
Number of pages15
JournalHuman molecular genetics
Volume24
Issue number16
DOIs
StatePublished - Apr 28 2015
Externally publishedYes

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics
  • Genetics(clinical)

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    Wert, K. J., Bassuk, A. G., Wu, W. H., Gakhar, L., Coglan, D., Mahajan, M. A., Wu, S., Yang, J., Lin, C. S., Tsang, S. H., & Mahajan, V. B. (2015). CAPN5 mutation in hereditary uveitis: The R243L mutation increases calpain catalytic activity and triggers intraocular inflammation in a mouse model. Human molecular genetics, 24(16), 4584-4598. https://doi.org/10.1093/hmg/ddv189