Genetic and Functional Analysis of Calpain-14 in Eosinophilic Esophagitis

Bibliographic Details
Main Author:	Davis, Benjamin
Language:	English
Published:	University of Cincinnati / OhioLINK 2015
Subjects:	Biology Animals Physiology Medicine calpain eosinophilic esophagitis desmoglein allergy eosinophil epithelium
Online Access:	http://rave.ohiolink.edu/etdc/view?acc_num=ucin1447688593

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spelling	ndltd-OhioLink-oai-etd.ohiolink.edu-ucin14476885932021-08-03T06:33:43Z Genetic and Functional Analysis of Calpain-14 in Eosinophilic Esophagitis Davis, Benjamin Biology Animals Physiology Medicine calpain eosinophilic esophagitis desmoglein allergy eosinophil epithelium Eosinophilic esophagitis (EoE) is a complex, allergic inflammatory disease of the esophagus, caused by both genetic and environmental factors, and featuring increased esophageal interleukin 13 (IL-13) levels and impaired barrier function. We interrogated >1.5 million genetic variants in European EoE cases and control subjects. We identified an association at 2p23 (encoding CAPN14, p = 2.5×10-10). CAPN14 was specifically expressed in the esophagus, dynamically upregulated as a function of disease activity and genetic haplotype and after exposure of epithelial cells to IL-13, and located in an epigenetic hotspot modified by IL-13. Next, we showed that recombinant CAPN14 possessed calpain activity and endogenous CAPN14 was specifically induced in esophageal epithelium following IL13 treatment in a kinetic fashion. Esophageal epithelial cells engineered to overexpress CAPN14 displayed membranous, cytoplasmic, and nuclear staining and demonstrated disrupted epithelial cell architecture and impaired epithelial cell barrier following ALI culture. Whereas, epithelial cells with gene silenced CAPN14 demonstrated increased dilated intercellular spaces following IL-13 exposure. CAPN14 overexpression resulted in loss of DSG1 expression, which was specifically blocked by CAPN14 gene silencing. We then identified the first reported CAPN14 mutation, a leucine to phenylalanine mutation at amino acid position 417 (L417F), reported in association with a familial case of EoE inherited in an autosomal dominant fashion. Thus far, the analysis of the mutant demonstrates similar activity compared to wild type CAPN14 as measured by western blot expression analysis, calpain protease activity, immunofluorescent staining, hematoxylin and eosin stain, barrier function analysis, and effect on DSG-1 expression.As an extension of the GWAS data collected in this work, we also report the association of eosinophilic esophagitis (EoE) and hypertrophic cardiomyopathy (HCM) and provide genetic data indicating a linkage between EoE and HCM. We began with an index patient diagnosed with both EoE and HCM and found to have a known genetic mutation for HCM. We then identify an odds ratio of nearly 8 for having both EoE and HCM following review of an electronic medical record with >1,000,000 individuals. Finally, via a candidate gene approach we identified significant association of HCM gene variants in a cohort of EoE patients versus control subjects. Collectively, we have identified a putative interaction between EoE and HCM with clinical and pathogenic implications.Together, these studies have identified new genetic, molecular etiology, and clinical implications in EoE. Importantly, we have identified the CAPN14 genetic locus as the highest associated locus in EoE and have described, for the first time, function of its gene product, CAPN14. This function is integral to epithelial integrity and barrier function of esophageal epithelium; physiologic structure and processes that are integral in EoE pathophysiology 2015 English text University of Cincinnati / OhioLINK http://rave.ohiolink.edu/etdc/view?acc_num=ucin1447688593 http://rave.ohiolink.edu/etdc/view?acc_num=ucin1447688593 unrestricted This thesis or dissertation is protected by copyright: all rights reserved. It may not be copied or redistributed beyond the terms of applicable copyright laws.
collection	NDLTD
language	English
sources	NDLTD
topic	Biology Animals Physiology Medicine calpain eosinophilic esophagitis desmoglein allergy eosinophil epithelium
spellingShingle	Biology Animals Physiology Medicine calpain eosinophilic esophagitis desmoglein allergy eosinophil epithelium Davis, Benjamin Genetic and Functional Analysis of Calpain-14 in Eosinophilic Esophagitis
author	Davis, Benjamin
author_facet	Davis, Benjamin
author_sort	Davis, Benjamin
title	Genetic and Functional Analysis of Calpain-14 in Eosinophilic Esophagitis
title_short	Genetic and Functional Analysis of Calpain-14 in Eosinophilic Esophagitis
title_full	Genetic and Functional Analysis of Calpain-14 in Eosinophilic Esophagitis
title_fullStr	Genetic and Functional Analysis of Calpain-14 in Eosinophilic Esophagitis
title_full_unstemmed	Genetic and Functional Analysis of Calpain-14 in Eosinophilic Esophagitis
title_sort	genetic and functional analysis of calpain-14 in eosinophilic esophagitis
publisher	University of Cincinnati / OhioLINK
publishDate	2015
url	http://rave.ohiolink.edu/etdc/view?acc_num=ucin1447688593
work_keys_str_mv	AT davisbenjamin geneticandfunctionalanalysisofcalpain14ineosinophilicesophagitis
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Genetic and Functional Analysis of Calpain-14 in Eosinophilic Esophagitis

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