Genetic Background is a Key Determinant of Glomerular Extracellular Matrix Composition and Organization

Abstract

Glomerular disease often features altered histologic patterns of extracellular matrix (ECM). Despite this, the potential complexities of the glomerular ECM in both health and disease are poorly understood. To explore whether genetic background and sex determine glomerular ECM composition, we investigated two mouse strains, FVB and B6, using RNA microarrays of isolated glomeruli combined with proteomic glomerular ECM analyses. These studies, undertaken in healthy young adult animals, revealed unique strain- and sexdependent glomerular ECM signatures, which correlated with variations in levels of albuminuria and known predisposition to progressive nephropathy. Among the variation, we observed changes in netrin 4, fibroblast growth factor 2, tenascin C, collagen 1, meprin 1-a, and meprin 1-b. Differences in protein abundance were validated by quantitative immunohistochemistry and Western blot analysis, and the collective differences were not explained by mutations in known ECM or glomerular disease genes. Within the distinct signatures, we discovered a core set of structural ECM proteins that form multiple protein–protein interactions and are conserved from mouse to man. Furthermore, we found striking ultrastructural changes in glomerular basement membranes in FVB mice. Pathway analysis of merged transcriptomic and proteomic datasets identified potentialECMregulatory pathways involving inhibitionofmatrixmetalloproteases, liverXreceptor/retinoidX receptor, nuclear factor erythroid 2-related factor 2, notch, and cyclin-dependent kinase 5. These pathways may therefore alter ECM and confer susceptibility to disease.

Description

Michael J. Randles,*† Adrian S. Woolf,† Jennifer L. Huang,‡ Adam Byron,§ Jonathan D. Humphries,* Karen L. Price,‡ Maria Kolatsi-Joannou,‡ Sophie Collinson,* Thomas Denny,*† David Knight,* Aleksandr Mironov,* Toby Starborg,* Ron Korstanje,| Martin J. Humphries,* David A. Long,‡ and Rachel Lennon*† *Wellcome Trust Centre for Cell-Matrix Research, Faculty of Life Sciences, University of Manchester, Manchester, United Kingdom; †Institute of Human Development, Faculty of Medical and Human Sciences, University of Manchester, Manchester, United Kingdom; ‡Developmental Biology and Cancer Program, Institute of Child Health, University College London, London, United Kingdom; §Edinburgh Cancer Research United Kingdom Centre, Institute of Genetics and Molecular Medicine, University of Edinburgh, Edinburgh, United Kingdom; and |The Jackson Laboratory, Bar Harbor, Maine This work was supported by a Wellcome Trust Intermediate Fellowship award (090006) to R.L., a Kids KidneyResearch grant awarded to R.L. and A.S.W to support a studentship for M.J.R, a Kidney Research UK Senior Non-Clinical Fellowship (SF1/2008) and Medical Research Council New Investigator Award (MR/J003638/1) both to D.A.L., aWellcome Trust grant (092015) toM.J.H. and by theWellcome Trust (097820/Z/11/B) support for the bioinformatics department. The mass spectrometer and microscopes used in this study were purchased with grants from the Biotechnology and Biological Sciences Research Council, Wellcome Trust and the University of Manchester Strategic Fund. Mass spectrometry was performed in the Biomolecular Analysis Core Facility, Faculty of Life Sciences, University of Manchester, and we thank Stacey Warwood for advice and technical support and Julian Selley for bioinformatic support.

Keywords

ECM, Geneics, Glomerulus

Citation

Randles et al. (2015) Genetic Background is a Key Determinant of Glomerular Extracellular Matrix Composition and Organization. Journal of the American Society of Nephrology, 26 (12), pp. 3021-3034

Rights

Research Institute

Institute for Allied Health Sciences Research