Glomerular cell cross-talk influences composition and assembly of extracellular matrix

Adam Byron, Michael J Randles, Jonathan D Humphries, Aleksandr Mironov, Hellyeh Hamidi, Shelley Harris, Peter W Mathieson, Moin A Saleem, Simon C Satchell, Roy Zent, Martin J Humphries, Rachel Lennon

Research output: Contribution to journalArticlepeer-review

Abstract

The glomerular basement membrane (GBM) is a specialized extracellular matrix (ECM) compartment within the glomerulus that contains tissue-restricted isoforms of collagen IV and laminin. It is integral to the capillary wall and therefore, functionally linked to glomerular filtration. Although the composition of the GBM has been investigated with global and candidate-based approaches, the relative contributions of glomerular cell types to the production of ECM are not well understood. To characterize specific cellular contributions to the GBM, we used mass spectrometry-based proteomics to analyze ECM isolated from podocytes and glomerular endothelial cells in vitro. These analyses identified cell type-specific differences in ECM composition, indicating distinct contributions to glomerular ECM assembly. Coculture of podocytes and endothelial cells resulted in an altered composition and organization of ECM compared with monoculture ECMs, and electron microscopy revealed basement membrane-like ECM deposition between cocultured cells, suggesting the involvement of cell-cell cross-talk in the production of glomerular ECM. Notably, compared with monoculture ECM proteomes, the coculture ECM proteome better resembled a tissue-derived glomerular ECM dataset, indicating its relevance to GBM in vivo. Protein network analyses revealed a common core of 35 highly connected structural ECM proteins that may be important for glomerular ECM assembly. Overall, these findings show the complexity of the glomerular ECM and suggest that both ECM composition and organization are context-dependent.

Original languageEnglish
Pages (from-to)953-66
JournalJournal of the American Society of Nephrology
Volume25
Issue number5
Early online date16 Jan 2014
DOIs
Publication statusPublished - May 2014

Keywords

  • Cells, Cultured
  • Coculture Techniques
  • Culture Media, Conditioned
  • Extracellular Matrix
  • Extracellular Matrix Proteins
  • Humans
  • Kidney Glomerulus
  • Phenotype
  • Podocytes
  • Protein Interaction Maps
  • Receptor Cross-Talk

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