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GPCRomics of cardiac fibroblasts and cardiac fibrosis
Author(s) -
Sriram Krishna,
Michkov Alexander V.,
Aroonsakool Nakon,
Insel Paul A.
Publication year - 2016
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.30.1_supplement.709.9
Subject(s) - g protein coupled receptor , receptor , biology , fibrosis , phenotype , microbiology and biotechnology , computational biology , bioinformatics , medicine , gene , genetics
G‐protein coupled receptors (GPCRs) are the largest category of cell surface receptors, serving as the most widely used set of targets for pharmacological intervention. Seeking to identify novel targets for cardiac fibrosis, we have used mRNA‐based (GPCRomic) methods to detect and quantify (i.e., profile) non‐chemosensory “endo‐GPCRs” in cardiac fibroblasts (CFs) and, complemented these results by verification of GPCR expression/function using antibody‐based and functional analyses. We used 2D, low passage (passage 1 or 2) cultures of primary isolates of CFs and myofibroblasts (MyoFs) from humans, rats and mice. RESULTS We found that CFs express ~120 GPCRs, including a large number (~40–50) of orphan GPCRs, and multiple highly expressed adhesion GPCRs. Biological replicates of human CFs correlate closely in terms of identity of GPCRs expressed and the magnitude of their expression. Further, our data show that MyoFs possess a smaller (number and average expression) of GPCRs but a distinct subset of GPCRs have increased expression in MyoFs compared to CFs. Growth of CFs on soft substrates (which minimize the spontaneous transformation of cultured CFs into MyoFs) produces relatively small effects on the profile of GPCRs expressed by CFs. A subset of GPCRs remains highly and robustly expressed in CFs, regardless of substrate stiffness, pro‐fibrotic stimulation, duration of culture, or growth under hypoxic conditions. The GPCR profiles of mouse and rat CFs closely mimic that of human CFs as do the GPCR profiles of rat and human MyoFs. Further, we find that the GPCR expression profile of rat CFs is very different from that of freshly isolated rat cardiomyocytes, implying that there are numerous potential GPCR targets for cardiac fibrosis, which if used therapeutically, are predicted to have minimal on‐target effects on cardiomyocytes. We conclude that a GPCRomic approach provides novel insights regarding the regulation of CFs and MyoFs and identifies numerous potential targets for cardiac fibrosis. Support or Funding Information Supported by Bristol‐Myers Squibb