title: Extracellular matrix secretion by cardiac fibroblasts: role of microRNA-29b and microRNA-30c.
abstract: BACKGROUND
  MicroRNAs (miRNAs), in particular miR-29b and miR-30c, have been implicated as important regulators of cardiac fibrosis.
  
  
  OBJECTIVE
  To perform a proteomics comparison of miRNA effects on extracellular matrix secretion by cardiac fibroblasts.
  
  
  RESULTS
  Mouse cardiac fibroblasts were transfected with pre-/anti-miR of miR-29b and miR-30c, and their conditioned medium was analyzed by mass spectrometry. miR-29b targeted a cadre of proteins involved in fibrosis, including multiple collagens, matrix metalloproteinases, and leukemia inhibitory factor, insulin-like growth factor 1, and pentraxin 3, 3 predicted targets of miR-29b. miR-29b also attenuated the cardiac fibroblast response to transforming growth factor-β. In contrast, miR-30c had little effect on extracellular matrix production but opposite effects regarding leukemia inhibitory factor and insulin-like growth factor 1. Both miRNAs indirectly affected cardiac myocytes. On transfection with pre-miR-29b, the conditioned medium of cardiac fibroblasts lost its ability to support adhesion of rat ventricular myocytes and led to a significant reduction of cardiac myocyte proteins (α-actinin, cardiac myosin-binding protein C, and cardiac troponin I). Similarly, cardiomyocytes derived from mouse embryonic stem cells atrophied under pre-miR-29 conditioned medium, whereas pre-miR-30c conditioned medium had a prohypertrophic effect. Levels of miR-29a, miR-29c, and miR-30c, but not miR-29b, were significantly reduced in a mouse model of pathological but not physiological hypertrophy. Treatment with antagomiRs to miR-29b induced excess fibrosis after aortic constriction without overt deterioration in cardiac function.
  
  
  CONCLUSIONS
  Our proteomic analysis revealed novel molecular targets of miRNAs that are linked to a fibrogenic cardiac phenotype. Such comprehensive screening methods are essential to define the concerted actions of miRNAs in cardiovascular disease.
authors.affiliation: From the King's British Heart Foundation Centre, King's College London, London, United Kingdom.
authors.forename: Mélanie
authors.initials: M
authors.lastname: Abonnenc
authors.forename: Adam A
authors.initials: AA
authors.lastname: Nabeebaccus
authors.forename: Ursula
authors.initials: U
authors.lastname: Mayr
authors.forename: Javier
authors.initials: J
authors.lastname: Barallobre-Barreiro
authors.forename: Xuebin
authors.initials: X
authors.lastname: Dong
authors.forename: Friederike
authors.initials: F
authors.lastname: Cuello
authors.forename: Sumon
authors.initials: S
authors.lastname: Sur
authors.forename: Ignat
authors.initials: I
authors.lastname: Drozdov
authors.forename: Sarah R
authors.initials: SR
authors.lastname: Langley
authors.forename: Ruifang
authors.initials: R
authors.lastname: Lu
authors.forename: Konstantina
authors.initials: K
authors.lastname: Stathopoulou
authors.forename: Athanasios
authors.initials: A
authors.lastname: Didangelos
authors.forename: Xiaoke
authors.initials: X
authors.lastname: Yin
authors.forename: Wolfram-Hubertus
authors.initials: WH
authors.lastname: Zimmermann
authors.forename: Ajay M
authors.initials: AM
authors.lastname: Shah
authors.forename: Anna
authors.initials: A
authors.lastname: Zampetaki
authors.forename: Manuel
authors.initials: M
authors.lastname: Mayr
mesh_terms: D000818:Animals; D002097:C-Reactive Protein / Q000378:metabolism; D002478:Cells, Cultured; D003094:Collagen / Q000378:metabolism; D005109:Extracellular Matrix / Q000378:metabolism*; D005347:Fibroblasts / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D005355:Fibrosis; D007334:Insulin-Like Growth Factor I / Q000378:metabolism; D053605:Leukemia Inhibitory Factor / Q000378:metabolism; D008297:Male; D020782:Matrix Metalloproteinases / Q000378:metabolism; D051379:Mice; D008810:Mice, Inbred C57BL; D035683:MicroRNAs / Q000502:physiology*; D023421:Models, Animal; D009206:Myocardium / Q000378:metabolism* / Q000473:pathology; D040901:Proteomics; D000683:Serum Amyloid P-Component / Q000378:metabolism; D016212:Transforming Growth Factor beta / Q000494:pharmacology; D000099153:Pentraxins
chemical_list: D002097:C-Reactive Protein; D003094:Collagen; D007334:Insulin-Like Growth Factor I; D053605:Leukemia Inhibitory Factor; D020782:Matrix Metalloproteinases; D035683:MicroRNAs; D000683:Serum Amyloid P-Component; D016212:Transforming Growth Factor beta; D000099153:Pentraxins; C547014:MIRN29 microRNA, mouse; C534715:Mirn30d microRNA, mouse
keywords: animal models; cardiovascular disease; fibroblasts; fibrosis; microRNAs; proteomics