Comparative proteomics profiling of a phospholamban mutant mouse model of dilated cardiomyopathy reveals progressive intracellular stress responses.
Defective mobilization of Ca2+ by cardiomyocytes can lead to cardiac insufficiency, but the causative mechanisms leading to congestive heart failure (HF) remain unclear. In the present study we performed exhaustive global proteomics surveys of cardiac ventricle isolated from a mouse model of cardiomyopathy overexpressing a phospholamban mutant, R9C (PLN-R9C), and exhibiting impaired Ca2+ handling and death at 24 weeks and compared them with normal control littermates. The relative expression patterns of 6190 high confidence proteins were monitored by shotgun tandem mass spectrometry at 8, 16, and 24 weeks of disease progression. Significant differential abundance of 593 proteins was detected. These proteins mapped to select biological pathways such as endoplasmic reticulum stress response, cytoskeletal remodeling, and apoptosis and included known biomarkers of HF (e.g. brain natriuretic peptide/atrial natriuretic factor and angiotensin-converting enzyme) and other indicators of presymptomatic functional impairment. These altered proteomic profiles were concordant with cognate mRNA patterns recorded in parallel using high density mRNA microarrays, and top candidates were validated by RT-PCR and Western blotting. Mapping of our highest ranked proteins against a human diseased explant and to available data sets indicated that many of these proteins could serve as markers of disease. Indeed we showed that several of these proteins are detectable in mouse and human plasma and display differential abundance in the plasma of diseased mice and affected patients. These results offer a systems-wide perspective of the dynamic maladaptions associated with impaired Ca2+ homeostasis that perturb myocyte function and ultimately converge to cause HF.
Gramolini
Anthony O
AO
Banting and Best Department of Medical Research, University of Toronto, Toronto, Ontario M5G 1L6, Canada. anthony.gramolini@utoronto.ca
Kislinger
Thomas
T
Alikhani-Koopaei
Rasoul
R
Fong
Vincent
V
Thompson
Natalie J
NJ
Isserlin
Ruth
R
Sharma
Parveen
P
Oudit
Gavin Y
GY
Trivieri
Maria G
MG
Fagan
Ailís
A
Kannan
Anitha
A
Higgins
Desmond G
DG
Huedig
Hendrik
H
Hess
George
G
Arab
Sara
S
Seidman
Jonathan G
JG
Seidman
Christine E
CE
Frey
Brendan
B
Perry
Marc
M
Backx
Peter H
PH
Liu
Peter P
PP
MacLennan
David H
DH
Emili
Andrew
A
D000818:Animals; D015415:Biomarkers / Q000097:blood; D002135:Calcium-Binding Proteins / Q000235:genetics*; D002311:Cardiomyopathy, Dilated / Q000097:blood / Q000000981:diagnostic imaging / Q000378:metabolism* / Q000503:physiopathology; D004195:Disease Models, Animal; D005260:Female; D005786:Gene Expression Regulation; D006333:Heart Failure; D006439:Hemodynamics; D006801:Humans; D008297:Male; D053858:Metabolic Networks and Pathways; D051379:Mice; D008822:Mice, Transgenic; D009154:Mutation / Q000235:genetics*; D009206:Myocardium / Q000473:pathology; D020411:Oligonucleotide Array Sequence Analysis; D010641:Phenotype; D040081:Protein Array Analysis; D040901:Proteomics / Q000379:methods*; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D015203:Reproducibility of Results; D013312:Stress, Physiological / Q000378:metabolism*; D013997:Time Factors; D014463:Ultrasonography; D000099211:Phospholamban
D015415:Biomarkers; D002135:Calcium-Binding Proteins; D012333:RNA, Messenger; D000099211:Phospholamban