Gene profiling of cathepsin K deficiency in atherogenesis: profibrotic but lipogenic.
Recently, we showed that cathepsin K deficiency reduces atherosclerotic plaque progression, induces plaque fibrosis, but aggravates macrophage foam cell formation in the ApoE -/- mouse. To obtain more insight into the molecular mechanisms by which cathepsin K disruption evokes the observed phenotypic changes, we used microarray analysis for gene expression profiling of aortic arches of CatK -/-/ApoE -/- and ApoE -/- mice on a mouse oligo microarray. Out of 20 280 reporters, 444 were significantly differentially expressed (p-value of < 0.05, fold change of > or = 1.4 or < or = - 1.4, and intensity value of > 2.5 times background in at least one channel). Ingenuity Pathway Analysis and GenMAPP revealed upregulation of genes involved in lipid uptake, trafficking, and intracellular storage, including caveolin - 1, - 2, - 3 and CD36, and profibrotic genes involved in transforming growth factor beta (TGFbeta) signalling, including TGFbeta2, latent TGFbeta binding protein-1 (LTBP1), and secreted protein, acidic and rich in cysteine (SPARC), in CatK -/-/ApoE -/- mice. Differential gene expression was confirmed at the mRNA and protein levels. In vitro modified low density lipoprotein (LDL) uptake assays, using bone marrow derived macrophages preincubated with caveolae and scavenger receptor inhibitors, confirmed the importance of caveolins and CD36 in increasing modified LDL uptake in the absence of cathepsin K. In conclusion, we suggest that cathepsin K deficiency alters plaque phenotype not only by decreasing proteolytic activity, but also by stimulating TGFbeta signalling. Besides this profibrotic effect, cathepsin K deficiency has a lipogenic effect owing to increased lipid uptake mediated by CD36 and caveolins.
Lutgens
S P M
SP
Departments of Pathology, Cardiovascular Research Institute Maastricht (CARIM), University of Maastricht, P. Debyelaan 25, Maastricht, The Netherlands.
Kisters
N
N
Lutgens
E
E
van Haaften
R I M
RI
Evelo
C T A
CT
de Winther
M P J
MP
Saftig
P
P
Daemen
M J A P
MJ
Heeneman
S
S
Cleutjens
K B J M
KB
D000818:Animals; D001057:Apolipoproteins E / Q000235:genetics; D050197:Atherosclerosis / Q000235:genetics*; D018955:CD36 Antigens / Q000235:genetics; D056657:Cathepsin K; D002403:Cathepsins / Q000172:deficiency* / Q000235:genetics; D022461:Caveolins / Q000235:genetics; D005355:Fibrosis / Q000235:genetics; D020869:Gene Expression Profiling / Q000379:methods*; D005786:Gene Expression Regulation / Q000235:genetics; D007150:Immunohistochemistry / Q000379:methods; D053779:Latent TGF-beta Binding Proteins / Q000235:genetics; D050356:Lipid Metabolism / Q000235:genetics; D008077:Lipoproteins, LDL / Q000378:metabolism; D008264:Macrophages / Q000502:physiology; D051379:Mice; D008810:Mice, Inbred C57BL; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods; D010641:Phenotype; D012333:RNA, Messenger / Q000032:analysis; D016212:Transforming Growth Factor beta / Q000235:genetics; D015854:Up-Regulation / Q000235:genetics
D001057:Apolipoproteins E; D018955:CD36 Antigens; D022461:Caveolins; D053779:Latent TGF-beta Binding Proteins; D008077:Lipoproteins, LDL; C509033:Ltbp1 protein, mouse; D012333:RNA, Messenger; D016212:Transforming Growth Factor beta; D002403:Cathepsins; D056657:Cathepsin K; C534132:Ctsk protein, mouse