Silencing of CD40 in vivo reduces progression of experimental atherogenesis through an NF-κB/miR-125b axis and reveals new potential mediators in the pathogenesis of atherosclerosis.
CD40/CD40L signaling exerts a critical role in the development of atherosclerosis, and microRNAs (miRNAs) are key regulators in vascular inflammation and plaque formation. In this work, we investigated mRNA/miRNA expression during progression of atherosclerotic lesions through CD40 silencing.



We silenced CD40 with a specific siRNA in ApoE-/-mice and compared expression of mRNA/miRNA in ascending aorta with scrambled treated mice.



siRNA-CD40 treated mice significantly reduced the extension and severity of atherosclerotic lesions, as well as the number of F4/80+, galectin-3+macrophages and NF-κB+cells in the intima. Genome-wide mRNA/miRNA profiling allowed the identification of transcripts, which were significantly upregulated during atherosclerosis; among them, miR-125b and miR-30a, Xpr1, a regulator of macrophage differentiation, Taf3, a core transcription factor and the NF-κB activator Ikkβ, whereas, the NF-κB inhibitor Ikbα was downregulated during disease progression. All those changes were reversed upon CD40 silencing. Interestingly, TAF3, XPR1 and miR-125b were also overexpressed in human atherosclerotic plaques. Murine Taf3 and Xpr1 were detected in the perivascular adipose tissue (PVAT), and Taf3 also in intimal foam cells. Finally, expression of miR-125b was regulated by the CD40-NF-κB signaling axis in RAW264.7 macrophages.



CD40 silencing with a specific siRNA ameliorates progression of experimental atherosclerosis in ApoE-/-mice, and evidences a role for NF-κB, Taf3, Xpr1, and miR-125b in the pathogenesis of atherosclerosis.
Department of Nephrology, Hospital Universitari Bellvitge, and Bellvitge Research Institute (IDIBELL), L'Hospitalet de Llobregat, Spain. Electronic address: mhueso@idibell.cat.
Miguel
M
Hueso
Laboratory of Experimental Nephrology, Bellvitge Research Institute (IDIBELL), L'Hospitalet de Llobregat, Spain.
Laura
L
De Ramon
Laboratory of Molecular Oncology, Bellvitge Research Institute (IDIBELL), L'Hospitalet de Llobregat, Spain.
Estanislao
E
Navarro
Laboratory of Experimental Nephrology, Bellvitge Research Institute (IDIBELL), L'Hospitalet de Llobregat, Spain.
Elia
E
Ripoll
Department of Nephrology, Hospital Universitari Bellvitge, and Bellvitge Research Institute (IDIBELL), L'Hospitalet de Llobregat, Spain.
Josep M
JM
Cruzado
Department of Nephrology, Hospital Universitari Bellvitge, and Bellvitge Research Institute (IDIBELL), L'Hospitalet de Llobregat, Spain.
Josep M
JM
Grinyo
Department of Nephrology, Hospital Universitari Bellvitge, and Bellvitge Research Institute (IDIBELL), L'Hospitalet de Llobregat, Spain.
Joan
J
Torras
D000818:Animals; D001011:Aorta / Q000378:metabolism* / Q000473:pathology; D001018:Aortic Diseases / Q000235:genetics / Q000378:metabolism / Q000473:pathology / Q000517:prevention & control*; D001057:Apolipoproteins E / Q000172:deficiency / Q000235:genetics; D050197:Atherosclerosis / Q000235:genetics / Q000378:metabolism / Q000473:pathology / Q000517:prevention & control*; D019013:CD40 Antigens / Q000172:deficiency* / Q000235:genetics; D004195:Disease Models, Animal; D018450:Disease Progression; D005260:Female; D020022:Genetic Predisposition to Disease; D018398:Homeodomain Proteins / Q000235:genetics / Q000378:metabolism; D006801:Humans; D008264:Macrophages / Q000378:metabolism; D051379:Mice; D018345:Mice, Knockout; D035683:MicroRNAs / Q000235:genetics / Q000378:metabolism*; D016328:NF-kappa B / Q000235:genetics / Q000378:metabolism*; D010641:Phenotype; D058226:Plaque, Atherosclerotic; D000067996:RAW 264.7 Cells; D034622:RNA Interference; D043562:Receptors, G-Protein-Coupled / Q000235:genetics / Q000378:metabolism; D011991:Receptors, Virus / Q000235:genetics / Q000378:metabolism; D015398:Signal Transduction; D035381:TATA-Binding Protein Associated Factors; D013997:Time Factors; D035362:Transcription Factor TFIID / Q000235:genetics / Q000378:metabolism; D059467:Transcriptome; D000089842:Xenotropic and Polytropic Retrovirus Receptor
D001057:Apolipoproteins E; D019013:CD40 Antigens; D018398:Homeodomain Proteins; C531888:MIRN125 microRNA, human; D035683:MicroRNAs; C527468:Mirn125 microRNA, mouse; C534715:Mirn30d microRNA, mouse; D016328:NF-kappa B; D043562:Receptors, G-Protein-Coupled; D011991:Receptors, Virus; C432689:TAF3 protein, human; D035381:TATA-Binding Protein Associated Factors; C532029:Taf3 protein, mouse; D035362:Transcription Factor TFIID; C000715267:XPR1 protein, human; D000089842:Xenotropic and Polytropic Retrovirus Receptor; C000715268:Xpr1 protein, mouse
Animal model; Biomarkers; CD40; Vascular biology