Cell division autoantigen 1 plays a profibrotic role by modulating downstream signalling of TGF-beta in a murine diabetic model of atherosclerosis.
OBJECTIVE
Excess accumulation of vascular extracellular matrix (ECM) is an important pathological process in cardiovascular diseases including diabetes-associated atherosclerosis. We explored how a recently identified molecule, cell division autoantigen 1 (CDA1), influences the profibrotic TGF-beta pathway leading to vascular ECM accumulation.


METHODS
Expression levels of genes encoding for CDA1, TGF-beta and connective tissue growth factor (CTGF) were examined in aorta from Apoe(-/-) mice with or without diabetes. We used retroviral and adenoviral constructs to knockdown or overexpress Tspyl2, the gene encoding CDA1, in mouse vascular smooth muscle cells (VSMCs) with or without TGF-beta treatment in order to demonstrate the role of CDA1 in TGF-beta signalling.


RESULTS
In vivo studies indicated that the mRNA levels of CDA1-encoding gene Tspyl2 and protein levels of CDA1 were elevated in the aorta of diabetic Apoe(-/-) mice, accompanied by increased levels of Tgf-beta (also known as Tgfb1), Ctgf and ECM accumulation. In vitro studies in vascular cells showed that TGF-beta treatment rapidly increased CDA1 protein levels, which then amplified TGF-beta signalling leading to upregulation of ECM genes. Knockdown of CDA1-encoding gene Tspyl2 to reduce cellular CDA1 level markedly attenuated TGF-beta-stimulated MAD homologue 3 (drosophila; SMAD3) phosphorylation and transcriptional activities. CDA1 overproduction increased and Tspyl2 knockdown decreased expression of TGF-beta receptor type I, TbetarI (also known as Tgfbr1), but not TGF-beta receptor type II, TbetarII (also known as Tgfbr2), providing a mechanism for CDA1's action in modulating TGF-beta signalling. Knockdown of CDA1-encoding gene Tspyl2 also blocked the profibrotic effect of TGF-beta in VSMCs.


CONCLUSIONS
CDA1 plays an important role in vascular ECM accumulation by amplifying TGF-beta signalling. This is critical for the profibrotic effect of TGF-beta in the vasculature. CDA1 is therefore a potential target for attenuating vascular ECM accumulation caused by enhanced TGF-beta action, as seen in diabetic atherosclerosis.
Diabetes and Metabolism Division, Baker IDI Heart and Diabetes Institute, Melbourne, Victoria 3004, Australia.
Y
Y
Pham
Y
Y
Tu
T
T
Wu
T J
TJ
Allen
A C
AC
Calkin
A M
AM
Watson
J
J
Li
K A
KA
Jandeleit-Dahm
B-H
BH
Toh
Z
Z
Cao
M E
ME
Cooper
Z
Z
Chai
D000818:Animals; D001011:Aorta / Q000502:physiology; D050197:Atherosclerosis / Q000503:physiopathology*; D001324:Autoantigens / Q000235:genetics / Q000502:physiology*; D001786:Blood Glucose / Q000378:metabolism; D055513:Connective Tissue Growth Factor / Q000502:physiology; D003921:Diabetes Mellitus, Experimental / Q000150:complications / Q000503:physiopathology*; D003925:Diabetic Angiopathies / Q000503:physiopathology*; D005109:Extracellular Matrix / Q000502:physiology*; D005786:Gene Expression Regulation; D017930:Genes, Reporter; D006442:Glycated Hemoglobin / Q000378:metabolism; D008074:Lipoproteins / Q000097:blood; D008156:Luciferases / Q000235:genetics; D051379:Mice; D012333:RNA, Messenger / Q000235:genetics; D034741:RNA, Small Interfering / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015398:Signal Transduction / Q000502:physiology; D016212:Transforming Growth Factor beta / Q000502:physiology*
D001324:Autoantigens; D001786:Blood Glucose; D055513:Connective Tissue Growth Factor; D006442:Glycated Hemoglobin; D008074:Lipoproteins; D008156:Luciferases; D012333:RNA, Messenger; D034741:RNA, Small Interfering; D016212:Transforming Growth Factor beta; C526318:CCN2 protein, mouse; C436320:cell division autoantigen-1