Marjan Jahangiri, Manuel Mayr, Marika Fava, Cathy Shanahan, Stephen J White, Jonathan M Hill, Pawel E Buszman, Robert Hayward, Krzysztof Milewski, Tomasz Roleder, Ferheen Baig, Wojciech Wojakowski, Ursula Mayr, Marc Lynch, Piotr P Buszman, Pawel Gasior, Chris Molenaar, Gonca Suna, Javier Barallobre-Barreiro, Ruifang Lu, Xiaoke Yin
Background -Extracellular matrix (ECM) remodeling contributes to in-stent restenosis and thrombosis. Despite its important clinical implications little is known about ECM changes post-stent implantation. Methods -Bare-metal (BMS) and drug-eluting stents (DES) were implanted in pig coronary arteries with an overstretch under optical coherence tomography guidance. Stented segments were harvested 1, 3, 7, 14 and 28 days post-stenting for proteomics analysis of the media and neointima. Results -A total of 151 ECM and ECM-associated proteins were identified by mass spectrometry. After stent implantation, proteins involved in regulating calcification were upregulated in the neointima of DES. The earliest changes in the media were proteins involved in inflammation and thrombosis, followed by changes in regulatory ECM proteins. By day 28, basement membrane proteins were reduced in DES compared with BMS. In contrast, the large aggregating proteoglycan aggrecan was increased. Aggrecanases of the ADAMTS (a disintegrin and metalloproteinase with thrombospondin motifs) family contribute to the catabolism of vascular proteoglycans. An increase in ADAMTS-specific aggrecan fragments was accompanied by a notable shift from ADAMTS1 and ADAMTS5 to ADAMTS4 gene expression after stent implantation. Immunostaining in human stented coronary arteries confirmed the presence of aggrecan and aggrecan fragments, in particular at the contacts of the stent struts with the artery. Further investigation of aggrecan presence in the human vasculature revealed that aggrecan and aggrecan cleavage were more abundant in human arteries compared to veins. Also, aggrecan synthesis was induced upon grafting a vein into the arterial circulation, suggesting an important role for aggrecan in vascular plasticity. Finally, lack of ADAMTS-5 activity in mice resulted in an accumulation of aggrecan and a dilation of the thoracic aorta, confirming that aggrecanase activity regulates aggrecan abundance in the arterial wall and contributes to vascular remodeling. Conclusions -Significant differences were identified by proteomics in the ECM of coronary arteries after BMS and DES implantation, most notably an upregulation of aggrecan, a major ECM component of cartilaginous tissues that confers resistance to compression. The accumulation of aggrecan coincided with a shift in ADAMTS gene expression. This study provides the first evidence implicating aggrecan and aggrecanases in the vascular injury response after stenting.
