Altered ubiquitin/proteasome expression in anastomotic intimal hyperplasia.

Objective: Anastomotic intimal hyperplasia remains a leading mechanism of prosthetic arterial graft failure. Recent studies using messenger RNA differential display have demonstrated altered proteasome gene expression at the anastomoses in an expanded polytetrafluoroethylene canine carotid model. However, this technique is technically limited because of a paucity of available hyperplastic tissue at early time periods after arterial injury. Microarray gene chip technology offers a new and sensitive technique to assay early gene expression, requiring far less tissue for analysis. The purpose of this study was to screen for altered proteasome gene expression at 48 hours and 14 days after prosthetic arterial grafting.

Methods: Expanded polytetrafluoroethylene grafts (6-mm diameter, n = 9) were implanted into 25-kg mongrel dogs. The normal intervening carotid artery was used as control. At 48 hours and 14 days, RNA was extracted from the perianastomotic tissue and compared with RNA from the control carotid. Messenger RNA was then hybridized to microarray genomes screening for differential gene expression.

Results: Two 26S proteasome genes and five ubiquitin pathway genes were significantly underexpressed at 48 hours, among several hundred significantly expressed clones. The two 26S proteasome genes were 26S proteasomal subunit p55 (0.26), and 26S proteasomal subunit p40.5 (0.13). The underexpressed ubiquitin genes included ubiquitin (0.31), Nedd-4-like ubiquitin-protein ligase (0.30), ubiquitin conjugating enzyme UbcH2 (0.25), putative ubiquitin C-terminal hydrolase UHX1 (0.11), and ubiquitin-conjugating enzyme UbcH7 (0.12). At 14 days, six ubiquitin genes were underexpressed, and 17 26S proteasome genes were significantly downregulated.

Conclusions: This study shows decreased expression of the ubiquitin/proteasome pathway 48 hours after graft implantation and similar diminished expression patterns after 14 days. This early and sustained underexpression after arterial bypass may lead to altered cell cycle control and matrix protein signaling, contributing to the unregulated proliferation of smooth muscle cells and extracellular matrix in anastomotic intimal hyperplasia after prosthetic arterial grafting.