Reduction of Transplant Vasculopathy by Intraoperative Nucleic Acid-based Therapy in a Mouse Aortic Allograft Model

Abstract

Background Transplant vasculopathy (TV) is the main limiting factor for long-term graft survival characterized by fibrosis, myofibroblast, and smooth muscle cell (SMC) proliferation. Decoy oligodeoxynucleotide (dODN) against the transcription factor activator protein-1 (AP-1) might interfere with the expression of AV-related genes that govern neointima formation. Methods Aortic allografts from DBA/2 mice were incubated with control buffer, consensus, or mutated control AP-1 dODN and were transplanted into the infrarenal aorta of C57BL/6 mice. Cyclosporine A (10 mg/kg body weight [BW]) was administered daily. Explantation and histomorphometric and immunohistochemical evaluation was performed after 30 days. Matrix metalloproteinase (MMP) activity was visualized by gelatin in situ zymography. Results Intima-to-media (I/M) ratio and neointima formation were significantly reduced in the consensus AP-1 dODN treatment group by 37% (p < 0.05) and 67% (p < 0.01), respectively. SMC α-actin-2 staining and macrophage marker expression revealed a marked reduction in the neointima. I/M ratio was found to correlate with the number of tissue macrophages (p < 0.05). MMP and fibrosis marker expression were not significantly altered. Conclusion Intraoperative AP-1dODN utilization might be a strategy to preserve graft function after transplantation.

AAV-Mediated Expression of AP-1-Neutralizing RNA Decoy Oligonucleotides Attenuates Transplant Vasculopathy in Mouse Aortic Allografts

“…15,34 Moreover, we have recently shown that AP-1 dODN application to tissue grafts prior to aortic transplantation can significantly reduce the development of TV in the same animal model by reducing SMC migration and decreasing the infiltration of inflammatory monocytes into the neointima. 35 Although local application of decoy ODNs has been proven effective, our approach provides the possibility to achieve a long-term formation of AP-1 decoy ONs in aortic smooth muscle cells after a single short-time incubation period.…”

Effects of Rapamycin on the Expression of Redox Enzymes in Aortic Vascular Smooth Muscle Cells from Marfan Syndrome Mice

“…Murine aortic SMCs were obtained by explant culture from 6- to 9-week-old homozygous male or female mgR/mgR mice and stained positive for smooth muscle α-actin (ab5694; Abcam, Cambridge, UK) as previously published [11]. In brief, the aorta was dissected between the aortic root and the origin of the renal arteries, and the adventitia was carefully removed.…”