AP-1 Oligodeoxynucleotides Reduce Aortic Elastolysis in a Murine Model of Marfan Syndrome

Abstract

Marfan syndrome is characterized by high expression of matrix metalloproteinases (MMPs) in aortic smooth muscle cells (AoSMCs) associated with medial elastolysis and aortic root aneurysm. We aimed to reduce aortic elastolysis through decrease of MMP expression with decoy oligodeoxynucleotides (dODNs) neutralizing the transcription factor activating factor-1 (AP-1). AP-1 abundance in nuclear extracts as well as MMP-2 and MMP-9 expression were significantly increased in isolated mAoSMC of mgR/mgR Marfan mice compared to wild-type cells. Exposure to AP-1 neutralizing dODNs resulted in a significant reduction of basal and interleukin-1β-stimulated MMP expression and activity in mAoSMCs. Moreover, increased migration and formation of superoxide radical anions was substantially decreased in mAoSMCs by AP-1 dODN treatment. Aortic grafts from donor Marfan mice were treated with AP-1- dODN ex vivo and implanted as infrarenal aortic interposition grafts in mgR/mgR mice. Pretreatment of aortic grafts with AP-1 dODN led to reduced elastolysis, macrophage infiltration, and MMP activity. Permeability of the endothelial monolayer was increased for dODN in mgR/mgR aortae with observed loss of tight junction proteins ZO-1 and occludin, enabling dODN to reach the tunica media. Targeting AP-1 activity offers a new potential strategy to treat the vascular phenotype associated with Marfan syndrome.

The reimplantation valve-sparing aortic root replacement technique for patients with Marfan syndrome: A single-center experience

“…Noteworthy is the observation that patients with MFS tend to develop at least one episode of postoperative blood-culture negative fever, without elevation of infectious parameters. This finding may be related to the enhanced inflammatory aspect of the aortic wall in MFS and is currently debated 22,23 . This phenomenon has been described after aortic surgery and may also be related to other factors such as endothelialization of the prosthesis 24 .…”

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

“…The effectiveness can be explained by functional AP-1 binding sites in the promoter of this cell-cycle regulator protein 37 . In addition, AP-1 decoy ODN has been described to significantly decrease SMC migration and arterial tissue remodeling 38 …”

“…The hairpin AP-1 dODN was designed as a single-stranded homoduplex molecule consisting of a small loop and a long beacon stem that contains the transcription factor binding site. The initially single-stranded hairpin AP-1 dODN quickly hybridizes to itself representing the active double-stranded ODN subsequently modulating effective gene expression 38 , 57 …”

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

“…The effectiveness of the on-time treatment can be explained, at least in part, by the observation that the AP-1 dODN significantly decrease SMC migration, proliferation, and arterial tissue remodeling. 15 Targeting of transcription factors known to activate expression of disease-related genes by using doublestranded transcription factor dODNs represents a pivotal therapeutic treatment option which already moved into clinical trials. 11 In line with our results, AP-1 dODNs have been used so far in preclinical trials as co-medication and significantly reduced neointima formation and restenosis after PTCA in hypercholesterolemic minipigs, 8 venous bypass graft grafting in rabbits 22 and CAV in a fully allogenic rat heart transplantation model.…”

“…Hairpin AP-1 dODN (purity !95%) were purchased from Biomers (Ulm, Germany) and prepared by self-annealing of single-stranded and self-complementary phosphorothionated dODNs as described previously. 15 A mutated form of AP-1 dODN was used as a control for the nonspecific actions of dODN administration. The sequences of the singlestranded ODNs were as follows (bold letters denote phosphorothioate-bonded bases, mutations are set in italics).…”