c-Myc antisense oligonucleotides preserve smooth muscle differentiation and reduce negative remodelling following rat carotid arteriotomy
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Updated: Aug 18, 2026
Isolation of Primary Patient-specific Aortic Smooth Muscle Cells and Semiquantitative Real-time Contraction Measurements In Vitro
Published on: February 15, 2022
Amalia Forte1, Umberto Galderisi, Marisa De Feo
1Excellence Research Center for Cardiovascular Diseases, Department of Experimental Medicine, Second University of Naples, Via Constantinopoli 16, IT-80138 Naples, Italy.
Objectives:
The vascular biology of restenosis is complex and not fully understood, thus explaining the lack of effective therapy for its prevention in clinical settings. The role of c-Myc in arteriotomy-induced stenosis, smooth muscle cell (SMC) differentiation and apoptosis was investigated in rat carotids applying full phosphorothioate antisense (AS) oligonucleotides (ODNs).
Methods:
Carotid arteries from WKY rats were submitted to arteriotomy and to local application of ODNs through pluronic gel. Apoptosis (deoxynucleotidyl transferase-mediated dUTP nick end-labelling), SMC differentiation (SM22 immunofluorescence) and vessel morphology and morphometry (image analysis) were determined 2, 5 and 30 days after injury, respectively.
Results:
AS ODNs induced a 60% decrease of target c-Myc mRNA 4 h after surgery in comparison to control sense (S) and scrambled ODN-treated carotids (p < 0.05). A significant 37 and 50% decrease in SM22 protein in the media of S ODN-treated and untreated carotids was detected when compared to uninjured contralateral arteries (p < 0.05). This reduction in SM22 expression was prevented in AS ODN-treated carotids. Stenosis was mainly due to adventitial constrictive remodelling. Lumen area in AS ODN-treated carotids was 35% greater than in control arteries 30 days after surgery (p < 0.05). TUNEL assay revealed increased apoptosis in AS ODN-treated carotids (p < 0.05).
Conclusions:
c-Myc AS ODNs reduce arteriotomy-induced negative remodelling. This is accompanied by maintained SMC differentiation and greater apoptosis. The combination of reduced c-Myc-induced proliferation and increased apoptosis may thus underlie the less severe remodelling upon treatment with c-Myc mRNA AS ODN.
Insights
Antisense oligonucleotides targeting c-Myc (AS ODNs) reduced arterial stenosis after injury by maintaining smooth muscle cell differentiation and increasing apoptosis. This suggests a potential therapeutic strategy for preventing restenosis.
Area of Science:
- Vascular Biology
- Molecular Medicine
- Regenerative Medicine
Background:
- Restenosis following vascular injury remains a clinical challenge due to complex underlying mechanisms.
- Effective therapeutic strategies for preventing restenosis are limited.
- The role of c-Myc in vascular smooth muscle cell (SMC) behavior during restenosis is not fully elucidated.
Purpose of the Study:
- To investigate the role of c-Myc in arteriotomy-induced stenosis.
- To evaluate the effect of c-Myc antisense oligonucleotides (AS ODNs) on SMC differentiation and apoptosis.
- To assess the therapeutic potential of targeting c-Myc in preventing vascular remodeling.
Main Methods:
- WKY rats underwent carotid artery injury (arteriotomy).
- Local administration of phosphorothioate antisense (AS) oligonucleotides targeting c-Myc or control oligonucleotides via pluronic gel.
- Assessment of apoptosis (TUNEL assay), SMC differentiation (SM22 immunofluorescence), and vessel morphology/morphometry at 2, 5, and 30 days post-injury.
Main Results:
- AS ODNs significantly reduced c-Myc mRNA levels by 60% within 4 hours post-surgery.
- AS ODNs prevented the decrease in SM22 protein expression, indicating maintained SMC differentiation.
- Arteries treated with AS ODNs exhibited a 35% greater lumen area and increased apoptosis compared to controls at 30 days post-surgery.
- Stenosis was primarily attributed to adventitial constrictive remodeling, which was reduced by AS ODNs.
Conclusions:
- c-Myc AS ODNs effectively reduce arteriotomy-induced negative vascular remodeling.
- Treatment with c-Myc AS ODNs maintains SMC differentiation and promotes apoptosis.
- The combined effect of reduced c-Myc-driven proliferation and enhanced apoptosis underlies the observed therapeutic benefit in mitigating restenosis.
