MMP-2 gene silencing attenuates age-dependent carotid stiffness via reduction of elastin degradation and increased eNOS activation

Background and aims Arterial stiffness is a hallmark of vascular aging. Being characterized by a loss of elasticity of large arterial walls, arterial stiffness is associated with an increased risk of cardiovascular disease (CVD). The age-dependent arterial stiffness is primarily attributed to alterations in the elastic and collagen deposition that is regulated by a number of enzymes, including matrix metalloproteinase-2 (MMP-2). Nevertheless, the mechanistic link between age-dependent arterial stiffness and MMP-2 remains unclear. In this study, we investigated the effect and efficacy of therapeutic MMP-2 knockdown using small interfering RNA (siRNA) on age-dependent arterial stiffness. Methods Pulse wave velocity (PWV) was assessed in the right carotid artery of wild-type (WT) mice of different age groups. MMP-2 levels and activity in the carotid artery and plasma of young (3 months) and aged (20–25 months) WT mice were determined. Old WT mice (18–21 months) were treated for 4 weeks with either MMP-2 or scrambled siRNA, in which carotid PWV was assessed at baseline, 2 and 4 weeks after the start of the treatment. Elastin to collagen ratio, desmosin (DES) level, and endothelial nitric oxide synthase (eNOS) pathways were also evaluated and compared. Lastly, levels of circulating MMP-2 and DES, the breakdown product of elastin, were measured in a human cohort (23–86 years old), in whom carotid-femoral PWV was assessed. Results Carotid PWV, as well as both vascular and circulating MMP-2 levels, were elevated with increasing age in WT mice (Figure 1). Therapeutic MMP-2 knockdown in aged WT mice reduced the vascular MMP-2 expression and attenuated age-dependent carotid stiffness. Increased elastin to collagen ratio and a lower plasma DES level were observed on MMP-2 silenced treated animals (Figure 2). Moreover, siMMP-2 treated mice showed enhanced eNOS phosphorylation on Ser1177. A direct interaction between MMP-2 and eNOS was also observed, which, interestingly, is augmented with age. Finally, collected human data showed a higher level of circulating MMP-2 levels on the elderly subjects. In addition, plasma DES level is positively correlated with age and aortic PWV, indicating the involvement of vascular elastin catabolism on arterial stiffness. Conclusions Therapeutic MMP-2 gene silencing, specifically targeting vascular MMP-2, attenuates age-dependent carotid stiffness. This effect is mediated by augmenting eNOS activation and reducing elastin degradation. Thus, our findings indicate MMP-2 as a potential therapeutic target to mitigate age-dependent arterial stiffness and CVD. FUNDunding Acknowledgement Type of funding sources: Foundation. Main funding source(s): Swiss National Science Foundation,Foundation for Cardiovascular Research–Zurich Heart House Figure 1

Arterial stiffness and carotid distensibility following acute formaldehyde exposure in female adults

Formaldehyde (FA) is a ubiquitous organic preservative used in several industries and represents an occupational health hazard. Short-term exposure to FA can increase oxidative stress and cause a decrease in conduit vessel function. These decrements in vascular function may extend to the arterial architecture, predisposing individuals to increased risk of cardiovascular disease. The purpose of this study was to investigate the impact of an acute 90-minute FA exposure period (259 ± 95 ppb) on indices of arterial architecture. Arterial stiffness and carotid distensibility as determined by central pressures, augmentation index (AIx), and carotid-femoral pulse wave velocity (cfPWV) ( n=13F, 24 ± 1 year) as well as carotid stiffness and intima media thickness (IMT) ( n = 9F, 23 ± 1 year) were assessed prior to (Pre-FA) and immediately following (Post-FA) exposure to FA in human cadaver dissection laboratories. Central pressures and cfPWV (Pre-FA: 5.2 ± 0.8 m.s−1, Post-FA: 5.2 ± 1.1 m s−1) were unchanged by acute FA exposure ( p > 0.05). Carotid stiffness parameters and distension were unchanged by acute FA exposure ( p > 0.05), although distensibility (Pre-FA: 33.9 ± 10.5[10–3*kPa−1], Post-FA: 25.9 ± 5.5[10–3*kPa-1], p < 0.05), and IMT (Pre-FA: 0.42 ± 0.05 mm, Post-FA: 0.51 ± 0.11 mm, p < 0.05) decreased and increased, respectively. Individual Pre- to Post-FA changes in these markers of arterial architecture did not correlate with levels of FA exposure ([FA]: 20–473 ppb) ( p > 0.05). Our group previously found vascular function decrements following acute FA exposure in human cadaver laboratories; here we found that carotid distensibility and intima media thickness are altered following FA exposure.

Arterial stiffness as an ultrasound biomarker of radiation-induced carotid artery disease

Summary: Background: Radiation-induced carotid artery disease (RICAD) is an important issue in head and neck cancer (HNC) survivors after radiotherapy (RT). The risk of cerebrovascular disease in these patients is doubled. The aim of this study was to assess the effect of RT on carotid artery stiffness in HNC patients. Patients and methods: Conventional arterial stiffness parameters were measured in a total of 50 HNC survivors treated with RT for at least 5 years and compared to 50 unirradiated HNC patients. Elastic modulus (Ep) and Beta stiffness index (β) were measured in proximal, mid and distal common carotid artery (CCA). Results: The mean age of the subjects was 68±9 years (range: 44–84) in the irradiated group and 67±10 years (range: 45–85) in the control group. The RT group was treated with a mean radiation exposure of 60.3±6.7 Gy (range: 44–72) in the neck. Carotid stiffness parameters showed significant group differences: Ep in the RT group was 2.329±1.222 vs 1.742±828 in the non-RT group (p=0.006) and β index in the RT group was 23±11 vs 15±8 in the non-RT group (p<0.001). Radiation-induced carotid stiffness was quantified and cervical exposure to RT increased Ep in 575 kPa (p=0.014) and β in 7 units (p<0.003). Conclusions: Ep and β index could be suitable ultrasound biomarkers of radiation-induced atherosclerosis in HNC survivors. Further prospective studies are needed to feature RICD in this setting.