Amber van Broekhoven

Background: Low‐dose colchicine reduces cardiovascular events in patients with chronic coronary disease. However, side effects may compromise compliance and lead to discontinuation of treatment. Therefore, we aimed to assess the incidence of side effects leading to discontinuation of treatment in the LoDoCo2 (Low‐Dose Colchicine 2) trial. Methods: The LoDoCo2 trial was a randomized controlled trial evaluating colchicine versus placebo in patients with chronic coronary disease. Participants entered a 30‐day open‐label run‐in phase during which all participants received colchicine 0.5 mg once daily, followed by randomization to colchicine or placebo. Tolerability‐related side effects leading to treatment discontinuation were evaluated at the end of the 30‐day run‐in phase and during the randomized phase (median follow‐up, 29.5 [interquartile range, 21.6–47.4] months). Cumulative incidence of side effects was presented using Kaplan–Meier curves and compared by treatment arm using Cox proportional hazards models. Results: During the 30‐day run‐in phase, 618 of 6519 (9.5%) participants reported side effects. Women had a higher risk of side effects than men during the 30‐day run‐in phase (odds ratio, 1.53 [95% CI, 1.23–1.89]). After randomization, 114 participants (4.2%) in the colchicine group and 120 (4.4%) in the placebo group discontinued treatment due to side effects (hazard ratio [HR], 0.95 [95% CI, 0.73–1.22]). Gastrointestinal symptoms were most common (colchicine 2.0% versus placebo 1.7%) followed by myalgia (colchicine 1.2% versus placebo 1.6%). No differences in the risk of side effects between colchicine and placebo were seen for any of the subgroups. Women did have a higher risk of reporting side effects compared with men, regardless of assigned treatment (HR, 1.66 [95% CI, 1.22–2.25]). Conclusions: In patients with chronic coronary disease that tolerated initial treatment with low‐dose colchicine, long‐term discontinuation due to side effects was uncommon and did not differ between colchicine and placebo. Registration: URL: https://www.anzctr.org/au ; Unique identifier: ACTRN12614000093684.

Backgrounds and Aims: Aortic stenosis (AS) involves fibrocalcific aortic valve remodelling, leading to left ventricular outflow obstruction. Innate immune responses can contribute to AS by promoting inflammation, fibrotic, and calcific remodelling. AS presents differently between sexes: males typically show a calcific phenotype, whereas females display more fibrosis. The mechanisms underlying these differences remain unclear. Therefore, this study aimed to investigate sex differences in innate immune phenotype and function in AS. Methods and Results: This cross-sectional study included 119 patients with tricuspid AS (70% male) and 65 healthy controls (52% male). Aortic valves from 23 surgical aortic valve replacement (SAVR) patients (61% male) were examined histologically to assess fibrocalcific remodelling patterns. Blood cell composition, circulating inflammatory markers, and monocyte phenotypes were analysed, with cytokine production evaluated after peripheral blood mononuclear cell (PBMC) stimulation. Males with AS exhibited greater valve calcification, while females demonstrated higher proteoglycan deposition and fibrosis. Males had higher circulating monocyte numbers and percentages, while females had higher lymphocyte percentages. PBMC stimulation induced higher cytokine responses (IL-1β, IL-1Ra) in males, predominantly TLR-3-driven, compared to females. Both sexes displayed higher systemic inflammation versus controls, but with distinct patterns: females had lower platelet counts, higher monocyte CD41 and lower HLA-DR membrane expression, whereas males showed higher cytokine production capacity. Conclusion: This study suggests sex-dependent inflammatory mechanisms in AS. Males exhibited a calcific valve phenotype, while females had more fibrosis. Males had higher monocyte numbers and a more pronounced innate immune response, whereas females showed higher lymphocyte percentages and, compared to controls, an altered monocyte phenotype. ClinicalTrials.gov ID: NCT04717219

BackgroundIn atherosclerotic cardiovascular disease (ASCVD), acute systemic inflammatory insults such as infections or major surgery are known to trigger cardiovascular events. However, it remains unclear whether such systemic triggers also modulate local inflammatory and degenerative processes within the aortic valve (AV). We therefore investigated the effect of orthopedic surgery on AV remodeling in an atherosclerosis-prone setting.MethodsApoE-/- mice subjected to orthopedic surgery, ApoE-/- controls, and non-atherosclerotic controls were analyzed at 5 and 15 days post-intervention. AVs were assessed for valve thickness, fibrosis, glycosaminoglycans (GAGs), lipid accumulation, calcification, iron deposition, and inflammatory cell infiltration.ResultsAtherosclerotic mice exhibited degenerative AV changes compared to non-atherosclerotic controls. Orthopedic surgery did not affect valve thickness or inflammatory cell infiltration, but induced compositional changes, including increased GAG content at day 5 (56.7 vs. 27.3; p=0.016), a trend toward increased fibrosis at day 15 (41.3 vs. 24.9; p=0.068), and reduced lipid content at day 5 (11.3 vs. 37.3; p=0.008). No differences were observed in calcification or iron deposition.ConclusionOrthopedic surgery induces early compositional remodeling of atherosclerotic aortic valves without increasing inflammatory cell infiltration or valve thickening. These findings suggest a role for systemic inflammatory mediators and matrix-driven remodeling, rather than local inflammatory cell influx, in early valve degeneration.

Recent landmark trials showed that colchicine provides a substantial benefit in reducing major cardiovascular events in patients with coronary artery disease. Yet, its exact mechanism of action is still poorly understood. This study aimed to unravel the effect of colchicine on monocyte and neutrophil phenotype and function. A randomized double‐blind placebo‐controlled cross‐over intervention study was executed in patients with a history of myocardial infarction. In neutrophils, colchicine treatment decreased CD62L expression and NGAL release upon ex vivo stimulation and increased PMA‐induced ROS production. The effects of colchicine on monocytes were limited to a decrease in HLA‐DR expression in the intermediate and nonclassical monocytes. Also, on the level of RNA expression, colchicine did not affect monocyte phenotype, while affecting various immunomodulating genes in neutrophils. Overall, our study suggests that treatment with colchicine affects neutrophil function, particularly by reducing neutrophil recruitment, lowering concentrations of NGAL, and changing the expression of various genes with immunomodulatory potential, whereas the effect on monocytes is limited.

Background: Recently, it was shown that 12 weeks of lipopolysaccharide (LPS) administration to nonatherosclerotic mice induced thickening of the aortic heart valve (AV). Whether such effects may also occur even earlier is unknown. As most patients with AV stenosis also have atherosclerosis, we studied the short‐term effect of LPS on the AVs in an atherosclerotic mouse model.Methods: ApoE*3Leiden mice, on an atherogenic diet, were injected intraperitoneally with either LPS or phosphate buffered saline (PBS), and sacrificed 2 or 15 days later. AVs were assessed for size, fibrosis, glycosaminoglycans (GAGs), lipids, calcium deposits, iron deposits and inflammatory cells.Results: LPS injection caused an increase in maximal leaflet thickness at 2 days (128.4 µm) compared to PBS‐injected mice (67.8 µm; P = 0.007), whereas at 15 days this was not significantly different. LPS injection did not significantly affect average AV thickness on day 2 (37.8 µm), but did significantly increase average AV thickness at day 15 (41.6 µm; P = 0.038) compared to PBS‐injected mice (31.7 and 32.3 µm respectively). LPS injection did not affect AV fibrosis, GAGs and lipid content. Furthermore, no calcium deposits were found. Iron deposits, indicative for valve haemorrhage, were observed in one AV of the PBS‐injected group (a day 2 mouse; 9.1%) and in five AVs of the LPS‐injected group (both day 2‐ and 15 mice; 29.4%). No significant differences in inflammatory cell infiltration were observed upon LPS injection.Conclusion: Short‐term LPS apparently has the potential to increase AV thickening and haemorrhage. These results suggest that systemic inflammation can acutely compromise AV structure.

The incidence of heart valve hemangioma is very low and is mostly observed in the mitral and tricuspid valve. In 2006, two cases of aortic valve hemangioma were reported for the first time, including one with calcifying aortic valve stenosis. We now present a case of aortic valve hemangioma in a patient suffering from aortic valve insufficiency combined with atherosclerotic thickening.

Objective: N(ε)‐(carboxymethyl)lysine (CML) is one of the major advanced glycation end products in both diabetics and nondiabetics. CML depositions in the microvasculature have recently been linked to the aetiology of acute myocardial infarction and cognitive impairment in Alzheimer's disease, possibly related to local enhancement of inflammation and oxidative processes. We hypothesized that CML deposition in the microvasculature of the heart and brain is age‐induced and that it could be inhibited by a diet intervention with docosahexaenoic acid (DHA), an omega‐3 fatty acid known for its anti‐inflammatory and antioxidative actions.Materials and methods: ApoE^−/− mice (n = 50) were fed a Western diet and were sacrificed after 40, 70 and 90 weeks. Part of these mice (n = 20) were fed a Western diet enriched with DHA from 40 weeks on. CML in cardiac and cerebral microvessels was quantified using immunohistochemistry.Results: Cardiac microvascular depositions of CML significantly increased with an immunohistochemical score of 11·85 [5·92–14·60] at 40 weeks, to 33·17 [17·60–47·15] at 70 weeks (P = 0·005). At the same time points, cerebral microvascular CML increased from 6·45; [4·78–7·30] to 12·99; [9·85–20·122] (P = 0·003). DHA decreased CML in the intramyocardial vasculature at both 70 and 90 weeks, significant at 70 weeks [33·17; (17·60–47·15) vs. 14·73; (4·44–28·16) P = 0·037]. No such effects were found in the brain.Conclusions: Accumulation of N(ε)‐(carboxymethyl)lysine in the cerebral and cardiac microvasculature is age‐induced and is prevented by DHA in the intramyocardial vessels of ApoE^−/− mice.

Objective—: In aortic aneurysms the arterial vessel wall is dilated because of destruction of its integrity, which may lead to lethal vessel rupture. Chronic infiltration of inflammatory cells into the vessel wall is fundamental to aneurysm pathology. We aim to limit aneurysm growth by inhibition of inflammation and reducing endothelial cell (EC) activation with immunosuppressive drug azathioprine (Aza). Approach and Results—: Aza and its metabolite 6-mercaptopurine have anti-inflammatory effects on leukocytes. We here demonstrate that treatment of ECs with 6-mercaptopurine inhibits cell activation as illustrated by reduced expression of interleukin-12, CCL5, CCL2, and vascular cell adhesion molecule-1 and inhibition of monocyte–EC adhesion. The underlying mechanism of 6-mercaptopurine involves suppression of GTPase Rac1 activation, resulting in reduced phosphorylation of c-Jun-terminal-N-kinase and c-Jun. Subsequently, the effect of Aza was investigated in aneurysm formation in the angiotensin II aneurysm mouse model in apolipoprotein E–deficient mice. We demonstrated that Aza decreases de novo aortic aneurysm formation from an average aneurysm severity score of 2.1 (control group) to 0.6 (Aza group), and that Aza effectively delays aorta pathology in a progression experiment, resulting in a reduced severity score from 2.8 to 1.7 in Aza-treated mice. In line with the in vitro observations, Aza-treated mice showed less c-Jun-terminal-N-kinase activation in ECs and reduced leukocyte influx in the aortic wall. Conclusions—: The immunosuppressive drug Aza has an anti-inflammatory effect and in ECs inhibits Rac1 and c-Jun-terminal-N-kinase activation, which may explain the protective effect of Aza in aneurysm development and, most importantly for clinical implications, aneurysm severity.