Plaque Inflammation Research Articles

Coronary plaque burden predicts perioperative cardiovascular events after coronary endarterectomy.

The risk factors of perioperative and long-term cardiovascular events in patients undergoing coronary artery bypass grafting (CABG) with adjunctive coronary endarterectomy (CE) are not well determined. This study evaluated the clinical value of coronary plaque burden, coronary anatomic stenosis, and serum biomarkers for predicting perioperative cardiovascular events after off-pump CABG + CE. This retrospective cohort single-center study enrolled 125 patients undergoing off-pump CABG + CE between February 2018 and September 2021 in China. Coronary plaque burden was reflected by the length of plaque removed by CE. Plaque length-max, which represents the plaque length in patients undergoing single-vessel CE and the maximum plaque length in patients undergoing multivessel CE, was calculated. The primary endpoint was perioperative myocardial infraction (PMI). Plaque length-max was significantly higher in patients with PMI than in those without PMI (2.4 ± 1.5 vs. 1.6 ± 0.9, p = .001). A threshold plaque length-max of 1.15 cm was an independent predictor of PMI (area under the curve: 0.67; sensitivity 87.9%; specificity 59.8%; p = .005). Patients with plaque length-max ≥1.15 had a > 5-fold increase in PMI after adjusting for confounding factors (odds ratio = 5.89; p = .002). Furthermore, interleukin-6 (Beta = .32: p = .028), CD68 (Beta = .34; p = .045), and osteopontin (Beta = .43; p = .008) were significantly correlated with plaque length-max. Plaque length-max was superior to clinical cardiovascular risk factors in predicting PMI occurrence after off-pump CABG + CE, which might be associated with systemic and plaque inflammation state.

Blockade of IL-6 signaling alleviates atherosclerosis in Tet2-deficient clonal hematopoiesis.

Clonal hematopoiesis (CH) increases the risk of atherosclerotic cardiovascular disease possibly due to increased plaque inflammation. Human studies suggest that limitation of interleukin-6 (IL-6) signaling could be beneficial in people with large CH clones, particularly in TET2 CH. Here we show that IL-6 receptor antibody treatment reverses the atherosclerosis promoted by Tet2 CH, with reduction of monocytosis, lesional macrophage burden and macrophage colony-stimulating factor 1 receptor (CSF1R) expression. IL-6 induces expression of Csf1r in Tet2-deficient macrophages through enhanced STAT3 binding to its promoter. In mouse and human Tet2-deficient macrophages, IL-6 increases CSF1R expression and enhances macrophage survival. Treatment with the CSF1R inhibitor PLX3397 reversed accelerated atherosclerosis in Tet2 CH mice. Our study demonstrates the causality of IL-6 signaling in Tet2 CH accelerated atherosclerosis, identifies IL-6-induced CSF1R expression as a critical mechanism and supports blockade of IL-6 signaling as a potential therapy for CH-driven cardiovascular disease.

Monocyte Differentiation‐Surveilling Nano‐Shuttles for Activatable Targeted Inhibition of Atherosclerotic Plaque Progression

AbstractAtherosclerosis can form intimal plaques in the arteries and undergo plaque rupture with followed stroke or myocardial infarction under certain pathological conditions. However, suboptimal intraplaque accumulation, along with severe off‐target effects, remain formidable obstacles for efficient pharmacotherapy of atherosclerosis. A characteristic feature of the progression of atherosclerosis is the continuous recruitment of inflammatory monocytes that differentiate into foamy macrophages with the generation of legumain in the plaques. In this study, a bio‐activatable nano‐shuttle based on human serum albumin is conceived that contained curcumin to allow the precise inhibition of atherosclerosis progression. Following intravenous injection, the nano‐shuttles are almost exclusively engulfed by inflammatory monocytes in an efferocytosis‐mimetic manner and selectively co‐migrated with the monocytes towards the plaques. The nano‐shuttles are readily activated to release the initially arrested curcumin in response to the generation of legumain upon the differentiation of monocytes into foamy macrophages within the plaques. Treatment assessments on atheroprone apolipoprotein‐E‐deficient mice indicated that the nano‐shuttles substantially inhibit the progression of atherosclerosis by attenuating the plaque inflammation burden without any obvious adverse effects. The integration of highly selective accumulation and specific drug activation of the nano‐shuttles in plaques can arouse an advanced therapeutic strategy to prevent atherosclerotic cardiovascular disease.

Air pollution and plaque healing in acute coronary syndromes.

Plaque healing plays a key role in coronary atherosclerosis1 and represents ‘the second hit’ required to cause an acute coronary syndrome (ACS) after plaque destabilization, this latter being the ‘first hit’.1–3 Moreover, an impaired plaque healing has been found in patients with recurrent ACS, suggesting its importance in preventing repeated symptomatic coronary thrombosis.3 Nonetheless, mechanisms modulating plaque healing are unknown. Air pollution is an important yet overlooked cardiovascular risk factor.4–6 We recently demonstrated that increased long-term particulate matter 2.5 (PM2.5) exposure was associated with coronary plaque inflammation, vulnerability and rupture at optical coherence tomography (OCT) analysis in ACS patients.4 However, the relationship between plaque healing and air pollution has not been yet determined. We investigated the association between air pollution and healed plaques in patients with ACS, stratified by first or recurrent ACS, and whether the previous observation that PM2.5 exposure is associated with coronary plaque vulnerability is valid also in patients with recurrent ACS.5

Systems immunology-based drug repurposing framework to target inflammation in atherosclerosis

The development of new immunotherapies to treat the inflammatory mechanisms that sustain atherosclerotic cardiovascular disease (ASCVD) is urgently needed. Herein, we present a path to drug repurposing to identify immunotherapies for ASCVD. The integration of time-of-flight mass cytometry and RNA sequencing identified unique inflammatory signatures in peripheral blood mononuclear cells stimulated with ASCVD plasma. By comparing these inflammatory signatures to large-scale gene expression data from the LINCS L1000 dataset, we identified drugs that could reverse this inflammatory response. Ex vivo screens, using human samples, showed that saracatinib—a phase 2a-ready SRC and ABL inhibitor—reversed the inflammatory responses induced by ASCVD plasma. In Apoe−/− mice, saracatinib reduced atherosclerosis progression by reprogramming reparative macrophages. In a rabbit model of advanced atherosclerosis, saracatinib reduced plaque inflammation measured by [18F]fluorodeoxyglucose positron emission tomography–magnetic resonance imaging. Here we show a systems immunology-driven drug repurposing with a preclinical validation strategy to aid the development of cardiovascular immunotherapies.

Puerarin-Tanshinone IIA Suppresses atherosclerosis inflammatory plaque via targeting succinate/HIF-1α/IL-1β axis

Ethnopharmacological relevanceInflammatory injury is an important pathological factor for the formation of atherosclerotic plaque. It is well known that Puerarin and Tanshinone IIA (Pue-Tan) can significantly reduce interleukin-1β (IL-1β) levels and delay the atherosclerosis (AS) process clinically in China. Previous evidence has shown that the Succinate/HIF-1α/IL-1β inflammatory signaling axis (Succinate axis) promotes the progression of atherosclerotic inflammatory plaques. It is not clear whether Pue-Tan inhibits inflammatory plaques by reducing the level of IL-1β through the succinate signaling axis. Aim of studyFind out the interaction between Pue-Tan targets and the succinate axis by means of network pharmacology and bioinformatics analysis and to further confirm whether Pue-Tan can inhibit vascular inflammation and delay the formation of atherosclerotic inflammatory plaques by targeting the succinate signaling axis. Materials and methodsFirstly, animal experiments were conducted to verify the changing relationship between Succinate and IL-1β under Pue-Tan intervention. Secondly, network pharmacology approach was employed to uncover the specific targets of Pue-Tan in the intervention of AS from multiple levels of components, proteins, and pathways, and at the same time, the target must be a key factor of the succinate signaling axis. Autodock vina1.5.6 was applied to molecular docking for Pue-Tan and target protein. Subsequently, cells experiment and animal experiment were performed to verify Pue-Tan inhibiting the inflammatory progression of atherosclerosis by targeting succinate signaling axis. ResultsFirstly, we first found that the reduction of IL-1β was positively correlated with succinate in the serum of Pue-Tan-treated mice. Secondly, network pharmacology compared with molecular docking showed that hypoxia-induced factor-1α (HIF-1α) was the key target of Pue-Tan and the key node of succinate singling axis. Finally, in vitro study, Pue-Tan significantly reduced the factors of succinate axis just as HIF-1α siRNA; in vivo study, we confirmed a decreased expression of succinate axis and ICAM-1 in the aorta of ApoE−/− mice under Pue-Tan intervention, which was consistent with the in vitro results. ConclusionThis study confirmed that Pue-Tan blocked the succinate axis by targeting HIF-1α to prevent the formation of atherosclerotic inflammatory plaques and delay the pathological process of AS. Network Pharmacology, Bioinformatics of Molecular Docking, and Molecular Biology Validation can be used as a effective way to discover and verify the pharmacological mechanism of TCM.

The impact of ischaemic stroke risk factors on imaging changes – a current literature review

Stroke is the leading cause of acquired disability in adults. Although all age groups may be affected, the risk of fatal outcome increases with age. Diagnostic imaging, including computed tomography and magnetic resonance imaging, is used in order to promptly initiate effective treatment. Imaging modalities make it possible to determine the type of stroke as well as the location and extent of damage. They are also used for treatment monitoring. It is essential to make an early diagnosis, identify the type of stroke and implement appropriate management. After the introduction of modern treatment modalities, such as thrombolytic therapy, time windows, i.e. periods of time during which a given intervention may bring benefits to the patient, were established for ischaemic stroke. The performance and life quality in a person after a stroke depends on the duration of neural ischaemia. The consequences of stroke also depend on the baseline condition of the patient. The incidence of stroke in the general population is shaped by risk factors that are often cumulative in nature. Scientific reports most often point to age, race, sex, blood glucose and diabetes, obesity, smoking and atherosclerosis. Ischaemic stroke is frequently caused by arterial stenosis or obstruction. According to one hypothesis, the activity of immune cells, including monocytes and basophils, in atherosclerosis leads to inflammation and formation of atherosclerotic plaques. As a result, wall thickening, atherosclerotic plaque rupture, regenerative responses and stenosis occur. The aim of the paper is to summarise the relationships between vasculitis, bacterial and viral infections or autoimmune disorders and stroke, in the etiopathogenesis of which, according to current research, they are involved. The paper also shows the importance of diagnostic imaging and attempts to explain the relationship between changes in computed tomography and magnetic resonance imaging and risk factors for stroke.

(+)-CatechinAttenuatesMultipleAtherosclerosis-Associated Processes In Vitro, Modulates Disease-Associated Risk Factors in C57BL/6J Mice and Reduces Atherogenesis in LDL Receptor Deficient Mice by Inhibiting Inflammation and Increasing Markers of Plaque Stability.

A prospective study of 34492 participants shows an inverse association between (+)-catechin intake and coronary heart disease. The effects of (+)-catechin on atherosclerosis and associated risk factors are poorly understood and are investigated. (+)-Catechin attenuates reactive oxygen species production in human macrophages, endothelial cells and vascular smooth muscle cells, chemokine-driven monocytic migration, and proliferation of human macrophages and their expression of several pro-atherogenic genes. (+)-Catechin also improves oxidized LDL-mediated mitochondrial membrane depolarization in endothelial cells and attenuates growth factor-induced smooth muscle cell migration. In C57BL/6J mice fed high fat diet (HFD) for 3 weeks, (+)-catechin attenuates plasma levels of triacylglycerol and interleukin (IL)-1β and IL-2, produces anti-atherogenic changes in liver gene expression, and reduces levels of white blood cells, myeloid-derived suppressor cells, Lin- Sca+ c-Kit+ cells, and common lymphoid progenitor cells within the bone marrow. In LDL receptor deficient mice fed HFD for 12 weeks, (+)-catechin attenuates atherosclerotic plaque burden and inflammation with reduced macrophage content and increased markers of plaque stability; smooth muscle cell and collagen content. This study provides novel, detailed insights into the cardio-protective actions of (+)-catechin together with underlying molecular mechanisms and supports further assessments of its beneficial effects in human trials.

(153) Investigating the Role of NELL1 as a Gene of Interest in Peyronie’s Disease

Abstract Introduction Peyronie’s Disease (PD) is a superficial fibrosing disorder characterized by inelastic fibrous plaques within the tunica albuginea. We previously performed high-resolution array comparative genomic hybridization analysis of genomic DNA from men with both PD and Dupuytren’s Disease (DD), yielding candidate genes, including NELL1, with potential involvement in these conditions. NELL1 is a secreted growth factor that enhances osteoblastic differentiation and chondrogenesis and may contribute to calcification and inflammation in fibrotic plaques via its interaction with bone morphogenetic protein (BMP) within the TGF-β signaling pathway. Here, we describe the creation of lentiviral gRNA vectors for the up- and down-regulation of NELL1 in an in vitro setting, as well as PCR results for NELL1 regulation and preliminary results from RNA sequencing following CRISPR modification. Objective To determine the influence of NELL1 on extracellular matrix (ECM) deposition and downstream gene regulation. Methods For down-regulation of NELL1, gRNAs were delivered to HEK293 cells with a nuclease deficient (d) dCas9-KRAB system adapted from Streptococcus pyogenes. For upregulation, gRNAs were delivered to HEK293 cells expressing the dCas9-VPR system adapted from Streptococcus pyogenes. Gene regulation via these systems was assessed using qRT-PCR. ECM deposition was evaluated using primary antibodies again Collagen I, Collagen III, and Elastin, and the appropriate secondary antibodies. RNA sequencing and analysis was performed by the High-Throughput Genomics and Bioinformatic Analysis core at the University of Utah. Results For up-regulation, four vectors were tested, with the most effective vector demonstrating a 40-fold increase in NELL1 expression via PCR testing. For down-regulation, three vectors were tested, with the most effective demonstrating a 95% reduction in expression via PCR testing (Figure 1). Non-target vectors did not change NELL1 expression levels. RNA sequencing results found over 10,000 differentially expressed genes between up-regulated and non-target cell lines, with significant changes in multiple pathways, including TNF-α signaling. Similarly, over 5,000 differentially expressed genes were found between down-regulated and non-target cell lines, with significant changes in inflammatory and G2-M checkpoint pathways, among others. Ongoing experiments are also testing the amount of extra-cellular matrix (ECM) deposited by up- and down-regulated cells and their non-target controls, in order to determine if changes in NELL1 expression influence ECM deposition. Conclusions We have demonstrated the creation of effective guides for the up- and down-regulation of NELL1, a gene of interest in Peyronie’s disease. We have also demonstrated, through RNA sequencing, that changes in NELL1 lead to large changes in gene and pathway expression. Ongoing experiments will determine if changes in NELL1 expression also affect ECM deposition. Disclosure Any of the authors act as a consultant, employee or shareholder of an industry for: Vault Health.

Effects of blue-light LED toothbrush on reducing dental plaque and gingival inflammation in orthodontic patients with fixed appliances: a crossover randomized controlled trial

BackgroundPatients with fixed orthodontic appliances have higher plaque accumulation and gingival inflammation. Our aim was to compare the effectiveness of a light emitting diode (LED) toothbrush with a manual toothbrush in reducing dental plaque and gingival inflammation in orthodontic patients with fixed appliances, and to investigate the effect of the LED toothbrush on Streptococcus mutans (S. mutans) biofilm in vitro.MethodsTwenty-four orthodontic patients were recruited and randomly assigned into 2 groups: (1) started with manual and (2) started with LED toothbrushes. After a 28-day usage and 28-day wash-out period, the patients switched to the other intervention. The plaque and gingival indices were determined at baseline and 28 days after each intervention. The patients’ compliance and satisfaction scores were collected using questionnaires. For the in vitro experiments, S. mutans biofilm was divided into 5 groups (n = 6) with 15-, 30-, 60-, or 120-sec LED exposure, and without LED exposure as a control group.ResultsThere was no significant difference in the gingival index between the manual and LED toothbrush groups. The manual toothbrush was significantly more effective in reducing the plaque index in the proximal area on the bracket side (P = 0.031). However, no significant difference was found between the two groups in other areas around the brackets or on the non-bracket side. After LED exposure in vitro, the percentages of bacterial viability after LED exposure for 15–120 s were significantly lower compared with the control (P = 0.006).ConclusionClinically, the LED toothbrush was not more effective in reducing dental plaque or gingival inflammation than the manual toothbrush in orthodontic patients with fixed appliances. However, the blue light from the LED toothbrush significantly reduced the number of S. mutans in biofilm when it was exposed to the light for at least 15 s in vitro.Clinical Trial RegistrationThai Clinical Trials Registry (TCTR20210510004). Registered 10/05/2021.

Lack of Rab27a attenuates foam cell formation and macrophage inflammation in uremic apolipoprotein E knockout mice.

As the most common cardiovascular disease, atherosclerosis (AS), is a leading cause of high mortality in patients with chronic renal failure. Rab27a has been reported to regulate the progression of cardiovascular and renal diseases. Nevertheless, little studies investigated the role and mechanism of Rab27a in uremic-accelerated AS (UAAS). An animal model of UAAS was established in apolipoprotein E knockout (apoE-/-) mice using 5/6 nephrectomy (NX). We conducted in vitro and in vivo functional experiments to explore the role of Rab27a in UAAS, including the presence of oxidized low-density lipoprotein (ox-LDL). Rab27a expression was upregulated in the plaque tissues of NX apoE-/- mice. The knockout of Rab27a (Rab27a-/-) reduced AS-induced artery injury, as manifested by the reductions of plaque area, collagen deposition, inflammation and lipid droplet. Besides, cholesterol efflux was increased, while the expression of lipid metabolism-related proteins and the secretions of pro-inflammatory factors were decreased in ox-LDL-induced NX Rab27a-/- apoE-/-mice group. Further, Rab27a deletion inhibited the activation of nuclear factor κB (NF-κB) pathway. In conclusion, our study indicated that Rab27a deficiency attenuated foam cell formation and macrophage inflammation, depending on the NF-κB pathway activation, to inhibit AS progression in uremic apoE-/- mice. This finding may provide a new targeting strategy for UAAS therapy.

Erisipelas and its Management in a Tertiary Care Hospital

Erysipelas is an acute dermo-hypodermic (non-necrotizing) infection of bacterial origin, mainly group A beta-hemolytic streptococcus. The lower limbs are affected in more than 80% of cases and the risk factors identified are skin barrier breakdown, lymphedema and obesity. Diagnosis is clinical and is based on the association of an acute inflammatory plaque with fever, lymphangitis, lymphadenopathy and leukocytosis. Bacteriology is generally not useful due to low sensitivity or late positivity. In atypical forms, erysipelas should be distinguished from necrotizing fasciitis and acute venous thrombosis. Penicillin remains the gold standard treatment, although new drugs may be used, given their pharmacodynamic profile. Recurrence is the main complication, being crucial the correct treatment of risk factors1.

IL‑37 suppresses macrophage ferroptosis to attenuate diabetic atherosclerosis via the NRF2 pathway.

IL-37 is a newly discovered inflammatory factor. However, the protective effect and underlying mechanisms of IL-37 on atherosclerosis remain unclear. In the present study, IL-37 was used for intraperitoneal injection in diabetic ApoE-/- mice caused by streptozotocin. High glucose (HG)/ox-LDL was used to stimulate THP-1 original macrophage followed by IL-37 pretreatment in vitro. The atheromatous plaque area, oxidative stress and inflammation levels in ApoE-/- mice were evaluated, and the level of macrophage ferroptosis was detected in vivo and in vitro. It was identified that IL-37 treatment significantly decreased plaque area in diabetic ApoE-/- mice. IL-37 not only improved blood lipid levels in mice, but also reduced serum levels of inflammatory factors including IL-1β and IL-18. Furthermore, IL-37 increased GPX4 and nuclear factor erythroid 2-related factor 2 (NRF2) in the aorta of diabetic mice. In vitro experiment revealed that IL-37 inhibited HG/ox-LDL-induced ferroptosis in macrophages, as evidenced by improved cell membrane oxidation, reduced malondialdehyde production and increased GPX4 expression. Moreover, it was also found that IL-37 enhanced the nuclear translocation of NRF2 in macrophages, while ML385, a specific NRF2 inhibitor, significantly attenuated the protective effect of IL-37 on macrophage ferroptosis caused by HG/ox-LDL. In conclusion, IL-37 suppressed macrophage ferroptosis to attenuate atherosclerosis progression via activating the NRF2 pathway.

Unmasking Arterial-resident Autoreactive B cells involved in Atherosclerosis and Peripheral Artery Disease Progression

Abstract Atherosclerosis activates the immune system, including B cells which secrete antibodies that bind antigens released by necrotic cells within plaques and may promote inflammation. To identify these antibodies, we isolated activated B cells from atherosclerotic ApoE-deficient mice aortas and human tibial arteries with peripheral artery disease post-amputation. Single-cell sequencing of the B cell receptors identified 35 mouse and 48 human overrepresented disease-relevant heavy and light chain pairings, which were sequentially expressed as antibody. Epitope determination was performed by human proteome array, and 14 mouse and 21 human protein targets were found with significant affinity and specificity. Mouse antigens were scrutinized by immunohistochemistry to confirm presence in inflamed plaques, and expression of these proteins were used to interrogate autoreactive antibody titers in diseased sera by ELISA. ApoE-deficient mice were immunized with the 6 most promising antigens. Serum antibody measurement showed strong, rapid, and early onset of antigen-specific antibody responses in the immunized mice compared to an adjuvant only control group. Strikingly, we saw decreased plaque accumulation in the immunized animals, suggesting antigen responses may clear pathogenic antigens before disease develops. Likewise, human antigen ELISAs comparing antigen-specific serum antibody titers to patient Framingham score for cardiovascular risk supported a role for two antigens in disease. To confirm these antigens as potential biomarkers, we will characterize the emergence of their antibody counterparts using a historical library of serum samples from individuals enrolled in the Baltimore Longitudinal Study on Aging. This work is directly funded by the National Institutes of Health – National Institute on Aging’s Intramural Research Program in the Lab of Molecular Biology and Immunology

Abstract 311: Fatty Acid Synthase Targeting Reduces Aortic Atherosclerosis And Inflammation

Fatty acid synthase (FAS) catalyzes the elongation of saturated fatty acids. FAS is elevated in liver and adipose tissue in the setting of diabetes, and serum circulating FAS (cFAS) is a marker of peripheral arterial disease. The natural antibiotic Platensimycin (PTM) competitively inhibits FAS. However, the impact of chemical FAS inhibition on atherosclerosis is unknown. We hypothesized that PTM and a liver-specific knockdown of FAS would reduce the burden of aortic atherosclerosis. ApoE-/- mice were placed on a 42% high-fat diet (HFD) with or without PTM (100mpk) for 16 weeks. Similarly, ApoE-/- mice with or without liver-specific knockdown of FAS ( ApoE-/- FasL-/- vs ApoE-/- FasL+/+ ) were placed on a 42% HFD for 16 weeks (Fig. A). Tissue FAS, cFAS, and serum triglycerides (TG) content were measured. FAS/cFAS activity was evaluated with a modified measurement of malonyl-CoA consumption of NADPH, and normalized to content. At 16 weeks, mouse aortas were harvested and stained with oil-red O and CD68 to assess the burden of atherosclerosis and inflammation. Comparisons between murine groups were analyzed using Mann-Whitney and Student t-tests. ApoE-/- mice treated with PTM, significantly reduced tissue FAS and serum cFAS activity (p<0.05), and significantly reduced serum TG (p<0.01). ApoE-/- FasL-/- mice demonstrated increased adipose FAS content and activity (p<0.05), reduced cFAS activity (p<0.05), and increased serum TG (p<0.05). Targeting FAS/cFAS with PTM or in FasL-/- led to decrease in overall total aortic plaque, particularly in the low-resistance thoracic aorta (B; p<0.05).Targeting FAS/cFAS also demonstrated reduced aortic root plaque, and macrophage content(C; p<0.05).This study reveals that selective targeting of FAS/cFAS either with PTM of via liver knockdown, appear to significantly alter serum lipid profiles, aortic atherosclerosis, and plaque inflammation. Thus, FAS/cFAS may be an important therapeutic target to inhibit atheroprogression.

Abstract 673: CHROMR Governs Interferon Signaling And Cholesterol Homeostasis In Human Atherosclerotic Plaque

Long noncoding RNAs (lncRNA) have emerged as critical regulators of gene expression, yet their potential contribution to human disease remains poorly understood. We previously identified the primate-specific lncRNA CHROMR as a key regulator of cholesterol homeostasis. Here, we report a novel function for CHROMR in controlling interferon signaling, which is known to regulate innate and adaptive immune responses that drive atherosclerosis. Using single cell RNA-sequencing of human atheroma, we observed robust expression of CHROMR in plaque immune cell populations, which correlated strongly with the expression of two subsets of interferon-stimulated genes (ISG). Interrogation of these genesets using DisGeNET, a comprehensive catalogue of genes and variants associated to human diseases, revealed associations with atherosclerosis and coronary artery disease. Differential expression analysis of CHROMR -expressing versus non-expressing immune cells showed higher levels of ISGs particularly in the monocyte-macrophage cluster. To assess the impact of CHROMR depletion on inflammatory gene expression, we knocked down CHROMR in THP1 macrophages using GapmeR antisense oligonucleotides (or controls) and treated with the TLR3 agonist poly(I:C) to stimulate the interferon response. Transcriptome profiling and unsupervised hierarchical clustering of genes differentially expressed in CHROMR -sufficient and -depleted macrophages revealed that CHROMR silencing markedly repressed expression of key ISGs, including MX1 , IFI44L , STAT1 , and CXCL and CCL family chemokines ( CXCL10 , CXCL11, CCL2 ). Conversely, overexpression of CHROMR in THP1 macrophages increased expression of these ISGs. Furthermore, CHROMR depletion in macrophages reduced histone acetylation at regulatory regions of ISG loci. Mechanistically, we showed that CHROMR sequesters the interferon regulatory factor (IRF)-2-dependent transcriptional co-repressor IRF2BP2, thereby licensing IRF-dependent signaling and transcription of the ISG network. Taken together, our work unveils a critical role for CHROMR in coordinating interferon signaling responses in macrophages and suggests that targeting this lncRNA may reduce inflammation in atherosclerotic plaques.

Abstract 126: Effect Of Icospaent Ethyl On In Vivo Atheroma Progression And Inflammatory Protease Activity

Background: Icosapent ethyl (IPE), a high purity ethyl ester of eicosapentaenoic acid (EPA), is FDA-approved to reduce cardiovascular events among patients with or at risk for cardiovascular disease with elevated triglyceride levels. However, in vivo mechanisms underlying these findings remain unclear. Here we investigated whether IPE reduces plaque progression and inflammation using intravascular molecular-structural imaging. Methods: Cholesterol-fed balloon-injured rabbits (N=16) bearing aortic atherosclerosis underwent intravascular ultrasound (IVUS) analysis of plaque burden and near-infrared fluorescence-optical coherence tomography (NIRF-OCT) molecular imaging of plaque inflammatory cathepsin protease activity (ProSense VM110 NIRF agent, 4 mg/kg/iv) at 6 weeks. Next, rabbits were randomized to control (N=8) or IPE (Vascepa, Amarin, NJ) treated group (N=8, 1 g/kg/day) milled in the diet. Four weeks later, imaging was repeated, animals were sacrificed, and histopathology analysis of dissected plaque was employed. Results: Quantitative IVUS analyses (40mm segment, 3200 slices/16 rabbits) revealed that IPE significantly suppressed atheroma progression (ΔPAV, 5.2% [2.36-12.98] vs. 9.1% [4.26-12.25] control, p=0.03). Inflammatory plaque cathepsin activity detected by NIRF-OCT was similar (ΔNIRF average plaque 17.6 [6.2- 51.5] vs. 30.5 [4.9- 55.5] nM control, p=0.19)). On fluorescence microscopy, however, plaques from IPE-treated animals exhibited reduced NIRF signal (1.16% [0.03-3.94] vs. 3.13% [0.04-12.92], p=0.005)) and fewer RAM11+ macrophages (23.5±15.6 vs. control 32.9±12.6 macrophages/section, 25 sections/group, p=0.04). In IPE-treated animals, plasma EPA levels were elevated (p<0.05), and EPA metabolite levels, particularly 5,6-diHETE, were also elevated in resected plaques (p<0.05). Conclusion: IPE reduces in vivo plaque progression and plaque cathepsin protease activity and plaque macrophages, providing novel experimental evidence supporting the anti-atherosclerosis effects of IPE.

Comparative Evaluation of Novel Herbal Mouthwash with 12.5% Xylitol Mouthwash and 0.7 M Saline Mouthwash on Plaque Control: An Interventional Study

Background and Aim: Among the wide array of herbal plants possessing medicinal values, Andrographis paniculata (Nilavembu) stands out as a time-tested premiere medicinal herb. A. paniculata (Nilavembu) extracts exhibited varying degrees of inhibitory activity against the growth of all the microorganisms. Hence, this research was made to assess and compare the effect of Nilavembu extract (A. paniculata) with 12.5% xylitol mouthwash and control group (0.7 M saline mouthwash) on dental plaque and gingival inflammation. Materials and Methods: A double-blinded parallel-arm randomized clinical trial was done among 40 participants aged 20–30 years. Participants were randomly allotted into four groups: Group 1 - Nilavembu extract with 0.7 M saline wash; Group 2 - Nilavembu extract with 12.5% xylitol mouthwash; Group 3-0.7 M saline mouthwash; and Group 4-12.5% xylitol mouthwash. The plaque and gingival status were assessed using oral hygiene simplified index and Loe and Silness index at baseline and end of the study (7th day). Statistical analysis was carried out to compare the effect of all four treatments groups using the Kruskal–Wallis test, Wilcoxon sign rank test, and post hoc test; P ≤ 0.05 was considered statistically significant. Results: Among four groups, Group 2 showed a greater mean reduction followed by Group 1 in oral hygiene index score and gingival score. There was a statistically significant difference between the scores of the four groups. Conclusion: It can be concluded that A. paniculata (Nilavembu extract) could serve as a potential herbal alternative in the therapeutic management of dental plaque and gingivitis.

Creeping Attachment Observed in RT2 following Labial Frenectomy: A Case Report

Introduction: Aberrant frenal attachment and shallow vestibule in the anterior mandible facilitates plaque accumulation, inflammation, and root exposure of the mandibular central incisors, especially where the “pull syndrome” is noted. Case Report: A 22 years old systemically healthy female patient complained of slight mobility and increased spacing in relation to lower front teeth. Clinical examination revealed Cairo recession type 2 (RT2) gingival recession in relation to mandibular central incisors and hyperplastic frenum with fan shaped attachment to gingival margin of mandibular central incisors. Frenectomy was performed using conventional surgical technique. After 14 days of surgery 0.5 mm of creeping attachment as well as reduction in recession width at apical level of recession was observed in relation to mandibular central incisors. The achieved clinical results remained stable till 6 months of follow-up. Conclusion: This case report suggests that frenectomy using conventional surgical technique may yield acceptable clinical outcomes and esthetics in RT2 gingival recession.

Atherosclerosis and Inflammation: Insights from the Theory of General Pathological Processes.

Recent advances have greatly improved our understanding of the molecular mechanisms behind atherosclerosis pathogenesis. However, there is still a need to systematize this data from a general pathology perspective, particularly with regard to atherogenesis patterns in the context of both canonical and non-classical inflammation types. In this review, we analyze various typical phenomena and outcomes of cellular pro-inflammatory stress in atherosclerosis, as well as the role of endothelial dysfunction in local and systemic manifestations of low-grade inflammation. We also present the features of immune mechanisms in the development of productive inflammation in stable and unstable plaques, along with their similarities and differences compared to canonical inflammation. There are numerous factors that act as inducers of the inflammatory process in atherosclerosis, including vascular endothelium aging, metabolic dysfunctions, autoimmune, and in some cases, infectious damage factors. Life-critical complications of atherosclerosis, such as cardiogenic shock and severe strokes, are associated with the development of acute systemic hyperinflammation. Additionally, critical atherosclerotic ischemia of the lower extremities induces paracoagulation and the development of chronic systemic inflammation. Conversely, sepsis, other critical conditions, and severe systemic chronic diseases contribute to atherogenesis. In summary, atherosclerosis can be characterized as an independent form of inflammation, sharing similarities but also having fundamental differences from low-grade inflammation and various variants of canonical inflammation (classic vasculitis).