Aortic Aneurysm Formation Research Articles

Alternative therapeutic strategy for existing aortic aneurysms using mesenchymal stem cell–derived exosomes

ABSTRACT Background Several studies demonstrated the therapeutic potential of mesenchymal stem cell–derived exosomes (MSC-exs) based on their anti-inflammatory properties. The objective was to determine the therapeutic effects of MSC-exs on aortic aneurysms (AAs) caused by atherosclerosis. Research design and methods Apolipoprotein E knockout mice with AAs induced by angiotensin II were injected with MSC-exs or saline as a control. The change in the diameter of the aorta was measured. The expression of AA-related proteins and the histology of the aortic wall were investigated at 1 week after treatment. MicroRNA and protein profiles of MSC-exs were examined. Results MSC-exs significantly attenuated AA progression (2.04 ± 0.20 mm in the saline group and 1.34 ± 0.13 mm in the MSC-ex group, P = 0.004). In the MSC-ex group, the expression of IL-1β, TNF-α and MCP-1 decreased, and expression of IGF-1 and TIMP-2 increased. MSC-ex induced the M2 phenotype in macrophages and suppressed the destruction of the elastic lamellae in the aortic wall. MSC-exs contained high levels of 10 microRNAs that inhibit AA formation and 13 proteins that inhibit inflammation and promote extracellular matrix synthesis. Conclusions MSC-ex might be a novel alternative therapeutic tool for treatment of existing AAs.

Abstract 9713: Ketosis Decreases Murine Plaque and Aortic Aneurysm Formation

Introduction: Atherosclerosis and aortic aneurysm (AA) are associated with leading causes of mortality worldwide. There is no medical therapy for the treatment of AA, and though diet modification is essential for the management of cardiovascular disease. A ketogenic diet has been accepted as an effective complementary treatment for hypertension and dyslipidemia. Hypothesis: We sought to evaluate if a high fat, low carbohydrate diet could attenuate plaque and AA formation in a mouse model. Methods: Apo E -/- mice (N=30) 12 weeks old, underwent Angiotensin II pump administration, with daily provision of β-aminopropionitrile (BAPN) over 28 days. Control mice (StD, N=15) received a standard diet (fat 42%, carbohydrates 42.7%, proteins 15.2%); while mice undergoing ketosis induction (KD, N=15) received a high fat, low carbohydrate diet (fat 90.5%, carbohydrates 0.3%, proteins 9.2%). Ketosis was achieved by a β-hydroxybutyrate (BHB) whole blood level >0.5 mmol/L. After 28 days, aortic diameters were measured by surgical microscopy. Isolated whole aortas were en-faced and stained with oil red O (ORO) to assess the grade of lesion area (plaque area + aneurysm degeneration area / whole aorta’s area) and plaque content (plaque area / whole aorta’s area). Results: KD mice remained in a ketotic state throughout the experiment. Thoracic and infrarenal AAs were identified in 60% and 74% of StD mice, respectively; however, only 13% and 33% of KD mice developed thoracic and infrarenal AAs, respectively. ORO staining (Fig. 1C and G) demonstrated a significantly smaller lesion area in KD mice (p = 0.04), (Fig. 1D). Furthermore, KD plaque content was attenuated in each segment of the aorta yet only significant in the thoracic aorta (p = 0.04), (Fig. 1H). Conclusions: Our results demonstrate that a ketogenic diet can attenuate the development of atherosclerotic plaque and AA degeneration in mice.

Abstract 9860: P38MAPK Deficiency in VSMCs Mitigates Abdominal Aortic Aneurysm and Atherosclerosis Progression via Promoting MKL1 Ubiquitin Proteasome Degradation

Cardiovascular diseases (CVD) are the leading cause of death in developed countries. Two major manifestations of CVD are aortic aneurysm and atherosclerosis, which are devastating and lack effective pharmacological therapy. The stress responsive pathway, p38 mitogen-activated protein kinase (MAPK), has been reported as a pivotal regulator in diverse processes, including cell proliferation, migration, inflammation, and apoptosis, contributing to cardiovascular development and pathology. However, a detailed understanding of the mechanistic role of p38MAPK in vascular smooth muscle cells (VSMCs) in the pathogenesis of aortic aneurysm and atherosclerosis has yet to be elucidated. Using a VSMC-specific lineage tracing reporter system, we showed that the phosphorylated form of p38MAPK (p-p38MAPK) was markedly elevated in VSMCs of aneurysmal vessels. VSMC-specific p38α ( Mapk14 ) knockout mice displayed attenuated abdominal aortic aneurysm (AAA) and atherosclerotic lesion formation compared to wild-type control mice. Further, loss of MAPK14 in VSMCs abolished the protein expression of multiple ER stress markers, including BIP, Protein Disulfide Isomerase (PDI), and ATF4, which was accompanied by the downregulation of the cleaved Caspase-3 and upregulation of BCL2 protein. This indicates a crucial role of p38MAPK in potentiating ER stress and apoptosis. Interestingly, aortas from VSMC-MAPK14 null mice displayed dramatically reduced protein levels of MKL1, a vital transcription factor with a well-recognized role in promoting AAA and atherosclerosis progression via regulating vascular apoptosis. In vitro, deficiency of p38MAPK in VSMCs via siRNA or SB203580, a selective inhibitor of p38MAPK pathway, downregulated whereas forced expression of MKK6 (activates p38MAPK) increased MKL1 protein expression. Finally, activation of p38MAPK inhibited MKL1 ubiquitination, which was partially rescued by SB203580. Collectively, our data demonstrated a critical role of VSMC-p38MAPK in promoting AAA and atherosclerosis progression, likely through stabilization of MKL1 via p38MAPK-dependent inhibition of MKL1 ubiquitin proteasome degradation.

Abstract 13417: Fluid-Structure Interaction Simulations of Bicuspid Aortic Valve Disease

Introduction: Aortic dilation and aneurysm formation is common in patients with a bicuspid aortic valve. Hemodynamics and genetics are both believed to cause dilation, but the relative contributions remain controversial. Further, retrospective studies assess hemodynamics when dilation has already set in. We simulate flows through a tricuspid and three bicuspid aortic valves and study their hemodynamics from a baseline non-dilated aorta. Hypothesis: We hypothesize that the bicuspid phenotype determines flow in the ascending aortic, and high velocity jets will correlate with known regions of dilation in patients with bicuspid valves. Methods: Using methods we recently developed, we constructed a tricuspid valve and bicuspid valves with left/right (LR), right/non (RN) and non/left (NL) coronary cusp fusion. To isolate the effect of leaflet fusion phenotype, we use one healthy, patient-specific aortic geometry throughout. Fluid-structure interaction simulations were performed. Results: Dramatic differences in flow occur between all cases. The tricuspid case shows a relatively uniform flow profile, velocities of ~200cm/s, normalized streamwise momentum <1.5, normalized tangential flow strength of <1.6, <20% reverse flow away from the sinus and sustained pressure gradients <2 mmHg. All bicuspid cases show a localized jet, peak velocities >300cm/s, normalized streamwise momentum >2.0, normalized tangential flow strength >1.0, >50% transient reverse flow and sustained pressure gradients >18 mmHg. With LR fusion, the jet hugs the outer curvature of the aorta from the sinotubular junction through the ascending aorta. With RN fusion, the jet moves from the inner to outer curvature of the aorta. Conclusions: With LR and RN cusp fusion, high flow rate jets occur at regions that generally correlate with dilation in prior studies, suggesting that hemodynamic factors alone may be sufficient to cause aortic dilation. This hypothesis should be tested in an animal model.

Abstract 11622: Resolution of Inflammation via Rvd1/fpr2 Signaling in Abdominal Aortic Aneurysms

Introduction: Resolvin D1 (RvD1) is derived from a ω-3 polyunsaturated fatty acid and is involved in the resolution phase of inflammation during abdominal aortic aneurysm (AAA) formation. Hypothesis: The aim of this study was to decipher the protective role of RvD1 via formyl peptide receptor 2 (FPR2) receptor signaling in attenuating AAA formation. Methods: C57BL/6 (WT) and FPR2 -/- mice (8-12 week old) underwent treatment with topical elastase or heat-inactivated elastase (control). Separate groups of mice were administered RvD1 (100ng/kg) or saline i.p. daily from day(d)1 to d13 and aortic diameter was measured by video micrometry on d14. mRNA quantification of FPR2 expression in aortic tissue was performed by RT-PCR on d7, 14 and 21. Human aortic tissue from AAA patients and controls (organ transplant donors) were analyzed for quantification of FPR2 expression. Data is presented as mean ± SEM and statistical analysis was performed by using Mann-Whitney test for phenotype or Wilcoxon Signed Rank Test for gene expression data. Results: Elastase-treated WT mice had significantly increased aortic diameter compared to controls (127±8% vs. 1± 0.6%; p<0.0001). Importantly, elastase-treated FPR2 -/- had a significant increase in aortic diameter compared to elastase-treated WT mice (165±16% vs. 127± 8%; p=0.03). RvD1 treatment attenuated AAA formation in WT mice, but not FPR2 -/- mice, on d14 (56± 10% vs.121± 8%; p<0.0001). Immunohistology documented decreased elastin and smooth muscle cell α-actin expression, as well as increased neutrophil and macrophage infiltration in FPR2 -/- mice compared to WT mice, after respective treatments with RvD1. Elastase-treated WT mice had significantly increased FPR2 receptor mRNA expression compared to respective controls on d7, d14, and d21. Human AAA tissue demonstrated significantly decreased FPR2 mRNA expression compared to controls (p=0.02). Conclusions: This data identifies the role of FPR2 in dysregulation of inflammation-resolution during AAA pathogenesis. Immunomodulation of FPR2 signaling by exogenous RvD1 identifies a novel mechanism in resolution of aortic inflammation and vascular remodeling during AAA pathogenesis.

MFAP4 Deficiency Attenuates Angiotensin II-Induced Abdominal Aortic Aneurysm Formation Through Regulation of Macrophage Infiltration and Activity.

Objective: Abdominal aortic aneurysm (AAA) is a common age-related vascular disease characterized by progressive weakening and dilatation of the aortic wall. Microfibrillar-associated protein 4 (MFAP4) is an extracellular matrix (ECM) protein involved in the induction of vascular remodeling. This study aimed to investigate if MFAP4 facilitates the development of AAA and characterize the underlying MFAP4-mediated mechanisms.Approach and Results: Double apolipoprotein E- and Mfap4-deficient (ApoE−/−Mfap4−/−) and control apolipoprotein E-deficient (ApoE−/−) mice were infused subcutaneously with angiotensin II (Ang II) for 28 days. Mfap4 expression was localized within the adventitial and medial layers and was upregulated after Ang II treatment. While Ang II-induced blood pressure increase was independent of Mfap4 genotype, ApoE−/−Mfap4−/− mice exhibited significantly lower AAA incidence and reduced maximal aortic diameter compared to ApoE−/− littermates. The ApoE−/−Mfap4−/− AAAs were further characterized by reduced macrophage infiltration, matrix metalloproteinase (MMP)-2 and MMP-9 activity, proliferative activity, collagen content, and elastic membrane disruption. MFAP4 deficiency also attenuated activation of integrin- and TGF-β-related signaling within the adventitial layer of AAA tissues. Finally, MFAP4 stimulation promoted human monocyte migration and significantly upregulated MMP-9 activity in macrophage-like THP-1 cells.Conclusion: This study demonstrates that MFAP4 induces macrophage-rich inflammation, MMP activity, and maladaptive remodeling of the ECM within the vessel wall, leading to an acceleration of AAA development and progression. Collectively, our findings suggest that MFAP4 is an essential aggravator of AAA pathology that acts through regulation of monocyte influx and MMP production.

Rosmarinic Acid Suppresses Abdominal Aortic Aneurysm Progression in Apolipoprotein E-deficient Mice.

An abdominal aortic aneurysm is a life-threatening cardiovascular disorder caused by dissection and rupture. No effective medicine is currently available for the > 90% of patients whose aneurysms are below the surgical threshold. The present study investigated the impact of rosmarinic acid, salvianolic acid C, or salvianolic acid B on experimental abdominal aortic aneurysms. Abdominal aortic aneurysms were induced in apolipoprotein E-deficient mice via infusion of angiotensin II for 4 wks. Rosmarinic acid, salvianolic acid C, salvianolic acid B, or doxycycline as a positive control was provided daily through intraperitoneal injection. Administration of rosmarinic acid was found to decrease the thickness of the aortic wall, as determined by histopathological assay. Rosmarinic acid also exhibited protection against elastin fragmentation in aortic media and down-regulated cell apoptosis and proliferation in the aortic adventitia. Infiltration of macrophages, Tlymphocytes, and neutrophils in aortic aneurysms was found, especially at the aortic adventitia. Rosmarinic acid, salvianolic acid C, or salvianolic acid B inhibited the infiltration on macrophages specifically, but these compounds did not influence T lymphocytes and neutrophils. Expression of matrix metalloproteinase 9 and macrophage migration inhibitory factor significantly increased in aortic aneurysms. Rosmarinic acid and salvianolic acid C decreased the expression of matrix metalloproteinase-9 in media, and rosmarinic acid also tended to reduce migration inhibitory factor expression. Further then, partial least squares-discriminate analysis was used to classify metabolic changes among different treatments. Rosmarinic acid affected most of the metabolites in the biosynthesis of the citrate cycle, fatty acid pathway significantly. Our present study on mice demonstrated that rosmarinic acid inhibited multiple pathological processes, which were the key features important in abdominal aortic aneurysm formation. Further study on rosmarinic acid, the novel candidate for aneurysmal therapy, should be undertaken to determine its potential for clinical use.

Vitamin B Mitigates Thoracic Aortic Dilation in Marfan Syndrome Mice by Restoring the Canonical TGF-β Pathway.

Thoracic aortic aneurysm (TAA) formation is a multifactorial process that results in diverse clinical manifestations and drug responses. Identifying the critical factors and their functions in Marfan syndrome (MFS) pathogenesis is important for exploring personalized medicine for MFS. Methylenetetrahydrofolate reductase (MTHFR), methionine synthase (MTR), and methionine synthase reductase (MTRR) polymorphisms have been correlated with TAA severity in MFS patients. However, the detailed relationship between the folate-methionine cycle and MFS pathogenesis remains unclear. Fbn1C1039G/+ mice were reported to be a disease model of MFS. To study the role of the folate-methionine cycle in MFS, Fbn1C1039G/+ mice were treated orally with methionine or vitamin B mixture (VITB), including vitamins B6, B9, and B12, for 20 weeks. VITB reduced the heart rate and circumference of the ascending aorta in Fbn1C1039G/+ mice. Our data showed that the Mtr and Smad4 genes were suppressed in Fbn1C1039G/+ mice, while VITB treatment restored the expression of these genes to normal levels. Additionally, VITB restored canonical transforming-growth factor β (TGF-β) signaling and promoted Loxl1-mediated collagen maturation in aortic media. This study provides a potential method to attenuate the pathogenesis of MFS that may have a synergistic effect with drug treatments for MFS patients.

Metformin therapy is not associated with the lower prevalence of ascending aortic aneurysm in diabetic patients.

Metformin therapy has previously been associated with reduced abdominal aortic aneurysm growth rate in diabetic patients and shown to suppress the formation and progression of abdominal aortic aneurysm in normoglycemic mice. Here, we investigated the association between Metformin treatment and prevalence of aneurysm in the ascending aorta (AscAA). A total of 734 patients undergoing open-heart surgery for AscAA and/or aortic valve disease were studied. Diabetes status and medication use were self-reported by the patients in a systematic questionnaire. Aortic dilatation was defined as an aortic root or ascending aortic diameter ≥4.0 cm. High-sensitivity C-reactive protein levels were assessed as a measure of systemic inflammation. We could confirm the inverse association between diabetes and AscAA prevalence (16% vs 43.9%, for diabetic and non-diabetic patients, respectively; Odds ratio 0.243; 95% CI, 0.129-0.460, P < 0.001). Furthermore, in diabetic patients, Metformin treatment was associated with lower high-sensitivity C-reactive protein levels. There was, however, no difference in the prevalence of AscAA among diabetic patients with and without Metformin treatment (16% vs 16% for treated and non-treated patients, respectively; OR 1.039; 95% CI 0.26-4.19, P = 0.957). Our data do not support a protective effect of Metformin therapy in AscAA formation. 161, 173.

The Histone Methyltransferase Setdb2 Regulates Abdominal Aortic Aneurysm Formation

The Histone Methyltransferase Setdb2 Regulates Abdominal Aortic Aneurysm Formation

From the Literature

From the Literature

PI 3-kinase isoform PI3Kalpha controls smooth muscle cell functionality and protects against aortic aneurysm formation

Abstract Background Class I PI 3-kinase isoform PI3Kα is a lipid kinase and signals downstream of receptor tyrosine kinases. Smooth muscle cells (SMCs) lacking PI3Kα are characterized by impaired proliferation, migration and survival. Mice, harbouring a smooth muscle specific PI3Kα deficiency (SM-PI3Kα−/−), display reduced vascular wall thickness and impaired vascular remodeling in response to vessel injury. We hypothesize that SM-PI3Kα−/− mice are prone to aortic aneurysm (AA) formation due to impaired SMC functions. Herein, we investigated, how PI3Kα-dependent signaling in SMCs affect aortic aneurysm (AA) formation, aortic wall structure, and expression of extracellular matrix (ECM) components. Methods and results AA formation in SM-PI3Kα−/− mice and wild-type littermates was examined by means of the “porcine pancreatic elastase” (PPE) AA model. PPE was infused into the infrarenal aorta to induce AA formation. Ultrasound examination revealed a significantly increased aortic diameter in SM-PI3Kα−/− mice (1.22±0.12 mm) compared to wild-type animals (0.96±0.02 mm, p=0.014). These data indicate a protective function of SM-PI3Kα in AA formation. In addition, the media thickness in the abdominal aorta was significantly reduced in SM-PI3Kα−/− mice (29.0±3.1 vs. 42.5±4.1 μm). Ultrastructural analysis of aortic wall morphology in SM-PI3Kα−/−mice using transmission electron microscopy (TEM) showed a deranged tunica media with detached SMCs and increased apoptotic cell death. Consequently, SM-PI3Kα deficiency significantly diminished responsiveness of aortic rings to vasodilator acetylcholine and NO-donor nitroglycerin, further indicating impaired aortic wall structure. Western blots demonstrated a reduced elastin and fibrillin expression in SMCs from SM-PI3Kα−/− mice. Furthermore, immunofluorescence stainings of PI3Kα−/− and wild-type SMCs, cultured for seven days under 10% fetal calf serum containing DMEM medium, showed significantly disturbed structures of elastin-, fibrillin-1- and collagen-1-fibers. These data indicate that PI3Kα signaling contributes to elastic fiber homeostasis thus affecting SMC phenotypic modulation. Immunoblots demonstrated that PDGF and insulin induced phosphorylation and inactivation of key regulators of SMC differentiation and dedifferentiation including FoxO1, FoxO3a, Foxo4, and GSK3b, respectively, were reduced or even abrogated in PI3Kα−/− SMCs. Conclusion These data show that deficiency of PI3Kα in SMCs promotes the formation and progression of AA. Causative is a deranged aortic structure of SM-PI3Kα−/− aortae which can likely be attributed to an impaired production of elastic fiber components by PI3Kα−/− SMCs. Funding Acknowledgement Type of funding sources: Public grant(s) – National budget only. Main funding source(s): Deutsche Forschungsgemeinschaft

Basigin genetic depletion reduces thoracic aortic aneurysm formation in marfan syndrome mice

Abstract Background Marfan Syndrome (MFS) is a rare genetic disease due to mutations in fibrillin 1 gene [1]. It is characterized by thoracic aortic aneurysm (TAA), whose dissection is the major cause of morbidity and mortality in MFS patients [2]. While pathological mechanisms underlying MFS extracellular matrix remodelling have started to be elucidated, an etiological treatment is not available yet [3,4]. Recently, EMMPRIN has been proposed as new actor in pro-fibrotic processes associated with TAA development in MFS patients [4,5]. Purpose To confirm the role of EMMPRIN (i.e. Basigin in mice, Bsg) in TAA formation and progression in a preclinical setting, we assessed whether Bsg genetic depletion in a mouse model of MFS (Fbn1C1039G/+) [6] had an impact on aortic phenotype. Methods We generated a double transgenic mouse model, Fbn1C1039G/+-Bsg+/−, by crossing commercially available Fbn1C1039G/+ with Bsg+/− mice (provided by Dr. Kadomatsu) [7] and we prospectively measured aortic root size in 12 Fbn1C1039G/+-Bsg+/−, 9 Fbn1C1039G/+, and 15 WT mice every 4 weeks from 12 to 20 weeks of age by 2D-echocardiography. Both qRT-PCR and Western Blot were used to evaluate EMMPRIN downstream pro-fibrotic pathway activation in thoracic aortic tissue obtained from mice sacrificed at 12 weeks. Plasma soluble EMMPRIN (sEMMPRIN) levels were assessed by ELISA. Results Echocardiographic assessment at 12 and 20 weeks of age confirmed significantly dilated aortic root in Fbn1C1039G/+ vs WT (mean±SD at 12 weeks: 1.772±0.081 vs 1.394±0.068 mm, respectively, p&amp;lt;0.0001; 20 weeks: 1.899±0.103 vs 1.520±0.083 mm, respectively, p&amp;lt;0.0001), and revealed that Fbn1C1039G/+-Bsg+/− mice had lower aortic root diameter extent compared to Fbn1C1039G/+ (12 weeks: 1.667±0.109 vs 1.772±0.081 mm, respectively, p=0.026; 20 weeks: 1.752±0.129 vs 1.899±0.103 mm, respectively, p=0.011). Pro-fibrotic gene profiling in a subset of 12 weeks old mice showed that Col1A1, Col3A1 and FN1 genes were more expressed in Fbn1C1039G/+ than in WT mice and that the up-regulation of these genes was mitigated in Fbn1C1039G/+ by Bsg depletion (ANOVA p=0.048, p=0.055, and p=0.0003, respectively). Moreover, we found a significant activation of the EMMPRIN downstream pro-fibrotic ERK1/2 pathway in Fbn1C1039G/+ compared to WT mice which was attenuated in the Fbn1C1039G/+-Bsg+/− model (p=0.030). Plasma sEMMPRIN levels were lower in Fbn1C1039G/+ than in WT mice, consistent with what we previously observed in MFS patients [5], and decreased over time in both Fbn1C1039G/+ and WT mice (2-way ANOVA disease p=0.013, time p=0.012, interaction p=ns). Conclusions Basigin genetic depletion reduces dilatation of the aortic root and pro-fibrotic pathway activation in a preclinical MFS model, confirming previous proof-of-concept findings and paving the way for the development of innovative therapies for MFS patients. Plasma sEMMPRIN levels stand as promising tool to predict the aortic enlargement progression in MFS models and patients. Funding Acknowledgement Type of funding sources: Public Institution(s). Main funding source(s): Italian Ministery of Health

Degeneration and Regeneration of Smooth Muscle Cells in Two Different Abdominal Aortic Aneurysm Models in Rabbits

We aimed to investigate the formation and self-healing process of rabbit abdominal aortic aneurysm (AAA) by focus on the degeneration and regeneration of smooth muscle cells (SMCs) in elastase-induced AAA model and enlarging AAA model in rabbits. Sixty rabbits were equally divided into 2 aneurysm groups (Group A and Group B). Rabbits received a 10-min incubation of elastase in Group A (10 units/µL) and Group B (1 unit/µL). Rabbits underwent aortic stenosis above the incubated segment in Group B. Aortic diameter was measured and rabbits were sacrificed for histopathological and immunohistochemical studies. The incubated aorta dilated immediately and ran up to maxima by day 21 in Group A. All aneurysms formed by day 21 and enlarged progressively in Group B. SMCs content, elastin content and intima-media thickness decreased significantly by day 0 in Group A. SMCs and elastic fibers were destroyed gradually in Group B, however, SMCs content was significantly lower than Group A by day 70. Intimal thickness increased significantly by day 70 in the Aneurysm groups. MMP2 maintained moderate expression in Group A, which decreased significantly by day 3 in Group B. MMP9 and RAM11 expressions were higher by day 1, but decreased significantly by day 3 in Group B. Irreversible degeneration of SMCs is critical to a rapid formation of elastase-induced rabbit AAA model, and SMCs excessive regeneration accounts for the selfhealing process. SMCs degradation and regeneration remain relatively stable in an enlarging AAA model. SMCs should be the key target for studying the mechanism of AAA and intervention therapy.

The Histone Methyltransferase SETDB2 Regulates Abdominal Aortic Aneurysm Formation

The purpose of the present project was to establish whether the histone methyltransferase, SETDB2, regulates abdominal aortic aneurysm (AAA) development. We hypothesized that during AAA development, SETDB2 will be upregulated, resulting in the “turning off” of genes that are normally protective against pathologic vascular remodeling via trimethylation of lysine 9 of histone 3 (H3K9me3) at gene promoters. Specifically, during normal aortic wall existence, a balance is in place between matrix metalloproteinases (MMPs), which function to break down the aortic wall, and tissue inhibitors of matrix metalloproteinases (TIMPs), which prevent the actions of MMPs on the aortic wall.

Circular RNA Cdyl promotes abdominal aortic aneurysm formation by inducing M1 macrophage polarization and M1-type inflammation

Macrophage polarization plays a crucial role in regulating abdominal aortic aneurysm (AAA) formation. Circular RNAs (circRNAs) are important regulators of macrophage polarization during the development of cardiovascular diseases. How-ever, the roles of circRNAs in regulating AAA formation through modulation of macrophage polarization remain unknown. In the present study, we compared circRNA microarray data under two distinct polarizing conditions (M1 and M2 macrophages) and identified an M1-enriched circRNA, circCdyl. Loss- and gain-of-function assay results demonstrated that circCdyl overexpression accelerated angiotensin II (Ang II)- and calcium chloride (CaCl2)-induced AAA formation by promoting M1 polarization and M1-type inflammation, while circCdyl deficiency showed the opposite effects. RNA pulldown, mass spectrometry analysis, and RNA immunoprecipitation (RIP) assays were conducted to elucidate the underlying mechanisms by which circCdyl regulates AAA formation and showed that circCdyl promotes vascular inflammation and M1 polarization by inhibiting interferon regulatory factor 4 (IRF4) entry into the nucleus, significantly inducing AAA formation. In addition, circCdyl was shown to act as a let-7c sponge, promoting C/EBP-δ expression in macrophages to induce M1 polarization. Our results indicate an important role for circCdyl-mediated macrophage polarization in AAA formation and provide a potent therapeutic target for AAA treatment.

Oxidative Stress-Related Susceptibility to Aneurysm in Marfan’s Syndrome

The involvement of highly reactive oxygen-derived free radicals (ROS) in the genesis and progression of various cardiovascular diseases, including arrhythmias, aortic dilatation, aortic dissection, left ventricular hypertrophy, coronary arterial disease and congestive heart failure, is well-established. It has also been suggested that ROS may play a role in aortic aneurysm formation in patients with Marfan’s syndrome (MFS). This syndrome is a multisystem disorder with manifestations including cardiovascular, skeletal, pulmonary and ocular systems, however, aortic aneurysm and dissection are still the most life-threatening manifestations of MFS. In this review, we will concentrate on the impact of oxidative stress on aneurysm formation in patients with MFS as well as on possible beneficial effects of some agents with antioxidant properties. Mechanisms responsible for oxidative stress in the MFS model involve a decreased expression of superoxide dismutase (SOD) as well as enhanced expression of NAD(P)H oxidase, inducible nitric oxide synthase (iNOS) and xanthine oxidase. The results of studies have indicated that reactive oxygen species may be involved in smooth muscle cell phenotype switching and apoptosis as well as matrix metalloproteinase activation, resulting in extracellular matrix (ECM) remodeling. The progression of the thoracic aortic aneurysm was suggested to be associated with markedly impaired aortic contractile function and decreased nitric oxide-mediated endothelial-dependent relaxation.

Six Shades of Vascular Smooth Muscle Cells Illuminated by KLF4 (Krüppel-Like Factor 4).

Multiple layers of vascular smooth muscle cells (vSMCs) are present in blood vessels forming the media of the vessel wall. vSMCs provide a vessel wall structure, enabling it to contract and relax, thus modulating blood flow. They also play a crucial role in the development of vascular diseases, such as atherosclerosis and aortic aneurysm formation. vSMCs display a remarkable high degree of plasticity. At present, the number of different vSMC phenotypes has only partially been characterized. By mapping vSMC phenotypes in detail and identifying triggers for phenotype switching, the relevance of the different phenotypes in vascular disease may be identified. Up until recently, vSMCs were classified as either contractile or dedifferentiated (ie, synthetic). However, single-cell RNA sequencing studies revealed such dedifferentiated arterial vSMCs to be highly diverse. Currently, no consensus exist about the number of vSMC phenotypes. Therefore, we reviewed the data from relevant single-cell RNA sequencing studies, and classified a total of 6 vSMC phenotypes. The central dedifferentiated vSMC type that we classified is the mesenchymal-like phenotype. Mesenchymal-like vSMCs subsequently seem to differentiate into fibroblast-like, macrophage-like, osteogenic-like, and adipocyte-like vSMCs, which contribute differentially to vascular disease. This phenotype switching between vSMCs requires the transcription factor KLF4 (Krüppel-like factor 4). Here, we performed an integrated analysis of the data about the recently identified vSMC phenotypes, their associated gene expression profiles, and previous vSMC knowledge to better understand the role of vSMC phenotype transitions in vascular pathology.

Allosteric activation of PP2A inhibits experimental abdominal aortic aneurysm

Although extremely important, the molecular mechanisms that govern aortic aneurysm (AA) formation and progression are still poorly understood. This deficit represents a critical roadblock toward the development of effective pharmaceutical therapies for the treatment of AA. While dysregulation of protein phosphatase 2A (PP2A) is thought to play a role in cardiovascular disease, its role in aortic aneurysm is unknown. The objective of the present study is to test the hypothesis that PP2A regulates abdominal aortic aneurysm (AAA) progression in a murine model. In an angiotensin II-induced AAA murine model, the PP2A inhibitor, LB-100, markedly accelerated AAA progression as demonstrated by increased abdominal aortic dilation and mortality. AAA progression was associated with elevated inflammation and extracellular matrix fragmentation, concomitant with increases in both metalloproteinase activity and reactive oxygen species production. Conversely, administration of a novel class of small molecule activators of PP2A (SMAPs) resulted in an antithetical effect. SMAPs effectively reduced AAA incidence along with the corresponding pathologies that were increased with LB-100 treatment. Mechanistically, modulation of PP2A activities in vivo functioned in part via alteration of the ERK1/2 and NFκB signaling pathways, known regulators of AAA progression. These studies, for the first time, demonstrate a role of PP2A in AAA etiology and demonstrate that PP2A activation may represent a novel strategy for the treatment of abdominal aortic aneurysms.

Abstract MP20: Pd-1 Pathway Upregulation By Orchiectomy Attenuates The Aldosterone And High Salt Induced Aortic Aneurysms In Male Mice

Objective: Male sex is a well-established risk factor for abdominal aortic aneurysms (AAA) but the underlying mechanisms remain to be fully understood. Using an aldosterone and high salt (Aldo/salt) induced AAA mouse model, we have demonstrated that androgen and its receptor mediate the high susceptibility to Aldo/salt induced AAA. The current study further investigates the mechanisms downstream of androgen. Approaches and Results: - To dissect the mechanisms connecting androgen and AAA, aortas were collected for RNA sequencing from 3 groups of 10-month-old wild-type mice #1 intact mice; #2 orchiectomized mice; and #3 orchiectomized mice plus DHT. All mice were given Aldo/salt for 7 days. Differentially expressed genes were analyzed using DESeq2 in R. We filtered genes that were upregulated in group #2 compared to group #1 and the up-regulation was reversed in group #3 by the DHT, or vice versa (fold change &gt;1.5 and padj &lt;0.05). Selected genes were run for gene ontology analysis in Enrichr (database Bioplanet 2019). Many pathways related to T cell activity were significantly enriched, particularly PD-1 signaling was one of the top pathways upregulated in orchiectomy group #2. PD-1 is known for its role as an immune checkpoint, and inflammation is a major hallmark for AAA development. Therefore to explore the role of PD-1, we first confirmed the PD-1 mRNA changes in aorta by qPCR. Secondly, IHC staining also showed PD-1 protein was significantly increased in the spleen of orchiectomized mice compared to intact controls. Finally, to investigate the potential causal role of PD-1 in the androgen-mediated aortic aneurysms formation, we injected αPD-1 antibody or control IgG antibody to orchiectomized mice 3 days before and during the 8 weeks of Aldo/salt administration. Results showed that 5 out of 12 αPD-1 mice, while none of the 8 control mice developed aortic aneurysms (p&lt;0.05). Conclusions: PD-1 pathway is involved in the androgen associated high susceptibility of Aldo/salt induced aortic aneurysms in mice.