Risultati per: Come stress e attacchi di cuore sono collegati

Circulation, Volume 150, Issue Suppl_1, Page A4143599-A4143599, November 12, 2024. Background:The Steroids to Reduce Systemic Inflammation after Infant Heart Surgery (STRESS) trial randomized 1200 infants undergoing cardiac surgery with cardiopulmonary bypass to prophylactic intraoperative methylprednisolone (MP) versus placebo.Aims:Evaluate benefits and harms associated with MP in subset populations.Methods:STRESS participants were categorized based on STAT Mortality Category (1-3 and 4-5), age (neonate ≤ 30 days< non-neonate), prematurity (< 37 weeks gestation) and any chromosomal or syndromic diagnoses (CSD). Key postoperative outcomes included any steroid administration (< 72 hours after surgery), peak blood glucose (7 days), thrombosis, and infections.Results:The cohort consisted of 30% (364/1200) neonates, 16% (193/1197) premature, and 81% (969/1197) STAT 1-3 operations. Stratified analyses demonstrated notable beneficial effects with MP including reduced use of postoperative hydrocortisone in neonates (OR 0.39 [0.25-0.60]), those following STAT 1-3 (OR 0.65 [0.47-0.91]) and STAT 4-5 operations (OR 0.57 [0.34-0.97]), term infants (OR 0.62 [0.47-0.83]), and those without CSD (OR 0.63 [0.46-0.86]). MP associated with lower thrombosis occurrence among neonates (OR 0.37 [0.16-0.87]) and term infants (OR 0.38 [0.19-0.75]). Notable adverse associations with MP included increased postoperative peak blood glucose levels and insulin use (all subgroups, P

Circulation, Volume 150, Issue Suppl_1, Page A4142430-A4142430, November 12, 2024. Background:Cardiac allograft vasculopathy (CAV) is a major cause of morbidity and mortality following heart transplantation (OHT). Noninvasive methods to detect CAV and risk stratify OHT patients are needed. The value of fully quantitative stress cardiac magnetic resonance imaging has been recently validated and may be a promising technique for OHT surveillance. We aimed to evaluate the feasibility of quantitative stress CMR after OHT.Methods:We enrolled asymptomatic OHT recipients without coronary artery disease to undergo regadenoson stress CMR (1.5T GE HealthCare) with cine imaging, tissue mapping, and late gadolinium enhancement (LGE) imaging for routine CAV surveillance. Using the dual sequence technique, quantitative perfusion values were determined using Fermi deconvolution. Myocardial perfusion reserve (MPR) was calculated as the ratio of stress to rest myocardial blood flow (MBF).Results:Fifty-three subjects (mean age 47.06 ± 17.14 years) were included. OHT recipients (n=11, mean 6.77 ± 4.34 years post-transplant) were compared with healthy controls (n=43). No life-threatening adverse events, brief or prolonged atrioventricular block or other arrhythmias occurred with regadenoson. Coronary angiography was performed in 9 OHT patients before CMR, with an average of 1.99 ± 2.05 years between studies. No visual inducible ischemia was reported. Post OHT, rest MBF was significantly higher (1.69 ± 0.52 mL/g/min vs 1.01 ± 0.24 mL/g/min, p=0.004) and stress MBF was lower (2.33 ± 0.69 mL/g/min vs 2.95 ± 0.88 mL/g/min, p=0.02) compared to controls. MPR was significantly lower in OHT recipients compared to controls (1.46 ± 0.51 vs 3.11 ± 1.12, p

Circulation, Volume 150, Issue Suppl_1, Page A4142206-A4142206, November 12, 2024. Background:To apply human pluripotent stem cell-derived endothelial cells (hPSC-ECs) in regenerative medicine, exploring methods for highly purified ECs is desirable. Cell sorting is a versatile technique for isolating and purifying specific cell types, yet mechanical cell loss persists. Previously, we established a differentiation method for human induced pluripotent stem cell-derived ECs (hiPSC-ECs) based on lineage control using vascular endothelial growth factor (VEGF) and 8-Bromo cyclic adenosine monophosphate (cAMP). However, achieving high hiPSC-ECs purity without cell sorting has not yet been possible.Hypothesis:We speculated that applying digital rocker-generated shear stress during a specific period of hiPSC-EC induction would yield highly purified hiPSC-ECs without cell sorting.Methods:We applied cyclic share stress to the cultured cells using a digital rocker. To optimize the frequency and duration of digital rocker application, ECs purity on day 13 of differentiation (d13) was analyzed by flow cytometry for vascular endothelial cadherin (VE-Cadherin). Shear stress was measured using a simulation model. The functionality of ECs was evaluated through reverse transcriptase quantitative polymerase chain reaction (RT-qPCR) for endothelial nitric oxide synthase (eNOS) and angiogenesis assay.Results:The optimized protocol consisted of a rocking period from day 5 (representing the EC progenitor stage) to d13, at 30 cycles/min with 13° tilt (equivalent to 1.09 dyn/cm2), which significantly increased the purity of ECs (Control vs Rocking: VE-Cadherin; 69.25±17.43 vs 86.68±6.023 %, P = 0.0090). Examining the number of cells on d13 revealed rocking stimulation reduced both ECs and non-ECs. Non-ECs were nearly absent, suggesting EC purification occurs by removing non-ECs, indicating ECs are more resistant to being eliminated by the rocking stimulation. The rocking culture also led to increased eNOS mRNA expression on d13 (Control vs Rocking: 0.5574±0.4985 vs 1.056±0.1652, P = 0.0393). The angiogenesis assay showed a longer vascular structure length trend in the rocking group, indicating enhanced angiogenic capacity. (Control vs Rocking: 15407±2929 vs 18335±3568 Pixel, P= 0.4309).Conclusion:In this study, we developed a method where digital rocker-generated shear stress during a specific period of hiPSC-EC induction not only selectively purifies ECs without cell sorting, but also enhances endothelial function, demonstrating their therapeutic potential.

Circulation, Volume 150, Issue Suppl_1, Page A4141840-A4141840, November 12, 2024. BACKGROUND:Pulmonary arterial hypertension (PAH) is characterized by obliteration of distal pulmonary arteries (PA) in association with endothelial cell (EC) dysfunction, leading to smooth muscle proliferation. SOX17 is a transcription factor (TF) expressed in arterial EC that is critical in vascular development. Deleterious variants causing reduced expression ofSOX17are linked to PAH, particularly in congenital heart defects (CHD) that cause increased PA flow and high shear stress (HSS).HYPOTHESIS:SOX17 deficiency is additive to HSS in compromising PAEC homeostasis and in promoting severe PAH.METHODS:SOX17was reduced ( >70%) by siRNA in primary human PAEC cultured in chamber slides in the IBIDI perfusion system. Computational modeling of distal PA indicated pathological HSS of 100 dyn/cm2in CHD with PAH whereas physiological laminar shear stress (LSS) is 15 dyn/cm2. EC were preconditioned under LSS for 24h, followed by HSS or LSS for 24h.RESULTS:SOX17 expression was increased under LSS versus static condition, as were known SOX17 target genes (e.g.,GJA5,GJA4,CGNL1,JAG1, andCTNNB1). SOX17siRNA and HSS similarly reduced SOX17 target genes and when combiningSOX17siRNA with HSS they were further decreased owing to an interaction between SOX17 and ERG, a TF reduced by HSS. Indeed, SOX17 and ERG motifs marked most enhancer and promoter H3K27acetyl marks that were reduced under HSS. We then carried out RNAseq of PAEC to find genes co-regulated by SOX17 and ERG (reduced by siRNA for either TF under LSS), decreased under HSS and more-so with HSS +SOX17siRNA. Those downregulated included SOX17 targets (e.g.,CGNL1andGJA5) and others not previously described, with links to BMPR2 signaling,YAP1(inducer of BMP ligands and suppressor of NF-kB),SETBP1(inducer of BMPR2 co-receptorBMPR1b),andTMEM100andSULT1B1important in NOTCH signaling and EC specification. We also found novel extracellular matrix target genes, e.g., elastin (ELN,top DEG),HMCN1,TMTC1and chromatin remodeler (TOX). Among genes upregulated, wereNAMPT, FAS, LYN, HPSErelated to NF-kB activation and/or inflammation.CONCLUSION:HSS plus SOX17 deficiency profoundly compromises EC homeostatic genes, among which are those affecting BMPR2 and NOTCH pathways, ELN fiber assembly, and those promoting inflammation. This can explain whySOX17mutations are associated with severe PAH in HSS-related congenital heart defects.

Circulation, Volume 150, Issue Suppl_1, Page A4143744-A4143744, November 12, 2024. Introduction/Background:Hypertension and hypercholesterolemia are important risk factors of endothelial dysfunction and atherosclerosis. Previous studies suggested a crosstalk between an activated renin-angiotensin-aldosterone system (RAAS), reactive oxygen species (ROS) and oxidized low-density lipoproteins (oxLDL) in atherosclerosis, but the underlying molecular mechanisms are not well understood.Research Question/Hypothesis:Can we identify novel signaling pathways controlling the molecular crosstalk of the RAAS with ROS and oxLDL in endothelial dysfunction and atherosclerosis?Methods/Approach:The impact of AT1R blockade on oxLDL-induced superoxide anion formation and endothelial dysfunction was studied in human umbilical artery endothelial cells and aortic rings of wild-type mice by chemiluminescence and Mulvany myograph. We cloned 5’-terminal deletions of the AT1R promoter and assessed the luciferase activity in human endothelial cells. Oct-1 binding to the human AT1R promoter region was studied by EMSA. Further assays included real-time PCR, confocal microscopy, Western blotting, G protein reporter assays, phospho-ERK1/2 antibodies and specific siRNAs. The data were validated in heart of obese C57BL/6 mice and cardiac and aortic tissue of AT1a/AT1bdouble knockout micein vivo.Results/Data:AT1R promoter activation studies upon Ang II- and oxLDL-stimulation in endothelial cells revealed that Ang II and oxLDL activate AT1R signaling through G protein Gα12/13, followed by activation of ERK1/2 MAP kinases, and transcription and translation of Oct-1, resulting in up-regulation of AT1R, LOX-1 and NOX2 expression, which could be antagonized by specific inhibitors at each step of the identified signaling cascade. AT1R blockade improved oxLDL-induced endothelial dysfunction in aortic rings of wild-type mice. Male C57BL/6 mice fed a high-fat diet exhibited upregulation of Oct-1 levels in cardiac tissues, compared to normal controls, while AT1a/AT1bdouble knockout mice demonstrated downregulation of Oct-1, AT1R, LOX-1, and NOX2 on mRNA and protein level in cardiac and aorta tissue, thus confirming the identified signaling cascadein vivo.Conclusions:Oct-1 is an essential transcription factor for Ang II- and oxLDL-induced upregulation of AT1R and LOX-1 expression in endothelium, thus identifying a novel molecular cross-talk of oxLDL with ROS signaling and the RAAS contributing to development of endothelial dysfunction and atherosclerosis.

Circulation, Volume 150, Issue Suppl_1, Page A4138303-A4138303, November 12, 2024. Background:A higher stress hyperglycemic ratio (SHR) has been reported to be associated with adverse cardiac outcomes. However, the role of SHR in predicting clinical outcomes by comparing patients with and without diabetes mellitus is yet to be explored.Objective:To evaluate the prognostic value of the SHR for predicting major adverse cardiovascular (MACE) and all-cause mortality in ACS patients with and without diabetes mellitus.Methods:Per PRISMA guidelines, we comprehensively reviewed PubMed, Google Scholar, and SCOPUS for eligible studies reporting on SHR and its association with MACE (8 studies) and all-cause mortality (7 studies) in ACS patients. Pooled odds ratios (ORs) and 95% confidence intervals (CIs) were calculated using a binary random-effects model, with results displayed as forest plots. Heterogeneity was assessed using I2 statistics, and a leave-one-out sensitivity analysis was performed. P

Circulation, Volume 150, Issue Suppl_1, Page A4142510-A4142510, November 12, 2024. Background:An acute hyperglycemic status is reportedly associated with poor prognosis in patients with acute cardiovascular diseases. Although the stress hyperglycemia ratio (SHR) is a novel index to accurately represent the hyperglycemic condition on admission, relations between SHR and clinical outcomes are not fully evaluated in a setting of acute myocardial infarction (MI).Methods:This retrospective, multicenter registry study included 2,386 patients with acute MI undergoing percutaneous coronary intervention. SHR was calculated as a blood glucose level on admission divided by the estimated average glucose derived from a glycated hemoglobin level. The co-primary endpoints of this study included heart failure (HF)-related events (a composite of all-cause death and worsening and hospitalized HF) and major atherosclerotic cardiovascular events (MACE) (a composite of all-cause death, recurrent MI, and ischemic stroke), during the index hospitalization and after discharge.Results:Of the 2,386 patients, 890 (37.3%) had diabetes, and the median SHR was 1.17 [0.99, 1.45]. HF events and MACE occurred in 680 (28.5%) and 233 (9.8%) during hospitalization. SHR was identified as a factor significantly associated with both in-hospital HF events (adjusted odds ratio 1.65, 95% confidence interval 1.18-2.29, p=0.003) and MACE (adjusted odds ratio 1.50, 95% confidence interval 1.10-2.03, p=0.009). Among 2,017 patients who survived to discharge and had follow-up information, 195 (9.7%) and 214 (10.6%) experienced HF events and MACE during the median of 536 days after discharge. Patients with the high SHR ( >1.45, 4th quartile) had an increased risk of HF events than those with SHR ≤1.45, while the incidence of MACE after discharge did not differ significantly between the two groups (Figure). The multivariable analysis confirmed the association of SHR with long-term HF events.Conclusions:In patients with acute MI, SHR was predictive of in-hospital outcomes including HF events and MACE, while after discharge, the higher SHR was associated with a higher HF risk but not with MACE. Further studies are needed to elucidate the underlying mechanisms and potential incremental benefit of SHR in stratifying patient risks after MI.

Circulation, Volume 150, Issue Suppl_1, Page A4144217-A4144217, November 12, 2024. Background:The ISCHEMIA trial questioned revascularization in chronic coronary syndrome (CCS) patients, but excluding subjects with left main (LM) coronary artery disease (CAD). A widely available diagnostic method excluding LMCAD would expand the implementation of an initial noninvasive strategy.Objective:Assessing the ability of excluding LMCAD through clinical and ECG stress testing (EST) variables in patients undergoing coronary angiography (CAG) for CCS.Methods:In a multicenter retrospective case-control study we evaluated CCS subjects undergoing CAG after a maximal EST.Caseswere patients with angiographic ≥50% LM stenosis or ≥70% stenoses of both proximal left anterior descending and proximal circumflex arteries; we matched them with similar patients without LMCAD (Controls)in a 1:3 ratio. Models were internally validated through logistic regressions.Results:219Caseswere matched with 554Controls. The c-statistic was 0.80 (optimism-adjusted: 0.73). Assuming LMCAD prevalence of 5% and a misclassification cost ratio of 1:100 (ratio of the cost of performing CAG in a subject without LMCAD to the cost of not performing CAG in a patient with LMCAD), the negative predictive value was 98.6%, correctly classifying 84.5% ofCases. CAG could be spared in 57.0% of subjects, missing one LMCAD diagnosis every 70 CAGs spared in patients without LMCAD (Figure).Conclusions:Among patients with CCS, LMCAD can be predicted withacceptablediagnostic accuracy anda very highnegative predictive value through a model based on clinical and EST parameters, allowing an initial noninvasive management of most patients able to perform an EST, reducing the costs of routine coronary imaging. Such results should enlarge the applicability of the ISCHEMIA results when coronary computed tomography angiography, used in ISCHEMIA, is not available, limiting the referral to invasive CAG.

Circulation, Volume 150, Issue Suppl_1, Page A4147100-A4147100, November 12, 2024. Introduction:Perceived stress is known as the feeling of uncontrollability in one’s life and the inability to cope with the amount of stress one is experiencing due to one’s circumstances. There is evidence indicating that perceived stress may be a risk factor for cardiovascular disease (CVD). However, it is unclear whether perceived stress is associated with higher left ventricular mass index (LVMI), a measure of target organ damage and strong predictor of CVD.Aim:We examined the association between perceived stress and LVMI.Methods:The Masked Hypertension Study is a multi-site study that assessed the prevalence and predictors of masked hypertension in working adults from the New York Metropolitan area. A total of 1011 participants were recruited for the study; 826 participants completed the Perceived Stress Scale questionnaire and an echocardiogram. The perceived stress scale is a 14-item likert scale that assesses different situations that affects our perceived stress or our feelings in certain situations. It has a scale of 1 to 5 with 1 being never to 5 being very often. LVMI was determined according to the 2D method based on the American Society of Echocardiography (ASE) recommendations.Linear regression models were specified predicting LVMI from perceived stress. Covariates were sex, race/ethnicity, age, BMI, systolic bp, diastolic bp, and caregiving status.Results:Of the 826 participants, 40.7% were male, 6.9% were Black, 11.9% were Hispanic/Latinx , and 48.79% were caregivers. Mean (SD) age was 45.3 (10.3) years, mean body mass index was 27.6 (5.3) kg/m2, and mean SBP and DBP were 114.7 (12.0) and 74.8 (8.5). Mean perceived stress score was 21.8 (7.6) and mean LVMI was 63.3 (15.7) gm/m2.Contrary to our hypothesis, perceived stress was not significantly associated with LVMI, B = -0.00, 95% CI: -0.13, 0.13, p= 0.98.Conclusion:There was not an association between perceived stress and LVMI. Future research should examine whether other types of stresses/stressors are related to LVMI and the implications of this for patients and their health.

Circulation, Volume 150, Issue Suppl_1, Page A4140558-A4140558, November 12, 2024. Background:Single ventricle congenital heart disease such as hypoplastic left heart syndrome (HLHS) with a Fontan circulation constitute the largest group of children hospitalized with circulation failure, experiencing an in-hospital mortality rate of 20-50%. We investigated the mechanisms leading to circulation failure so as to identify novel therapeutic targets.Methods:Blood was collected from patients with HLHS s/p Fontan and controls with normal cardiac anatomy and function (N=6/group). Plasma microvesicles (MV) were isolated, and proteomics assessed using data independent acquisition mass spectroscopy. Dysregulated proteins with a fold change >1.5 or < -1.5, p

Circulation, Volume 150, Issue Suppl_1, Page A4143847-A4143847, November 12, 2024. Introduction:Our previous studies have shown that sustained activation of the DNA damage response (DDR) in cardiomyocytes leads to p53/p21 activation and cardiac dysfunction. Although the DDR generally involves molecules in DNA replication and repair pathways, the non-proliferative nature of cardiomyocytes suggests a cardio-specific DDR mechanism. However, our understanding of DDR in cardiomyocytes remains limited. Here, we aim to use CRISPR interference (CRISPRi) knockdown screens to identify genes critically involved in DDR regulation in human cardiomyocytes. We hypothesize that identifying these gene clusters may allow us to develop methods to prevent cardiac dysfunction by suppressing DDR in cardiomyocytes.Methods and Results:We established a human iPS cell line stably expressing dCas9-KRAB, which allows CRISPRi-mediated gene knockdown, and differentiated the cells into cardiomyocytes. The resulting human iPS cell-derived cardiomyocytes (hiPSCMs) showed the achievement of approximately 80% knockdown efficiency after gRNA transfection. We stimulated the hiPSCMs with H2O2and quantitatively evaluated the expression levels of the DDR markers γH2AX and p21 by immunostaining using the Operetta®high content imaging system. The DDR markers showed a significant concentration-dependent increase in response to H2O2administration. For arrayed CRISPRi screening, we constructed a gRNA library targeting 437 DDR-related genes. Using this library, we knocked down each DDR-related gene in hiPSCMs followed by H2O2stimulation. We quantified the expression levels of DDR markers by calculating the fluorescence intensity ratios relative to control after gene knockdown, and standardized them to calculate Z scores for all 437 genes. The screening successfully revealed the differential impact of each gene knockdown on γH2AX and p21 expression. We identified 71 genes that significantly affected their expression (Z-score < -1 or > 1). Mapping these genes to DDR pathways highlighted the differential impact of gene knockdown within the same pathway, and stratified their importance in cardiomyocytes.Conclusions:Arrayed CRISPR screening using hiPSCMs revealed differential functional significance of DDR-related genes in cardiomyocytes, identifying 71 genes of particularly significant importance. These findings provide a critical understanding of the cardio-specific DDR pathway and important clues for establishing an appropriate method to suppress DDR in the failing heart.

Circulation, Volume 150, Issue Suppl_1, Page A4142424-A4142424, November 12, 2024. Background:Risk assessment for chronic thromboembolic pulmonary hypertension (CTEPH) remain challenging. Stress hyperglycemia represents the regulation of glucose metabolism in response to stress. Meanwhile, stress hyperglycemia ratio (SHR) is recently found to reflect true acute hyperglycemic status. However, the relationship between SHR and adverse prognosis is uncertain. This study aimed to investigate the prognostic value of SHR in CTEPH patients.Methods:A total of 451 CTEPH patients with available baseline SHR measurement were enrolled between February 2014 to July 2023 at Fuwai hospital. The predictive values of SHR for adverse events were assessed.Results:During a median follow-up of 21 months, 89 (19.7%) CTEPH patients experienced adverse clinical outcomes. Kaplan-Meier curve analysis revealed that the cumulative adverse event rates were significant higher in the SHR≥0.747 with CTEPH patients, compared with patients in the SHR

Circulation, Volume 150, Issue Suppl_1, Page A4140462-A4140462, November 12, 2024. Background:Right ventricular pacing (RVP) can have adverse cardiac effects and cause pacing induced cardiomyopathy (PiCM). His bundle pacing (HBP)&Left Bundle Branch area pacing (LBBAP) mimic physiologic conduction (PhysioP) and maintain biventricular synchrony.Hypothesis and Aims:Reduced left ventricular (LV) systolic function reserve in the presence of normal baseline LV ejection fraction (EF) could precede development of RV PiCM. Our aim was to compare the effects of RVP vs. PhysioP on bicycle exercise cardiopulmonary performance in patients with normal LVEF who required pacing for bradyarrhythmias.Methods:Patients with sinus rhythm and RVP or PhysioP&ventricular pacing burden of >70% who completed cardiopulmonary exercise test and simultaneous stress echocardiography (SE) were included. Pulmonary gas exchange was calculated using Ventilation/CO2 production at rest and during exercise. Changes in LV size, EF, longitudinal strain and diastolic function and gas exchange parameters were compared post and pre exercise in the 2 groups.Results:25 of 29 patients completed the study [68 ± 23 yrs, 48% M; LVEF 56±5%, 11 RVP, 14 PhysioP]. There was no difference in baseline demographic&clinical variables, exercise duration, rest and peak heart rate and blood pressure between 2 groups. Pacing duration was 2.61±1.48 yrs in RVP vs. 0.84±0.67 yrs (p=0.003) in the Physio group. Resting echocardiographic parameters (Table 1A)were comparable. Compared to RVP, reduction in LV end-diastolic volume (EDV) 3.4±14.1 ml vs. -23.1±18.1ml, p=0.006)&LV end-systolic volume (ESV -5.7±11.6 ml vs. -18.0±9.5ml, p=0.01) was more pronounced in the PhysioP group. Changes in LVEF, LV strain&diastolic function were not different between the 2 groups (Table 1B). There were no significant differences in changes in pulmonary gas exchange parameters in the 2 groups.Conclusions:In patients with normal LVEF and pacemaker dependent, RVP is associated with impaired but PhysioP with preserved LV systolic function reserve, which can be detected by exercise SE. SE may help identify patients at risk for RV PiCM. Benefit of PhysioP needs to be determined by larger studies with longer follow-up.

Circulation, Volume 150, Issue Suppl_1, Page A4137735-A4137735, November 12, 2024. Background:Aortic aneurysms account for ~10,000 deaths annually in the USA. Oxidative stress is implicated in both abdominal (AAA) and thoracic (TAA) aneurysm formation, but the mechanisms are incompletely understood. We used a chemogenetic approach to modulate oxidative stress in the vascular wall by creating a transgenic mouse (DAAO-TGTie2) that expresses yeast D-amino acid oxidase (DAAO) driven by the endothelial Tie2 promoter. DAAO generates the reactive oxygen species hydrogen peroxide from D-amino acids. Vascular tissues contain L-amino acids, so yeast DAAO is quiescent until DAAO-TGTie2mice are provided with D-amino acids. Here we characterize the cellular and molecular consequences of vascular oxidative stress in DAAO-TGTie2mice.Hypothesis:Chronic oxidative stress in the vascular wall causes arterial dysfunction.Aim:To characterize the phenotype of DAAO-TGTie2mice after oxidative stress induction in vascular endothelium.To identify the mechanisms whereby vascular oxidative stress causes arterial dysfunction and hypertension.Methods:Systolic blood pressure and aortic sonography were measured weekly in D-alanine-fed DAAO-TGTie2. Proteomic analyses were used to identify mechanistic targets, which were validated using biochemical and immunohistochemical methods.Results:D-alanine-fed DAAO-TGTie2mice develop systolic hypertension and abdominal but not thoracic aortic aneurysms; treated mice die in >3 months with burst abdominal aortic aneurysms. Transgene expression is similar in abdominal and thoracic endothelium. Levels of oxidative stress markers (oxidized proteins, lipid, and DNA) were similar in thoracic and abdominal aorta. Proteomic analyses established phenotypic switching in abdominal but not thoracic aorta, and also revealed activation of the oxidant-activated kinase ASK1 and of the MAP kinase cascade in abdominal but not thoracic aorta. Immunoblot analyses showed a marked decrease in JNK1 phosphorylation by phosphatase DUSP3 and an increase in vascular KLF4, leading to phenotypic switching of contractile to synthetic VSMCs.Conclusion:Chronic chemogenetic oxidative stress induces hypertension and abdominal aortic aneurysm formation caused by KLF4-dependent VSMC phenotypic switching.

Circulation, Volume 150, Issue Suppl_1, Page A4138348-A4138348, November 12, 2024. Methods and Results:We describe a comprehensive characterization of the AVICs nucleus landscape as determined by transmission electron microscopy (TEM) of samples obtained from CAVD patients. Turbulence led to nuclear envelope integrity lose in AVICs cultured in shear stress experiments, with three different fluid conditions [static (ST), laminar stress (LS), and oscillatory stress (OS)], indicated by Western blot and immunofluorescence (IF). Silencing lamin A/C (LMNA) through small interfering RNA (siRNA), accelerated nuclear envelope damage , as indicated by Western blot, qPCR, and immunofluorescence (IF). The formation of Z-DNA and its co-localization with Z-DNA binding protein (ZBP1) was observed due to the nuclear envelope damage by IF. Western blot, qPCR, IHC and IF confirmed Z-DNA-induced inflammation in AVICs through the ZBP1-RIPK3-NF-κB signaling pathway. ZBP1 and RIPK3 knockdown with siRNA markedly reduced the protein level of osteogenic markers alkaline phosphatase (ALP), runt-related transcription factor 2 (RUNX2), and bone morphogenetic protein 2 (BMP2) in VICs. In vivo, aortic valve disease was constructed by direct wire injury (DWI), and we showed that overexpression of LMNA by adeno-associated virus significantly decelerated the progression of aortic valve lesion induced by DWI in mice.Conclusion:Excessive mechanical stress can induce damage to the nuclear envelope of AVICs by causing cytoskeletal remodeling, initiating the formation of Z-DNA, and hastening the calcification process in AVICs and CAVD.

Circulation, Volume 150, Issue Suppl_1, Page A4146930-A4146930, November 12, 2024. Background:Young adulthood (19–39 years) is the life stage of greatest declines in cardiovascular health (CVH). It is hypothesized that this decline may be related to competing demands (e.g., stressors) of this period of the lifecourse such as work and child-rearing. The AHA’s Life’s Essential 8 CVH framework identifies the scored domains (behavioral and clinical factors), as needing to be contextualized by the important construct of psychological health (including stress). However, scarce data are available to assess the relationship between CVH and reported stress – especially among YA.Purpose:The current study aims to be the first to use Cosmos electronic health record (EHR) data to assess the relationship between YA CVH and reported stress in a nationally representative, very large sample of YAs.Methods:Cosmos is a platform hosting de-identified Epic EHR data on >250 million patients. We assessed Cosmos data from May 2022 through April 2024 to identify all YAs with reported stress data (5-point scale from “not at all” to “very much”). We then compared trends of each CVH metric across stress categories.Results:1,397,375 individuals 19 – 39 years of age had reported stress data available. The sample was 62% White, 17% Black, 4% Asian, 12% Hispanic, and 63% female. Generally, stress levels were stable across YA age groups (Figure 1). For lifestyle behavior related domains (physical activity (PA), smoking, and BMI), the prevalence of “poor” CVH scores (worst categorization) increased in YA as reported stress amount increased (Figure 2). Prevalence of “poor” scores in clinical metrics (BP, HbA1c, nonHDL-C) were not associated with stress.Conclusion:In a very large sample of YA, greater reported stress was associated worse CVH for the behavioral domains of PA, smoking and BMI. Interventions aimed at reducing stress in YA may have the added benefit of improving CVH.