bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 15143: The Sphk1/S1P/S1PR2 Axis Regulates Pulmonary Vascular Remodeling by Activating a Mitochondrial ROS and NLRP3 Inflammasome Regulatory Loop
Authors: Bai Yang , Gomes Marta T , Prohaska Clare , Lockett Angelia , Machado Roberto F
Published: June 2023

Introduction:
Sphk1/S1P/S1PR2 axis regulated NLRP3 inflammasome and promoted pyroptosis by activating NF-κB in human PASMC. Abnormal or inappropriate ROS mediates the development of PAH modulating inflammation, proliferation, and energy metabolism. Mitochondrial ROS promotes deubiquitylation of NLRP3 and activate the inflammasome. However, the function and regulation of S1P axis mediated mitochondrial ROS and NLRP3 inflammasome in PAH remain unknown.

Hypothesis:
We hypothesized that S1P axis regulated pulmonary vascular remodeling by activated mitochondrial ROS and NLRP3 inflammasome.

Methods:
NLRP3 inflammasome and pyroptosis activation were detected in PASMC from patients with PAH. JTE013, PF543, MCC950 and MitoTEMPO were used to modulate S1P, NLRP3 inflammasome and mitochondrial ROS signaling, respectively. Activation of inflammasome and pyroptosis were detected by western blots (WB), real-time PCR or immunofluorescence (IF). IL-1β secretion was detected by ELISA. ROS were measured by Mitosox Red and DCFDA. Mitochondrial membrane potential was determined by JC-1 dye assay. PASMCs which were isolated from Sphk1
fl/fl Tagln Cre (+)
and Sphk1
fl/fl
mice were used to detect the role of S1P axis in vivo.

Results:
NLRP3 Inflammasome and pyroptosis were activated in PASMCs from patients with PAH. JTE013 or PF543 treatment inhibited NLRP3, ASC, N-GSDMD, HMGB1, cleaved IL-1β and IL-18 expression, and suppressed mitochondrial ROS, as well as mitochondrial hyperpolarization induced by S1P. Treatment with MitoTEMPO decreased S1P induced-NLRP3, ASC, N-GSDMD, HMGB1, cleaved IL-1β and IL-18 expression and inhibited IL-1β secretion. MCC950 controlled the activation of mitochondrial ROS, as well as mitochondrial hyperpolarization which were activated by S1P. NLRP3, cleaved IL-1β, ASC, HMGB1 and N-GSDMD expression were decreased in mPASMCs from Sphk1
fl/fl Tagln Cre (+)
mice.

Conclusions:
S1P induces activation of mitochondrial ROS and NLRP3 inflammasome, and that the Sphk1/S1P/S1PR2 signaling axis may be involved in this context. Activation of the mitochondrial ROS and NLRP3 inflammasome regulatory loop may be an important mechanism underlying PAH pathogenesis, targeting of which may be exploited for the treatment of PAH.
Abstract 15168: Plasma Metabolomic Profiling Identifies Markers of Cardiorespiratory Fitness Responsive to Endurance Exercise Training
Authors: Rao Prashant , Mi Michael , Barber Jacob , Deng Shuliang , Farrell Laurie , Bouchard Claude , Clish Clary , Sarzynski Mark A , Gerszten Robert E , Robbins Jeremy
Published: June 2023

Background:
High-throughput metabolite profiling allows for the assessment of thousands of metabolites that may participate in exercise-related pathways. Here, we characterize plasma metabolite changes in healthy individuals undergoing endurance exercise training (ET) to identify biochemical features of VO
2
max and/or those responsive to ET.

Methods:
We measured 444 known metabolites and 3221 non-targeted metabolite peaks using liquid chromatography tandem mass spectrometry and VO
2
max using CPET before and after 20 weeks of supervised, endurance ET in 654 sedentary participants (mean age=34y; 39% self-identified Black) from the HERITAGE Study. Multivariate linear regression assessed the relation between metabolites and baseline VO
2
max (ml·kg
–1
·min
–1
), adjusting for age, sex, and race. Changes after ET were determined using paired
t
tests. We used a false discovery rate (q)
<
0.05 to determine statistical significance.

Results:
494 metabolites were associated with VO
2
max, the majority (375/494) of which were non-targeted (unknown) metabolites with greater effect sizes than targeted peaks [β range: -29.66 - 17.80 and -22.76 - 12.87 for unknown and known metabolites]. We recapitulated known metabolite associations with VO
2
max including DMGV, branched chain amino acids, and uric acid (
Figure 1
). 110 of 560 metabolites that changed after ET were associated with VO
2
max, including non-targeted peaks that concordantly changed after ET (e.g. metabolite peaks positively associated with VO
2
max that increased with ET;
m/z
385.3056 and 695.5086; β for VO
2
max = 9.82 and -19.33, q= 5.56e-7 and 2.61e-17, and log fold-change = 0.03 and -0.01, q=9.58e-13 and 1.93e-4, respectively).

Conclusions:
We identified novel metabolite peaks associated with VO
2
max that concordantly changed after ET. Non-targeted metabolomic profiling reveals new markers related to VO
2
max and exercise response, motivating ongoing work to unambiguously identify these compounds.
Abstract 15179: Histology-Validated Delayed Enhancement Magnetic Resonance Imaging of Atrial Fibrillation Driver Substrate
Authors: Mylavarapu Uma , Buck Benjamin , Subr Megan , Hoenie Lisa , Kalyanasundaram Anuradha , Pan Yue , Kennelly James , LI Ning , Hansen Brian , Zhao Jichao , Simonetti Orlando , Hummel John , Fedorov Vadim V
Published: June 2023

Introduction:
Persistent (per) AF may be maintained by localized extra-PV drivers (DRs). Regional fibrosis can serve as a substrate for AF DRs, but its visualization by delayed enhancement (DE)-MRI lacks proper validation for driver substrates. The objective of this study is to evaluate if histology-validated normalized (HVN) DE-MRI thresholds can define arrhythmogenic fibrotic content in multielectrode mapping (MEM)-defined DRs from perAF patients’ (Pts) DE-MRI.

Methods:
PerAF Pts (n=14, 71% male; 63±10 y/o) had DE-MRI at 3T (0.63x0.63x0.63mm
3
, 0.2mmol/kg Gd) before their perAF ablation procedure. Chamber-specific DE-MRI intensity was normalized to the healthy atria (mean lowest 5% intensity, 0 AU) and mitral (LA) or tricuspid (RA) valve (100 AU). 3 control and 5 perAF canines had DE-MRI and regional histology to validate DE-MRI fibrotic thresholds for the LA and RA, which were defined as standard deviations (SD) above 40% of the mean healthy atrial tissue intensity (LA: 3.7 SD, RA: 2.7 SD). Fibrosis-enhancement of segmented MEM-defined DRs (LA: n=25, RA: n=11) were analyzed using conventional, non-normalized 3 SD and HVN thresholds and compared to 5 random regions (non-DRs) within the same anatomical region(s).

Results:
Only our HVN approach could reveal arrhythmogenic fibrotic substrates in LA DR regions, but not the conventional approach. Both approaches could not provide significant differences for RA DR vs non-DR fibrosis regions. The cut-off of 33.6% of LA DE-MRI fibrotic enhancement using the HVN approach allowed to distinguish LA DR vs. non-DR regions with 56% sensitivity and 70% specificity. At 12 months follow-up, 10 Pts remained free of AF.

Conclusion:
MEM-defined DRs within fibrotic regions, identified by HVN DE-MRI, may represent efficient targets for ablation of LA arrhythmogenic perAF substrates. Integration of MEM and MRI may lead to improved targeted ablation and long-term AF freedom.
Abstract 15183: From Insult to Injury: A Rare Consequence of Sodium Glucose Cotransporter 2 Inhibitors
Authors: Sharma Aakanksha , Sharma Apurva , Kornberg Robert
Published: June 2023

Introduction:
Emphysematous pyelonephritis (EPN) is a rare, life-threatening necrotizing infection associated with diabetes mellitus (DM), especially in older women. Patients can remain asymptomatic or present with severe sepsis and bacteremia due to E. Coli (58%) or K. pneumonia (21%). Herein, we present a unique case of recurrent life-threatening sepsis due to EPN in a male with Heart failure with reduced ejection fraction (HFrEF) and well controlled DM treated with a sodium-glucose cotransporter two inhibitor (SGLT2i).

Case:
A 74-year-old man with metabolic syndrome, HFrEF (EF 30%), chronic kidney disease (stage 2) who was started on empagliflozin by his cardiologist four days prior, presented to the emergency department with dizziness and fever for one day. Vitals were notable for temperature of 102.9F and blood pressure of 88/52 mmHg. The physical exam was unremarkable. His WBC was 16,000/dL, and lactate was 2.8. A sepsis workup revealed a urinalysis positive for cystitis along with a CT of the abdomen, which demonstrated evidence of acute right-sided emphysematous pyelonephritis (Fig. 01). Blood culture grew pan sensitive E coli. He was treated with IV Ceftriaxone 2g once daily for 14 days. His previous records unveiled that the patient was initially started on empagliflozin two months ago, followed by admission with EPN within ten days, after which the medication was held until its reintroduction before the current presentation.

Conclusion:
Emphysematous pyelonephritis as a complication of SGLTI2I use is remarkably under-reported. The median time of onset of disease from SGLT2i initiation is 45 days. The patient experienced his first episode of EPN within ten days and then four days after reintroduction of SGLT2i. This case illustrates the potential for severe UTIs with SGLT2i and the value of a thorough history emphasizing the history of UTIs, especially those with obstructive renal symptoms.
Abstract 15205: Cardiovascular Outcomes With Use of Primary Prevention Implantable Cardioverter Defibrillators in Patients With Non-Ischemic Cardiomyopathy: A Meta-Analysis and Meta-Regression Analysis
Authors: Lone Ahmad Naeem , Ghosh Priyanka , Zaidi Ali , Sporn Daniel , Deshmukh Pramod
Published: June 2023

Introduction:
Guidelines recommend use of implantable cardioverter defibrillator (ICD) in all patients with non-ischemic cardiomyopathy (NICM) and ejection fraction (EF) < 35% for primary prevention of sudden cardiac death (SCD) and hence reduction of overall mortality. Contemporary randomized clinical trials (RCTs) with longer follow up duration have thrown into question this beneficial effect of ICDs.

Hypothesis:
Prophylactic use of ICDs in addition to optimal medical therapy (OMT) does not reduce adverse outcomes in patients with NICM and EF < 35% compared to OMT alone.

Methods:
Six RCTs (n=4541) comparing ICD + OMT vs OMT alone were selected from MEDLINE and EMBASE databases from inception through March 2022. Meta-analysis was performed using random effects model. Meta-regression analysis was performed with inverse-variance weighting and Knapp and Hartung adjustments for all-cause mortality.

Results:
ICDs did not reduce risk of all-cause mortality (Relative Risk (RR) 0.87, [95% confidence interval, 0.75-1.00], cardiovascular mortality (RR, 0.80 [0.49-1.31]), non-cardiovascular mortality (RR, 0.66 [0.13-3.34]) or non-sudden cardiac death (RR, 1.03 [0.51-2.08]). ICDs reduced the risk of sudden cardiac death (HR, 0.49 [0.26-0.92]). Meta-regression analysis did not show any association between all-cause mortality and follow up duration (p=0.09).

Conclusions:
ICDs reduce risk of SCD however this does not translate into meaningful improvement in long term cardiovascular outcomes.
Abstract 15248: Clinical Outcomes of Transcatheter Aortic Valve Replacement With and Without Percutaneous Coronary Intervention - An Updated Meta-Analysis and Systematic Review
Authors: Yassen Mohammad , Moustafa Abdelmoniem
Published: June 2023

Hypothesis:
Transcatheter aortic valve replacement (TAVR) serves as a less-invasive treatment option for high-risk patients with severe aortic stenosis. Given the coexistence of obstructive coronary artery disease in patients with high-grade aortic stenosis (estimated to be 40-75%), there is inconsistent clinical data regarding potential mortality benefits of paired percutaneous coronary intervention (PCI) with TAVR procedures.

Methods:
We performed a literature search using PubMed, Embase, and Cochrane Library from inception through April 2022 to assess the mortality impact of preceding/concomitant PCI in patients undergoing transcatheter aortic valve replacement. The primary outcomes were 30-day all-cause mortality, 30-day cardiovascular mortality, and 6 months-1 year all-cause mortality. Secondary outcomes included 30-day myocardial infarction, stroke, major bleeding/vascular complications, and acute kidney injury.

Results:
11 studies (10 retrospective cohort studies, 1 randomized control trial) involving 2791 patients were included in the meta-analysis. Compared to patients undergoing TAVR alone, the TAVR+PCI group showed no significant difference in 30-day all-cause mortality (RR 0.90, CI 0.66, 1.22, p =0.49), 30-day cardiovascular mortality (RR 0.71 CI 0.44, 1.14, p =0.16), or 6 months-1 year all-cause mortality (RR 0.94, CI 0.75, 1.18, p =0.57). Regarding secondary outcomes, 30-day myocardial infarction was higher in the TAVR+PCI group compared to the TAVR group (RR 3.09, CI 1.26, 7.57, p =0.01), while no significant differences were found in rates of 30-day stroke (RR 1.14, CI 0.56, 2.33, p =0.72), major bleeding/vascular complications (RR 1.11, CI 0.79, 1.56, p =0.55), and acute kidney injury (RR 1.07, CI 0.75, 1.54, p =0.71).

Conclusion:
Concomitant/preceding percutaneous coronary intervention does not confer any additional mortality benefit, and may increase the risk of 30-day myocardial infarction, in patients with high-grade aortic stenosis undergoing transcatheter aortic valve replacement. Further trials with large sample sizes are needed to confirm our findings.
Abstract 15273: Impact of Lipid Lowering Therapy on Progenitor Endothelial Cell Phenotype and Function in Coronary Artery Disease
Authors: Jimenez-Trinidad Francisco , Bobi Joaquim , Ortega-Paz Luis G. , Rodr?guez Arias Juan J , Roque Merce , Sabate Manel , Brugaletta Salvatore , Dantas Ana Paula V
Published: June 2023

Introduction:
Early revascularization of infarcted zone is crucial to improve patient′s outcomes. Despite their primary actions, major lipid-lowering drugs may also reduce cardiovascular events by pleiotropic mechanisms.

Hypothesis:
Conventional (statin) and novel lipid-lowering therapies [(PSCK9-inhibitor (PCSK9i) and eicosatetraenoic acid (EPA)] can improve endothelial progenitor cells (EPC) phenotype and favor neovascularization.

Methods:
EPC were isolated from peripheral blood of patients with acute coronary syndrome (n=62) who were statin-naïve (n=34) or chronic statin treatment (n=28) at time of inclusion. A set of statin-naïve EPC and mature endothelial cells (HUVEC) were treated
in vitro
with statin (20nM), PCSKi (1μM) and EPA (3μM). EPC phenotype was determined as EPC colony number and time of appearance, proliferative rate, and ability to form capillary-like structures. Serum PCSK9 and markers of angiogenesis and endothelial function were determined by multiplexed protein assay.

Results:
Frequency of EPC appearance was higher in samples of statin group (78%) compared to naïve group (66%) (Fig A). They also appeared much earlier (Fig B), in greater quantity (4.7 vs 2.1 colony, p = 0.0009) and had higher proliferative rate than EPCs of naïve group. Besides they had increased potential of tube formation and shaped more branched vessel-like structures (Fig C). No differences in serum levels of biomarkers were found. In vitro treatment of EPC with statin, PCSK9i and EPA did not modify colonies behavior (Fig D). PCSK9 levels showed negative correlation with EPC proliferation (r -0.4150, p=0.004), and PCSK9i treatment increased angiogenic potential of HUVEC (Fig E).

Conclusions:
Statin and PCSK9i have pleiotropic effects on EPC, boosting their potential to form endothelial cells colonies and to induce angiogenesis. These effects could contribute to the progression and outcomes of coronary artery disease.
Funding:
ISCIII - FEDER-ERDF (PI19/00264)
Abstract 15278: Transcatheter Closure of Atrial Septal Defect Over the Age of 60
Authors: Balint Olga Hajnalka H , Kecskemeti Dorottya , Nagy Zsolt , Szabo Balazs , Temesvari Andras
Published: June 2023

Introduction:
The long-term benefits of transcatheter device closure of a secundum atrial septal defect (ASD) over 60 remain controversial. The aim of the study was to assess the predictors of death late after device closure in these elderly patients.

Methods:
This retrospective study enrolled 87 patients with successful ASD transcatheter closure over 60 and at least 1 year follow-up after closure. The mean age was 73.3±5 years, and 74 of them were female (85%). All patients had an isolated secundum ASD with a mean defect size of 18±4 (9-30) mm. At the time of closure 10 patients were known with pulmonary hypertension (11%), 52 with atrial arrhythmia (61%), 56 with heart failure (64%), and 15 with coronary artery disease (17%). Demographic, clinical and echo data were compared between values at the time of closure, and at the last follow-up. Predictors of death were analyzed with multivariate Cox regression.

Results:
After the mean follow-up of 5±4 years 11 patients developed high LV filling pressure, 15 patients had new atrial arrhythmia and 10 patients right heart failure. At the last FU a significant decrease in right ventricular (45±5 vs 38±8 mm, p=0.032) and right atrium dimension (62±10 vs. 59 ±10mm, p=0.045) were seen, together with right ventricular systolic pressure improvement (51±18 vs. 40±19 mmHg, p=0.038). The left atrium showed significant further dilatation after closure (60±9 vs. 62±9.5, p=0.048). The presence of pulmonary hypertension, right ventricular dilatation and tricuspid regurgitation correlated with the right heart failure after closure, and the atrial dimension, tricuspid regurgitation, and high filling pressure with new atrial arrhythmia. Fourteen patients died during follow-up. On multivariate analysis the main predictor of death was right heart failure (HR: 10; 95% CI 2,15-41).

Conclusions:
In terms of right heart remodeling, elderly patients still benefit from late ASD catheter closure. However, those who develop right heart failure after closure are at a high risk of death (HR:10). Diastolic dysfunction after the defect closure was not a predictor of death.
Abstract 15280: Lower Tricuspid Annular Plane Systolic Excursion/Pulmonary Artery Systolic Pressure Coupling is Associated With Longer Hospital Stay and Mortality in Patients With Pulmonary Hypertension
Authors: Oren Daniel , Moeller Cathrine Marie , Raphael Roy , Gelikas Shaul , Faierstein Kobi , Canetti Michal , Mouallem Meir , Mayan Haim , Shilo Noya
Published: June 2023

Introduction:
Heart failure (HF) is a leading cause of admissions to Internal Medicine (IM) wards worldwide and is associated with significant morbidity, mortality, and healthcare costs. Growing utilization of point-of-care cardiac ultrasound by IM physicians may help reveal overlooked markers of worse outcomes in the indexed hospitalization. The ratio of Tricuspid Annular Plane Systolic Excursion (TAPSE)/Echocardiographically measured Pulmonary Artery Systolic Pressure (PASP) may offer insight into right ventricular function and the course of HF decompensation hospitalization in patients with known Pulmonary Hypertension (PH).

Methods:
We retrospectively reviewed the files of 2500 in-patients adults >18 years old who were admitted to an IM ward in a single tertiary medical center, between 2010-2022. Inclusion criteria included a known prior diagnosis of HF and PH and an in-house echocardiogram performed at the time of admission. TAPSE/PASP coupling was assessed and the quotient was plotted on a logarithmic scale in patients who expired during the indexed hospitalization and those who did not. To assess the length of hospitalization we used a cox regression analysis to compare the probability of discharge based on the degree of TAPSE/PASP coupling.

Results:
Patients who survived the indexed hospitalization had a median TAPSE/PASP of -3.19 IQR [(-3.505) - (-2.853)] as opposed to -3.47 IQR [(-3.749) - (-3.188)], P <0.000, figure 1. Cox proportional hazard regression model used to assess if there was a difference in hospitalization duration given minimal vs. maximal TAPSE/PASP coupling had a likelihood ratio of 13.25 P = 0.0002, figure 2.

Conclusions:
Echocardiographically determined TAPSE/PASP is a non-invasive measure of Right Ventricular (RV)-arterial coupling and a good estimator of RV function. In our cohort lower TAPSE/PASP was associated with longer length of hospitalization and mortality.
Abstract 15303: Maternal Features for Live Births in Fontan Patients
Authors: Sankar Parvathy Ravi , Garraud Cassandra , Scandinaro Anna , Saef Joshua M , Ghobrial Joanna
Published: June 2023

Introduction:
As the population of women with congenital heart disease with prior Fontan palliation (PFPs) increases, doubling in size over the next 20 years, there will be a concordant need for multidisciplinary obstetrics care. There is an increased prevalence of miscarriage, maternal and fetal complications including arrhythmias, heart failure, premature birth, and neonatal death. We sought to review our single center experience in the context of an established cardio-obstetrics multidisciplinary team.

Methods:
Single-center retrospective cohort study of PFPs with live births who received care at our cardio-obstetrics clinic. A chart review was performed to identify maternal characteristics as well as fetal outcomes. Descriptive statistics were calculated using R Studio Statistical Software version 1.2.5033.Result: Ten women with PFPs and live births were identified. None had more than one live pregnancy. The average number of pregnancies was 2 (range 1-6). The mean gestational age was 31.2 weeks (range 23-37). Five patients were primigravida. Six of the pregnancies were planned and eight received preconception counseling. Baseline NYHA was 50% class I and 50% class II. The most common maternal comorbidity in patients with a live pregnancy was atrial arrhythmia, and the most common pre-pregnancy medication was warfarin. Prematurity, low birth weight, and associated comorbidities (see table 1) were common in this population. The average birthweight was 1,589g (range 460g-2210g). Indication for delivery was more commonly obstetric than maternal concern. No maternal strokes, thromboembolic events, cardiac arrests, maternal or infant deaths and rehospitalizations were noted.

Conclusion:
Maternal and fetal complications in mothers with PFP are common. A multidisciplinary approach in a specialized cardio-obstetrics clinic has the potential to improve outcomes. Larger cohort and long term outcomes are needed in this patient population.
Abstract 15337: Chronic Oral Finerenone Treatment Does Not Improve Left Ventricular Diastolic Compliance in Swine With Repetitive Pressure Overload-Induced HFpEF
Authors: Hudson Emily , Spinella Samantha , Nsumbu Dorcas , Palka Beth , Canty John M , Weil Brian R
Published: June 2023

Introduction:
The non-steroidal mineralocorticoid receptor antagonist finerenone has emerged as a potential treatment for heart failure with preserved ejection fraction (HFpEF) based, in part, on initial evidence of its anti-fibrotic properties. However, it is unclear whether chronic oral finerenone therapy can reverse established cardiac fibrosis. Accordingly, we tested the efficacy of chronic oral finerenone in a porcine model of HFpEF characterized by interstitial fibrosis and reduced left ventricular (LV) diastolic compliance in the absence of co-morbidities or anatomic LV hypertrophy.

Methods:
Swine (n=12) were subjected to daily (1 hr) episodes of repetitive LV pressure overload (RPO) for 2 weeks using phenylephrine (24 mg/hr) to transiently elevate LVEDP to ~30 mm Hg. After 2 weeks, RPO was discontinued and animals were followed for an additional 4 weeks, during which a subset received oral finerenone (20 mg/day; n=6). LV end-diastolic volume (EDV) and pressure (EDP) were measured at normal and elevated preload to assess LV compliance (ΔLVEDV/ΔLVEDP) and post-mortem picrosirius red staining was performed to quantify interstitial fibrosis vs. normal controls (n=8).

Results:
Four-weeks after cessation of RPO, untreated animals exhibited a significant increase in interstitial fibrosis vs. normal controls that was attenuated in finerenone-treated animals (
Figure A
). Despite this regression of interstitial collagen, a persistent reduction in LV diastolic compliance was observed after finerenone treatment (1.18 ± 0.33 mL/mmHg vs. 2.11 ± 0.22 mL/mmHg in normal controls; p=0.04) that was comparable to that of untreated animals after RPO (0.97 ± 0.22 mL/mmHg; p=0.61;
Figure B
).

Conclusion:
Four weeks of chronic oral finerenone treatment does not restore LV diastolic compliance in swine with RPO-induced HFpEF despite regression of interstitial collagen content, suggesting that other factors may contribute to diastolic stiffening in this model.
Abstract 15339: Er-Resident S1pr1 in Lung Endothelium Regulates Innate Immune Function and Mediates Inflammatory Lung Injury
Authors: Anwar Mumtaz , Balaji Ragunathrao Vijay Avin , Tauseef Mohammad , Joshi Jagdish Chandra C , Sheikh Rayees R , Joshi Bhagwati , Tsukasaki Yoshikazu , Chronis Konstantinos , Mehta Dolly
Published: June 2023

Endothelial-cell surface localized sphingosine 1 phosphate receptor 1 (S1PR1) is known to promote anti-inflammatory and barrier enhancing niche upon ligating S1P. Recently we showed that S1P and TNFα, later being a well-known inflammatory agonist, phosphorylate S1PR1 at tyrosine
143
(Y
143
) which functions as an endoplasmic reticulum (ER) import signal (Anwar et al, 2021). ER-retained S1PR1 instructs barrier disruptive signaling but the mechanism remains unclear. Here, we generated S1PR1 knock-in mice using CRISPR-Cas9 strategy to edit endogenous S1PR1 into Y
143
D-S1PR1 (phospho mimicking) or Y
143
F-S1PR1 (phosphodefective) to test the hypothesis that ER-localized S1PR1 subverts EC from anti-inflammatory to pro-inflammatory EC leading to vascular injury. Because EC constitutes about 50% of cells in the lungs, we assessed if knock-in of Y
143
D-S1PR1 impaired lung homeostasis. We show that editing of S1PR1 into Y
143
D- or Y
143
F-S1PR1 did not alter total S1PR1 expression. Interestingly, Y
143
D-S1PR1 knock-in mice showed marked vascular leak at homeostasis along with increased neutrophil influx and inflammatory cytokine generation including TNFα, IL1ß and MiP2 as compared to Y
143
F-S1PR1 or WT mice. We next challenge these mice with intratracheal LPS. LPS-induced non-resolvable vascular inflammatory injury in Y
143
D-S1PR1 mice. Surprisingly, Y
143
F-S1PR1 knock-in mice did not develop vascular inflammatory injury. Furthermore, NFκB activity, a predominant transcription factor inducing inflammatory EC phenotype, was increased in EC transducing Y
143
D-S1PR1 mutant as compared to WT. However, TNFα failed to induce NFkB activity in EC transducing Y
143
F-S1PR1 mutant. Together, these results show that ER-resident S1PR1 program endothelial niche into immune-active niche by activating NFkB pathway leading to irreparable lung injury. Further experiments are being done to assess epigenetic changes (ATACseq and ChIP-Seq) in EC to address the concept the ER-resident S1PR1 controls the fate of immune cells in the lungs. We believe that understanding how ER-resident S1PR1 programs EC into inflammatory phenotype would allow development of new targets for treating the inflammatory vascular diseases including lung injury, ARDS, and COVID-19
Abstract 15340: Association of Serum Lipoprotein (a) Levels and Lipid Burden by Near Infrared Spectroscopy in Intermediate Coronary Artery Disease
Authors: SOH Moon Seung , Lee Kwang-No , Seo Kyoung Woo , Park Jin-Sun , Yang Hyoung-Mo , Lim Hong Seok , Choi Byoung-Joo , Choi So-Yeon , Hwang Gyo-Seung , Shin Joon-Han , Tahk Seung-Jea , Yoon Myeong Ho
Published: June 2023

Introduction:
The relationship between elevated lipoprotein (a) (Lp(a)) and poor prognosis in coronary artery disease (CAD) has been addressed for decades. However, data of mechanism of increased risk with plaque lipid burden has not been established.

Hypothesis:
This study aimed to investigate the association between Lp(a) and lipid burden in patients with intermediate CAD by Near Infrared Spectroscopy (NIRS).

Methods:
We retrospectively analyzed 128 patients with intermediate CAD who performed intravascular ultrasound (IVUS) during coronary angiography and checked Lp(a) level. The lipid core burden index (LCBI) for the 4-mm segments at the target lesion was calculated by NIRS. We divided the patients into 3 groups by calculating baseline Lp(a) tertile level. A mixed model adjusted for baseline sex, age, weight, pre-minimal luminal area, lesion length, apolipoprotein (Apo) E, Apo A1, Apo B100, total cholesterol, triglyceride, high density lipoprotein (HDL), low density lipoprotein (LDL), fasting glucose, past history, current medication including statin use was used to calculate the risk-adjusted target LCBI.

Results:
We enrolled 128 patients (60 years; male, 114; mean Lp(a) 21.03±20.84 mg/dl, Figure 1) and NIRS was taken in 90 patients among them. Mean and median target LCBI by NIRS were 78.4±60.3 and 62.0. Target LCBI significantly higher in the intermediate Lp(a) group than low Lp(a) group in risk-adjusted analyses (Lp(a)<9; 45.7 versus 9≤Lp(a)<19; 111.48,
p
=0.026, Table 1). However, between intermediate and high Lp(a) groups, there was no statistical difference (9≤Lp(a)<19; 111.48 versus 19≤Lp(a); 86.41,
p
=0.774).

Conclusions:
Elevated Lp(a) level is independently associated with increased lipid burden of target lesion in intermediate CAD, and it’s important for treatment decision. Further investigation is needed to clarify the effect of Lp(a) in various type and severity of plaque.
Abstract 15352: Lung Endothelial Transport in Glypican 1 Knockout Mice
Authors: Black Stephen , Zemskov Evgeny A , Carvalho-de-Souza Joao , Chignalia Andreia
Published: June 2023

Introduction:
Pulmonary edema is a life-threatening condition that can result from the acute increase in hemodynamic forces in the lung vasculature. The mechanisms whereby pulmonary edema develops are unresolved but increased lung endothelial permeability may be important. The endothelial glycocalyx is a surface layer known to mediate the effects of mechanical forces in the lung vasculature by eNOS-dependent mechanisms. Among the components of the glycocalyx, glypican 1 (Gpc1) is considered a potential mechano-transducer due to its physical interaction with the cellular membrane. Herein we sought to investigate the mechanisms whereby Gpc1 regulates pulmonary permeability.

Hypothesis:
We hypothesize that Gpc1 regulates endothelial transport by eNOS-dependent mechanisms.

Methods:
Mouse lung endothelial cells (MLEC) isolated from male and female wild-type (WT) and Gpc1 knockout mice (Gpc1KO, 6-8 weeks old) were used. Endothelial transport was assessed by dextran permeability
in vitro,
lung barrier stability was assessed by TEER, eNOS pathway was assessed by immunoblotting. To assess the role of eNOS to Gpc1 effects on endothelial transport, MLEC were exposed to an eNOS decoy peptide.

Results:
Gpc1KO MLEC formed a tighter barrier in vitro when compared to WT MLEC (3.18±0.11 [Gpc1KO]
vs
2.34±0.11 [WT], normalized TEER, n=3/group). This was preserved during high pressure conditions. High pressure increased transcellular but not paracellular transport in both WT and Gpc1KO MLEC (1.8 fold in WT and 1.2 fold in Gpc1KO, n=6/group). Gpc1KO showed decreased eNOS expression (0.3 fold, n=4/group ) and activity during (1.947±0.32 [WT]
vs
1.316±0.13 [Gpc1KO], n=4/group). The eNOS decoy peptide increased transcellular transport only in Gpc1KO MLEC (3 fold, n=5/group).

Conclusions:
In conclusion, these data suggest that intracellular transport may be the main mechanism for pressure-induced changes in endothelial permeability in MLEC . The mechanisms whereby Gpc1 regulates transcellular transport seem to be associated with eNOS signaling. This may be important for the management of acute changes in pressure seen in clinical settings when NO donors are used as vasodilators to decrease blood pressure.
Abstract 15368: Outcomes of Anticoagulation in Patients With Transthyretin Cardiac Amyloidosis
Authors: Kolseth Clint , Prasad Mark , Chandrashekar Pranav , Kim Morris , Rashdan Lana , Burton Yunwoo , Masri Ahmad
Published: June 2023

Introduction:
Up to 70% of patients with transthyretin amyloid cardiomyopathy (ATTR-CM) have (AF) and require anticoagulation (AC). ATTR-CM is also associated with a higher bleeding risk due to advanced age, frailty, and comorbidities. Prior studies have shown that thrombi can be seen in any chamber of the heart. In the era of left atrial appendage occlusion (LAAO) devices, a reappraisal of AC outcomes in ATTR-CM is needed.

Hypothesis:
AC in patients with ATTR-CM is not associated with significantly different rate of bleeding as compared to patients who are not on AC.

Methods:
This observational study was performed on a single center of ATTR-CM patients (n=149) diagnosed between 2005-2022. Bleeding outcomes were stratified according to AF and AC status. The primary outcome was defined as a composite of all major bleeding. Bleeding episodes were further categorized into correctable or uncorrectable. HASBLED scores for each patient were also calculated.

Results:
Of the 149 patients reviewed, 94 (63%) had AF and 95 (64%) were on AC. In the overall population, 20 patients (13%) experienced a bleeding episode, with 30% (6
/20)
of the episodes classified as uncorrectable requiring indefinite AC cessation. During a medium (IQR) of 3.1 (2.6,4.9) years of follow-up, there was no significant difference in bleeding between those who were on AC compared to those who were not on AC (P = 0.105). There was also no significant difference in bleeding between those who had atrial fibrillation and those who did not (P = 0.464). Patients who experienced a bleeding episode had a significantly higher average HASBLED score compared to those who did not experience a bleed (2.7 vs 1.7, P<0.01) In total, 5 (5%) of ATTR-CM patients with AF would have been candidates for LAAO devices.

Conclusions:
AC is not associated with significantly increased risk of bleeding in patients with ATTR-CM compared to those who were not on AC. HASBLED scores may be useful in ATTR-CM. While further studies are needed to study the outcomes of LAAO devices in ATTR-CM, the incidence of requiring such an intervention in ATTR-CM appears to be low.
Abstract 15376: Nrf2 Agonist Omaveloxolone Improves Cardiac Function in Mice With Friedreich's Ataxia
Authors: Figueroa Francisco , Salinas Lili , Montgomery Claire , Thai Phung , Chiamvimonvat Nipavan , Cortopassi Gino , Dedkova Elena N
Published: June 2023

Introduction:
Friedreich’s ataxia (FA) is an inherited, life-shortening, degenerative neuromuscular disorder affecting ~1 in every 50,000 people in the USA and Europe. Initial symptoms, typically seen in children and teenagers, include loss of balance and coordination, muscle weakness placing these patients in wheelchairs. Early death is common and most of FA patients die in their 30s or earlier from cardiac failure, caused by deficient frataxin (FXN) expression in the heart. Omaveloxolone (OMAV), a known Nrf-2 activator, has been recently proposed as the first treatment of the neurological symptoms of FA; however less is known about OMAV effects on cardiac function in FA.

Aim:
To test the hypothesis that OMAV improves cardiac function in a mouse model of FA via Nfr2 signaling pathway activation.

Methods and Results:
Cardiospecific FXN knockout (FXN Mck-Cre KO) mice were used as a model of severe cardiomyopathy which is characteristic for the late stage of FA. Animals of either sex were randomly split in vehicle and OMAV-treated groups. Treatment with 24 mg/kg OMAV started at 3 weeks of age and continued for 5 weeks. Cardiac function was examined
in-vivo
by echocardiography. We found that FXN cKO mice developed severe heart failure with 45% reduction in ejection fraction, increased left ventricular (LV) mass (+80%) and diameter (+308%), decreased stroke volume (-26%) and cardiac output (-38%, n=10) as compared to WT mice (n=12). Cardiac deficits were more pronounced in FA males compared to females. OMAV (n=14) significantly decreased LV mass (-24%), LV diameter (-28%), and partially recovered a decrease in ejection fraction (+40%), stroke volume (+22%) and cardiac output (+35%). Gene expression of NADP(H) quinoline oxidoreductase-1 (NQO1), superoxide dismutase 2 (SOD2), and glutathione S-transferase were decreased by 60-90% in FA and recovered by 60-90% by OMAV. Aconitase activity, used as a surrogate measure of frataxin's iron-sulfur biogenesis function, was decreased in FA by 49% and recovered 33% by OMAV.

Conclusions:
This is the first study to demonstrate sexual dimorphism in FA hearts and show that OMAV can improve cardiac function in FA mice with severe cardiomyopathy.
Abstract 15377: Investigating the Role of Breast Cancer Susceptibility Gene 2 in High Glucose-Induced Endothelial Dysfunction
Authors: Nikfarjam Sepideh , Bu Shuhan , Nguyen Hien Chi , Rasheed Berk , Frisbee Jefferson C , Singh Krishna
Published: June 2023

Introduction and Hypothesis:
Mutations in the breast cancer susceptibility gene 2 (BRCA2) impair DNA damage repair which can eventually culminate in cancer, inflammation and/or apoptosis. Hyperglycemia-induced oxidative stress triggers DNA damage, inflammation and apoptosis, which are common contributors to endothelial dysfunction in diabetes. Here, we assessed the hypothesis that BRCA2 plays critical role in hyperglycemia-induced endothelial dysfunction.

Methods and Results:
Human umbilical vein endothelial cells (HUVECs) were exposed to high- (HG; 25 mM) or normal glucose (NG; 5 mM) and BRCA2 expression was measured. Our data showed significant BRCA2 upregulation in HG-treated HUVECs. Next, we successfully silenced BRCA2 in HUVECs, and measured the expression levels of p53 and p21 in HG- and NG-treated BRCA2-silenced HUVECs. We observed a significant upregulation of p53 and p21 in BRCA2-deficient HUVECs, which was further augmented by HG. Next, we measured the migratory and angiogenic potential of control and BRCA2-seilenced HUVECs under NG and HG. Our preliminary data showed that HG and loss of BRCA2 together inhibited the migratory capacity and angiogenic potential of HUVECs. To understand the mechanism behind the effect of HG on BRCA2-deficient ECs, we measured expression levels of endothelial nitric oxide synthase (eNOS). Both expression and activation of eNOS was reduced in HG-treated BRCA2-deficient HUVECs. To understand the effect of loss of BRCA2 on overall endothelial transcriptome and to understand mechanisms, we performed RNA sequencing on scrambled control- and siBRCA2-transfected HUVECs. Interestingly, a total of 3150 genes were differentially expressed in BRCA2-deficient HUVECs compared to the control group.

Conclusion:
Collectively, these findings reveal a novel role for BRCA2 as a regulator of endothelial function in context of HG treatment
in vitro
. Our RNA sequencing data shows theimportance of BRCA2 in regulating endothelial transcriptome and overall endothelial function. Further studies are underway; however, these results may provide important implications concerning the susceptibility of BRCA2 mutation carriers to HG-induced endothelial dysfunction as well as other CVDs.
Abstract 15379: Syncope and Pre-Syncope on the Exercise Test
Authors: Csecs Ibolya , Barillas Maria , Garzona-Navas Andres F , Bonikowske Amanda , Allison Thomas G
Published: June 2023

Background:
Data on the occurrence of syncope and presyncope with hemodynamic instability during graded exercise test (GXT) are limited.

Methods:
We reviewed the Mayo Integrated Stress Center database for non-imaging GXTs performed between January 2006 and December 2010 and registered syncopic/presyncopic episodes. Electronic medical records were reviewed for clinical information, outcomes, and mortality. SAS was used to provide background statistics.

Results:
During the study period, a total of 40,715 GXTs were performed on 33,885 unique patients, including 16,306 (40.0%) GXTs on patients with known cardiovascular disease (CVD). High risk for cardiac syncope is defined as prior cardiac arrest/ventricular tachycardia, ischemic cardiomyopathy, implanted defibrillator, history of atrial fibrillation, sotalol, amiodarone, LQTS, HOCM, aortic stenosis was identified in 3263 patients (6.0%), while risk factors for non-cardiac syncope (POTS, history of dizziness/spells/syncope, rest SBP < 100 mmHg, age ≥ 80 years, female age < 30 years, beta blocker use) were found in 15,925 patients (39.1%). Despite the presence of so many patients at risk, only 8 episodes of syncope occurred (0.020% per test; 0.023% per individual patient). Another 5 episodes of near-syncope with documented hypotension were aborted by the quick action of the testing staff (0.012% per test; 0.015% per individual patient). Only 1 patient (a 71yo M with severeCAD) required electrical cardioversion for ventricular fibrillation. None of the other patients, including all 6 women, presented with tachyarrhythmias. There were no deaths and no significant musculoskeletal trauma.

Conclusion:
Despite many patients with risk factors for cardiac and non-cardiac syncope in the cohort, syncope, and near-syncope with hemodynamic instability were very rare findings on guideline-directed GXTs.
Abstract 15382: Geriatric HFpEF: When It Isn’t Hypertensive Heart Disease
Authors: Gruen David S , Chan Marvyn Allen G , Teweldemedhin Bisrat , Oyenubi Olamide , Benjamin Patrick , Boy Gerald Kristoffer E , Jesmajian Stephen , Gitler Bernard
Published: June 2023

Introduction:
Transthyretin amyloid cardiomyopathy (ATTR) is thought to be a rare cause of HF, but recent studies showed this is the cause of 30% of HFpEF patients older than 75. With new treatments for amyloidosis, timely diagnosis has become critical.

Case:
A 78-female with HFpEF, HTN, and diabetes followed for progressive functional decline and hospitalizations for HFpEF exacerbations. EKG at baseline showed LBBB with normal QRS voltages. Serial echocardiogram (echo) showed an ejection fraction (EF) of 60-70%, mild LVH, and moderate TR with RVSP >50 mmHg. Latest echo showed progressive LA enlargement, MR, a new small pericardial effusion, and EF 50%. Regadenoson nuclear scan showed normal perfusion with preserved LVEF. Despite adequate control of her HTN, dyspnea continued to worsen. Because of this, the diagnosis of Cardiac amyloidosis

was considered. Further review revealed a history of Carpal Tunnel Syndrome (CTS) and speckled pattern within the ventricular wall on echo. A technetium pyrophosphate scintigraphy (
99m
Tc-PYP) confirmed ATTR with grade three uptake, AL amyloid was ruled out with normal SPEP/UPEP, and she was started on Tafamidis.

Discussion:
Cardiac amyloidosis should be considered in HFpEF patients with functional decline despite medical optimization. It can mimic LVH caused by HTN on echo and easily dismissed in patients with traditional risk factors, but presence of CTS or lumbar spinal stenosis can be supportive. Traditionally ATTR is associated with low-voltage EKG, however this patient presented with normal QRS voltages for years, making the diagnosis more elusive.

Conclusion:
Advances in noninvasive imaging and treatments allow for early diagnosis of cardiac amyloidosis, reducing morbidity and mortality. Diagnosis is often delayed or missed, as features could be subtle and mimic other more common cardiac disease. Screening 
99m
Tc-PYP for elderly patients with HFpEF could translate to improved outcomes
Abstract 15391: Transcriptomic Expression Changes in Embryonic Mouse Hearts Following Single-Dose Maternal Binge Alcohol Consumption: Implications for Congenital Heart Defects
Authors: Srivatsa Anagha , Sulkouskaya Darya , Shao Richard , Alvarez Jose D , Chordiya Shreya , Valencia Jesu R , Le Brandon , Yooseph Shibu , Pacher Pal , Ebert Steven N
Published: June 2023

Introduction:
Prenatal alcohol exposure is a risk factor for congenital heart defects, but it is unclear how alcohol exposure in utero leads to disruption of normal cardiovascular development. To gain insight into the transcriptomic pathways involved, we performed RNA-sequencing (RNA-seq) of embryonic mouse hearts 24h (E10.5) and 48h (E11.5) after a single dose of maternal alcohol.

Hypothesis:
Maternal binge alcohol consumption at E9.5 disrupts expression of specific embryonic genes important for heart development.

Methods:
Alcohol treatment (2.5g/Kg ethanol in saline) or control solution (0.9% saline) was administered to pregnant C57Bl/6 mice via single oral gavage at E9.5. Embryonic hearts were collected 24h and 48h later for RNA isolation and analysis by RNA-Seq and quantitative PCR (qPCR) methods. Reactome® pathway analysis was performed to determine which were most affected.

Results:
Of the nearly 20,000 genes expressed in the embryonic heart, we found remarkably few (less than 0.1%) showing significant alterations in expression. Most of the gene expression changes induced by alcohol occurred within 24h and were largely genes involved in transcriptional regulation or signaling pathways. In particular,
Wnt7a
showed significant (
p=0.013
) and substantial change (nearly 5-fold decrease relative to control expression, n=11 per group) in response to maternal binge alcohol 24h post-gavage. Similar results were observed for
Wnt7a
at 48h post-binge (p=0.043, n=8/group). Further analyses showed that
Wnt
-related genes,
Dkk2, Rtl1,Wnt3, Wnt6, and Wnt10a
showed significantly decreased expression and
Wnt11
showed significantly increased expression in the embryonic heart 24h post-binge alcohol.

Conclusions:
These results suggest that Wnt7a and related signaling pathway genes are disrupted by alcohol action in the embryonic heart, thereby opening new avenues for investigation into molecular underpinnings of alcohol-induced congenital heart defects.
Abstract 15402: P90 Ribosomal S6 Kinase Rescues Contractility of Myosin Light Chain Kinase Null Smooth Muscle
Authors: Kalra Jaspreet , Ayon Ramon J , Markowska Zaneta , Derewenda Urszula , Sonkusare Swapnil K , Somlyo Avril V
Published: June 2023

Introduction:
Smooth muscle (SM) contraction is regulated by an increase in [Ca
2+
]
,
activation of myosin light chain kinase (MLCK) and phosphorylation of myosin regulatory light chain (RLC
20
) activating myosin crossbridge cycling. MLCK null mice die at birth, but SM retains the ability to contract suggesting the presence of another kinase(s). Based on studies with WT and p90 ribosomal S-6 kinase (RSK2) null mice, we reported that RSK2 activity accounts for ~ 25% of maximal contractile force in wild type (WT) resistance arteries.

Hypothesis:
RSK2 confers contractility via RLC
20
activation in smooth muscle cells of MLCK null mice.

Methods:
Tension was measured on E18 aorta, bladder, and umbilical artery from WT and MLCK
-/-
embryos, in response to phenylephrine, carbachol, high potassium, SM myosin phosphatase, calyculinA (10 uM), as well as Ca
2+
following permeabilization with α-toxin. VSMCs were cultured from E18 MLCK
-/-
aortae and from WT and RSK2
-/-
mouse mesenteric arteries. Serum starved cells were stimulated with serum and LPA (0.5 uM), in the presence and absence of RSK2 inhibitor, LJH685 (10 μM). Protein-protein interactions were assessed by in-situ PLA assay and immunoprecipitation. On-going
in-vitro
studies using purified myosin and kinases address RSK2 direct regulation of MLCK activity.

Results:
Strips from bladder SM and umbilical artery from WT and MLCK
-/-
embryos (E18), contracted in response to agonists and high [K]. pCa-tension curves showed the same sensitivity to Ca
2+
in MLCK
-/-
and WT. Maximal force induced by calyculinA was also the same indicating an intact normal contractile apparatus in the MLCK
-/-
. Significant phosphorylation of RLC
20
at Ser19 occurred in LPA- and serum-stimulated MLCK
-/-
SM cells, which was significantly inhibited by the RSK2 inhibitor, LJH685 (n=3). PLA assay showed co-localization of RSK2 and MLCK (<40nm) (n=3). RSK2 is immunoprecipitated along with its activators ERK1/2, PDK1, suggestive of cross talk between RSK2 and MLCK on the actin filament (n=5).

Conclusions:
RSK2 signaling accounts for the contractile activity in the absence of MLCK. Furthermore, RSK2 and MLCK form a signaling complex on the actin filament, optimally positioning them for interaction with adjacent myosin heads.