bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 14797: Single Cell Multi-Modal Analyses Reveal Epigenomic and Transcriptomic Basis Underlying Cardiac Defects in Maternal Diabetes
Authors: Nishino Tomohiro , Ranade Sanjeev , Angelo Pelonero , van Soldt Benjamin J , Ye Lin , Alexanian Michael , Koback Frances , Huang Yu , Sadagopan Nandhini , Padmanabhan Arun , Thomas Reuben , van Bemmel Joke , Gifford Casey , Costa Mauro , Srivastava Deepak
Published: June 2023

Introduction:
Birth defects can be caused by combinations of genetic and environmental influences. Large-scale DNA sequencing efforts reveal genetic influences, yet the environmental contributions have been mainly limited to association studies with little mechanistic insight. Hyperglycemia in pre-gestational diabetic (PGDM) mothers are among the most frequent environmental contributors to congenital defects, such as congenital heart defects and craniofacial anomalies. However, the cell types involved and underlying mechanisms by which PGDM affects these regions are unknown.

Methods and Results:
To determine how PGDM affects the epigenomic and transcriptomic state within individual cells during cardio-pharyngeal development, we utilized multi-modal single cell analyses with streptozotocin-induced mouse PGDM model. Analysis of E10.5 embryos showed that PGDM specifically affects the epigenomic state of two small subsets of cardiac and craniofacial progenitors. In PGDM derived embryos, a newly recognized subpopulation of anterior heart field progenitors expressing
Alx3
acquires a more posterior-like identity in response to PGDM, based on gene expression and chromatin status. Similarly, a sub-population of neural crest-derived cells in the second pharyngeal arch, which contributes to craniofacial structures, also displayed patterning abnormalities. Analyses of differentially accessible chromatin regions revealed disrupted patterning was due to increased intrinsic retinoic acid signaling in affected cell types in PGDM.

Conclusions:
We showed the selective epigenomic vulnerability of a small subset of cells that help explain cardiac and craniofacial defects observed in PGDM despite all embryonic cells experiencing similar environmental exposure. We also showed that the selectivity was partly driven by a previously unrecognized dysregulation of retinoic acid signaling in PGDM. Discovery of the consequences of environmental factors on embryonic development using multi-modal single cell genomics will lead to potential gene-environment combinations that contribute to the complex and multifactorial diseases and preventive approaches to protect vulnerable cells.
Abstract 148: No Bystander Cardiopulmonary Resuscitation For Out-Of-Hospital Cardiac Arrest In Vietnam: Impact On The Outcomes
Authors: Do Son N , Luong Chinh Q , Manabe Toshie , Nguyen My H , Pham Dung T , Ton Tra T , Hoang Quoc T , Nguyen Dat T , Pham Thao T , Hoang Hanh T , Khuong Dai Q , Nguyen Quan H , Nguyen Tuan A , Dang Hien D , Hoang Hai B , Nguyen Chi V , Nguyen Anh D , McNally Bryan F , Ong Marcus E , Dao Co X
Published: June 2023

Introduction:
Bystander CPR is not often performed on OHCA patients, particularly in limited-resource regions. This study aimed to investigate the rate of OHCA patients who did not receive bystander CPR and its impact on the outcomes in an LMIC.

Hypothesis:
Understanding the reasons bystanders are reluctant to call EMS and how no bystander CPR impacts the outcomes of OHCA patients are crucial for improving survival in Vietnam.

Methods:
We performed a multicenter prospective cohort study of OHCA patients (≥18 years) presenting to three central hospitals in Vietnam from February 2014 to December 2018. We collected data on characteristics, management, and outcomes and compared these data between patients who did not receive bystander CPR and patients who did. Using logistic regression, we assessed factors associated with survival and good neurological function on discharge (a CPC score of 1 or 2).

Results:
Of 521 patients, 388 (74.5%) were male, and the mean age was 56.71 years (SD: 17.32). Although most cardiac arrests (68.7%; 358/521) occurred at home and 67.9% (353/520) were witnessed by bystanders, a high rate (77.9%, 406/521) of these patients did not receive bystander CPR. Only half of the patients were taken by EMS (8.1%, 42/521) or private ambulance (42.8%, 223/521); 50.8% (133/262) of whom were given resuscitation attempts by EMS or private ambulance. There was no significant difference in survival to admission (16.7%; 68/406 and 24.3%; 28/115; p=0.064) and survival to discharge (7.9%; 32/406 and 14.8%; 17/115; p=0.094) between patients who did not receive bystander CPR and patients who did. In contrast, the rate of good neurological function of patients who did not receive bystander CPR (4.7%, 19/406) was significantly lower than that of patients who received bystander CPR (12.2%, 14/115; p=0.004). Moreover, multivariate analysis showed that no bystander CPR (OR: 0.276; 95% CI: 0.124-0.614) was inversely and independently associated with good neurological function.

Conclusions:
In our study, poor outcomes emphasize the need for increasing bystander CPR performance, increasing the number of EMS ambulances and the utilization of private ambulances, and developing a standard emergency first-aid program for both healthcare personnel and the community.
Abstract 14821: Effect of Single Volatile Organic Compound Metabolites and Volatile Organic Compound Mixtures on Vascular Function
Authors: Stopforth Cameron , Sears Clara , Dials Justin , Srivastava Sanjay , Bhatnagar Aruni , Keith Rachel J
Published: June 2023

Volatile organic compounds (VOCs) are ubiquitous environmental toxicants with pre-clinical evidence of decreased vascular health. Research to date has focused on single VOC metabolites and data is lacking on the effects on human vascular health. Many VOCs are strongly correlated with similar exposure sources. Though the compounding effects are poorly defined, they are likely to increase the overall toxicity. Therefore, we examined single VOC metabolites and mixtures’ association with flow mediated dilation (FMD), an index of vascular health.

We recruited adult participants into a CV risk cohort, aged 25-70 years, from 2018 and 2019. A panel of urinary VOC metabolites were measured in 308 non-smoking participants who had completed brachial artery FMD. Regression analysis of 16 VOC analytes was tested for associations with %FMD and models were adjusted for covariates selected a priori including age, sex, diabetes and ACE inhibitors. We examined the joint effect of MU, 3HPMA, CEMA, CYMA, DHBMA, 2HPMA, 2MHA, and AAMA mixtures on log-transformed %FMD using quantile-based g-computation models adjusted for the covariates above and for correlated VOC metabolites that were not associated with FMD in single metabolite analyses.

The mean %FMD was 7.1 % ± 4.7%. VOC metabolites that were predictors of decreased %FMD in fully adjusted models were 3HPMA (β=-1.275; 95% CI= -2.402, -0.148; p=0.027), CEMA (β= -1.204; 95% CI= -2.051; -0.357 p=0.006), AAMA (β=-1.125; 95% CI= -2.156, -0.094; p=0.033), 2HPMA (β= -1.035; 95% CI= -1.847, -0.222 p=0.013), and 2MHA (β=-0.623; 95% CI= -1.123, -0.124; p=0.015). Most VOC metabolites had fair to moderate correlations in bivariate analyses (Spearman correlation from 0.06 to 0.60). We found that a quartile increase in the VOC mixture was associated with 20% lower percent FMD (ψ -0.18; 95%CI= -0.35, -0.02; p=0.027) in adjusted models. We found the overall mixture effect was driven primarily by 3HPMA, followed by CYMA, CEMA, 2MHA, and AAMA. Therefore, the main driver of the association between the VOC mixture and decreased vascular function is acrolein and to a lesser extent acrylonitrile. Future research could focus on identifying sources of exposure to these VOCs to promote better vascular health among non-smokers.
Abstract 14827: Clinical Validation of a Novel Cuffless Portable Blood Pressure Measurement Device
Authors: Reynolds Jeffery S , Agashe Harsha , Peterson Eric D , Zemel Marc I
Published: June 2023

Introduction:
Ambulatory blood pressure (BP) measurements using an arm-cuff can be both cumbersome and painful; limiting patients’ willingness to obtain routine measurements. A novel, simple, rapid, cuffless device (TouchBP™) calculates systolic and diastolic BP based on observed blood volume oscillations that result from the variable finger pressure on a custom sensor pad. The accuracy of this novel device relative to cuff based standards is not known.

Methods:
An independent physiology lab compared a blind TouchBP™ reading to reference SBP and DBP measurements made via an experienced auscultator. Subsequent data from 6 subjects was excluded due to signal artifacts resulting in 52 subjects in the analysis dataset. The Digitouch prediction algorithm was created using data from two previous clinical studies with different subject pools.

Results:
Among 52 subjects, SBP ranged from 95 to 167 mmHg, and DBP ranged from 58 to 106 mmHg. The SBP estimation error was 0.4 ± 7.9 mmHg, while DBP error was 0.3 ± 7.2 mmHg [bias ± precision]. (See Figure 1). The bias and precision results meet accuracy thresholds established by AAMI standard (bias< 5 mmHg and precision < 8mmHg).

Conclusion:
Digitouch’s TouchBP™, a novel cuffless mobile BP measurement system produces accuracy results like traditional oscillometric cuffs.
Abstract 14832: Prognostic Impacts of Prehospital Simple Risk Index in Patients With St Elevation Myocardial Infarction
Authors: Yumi Hirota , Moriwaki Keishi , Ito Hiromasa , Ishise Takuro , Masuda Jyun , Kawasaki Atsushi , Dohi Kaoru , Kurita Tairo , Fujimoto Naoki , Yamada Norikazu
Published: June 2023

Introduction:
Early identification of high-risk patients is the cornerstone of managing patients with ST elevation myocardial infarction (STEMI). The simple risk index (SRI), which is calculated by the formula: (HR х [age/10]2) / systolic blood pressure was shown to be able to stratify 30-day prognosis. However, prognostic impacts of prehospital SRI (Pre-SRI) in STEMI patients remain unknown.

Methods:
We performed retrospective analysis of 2047 STEMI patients who underwent emergency primary percutaneous coronary intervention (PCI) from January 2013 to December 2017. Pre-SRI was recorded by emergency services at the first contact with the patient before admission, and ER-SRI was recorded at emergency room. The primary end point was defined as 2 years all-cause death.

Results:
All-cause death was observed in 219 (11%) patients. The cut off value of Pre-SRI for predicting 2 years all cause death was 34 from ROC (AUC 0.800, P<0.001), and the cut off value of ER-SRI for predicting 2 years all cause death was also 34 from ROC analysis (AUC 0.809, P<0.001) According to the Kaplan-Meier survival analysis by combination of Pre-SRI and ER-SRI, the patients with Pre-SRI≧34 and ER-SRI≧34 showed significantly higher all-cause mortality compared to the patients with Pre-SRI≧34 and ER-SRI<34, the patients with Pre-SRI<34 and ER-SRI≧34, and the patients with Pre-SRI<34 and ER-SRI<34 (p<0.001).

Conclusions:
High Pre-SRI increase mortality and assessment of both high Pre-SRI and high ER-SRI enhance risk stratification in STEMI patients. Early recognizing high Pre-SRI may help us make better strategies and improve prognosis for high-risk STEMI patients.
Abstract 14838: Prevalence and Outcomes of Prosthetic Valve Endocarditis Post-Tavr: A Proportion Meta-Analysis
Authors: Varghese Jeffy Susan , Vadiyala Mounika Reddy , Ang Song Peng , Chia Jia Ee , Reddy Akhil , Ishak Angela , Yaqoob Sadia
Published: June 2023

Introduction:
Prosthetic valve endocarditis (PVE) may occur after trans-catheter aortic valve replacement (TAVR), resulting in bio-prosthetic valve dysfunction. We aim to assess the clinical characteristics and long-term outcomes post TAVR.

Methods:
We performed a systematic literature search in PubMed, Scopus, and Embase for relevant articles from inception until June 01, 2022. Primary outcome was all-cause mortality. Secondary outcomes include acute kidney injury, stroke, systemic embolisation, heart failure and valve re-intervention. All endpoints were pooled using a random-effect model.

Results:
A total of 33 studies were included, comprising 346,094 patients who underwent TAVR. Amongst the various co-morbidities, most common were diabetes (11%), chronic kidney disease (8%) and chronic obstructive pulmonary disease (COPD) (8%).The pooled incidence of PVE was 1% [95% CI (1%-2%)]. The most common approach was trans-femoral (n=15) (88% [95% CI (79%-95%)]) followed by trans-apical (n=9) (14% [95% CI(10%-18%)]). The most common organisms isolated were staphylococcus species (n=24) (34% [95% CI (30%-38%)]), followed by streptococcal species (n=23) in 23% and enterococcus species (n=22) in 24%. The leading complications reported were acute kidney injury (AKI) (30% [95% CI (19%-44%)]) (n=7), cerebrovascular accident (10% [95% CI (7%-14%)]) (n=13), embolisation (17% [95% CI (10%-26%)]) (n=11) and patients requiring a surgical valve replacement (12% [95% CI (8%-18%)] ) (n=14). The all-cause mortality was 49% [95% CI (41%-56%)], while the in-hospital mortality was 4% [95% CI (2%-9%)].

Conclusions:
Incidence of IE post TAVR is low (1%), but is associated with a higher rate of mortality (49%) and morbidity. As a result, aggressive management is needed to reduce post-TAVR complications.
Abstract 14872: Lactate Promotes Endothelial-to-Mesenchymal Transition via Snail1 Lactylation After Myocardial Infarction
Authors: Fan Min , Yang Kun , Wang Xiaohui , Gill P S , Ha Tuanzhu , Williams David , Li Chuanfu
Published: June 2023

Introduction:
High levels of lactate are positively associated with the prognosis and mortality in the patients with acute heart attack and heart failure. Endothelial-to-mesenchymal transition (EndMT) plays an important role in the progression of cardiac fibrosis in multiple cardiovascular diseases. We investigated whether lactate exerts a biological role in EndMT following myocardial infarction (MI).

Methods:
MI was induced in endothelial cell specific GFP labeled (TIE2GFP) mice by left anterior descending (LAD) coronary artery ligation. To suppress lactate production, 2-Deoxy-D-glucose (2-DG, 2g/kg body weight) was intraperitoneally injected daily for 7 days. Supplemental lactic acid (adjust to pH 6.8, 0.5 g/kg body weight) was administrated to mice either by i.p. injection every 7 days for 1 or 4 weeks or using osmotic mini-pumps after surgery.
In vitro
, endothelial cells were treated with 5 mM or 10 mM L-lactic acid followed by normoxic or hypoxic challenge in a hypoxia chamber.

Results:
Our data showed that inhibition of lactate production by 2-DG ameliorated MI induced EndMT, cardiac fibrosis and cardiac dysfunction. On the contrary, administration of supplemental lactate further promoted EndMT and worsened cardiac dysfunction post-MI.
In vitro
analysis shows that treatment of endothelial cells with lactate decreases the expression of endothelial cell markers and accelerates mesenchymal marker expression, as well as disrupts endothelial cell function and induces mesenchymal like function following hypoxic challenge via activating transforming growth factor-β (TGF-β)/Smad2 signaling pathway. Of note, lactate accelerated the lactylation of Snail1, a transcription factor of TGF-β, promoted its nuclear translocation to bind at the promoter of TGF-β, and upregulated TGF-β expression.

Conclusion:
In summary, we conclude that lactate acts as an important molecule that upregulates cardiac fibrosis after MI via induction of Snail1 lactylation and nuclear translocation to activate TGF-β/Smad2 pathway, resulting in EndMT.
Abstract 14889: Association of Calcium Channel Blockers With Mortality and Hospitalization Rate In Sars-Cov-2 Patients
Authors: Douen Aaron , Tan Samuel , Sukhoo-Pertab Marc , Siu Wenchy , Oh Jeremy , Romney Wesley , Ramdass Prakash , Echeverry Tatiana , Juang George , Toreli Aleksandre
Published: June 2023

Introduction:
Literature has shown that calcium metabolism mediates the body’s defense against viral pathogens. In particular, studies have found SARS-CoV-2 to use calcium to assist entry and replication in host cells.

Hypothesis:
The use of CCBs are associated with decreased mortality and hospitalization in SARS-CoV-2 positive patients.

Methods:
A retrospective chart review of patients 18 years and older with a positive SARS-CoV-2 polymerase chain reaction test between March 2020 and April 2021. Patients were categorized based on their use and non-use of CCB. The primary outcomes were all-cause mortality and hospitalization. Secondary outcomes were acute kidney injury, hypotension, intubation, myocardial infarction, systolic heart failure, ventricular tachycardia (VT), and ventricular fibrillation (VF). Multivariate logistic regression was used to adjust for baseline characteristics.

Results:
We identified 1,402 patients taking CCB and 21,674 patients not taking CCB who tested positive for SARS- CoV-2. The baseline characteristic incidence was 370 (26.4%) for coronary artery disease, 262 (18.7%) for heart failure, 1402 (100%) for hypertension in patients taking CCBs. Secondary outcome incidence was 85 (6.1%) for myocardial infarction, 257 (18.3%) for hypotension, and 100 (7.1%) for systolic heart failure in patients taking CCBs. After adjusting for comorbidities using logistic regression, OR was 0.605 (95% Confidence Interval [CI]: 0.519-0.705; p < 0.001) for all-cause mortality, and 27.641 (95% CI: 24.396-31.319; p<0.001) for hospitalization rate. The CCB group had significantly decreased adjusted OR for all secondary outcomes (p<0.001) except for VT (p<0.425) and VF (p <1.00).

Conclusions:
The incidence of cardiovascular outcomes and baseline cardiovascular risk factors were significantly greater in SARS-CoV-2 positive patients taking CCB. SARS-CoV-2 positive patients in the CCB group were associated with decreased all-cause mortality, myocardial infarction, and systolic heart failure. However, the CCB group had a significantly increased association with hospitalization. Therefore, when indicated, clinicians should consider using CCB in SARS-CoV-2 patients, especially those with cardiovascular risk factors.
Abstract 149: Epinephrine Administration First Versus Advanced Airway Management First For Adult Patients With Out-of-hospital Cardiac Arrest
Authors: Okubo Masashi , Komukai Sho , Izawa Junichi , Matsuyama Tasuku , Kiyohara Kosuke , Iwami Taku , Kitamura Tetsuhisa
Published: June 2023

Introduction:
Epinephrine administration and advanced airway management (AAM: ie, endotracheal intubation and supraglottic airway placement) have been commonly used for patients with out-of-hospital cardiac arrest (OHCA). However, the optimal sequence of these interventions remains unclear. We, therefore, compared epinephrine administration 1
st
strategy with AAM 1
st
strategy.

Methods:
We conducted a secondary analysis of a prospective, nationwide, population-based OHCA registry in Japan. We included emergency medical services (EMS)-treated adult (≥18 years) OHCA from 2014 through 2019, stratified into shockable or nonshockable rhythms. We excluded those who did not receive epinephrine or AAM. The primary outcome was 1-month survival and the secondary outcome was 1-month survival with favorable functional status, defined as a Cerebral Performance Category scale 1 or 2. To account for treatment selection bias between patients who received epinephrine before AAM and patients who received AAM before epinephrine, we calculated propensity score and used inverse probability treatment weighting analyses.

Results:
We included 8,854 patients with shockable and 79,687 patients with nonshockable rhythm. 1,825 patients (20.6%) of shockable rhythms and 11,769 patients (14.8%) of nonshockable rhythm received epinephrine before AAM. Among patients with shockable rhythm, epinephrine administration before AAM was associated with lower odds of 1-month survival (Odds ratio [OR] 0.83, 95% confidence interval [CI] 0.76 to 0.90) but not associated with favorable functional outcome (OR 0.94, 95% CI 0.82 to 1.70). Among patients with nonshockable rhythm, epinephrine administration before AAM was not associated with 1-month survival (OR 0.96, 95% CI 0.91 to 1.01) but was associated with higher odds of favorable functional status (OR 1.15, 95% CI 1.002 to 1.33).

Conclusions:
In this observational study, we found that epinephrine administration before AAM was associated with lower odds of 1-month survival among shockable rhythm but was associated with higher odds of 1-month survival with favorable functional status among nonshockable rhythm, suggesting that the optimal sequence of epinephrine administration and AAM may vary between initial rhythms.
Abstract 14908: Vascular Events Associated With Cisplatin Based Combination Therapy: An Analysis From Faers Database
Authors: Mann Chitsimann , Patel Brijesh , malla midhun , Almubarak Mohammad , Fang Wei
Published: June 2023

Introduction:Cisplatin is an alkylating agent that has been used to treat various cancer types in combination with other antineoplastic agents. It has been associated with vascular toxicity. However, data is lacking whether one combination is associated with increased risk of vascular events.

Methods:
We queried 2016-2020 FDA Adverse Reporting System (FAERS) database to analyze the association. We identified cases with the following four combinations: (1) cisplatin + etoposide, (2) cisplatin + gemcitabine, (3) Cisplatin + docetaxel/paclitaxel, and (4) cisplatin + 5-fluorouracil/capecitabine. The effect size is reported as Rate of Reporting (ROR) with 95% confidence intervals (CI). We aggregated various vascular events including pulmonary, coronary, cerebrovascular and peripheral vascular events including venous events. A lower limit of 95% CI > 1 suggests the drug is potentially associated with the event.

Results:
From 2016 to 2020, we identified 23513 cases of cisplatin combinations. More than 67% of cases had age over 50 years and 47%, 32.9% and 19.9% were males, females and gender not reported respectively. Most cases were reported from the United States (20%). The ROR (95% CI) for Cisplatin + etoposide was 1.55 (1.37-1.75), cisplatin + gemcitabine was 3.40 (3.03-3.82), cisplatin + docetaxel/paclitaxel was 1.10 (0.90-1.37), and cisplatin + 5FU/capecitabine was 2.34 (1.88-2.92). More than 25% had either a life-threatening condition or died.

Conclusion:
Cisplatin with Gemcitabine is associated the highest ROR amongst the studied Cisplatin -chemotherapy based combinations. The results obtained from the study emphasizes the role of multi-disciplinary care including cardio-oncology in order to minimize and effectively manage the vascular toxicity.
Abstract 14919: EN-RAGE is Associated With Improved Outcomes in Hospitalized COVID-19 Patients
Authors: Ahmad Javaria , Solorzano Juan , Jaisingh Keerthish , Thevuthasan Sindhu , Foster Addison , Watts Megan , Orr Wayne W , Kevil Chris , Kolluru Gopi Krishna , Dominic Paari
Published: June 2023

Background:
Inflammation has been implicated in the pathogenesis, severity, and outcomes of COVID-19. Extracellular newly identified receptor for advanced glycation end-product (EN-RAGE), a soluble EF-hand calcium-binding protein, causes integrin activation, activates inflammation, and upregulates the synthesis of vascular cell adhesion molecule-1 and intracellular adhesion molecule-1. While it is positively associated with atherosclerosis, high EN-RAGE levels have been shown to predict remission of auto-immune diseases. The role of EN-RAGE as a biomarker in COVID-19 is unknown.

Methods:
Serum EN-RAGE levels were measured from blood samples of 146 COVID-19 in-patients using Myriad® Cytokine MAP and compared between the subgroups based on race and outcomes. T-test, Chi-square, and multivariable regression analysis were performed to identify the correlation of EN-RAGE with race and outcomes.

Results:
Of 146 patients (mean age=59 years), 60% were female, 35% were White, and 60% were African Americans. 21 of the 146 (14%) patients died. Mortality was higher in Whites compared to African Americans (21% vs 10%) consistent with CDC data showing higher mortality in Whites compared to non-Whites. Surprisingly, African Americans had significantly higher EN-RAGE levels compared to Whites. (574.6 vs 360.9 respectively, p=0.002). Congruently, EN-RAGE levels were significantly higher in the patients who survived compared to the patients who died. (514.82 vs 336.29, p=0.039) suggesting that increased serum EN-RAGE levels may be associated with improved outcomes in hospitalized COVID-19 patients.

Conclusion:
Our study reveals a negative association of EN-RAGE levels with in-patient mortality associated with COVID-19 and could possibly be used as a biomarker of survival in hospitalized COVID-19 patients. EN-RAGE levels could partly explain mortality differences based on race. Further studies are required to confirm this association.
Abstract 14922: Combined Lineage Tracing and Scrna-Seq Reveals Unexpected First Heart Field Predominance of Human Ipsc Differentiation
Authors: Galdos Francisco X , Lee Carissa , Lee Soah , Goodyer William R , Paige Sharon , Escobar Gabriela V , Darsha Adrija , Beck Aimee , Xu Sidra , Bak Rasmus O , Porteus Matthew , Wu Sean M
Published: June 2023

Introduction:
During mammalian development, the left and right ventricles arise from early populations of cardiac progenitors known as the first and second heart fields, respectively. While these populations have been studied in non-human model systems, human induced pluripotent stem cells (hiPSCs) present an exciting alternative for modeling early human embryogenesis. A major question is whether hiPSCs can model cardiac progenitor development as seen in animal systems.

Hypothesis:
Dual TBX5/MYL2 lineage tracing and time course single cell RNA sequencing can uncover the identity of human cardiac progenitor lineage
in vitro.

Methods:
Using CRISPR/Cas9 genome editing we introduced a TBX5/MYL2 fluorescent reporter lineage tracing system into two independent hiPSC lines to identify FHF-progenitors and their descendants. Using a standard biphasic WNT modulation protocol, we differentiated hiPSCs into cardiomyocytes and characterized the expression of TBX5-lineage positive and MYL2 cardiomyocytes between day 3 and day 30 of differentiation. To further characterize the cell populations, present during differentiation, we conducted single cell RNA sequencing on differentiating hiPSCs across 12 timepoints with oligonucleotide-based sample multiplexing.

Results:
Our reporter system revealed the surprising finding that using a standard WNT modulation small molecule protocol across two cell lines yielded 93.2%±0.80% and 92.1%± 1.60% TBX5-lineage positive cardiomyocytes, respectively. Among MYL2 expressing ventricular cardiomyocytes, we observed 99.0%±0.21% and 93.2±0.84% as TBX5-lineage positive, respectively. ScRNA-seq analysis and trajectory inference revealed high expression of known FHF gene markers relative to murine developmental data and identified a clear bifurcation of cardiac progenitor populations into myocardial and epicardial lineage consistent with a FHF differentiation trajectory. Lastly, a comparison of our data with scRNA-seq data from a recently published 3D protocol reveals the identity of a SHF progenitor population within a 3D system.

Conclusions:
Our work provides a powerful new genetic lineage tracing approach, as well as a single cell transcriptomic atlas of hiPSCs undergoing cardiac differentiation.
Abstract 14927: Real-Time Super-Resolution Analysis of Sarcomere Motion Reveals Context-Dependent Stochastic Heterogeneity in Induced Pluripotent Stem Cell-Derived Cardiomyocytes
Authors: Haertter Daniel , Hauke Lara , Schmidt Christoph F , Zimmermann Wolfram H
Published: June 2023

Studies of sarcomere dynamics in cardiac myofibrils are key to understand how myosin-actin contractility translates to whole heart contractions. Such analyses require high spatial (nm) and temporal (ms) resolution of multiple sarcomeres in parallel. Here, we present a new imaging and computational method to analyze sarcomere structure and function in human induced pluripotent stem cell (iPSC)-derived cardiomyocytes. For this we (1) engineered a fluorescent z-band tag (ACTN2-Citrine) by CRISPR-editing, (2) developed micropatterned soft (7:1, 10 kPa) substrate cultures to mimic in vivo conditions, (3) made use of high-speed microscopy (66 fps), and (4) designed an automated AI-based sarcomere detection and analysis tool (SarcAsM - Sarcomere Analysis Multitool). Cardiomyocytes were differentiated from the ACTN2-Citrine iPSC-line and purified using a staged growth factor and metabolic selection protocol (>95% purity with a ventricular myocyte phenotype). Our analysis revealed highly heterogeneous single sarcomere motion, i.e., shortening and lengthening patterns during the same time of the contraction cycle. Notably, the observed rich dynamics on single sarcomere-level are not apparent from whole cell analyses. Interestingly, culture on stiffer substrates (>20 kPa; mimicking cardiac fibrosis) resulted in reduced cell-level contraction, but constant sarcomere level contractility with however enhanced heterogeneity. Analysis of a large data set (>1,200 cells) in conjunction with a biophysical model showed that heterogeneous sarcomere motion is not caused by static non-uniformity (e.g., strong/weak sarcomeres), but can be primarily attributed to the stochastic and non-linear nature of sarcomere dynamics and thus occurs intrinsically during cardiomyocyte beating. Our findings show that that sarcomere heterogeneity is an important feature of cardiomyocyte contractility. The introduced tools for unbiased high-throughput analysis of sarcomere-level dynamics can be exploited to gain a better understanding on how sarcomere level contractility translates to whole cardiomyocyte contractions under simulated physiological and pathological conditions.
Abstract 14938: Endothelial Cell-Targeted Nanoparticle Delivery of Crispr/Cas9 Plasmid Dna Disrupts Endothelial
Epas1
in Adult Rats and Inhibits Experimental Pah
Authors: Machireddy Narsa , Dai Jingbo , Peng Yi , Trewartha Shawn , Zhao Youyang
Published: June 2023

Introduction:
Pulmonary arterial hypertension (PAH) is a devastating disease characterized by obliterative vascular remodeling that leads to right heart failure and death. HIF-2α (hypoxia-inducible factor-2α) activation in lung endothelial cells is obligatory to obliterative pulmonary vascular remodeling in PAH patients and animal PAH models.

Hypothesis:
Endothelial cell-targeted nanoparticle can deliver plasmid DNA expressing the CRISPR/Cas9 system to disrupt
Epas1
(encoding HIF-2α) selectively in endothelial cells in adult rats and inhibit PAH in monocrotaline rats.

Methods:
To target endothelium for robust genome editing, we developed poly(ethylene glycol) methyl ether-
block
-poly(lactide-co-glycolide) (PEG-
b
-PLGA) copolymer-based nanoparticle formulated with polyethyleneimine. A single i.v. administration of mixture of nanoparticles and plasmid DNA expressing Cas9 controlled by rat
Cdh5
promoter and guide RNAs targeting
Epas1
driven by
U6
promoter induced highly efficient genome editing in endothelial cells in 6-weeks old SD rats. Rat
Cdh5
-Cas9 plasmid DNA with scramble guide RNA were injected as a control group. One week after nanoparticle administration, the rats were injected with a single dose of monocrotaline (MCT) to induce PAH. At 4 weeks post-MCT, hemodynamic measurement of right ventricular systolic pressure RVSP and weight ratio of right ventricular versus left ventricular plus septum (RV/LV+S), an indicator of RV hypertrophy were determined.

Results:
Single administration of mixture of nanoparticles and CRISPR

Cdh5

plasmid DNA caused diminished HIF-2α protein expression selectively in endothelial cells indicating highly efficient genome editing. Genomic editing-mediated disruption of
Epas1
in endothelial cells reduced RVSP and right ventricular hypertrophy as well as obliterative pulmonary vascular remodeling in MCT-induced PAH rats.

Conclusions:
These findings demonstrate robust genome editing of
Epas1
via EC-targeted nanoparticle delivery of CRISPR-Cas9 plasmid DNA in adult rats leading to attenuated PAH in MCT rats, suggesting EC-targeted nanoparticle delivery of genome editing system to disrupt PAH-causing gene(s) in ECs is a promising precision gene therapy of PAH in patients.
Abstract 14945: Geographic Disparity in Mortality From Aortic Diseases in the United States
Authors: Tanaka Yoshihiro , Greenland Philip
Published: June 2023

Introduction:
Aortic diseases (AD) have a high mortality rate. Previous studies have reported that winter season and/or lower temperature climates were associated with a higher incidence of hospitalization due to AD. It remains unclear whether this observation can be translated into higher mortality rate in the country level.

Hypothesis:
We hypothesized that AD mortality might be higher in areas of the United States with colder climates or longer winter seasons.

Methods:
The underlying cause of death file in the CDC WONDER online database was queried between 2015 and 2019. AD deaths were identified by listing aortic aneurysm and dissection (ICD-10 code: I71) as underlying cause of death among persons aged ≥ 35 years. We calculated age-adjusted mortality rate (AAMR) at the Census region level and stratified by sex and race-ethnicity groups (non-Hispanic White, non-Hispanic Black, and Hispanic) per 100,000 population. Adjusted linear regression model by sex and race/ethnicity groups was used to examine the association between AAMR and Census region. We also calculated state-level AAMR of AD, and compared AAMRs in top-5 warmest states with those in the top-5 coldest states in winter.

Results:
A total of 46,708 AD deaths were identified between 2015 and 2019. AAMR in Midwest was the highest (4.7), followed by West (4.0), Northeast (3.8), and South (3.7). In comparison with South as a referent, Midwest was significantly associated with higher AAMR after adjusting for gender and race/ethnicity groups (beta coefficient: 0.87 [95% confidence interval: 0.37 - 1.37], p-value = 0.002). State-level AAMR seemed higher in northern part of the US and AAMRs were significantly higher in the coldest states compared with the warmest states in winter (p < 0.05) (Figure).

Conclusion:
The present study showed that the states with colder climates in winter season was significantly associated with higher AAMR in, suggesting that geographical factors are likely to affect the mortality rate in AD.
Abstract 14957: Validation of Echocardiography Color Doppler Velocity Reconstruction Against 4d Flow Mri in Assessment of Right Ventricular Intracardiac Flow
Authors: Meyers Brett , Nyce Jonathan , Zhang Jiacheng , Loke Yue-Hin , Vlachos Pavlos
Published: June 2023

Introduction:
Intracardiac flow imaging of the right ventricle [RV] may improve the management of congenital heart disease like repaired Tetralogy of Fallot [rTOF] or atrial shunts that cause RV dilation [RVD]. However, this is mostly performed via 4D flow magnetic resonance imaging [MRI], which has limited availability and is resource intensive. We propose the use of conventional echocardiography as a viable alternative to assess RV intracardiac flow.

Hypothesis:
We hypothesize that a velocity reconstruction algorithm implemented for conventional color Doppler echocardiography can resolve flow quantities with good agreement to 4D flow MRI.

Methods:
We retrospectively collected echos for children with rTOF/RVD and age-matched controls. Color Doppler echos of the RV were evaluated using an in-house algorithm called Doppler Velocity Reconstruction [DoVeR]. Measurements from 4D flow MRI and DoVeR were post-processed to collect hemodynamics parameters for relative pressure difference , vortex strength , total kinetic energy , and viscous energy loss . Agreement between modalities was performed by cross-correlation and L2-norm difference between indexed chamber-average timeseries measurements.

Results:
10 rTOF subjects, 3 RVD, and 7 age-matched controls were identified. There was qualitative agreement in spatial and time series measurements between DoVeR and 4D flow MRI (Figure 1). For timeseries measurements, there was strong correlation (median R > 0.85) with low differences (median L2 < 15%) between DoVer and 4D flow MRI.

Conclusions:
A color Doppler reconstruction method, DoVeR, can effectively measure hemodyanmics parameters using conventional color Doppler echocardiography, with strong agreement against 4D flow MRI.

Figure 1: Left – Flow field overlaid onto pressure reconstruction and pressure difference estimates between DoVeR and 4D flow modalities. Right – Correlation coefficient and difference between modality timeseries measurements.
Abstract 14972: Sestrin2 Prevents Excessive Cardiac Ischemia/Reperfusion Injury by Regulating Unfolded Protein Response
Authors: Li Xuan , Wang Zhen , Docarmo Jussara M , Da Silva Alexandre A , Mouton Alan J , Omoto Ana C , Hall John E
Published: June 2023

Introduction:
Cardiac ischemia/reperfusion (I/R) can alter redox and calcium homeostasis, causing proteins misfolded or unfolded in endoplasmic reticulum (ER) and activating unfolded protein response (UPR). Appropriate UPR is an adaptive response of the cell, however excessive and prolonged UPR can further deteriorate calcium overload and reactive oxygen species (ROS) overproduction. We have found that Sestrin2 (Sesn2), an endogenous inhibitor of the mammalian target of rapamycin complex 1 (mTORC1) is upregulated after cardiac I/R and lack of Sesn2 in hearts may cause more severe I/R injury.

Hypothesis:
Sesn2 maintains the ER homeostasis as an adaptive response to attenuate calcium overload and reduce ROS overproduction during myocardial I/R.

Methods:
Ligation/release of the left anterior descending coronary arteries (LAD) were performed in the C57BL/6J and cardiac specific Sesn2 knock out (Sens2

cKO

) mice to induce myocardial I/R injury.

Results:
We found that absence of Sesn2 in the hearts increased myocardial infarction by 49.4%, and decreased the systolic function of the heart by 22.9%, compared to wild type (WT) mice. I/R-induced UPR is a time-dependent process, and it reached its peak at 6 hours post reperfusion, but returned to normal after 24 hours of reperfusion in WT mice, however, Sesn2 deficiency resulted in higher and prolonged activation curve of UPR, as well as more myocardial apoptosis, compared to WT mice. We also found that Sesn2 deficiency impaired calcium transient and increased ROS after I/R. Furthermore, the activation of mTORC1 pathway after I/R was also significantly increased and prolonged in Sesn2

cKO

hearts. Inhibiting the mTORC1 by rapamycin after I/R reduced myocardial apoptosis in both WT and Sesn2

cKO

mice.

Conclusions:
During I/R, the regulation of the mTORC1 signaling pathway by Sesn2 may be a key factor in maintaining ER homeostasis to reduce cardiac I/R injury.
Abstract 14978: Association Between Microvascular Dysfunction and Heart Failure Related Outcomes in Ischemia With Non-Obstructive Coronary Arteries Based on a Novel Angiography-Derived Index
Authors: Leung Ka Lam Calvin , Lam Lok Yee , LI Kwan Yu , Yi Kei Stephanie Tse , Yu Shuk Yin , Feng Yundi , Huo Yunlong , Tse Hung Fat , Yiu Kai Hang
Published: June 2023

Introduction:
Coronary angiography-derived index of microvascular resistance (caIMR) is a index that quantifies coronary microvascular dysfunction (CMD), which obviates the need of wire manipulation compared to index of microvascular resistance (IMR). In patients with ischemia with non-obstructive coronary arteries (INOCA), CMD plays a key role and is associated with development of heart failure (HF). The significance of global CMD, reflected by the average of caIMR values obtained in the three coronary arteries was however not fully known. The aim of this study was to evaluate the association between HF related outcomes (HFRO) and global CMD in INOCA patients based on caIMR.

Methods:
Patients with clinical myocardial ischemia and without ≥50% diameter stenosis in any coronary arteries on coronary angiography were included. For every patient, the global-caIMR value was calculated by averaging caIMR values measured in the three major coronary arteries, and it reflects the extent of global CMD. Based on IMR threshold, patients were stratified into high-global-caIMR group (global-caIMR≥25U) and low-global-caIMR group (global-caIMR<25U). The primary endpoint was HFRO, defined by a composite of HF related hospitalization and outpatient diuretic intensification.

Results:
Among 325 patients included (mean age 63.4±11.0; male 57.2%), 105 and 220 patients were stratified into the high- and low-global-caIMR groups respectively. The rate of HFRO at 3 years was higher in the high-global-caIMR group compared to low-global-caIMR group (19.0% vs. 8.2%; P=0.004). In multivariable analysis adjusted for age, gender, Charlson Comorbidity Index and baseline HF, global-caIMR≥25U was associated with a higher risk of HFRO at 3 years (subdistribution hazard ratio [sHR], 2.65; 95% confidence interval [CI], 1.39-5.06; P=0.003). With the same multivariable adjustment, every 1U increase in global-caIMR was associated with an increased risk of HFRO at 3 years (sHR, 1.07; 95% CI, 1.05-1.09; P<0.001). Similar trends were observed at 5 years of follow-up.

Conclusions:
Global CMD, reflected by increased global-caIMR, was associated with a higher risk of HFRO at 3 and 5 years in INOCA patients. This study provides clinical data that support real-world application of caIMR.
Abstract 14988: Longitudinal Strain for Early Identification of Anthracycline-Induced Cardiomyopathy in Pediatrics
Authors: Ezeoke Ogochukwu , Husain Nazia , Reichek Jennifer L , Franklin Dana R , Acevedo Jennifer , Watanabe Kae
Published: June 2023

Introduction:
Anthracycline-induced cardiomyopathy (ACM) affects pediatric cancer survivors and echocardiogram (echo) surveillance is required. In adults, a decline in global longitudinal strain (LS) >15% is a pre-clinical marker of ACM. There is utility in determining change in strain associated with pre-clinical ACM in pediatrics.

Methods:
We queried our institutional database to identify patients <19 yo, with acute myeloid leukemia (AML), Ewing’s sarcoma or osteosarcoma, who received anthracyclines (1/1/2008 - 12/31/2020), and developed sub-acute or chronic ACM (LVEF <55% greater than 30 days post anthracycline completion). Echo at baseline (time of cancer diagnosis) and pre-ACM (within 2 years of first documented LVEF <55%) were compared for strain differences. Four-chamber LS, mid-chamber circumferential strain (CS) and left atrial strain (LAS) were analyzed using TomTec software.

Results:
Twenty-two patients were eligible for analysis from 428 meeting inclusion criteria (Figure 1). Demographics are shown in Table 1. There was a significant decline in LS from baseline to pre-ACM echo (-20.7 +/-2.98 vs -18.2 +/-3.74; p=0.026) amounting to a mean absolute LS decline of 2.5% and relative decline of 12.4% from baseline. Differences in CS (25.15 +/-4.89 vs 23.44 +/-5.11; p=0.35), LAS (43.09 +/-10.48 vs 45.23 +/-13.43; p=0.54) and heart rate (99.55 +/-19.88 vs 90.24 +/-26.47; p=0.09) were not significant.

Conclusions:
A significant decrease in LS occurs following anthracyclines pre-dating the diagnosis of subacute or chronic ACM in children. Prospective research with established imaging protocols may validate the role of strain in surveillance for pediatric ACM.
Abstract 14991: Local Prolongation of Repolarization Prevents Post Myocardial Infarction Ventricular Tachycardia
Authors: Ohara Ryan , Meijborg Veronique , Jelvehgaran Pouya , van der Waal Jeanne , Boink Gerard J , Trayanova Natalia , Coronel Ruben , Boukens Bastiaan J
Published: June 2023

Introduction:
Reentry is the predominant mechanism underlying ventricular arrhythmias in chronic myocardial infarction patients. Reentry can be prevented by prolonging effective refractory period, thereby blocking reexcitation of recently activated tissue. This is currently achieved with systemically administered class III antiarrhythmic drugs. These therapies are limited by their risk of provoking ventricular arrhythmias by off-target effects.

Hypothesis:
Local prolongation of repolarization in healthy myocardium adjacent to the exit site attenuates reentry and postinfarction arrhythmias.
Method and results:
We used a pig model and generated myocardial infarction by balloon occlusion of the left anterior descending coronary artery and performed epicardial mapping 6 weeks later during Langendorff-perfusion. Programmed stimulation induced reentry leading to ventricular premature beats and sustained ventricular tachycardia (VT). Local injection of Sotalol adjacent to the infarcted myocardium prolonged repolarization and prevented VT after programmed stimulation. To translate these experimental findings to the patient situation we used a virtual heart based on data from a patient suffering from postmyocardial infarction VTs. The patient underwent pre-ablation cardiac late gadolinium enhanced- magnetic resonance imaging followed by geometrical reconstruction of the left ventricle and computational electrophysiological modelling of VT induction. We used a rapid pacing protocol to induce VT and determined the site where the activation front exited the infarct zone and reactivated the adjacent normal myocardium. Next, we prolonged repolarization of the normal myocardium adjacent to the exit site with 50 ms and performed rapid pacing to induce VT. This only resulted in a single premature ventricular beat after which sinus rhythm restored.

Conclusion:
Our experimental and
in silico
findings show the potential of local prolongation of repolarization a novel potentially genetic anti-arrhythmic therapy in post-infarction patients.
Abstract 15001: Correlating Site of Origin in Ventricular Arrhythmias With Site of Inflammation in Patients With Underlying Myocardial Inflammation via FDG-PET Scan
Authors: Charate Rishi , Ahmed Adnan , Pothineni Naga , Gopinathannair Rakesh , Lakkireddy Dhanunjaya
Published: June 2023

Introduction:
Patients with underlying myocardial inflammation often present with otherwise unexplained ventricular arrhythmias. We seek to further elucidate if these arrhythmias originate from the areas of inflammatory FDG uptake, suggesting a direct arrhythmogenic substrate.

Hypothesis:
To determine if sites of inflammation on Fluorodeoxyglucose (FDG)-PET scan correlate with the site of origin on electroanatomic mapping for PVC/VT ablation in underlying myocardial inflammation.

Methods:
We retrospectively reviewed patients presenting with frequent ventricular arrhythmias

(PVC/VT) with positive FDG-PET scans indicative of underlying myocardial inflammation, and included patients who had persisting, symptomatic, burdensome arrhythmias despite multiple cycles of immunosuppressive therapy or resolution of inflammation that eventually required catheter ablation for PVC/VT. Patients were then reviewed to see if their site of origin (SOO) of PVCs or VT correlated with the site of inflammation (SOI) seen on FDG-PET scan.

Results:
We studied 42 patients with documented PVCs or VT, a positive FDG-PET scan and eventual PVC/VT ablation. Mean age was 58.9 years
+
14.5. Mean EF was 42.6
+
8.6%. Average PET SUV Max was 3.8
+
1.2, with an average involvement of 2.7
+
2.1 segments. Nearly all patients had multiple sites of FDG uptake (95.2%), with 69.% localized vs 31.0% diffuse distributions. The 42 patients had SOO in the following distribution: RVOT (30.1%), Inferoseptal (26.2%), RCC area, (19.0%), LVOT: (14.3%), Anteroseptal: (14.3%), Basal (11.9%), LV Summit (9.5%), and anterolateral (4%). 35.7% of patients had multiple SOO. Exact correlation between SOO-SOI was seen in 23/42 (54.8%), whereas partial correlation (matching sites with additional PVC locations vs uptake distributions, or additional uptake distributions vs PVC locations) was seen in 10/42 (23.8%). No correlation was seen in 9/42 (21.4%). Of the 19 that had partial or no correlation, the most frequent distributive area of uptake without PVC correlation was anterolateral, present in 7/19 (36.8%).

Conclusions:
In our retrospective study, patients undergoing cardiac FDG-PET scans to confirm myocarditis, had a positive correlation of site of PVC/VT origin and site of inflammation.