Risultati per: Probiotici: in vivo vs in vitro

Circulation, Volume 146, Issue Suppl_1, Page A15749-A15749, November 8, 2022. Introduction:Fluoroscopy-guided catheter ablation remains a common practice. With the presence of technologies such as intracardiac echocardiography (ICE) and electroanatomic mapping (EAM), it is possible to perform fluoro- and lead-free procedures to avoid the hazards of prolonged radiation exposure. Left atrial appendage electrical isolation (LAAEI) is a proven e!ective ablation strategy for atrial fibrillation (AF). We sought to assess the characteristics, complications, and outcomes of fluoroscopy-guided LAAEI and compare them to fluoroless LAAEI.Methods:We retrospectively analyzed patients with AF who underwent LAAEI as part of their ablation procedure. The study group included patients who underwent LAAEI either under fluoroscopy guidance or with ICE and EAM alone. We analyzed outcomes and procedural data, including procedural times, fluoroscopy times, radiation exposure (mGy), and radiofrequency times.Results:Overall, we analyzed 168 patients who underwent LAAEI; 110 under fluoroscopy guidance and 58 fluoroless. The mean procedural time was similar in both groups (248 ± 101 vs 264 ± 83 minutes, respectively, p=0.27). The mean RF time was longer in the fluoroscopy-guided group (50 ± 28.2 compared to 43 ± 17.1 minutes, respectively, p

Circulation, Volume 146, Issue Suppl_1, Page A14766-A14766, November 8, 2022. Introduction:The cardiovascular disease that causes most of the deaths is atherosclerosis. The current research to assess efficacy of treatments is based on animal models, but these present several limitations related to time and costs associated. Ex-vivo systems represent useful models to study vascular biology and to test medical devices. The main goal of this research was to culture coronary arteries in an ex-vivo vascular bioreactor in order to maintain them alive and functional by mimicking physiological conditions.Methods:Culture experiments were performed on porcine right coronary arteries (RCA). Duration was set to 2 (n=3), 4 (n=3), and 9 days (n=3). The RCAs were dissected from hearts harvested from a local slaughterhouse. The bioreactor containing the dissected RCA was placed in a 38°C incubator. The RCA was cultured in a blood-mimicking culture medium. To assess RCA morphology and diameter, ultrasound imaging was performed. Pressure and flow rate were monitored daily. The flow rate was set to ensure a physiological peak endothelial shear stress of around 0.8-1.8 Pa. After 2, 4 or 9 days of culture, the RCA was embedded in paraffin and cut into 4 μm sections. Histological stainings (H&E and RF) were performed on the sections.Results:During RCA cultures, the mean pressure was 60-100 mmHg, and the peak flow rate was around 60 ml/min. Ultrasound imaging showed that the morphology and structure of cultured arteries was maintained during the culture. Histological analyses showed preserved endothelium and extracellular matrix. After 2 days of culture, endothelial coverage was not complete while after 4 days it was completely restored. After 9 days of culture, a layer of newly formed neointima was visible.Conclusions:The cultured RCAs maintained their physiological morphology up to 9 days of culture. The investigated model represents a useful tool for testing vascular therapies and devices, supporting the translational phase between in-vitro and in-vivo studies.

Circulation, Volume 146, Issue Suppl_1, Page A11397-A11397, November 8, 2022. Introduction:TOTUM-070 is a patented polyphenol-rich compound which has shown hypocholesterolemic properties in preclinical studies. However, clinically validated approaches and further investigations on the mode of action at cellular level especially in humans are required for optimized care management. In this study, we designed an ex-vivo clinical innovative approach considering the metabolites produced by the digestive tract following the ingestion of TOTUM-070 in humans.Methods:Human serum was collected in healthy subjects before and following acute intake of 5g of TOTUM-070. Availability of circulating metabolites was confirmed and characterized by UPLC-MS. Human serum enriched with metabolites deriving from TOTUM-070 absorption was further incubated with human hepatocytes, pretreated or not with palmitate (250 μM). In such lipotoxic environment, hepatocyte behavior was monitored to determine whether and how TOTUM-070 metabolites may improve cholesterol metabolism in human.Results:In the presence of the human metabolites from TOTUM-070, human hepatocytes were protected from an induced lipotoxic stress. No effect on cell toxicity was detected in the presence of enriched sera. Hepatocyte protection was characterized by (1) the inhibition of both triglycerides (-41%, p

Circulation, Volume 146, Issue Suppl_1, Page A15119-A15119, November 8, 2022. Introduction:Marfan syndrome (MFS) carries a risk of aortic dissection and death. The current aneurysm repair guideline of diameter ≥5cm may neglect up to 15% of MFS patients who dissect at smaller diameters. Since dissection can occur biomechanically when wall stress exceeds tissue strength, patient-specific biomechanics may enhance risk stratification for MFS patients. The goal of this study was to compare aneurysm wall stresses between propensity score-matched MFS and non-MFS aneurysm patients.Methods:The cohort consisted of 5 MFS and 42 non-MFS patients with aortic aneurysm. Preoperative CTA or MR scans were used to create 3D hexahedral meshes. Finite element analysis was performed with MFS and nonMFS material properties to determine peak (99th percentile) diastolic stresses[TE1] . Propensity scores were calculated by logistic regression using age, sex, height, hypertension, diabetes, smoking history, valve type, and sinus diameter. Five pairs were formed by 1:1 optimal matching without replacement. Comparisons used the paired t-test.Results:Diameters were similar at the sinus but lower for MFS at the STJ (3.4±0.5 vs 3.9±0.7cm, p=0.003) and the ascending aorta (3.1±0.5 vs 4.3±0.5cm, p

Circulation, Volume 146, Issue Suppl_1, Page A12697-A12697, November 8, 2022. Introduction:The co-chaperone Bcl2-associated athanogene-3 (BAG3), along with its cofactor heat shock protein 70 (HSP70), is fundamental for protein quality control and cell survival in a healthy heart. However, elevated expression of BAG3 is also associated with metastasis in breast and other cancers, and small molecules such as JG-98 that disrupt BAG3-HSP70 binding have been shown to reduce cancer cell proliferation. We previously showed JG-98 is cardiotoxic in neonatal rat ventricular myocytes.Hypothesis:As BAG3-HSP70 is fundamental for autophagic protein turnover, JG-98 will reduce tumor size but have cardiotoxic effects.Methods:BT474 Resistant breast cancer cells were injected bilaterally into mammary fat pads of female athymic nude mice. When tumors reached 25 mm2, mice received a baseline echocardiography and were randomized to intraperitoneal injections of either PBS/DMSO vehicle or 3 mg/kg JG-98 twice weekly for 3 weeks (n = 4/group) or 6 weeks (n=10/group). Tumor growth was monitored using calipers and echocardiography was performed prior to sacrifice.Results:After 3 weeks, tumor volume increased by ~44% in Vehicle treated mice but decreased by 17% in JG-98 treated mice, effectively halting tumor growth. However, cardiac echocardiography did not reveal any functional or structural differences between the vehicle and JG-98 treated mice after either 3 or 6 weeks. JG-98 did result in dysregulation of BAG3 and its interactome in the LV. JG-98 slightly increased BAG3 levels in the LV at 3 weeks, but slightly decreased levels at 6 weeks. This agrees with data showing acute stress upregulates BAG3, but chronic stress downregulates it. Similarly, we found altered protein expression of the BAG3 binding partners HSBP8, HSPB5, and HSP70.Conclusions:JG-98 had no overt cardiotoxic effects, suggesting the adult heart may be resistant to disrupted BAG3-HSP70. However, we also observed dysregulated protein levels in the BAG3-mediated autophagy pathways that could precede dysfunction with more chronic treatment. Furthermore, mutations in BAG3 or altered expression levels could sensitize individuals to JG-98 treatment. Further work is necessary to understand the cardiac impact of cancer therapeutics that target BAG3.

Circulation, Volume 146, Issue Suppl_1, Page A13987-A13987, November 8, 2022. Background:Several studies have shown the cost-effectiveness of direct oral anticoagulants (DOACs), compared with warfarin, to prevent atrial fibrillation (AF) related complications. However, few have reported cost-effectiveness of DOACs in AF patients with intermediate stroke risk. Thus, we investigated the cost-effectiveness of DOACs vs. warfarin in non-valvular AF patients with intermediate stroke risk using national representative data.Methods:We identified 7,954 newly diagnosed non-valvular AF patients (≥18 years) with intermediate stroke risk (CHA2DS2-VASc score: 1 for men and 2 for women) using the national healthcare utilization data from August 1, 2016, to July 31, 2019. Annual incidence rate of AF-related composite outcomes (heat failure, myocardial infarction, ischemic stroke, intracerebral hemorrhage, and gastrointestinal bleeding) was estimated. Cost-effectiveness was estimated using a Markov chain model with the transition probability of 1 year. The willingness-to-pay (WTP) was set at $32,000 per quality-adjusted life-year (QALY) gained.Results:The total cost of warfarin, rivaroxaban, apixaban, dabigatran and edoxaban was $2,874, $5,761, $5,151, $5,761 and $5,851, respectively. The QALYs gained were 10.83, 10.95, 11.10, 10.49 and 10.99 years, respectively. The incremental cost-effectiveness ratio of rivaroxaban, apixaban, dabigatran and edoxaban was $29,743.99, $8,426.71, -$8,483.04 and $18,483.55, respectively. The WTP was set at $32,000. DOACs (except dabigatran) were more cost-effective compared with warfarin because they did not exceed the WTP in the base-case analysis.Conclusion:Our findings showed that DOACs were more cost-effective than warfarin in non-valvular AF patients with intermediate stroke risk.Figure 1. Cost-effectiveness acceptability curve of Warfarin vs. DOACs in AF patients with intermediate stroke risk.

Circulation, Volume 146, Issue Suppl_1, Page A15172-A15172, November 8, 2022. Introduction:Segmenting cardiac computed tomography (CT) to provide anatomic guidance for Atrial Fibrillation (AF) ablation is routinely applied, but is time-consuming and prone to error. Machine learning (ML) is a powerful approach that could automate this approach, but is hindered by the small size of available labeled datasets.Hypothesis:We hypothesized that a new computational pipeline, in which an ML model is trained mathematically in a small cohort (N=20) using geometrical heart avatars derived from computer graphics imaging (CGI), rather than on manually-segmented data, would enable rapid expert-level segmentation of raw cardiac CT scans.Methods:We first encoded anatomical knowledge with generic geometrical avatars and derived a “virtual dissection” method to geometrically parse the heart (Fig A). An ML model trained by virtual dissection using 20 cases was able to rapidly and accurately segment the pulmonary veins (PVs), left atrial appendage (LAA), and left atrium (LA) from cardiac CT scans (Fig B), which we tested in a retrospective study of N=100 patients (30% women, 64.7±10.1Y) and in a prospective clinical trial of N=42 patients (42.9% women, 65.2±10.8Y) undergoing AF ablation, against a panel of 3 experts (Fig C).Results:In a retrospective study (N=100), ML achieved median Dice scores of 96.6% (IQR: 95.1% to 97.5%), similar to experts (p

Circulation, Volume 146, Issue Suppl_1, Page A15237-A15237, November 8, 2022. Introduction:The MitraClip for treatment of mitral regurgitation (MR) is a generally safe and effective procedure. However, recent clinical trials have reported contradictory outcomes, and long-term efficacy remains almost completely unknown. Therefore, there is an urgent need for quantitative, patient-specific treatment optimization grounded on a robust understanding of the functional consequences of the MitraClip repair procedure on each patient-specific mitral valve (MV).Methods:Real-time 3D echocardiography images of ten MR patients’ MVs were acquired during the MitraClip procedure. These images were analyzed using our validated noninvasive strain estimation method to compute the pre-operative leaflet strain fields in the circumferential and radial directions. For the post-operative analysis, 3D MitraClip models were implemented in the corresponding size and number in the simulation to directly grasp the MV leaflets at echo-dictated positions.Results:The MV pre-surgical states were highly variable in both geometry and deformation, underscoring the heterogeneity of the MR in the patient population. There were also no consistent post-surgical global or local strain patterns between MVs. However, the pre- and post-surgical strain fields in the same MV were significantly better correlated than random pre- and post-surgical pairs (p < 0.005). This quantitative result implies that the post-surgical state for each patient can bepredictedfrom available pre-surgical data-derived modelling.Conclusions:Due to the highly variable pre-surgical state of regurgitant MVs, a patient-specific approach to MitraClip repair is crucial to improving long-term patient outcomes. Moreover, these findings suggest that the post-surgical state can be correlated to, and therefore predicted from, the pre-surgical state, which would lay the foundation for quantitative surgical planning, tailored patient selection, and ultimately, a more durable repair.

Circulation, Volume 146, Issue Suppl_1, Page A12656-A12656, November 8, 2022. Introduction:Pulmonary vein isolation (PVI) remains the cornerstone of atrial fibrillation (AF) ablation procedures. Because many patients have continued AF after PVI alone, posterior wall isolation (PWI) has been proposed as an adjunctive strategy to PVI. We conducted a meta-analysis of available data to assess the incremental impact of PWI based on whether patients had paroxysmal or persistent (PAF, PeAF) AF.Objective:To evaluate the incremental impact of PWI following PVI in patients with PAF and PeAF.Methods:The online databases (Pubmed, Medline, Embase) were searched using the terms “pulmonary vein isolation”, “posterior wall isolation”, or “posterior wall ablation”. Out of 1843 studies, 11 met our strict criteria (Table). Specifically, patients were undergoingde novoablation for treatment of PAF or PeAF using either cryoballoon or radiofrequency energy. Patients had PVI +/- PWI; studies that included additional ablations (except for cavotricuspid isthmus ablation) were excluded. The primary outcome was long-term freedom from recurrence of any atrial tachyarrhythmia (AT).Results:The cohort consisted of 1702 patients (65 ± 18 years, 75% male); 1404 (82%) pts with PeAF and 298 (18%) pts with PAF (Table). In the PeAF group, AT recurrence was significantly higher in patients who underwent PVI alone as compared to those who also had PWI (OR 1.85, 95% CI [1.23, 2.77], P = 0.003, Figure). In contrast, in the PAF group, outcome was similar in patients who did and did not undergo PWI in addition to PVI (OR 0.95, 95% CI [0.53, 1.72], P = 0.86, Figure).Conclusions:PWI was rarely used during thede novoablation procedure in PAF patients and did not seem to impart additional benefit. In contrast, PeAF patients seemed to derive favorable impact from the addition of PWI to PVI. Additional randomized studies are needed to determine the role of PWI in PeAF patient undergoing de novo ablation for management of AF.

Circulation, Volume 146, Issue Suppl_1, Page A15753-A15753, November 8, 2022. Epigenetic regulation is critical for cardiac electrophysiology and pathology. Epigenetic modulators, such as histone deacetylases (HDACs) and histone acetyltransferases (HATs) are known master regulators of gene expression. Recently, novel pharmacological agents, HDAC inhibitors, have been developed as treatments for cancer and immune diseases. The effects of HDAC inhibitors on cardiac ion channels (ICs) are of great interest. To exert specific gene modulation, we used small interfering RNAs against the known HDACs, including sirtuins, and deployed them in human induced pluripotent stem-cell-derived cardiomyocytes (hiPSC-CMs). Follow-up RNAseq data (n = 61) were compared to identically processed and normalized RNAseq data from human left ventricle (LV) from the GTEx database (n = 84). Gene expression of cardiac ICs displayed similar patterns, with some differences. For example, hiPSC-CMs showed upregulatedCACNA1C, SLC8A1and downregulatedKCNJ2andRYR2compared to the adult LV, most of which are known distinctions (Fig. 1A). Correlative analysis (Fig. 1B) and partial least square regression models helped visualize links between HDACs/HATs, key transcription factors (TFs) and cardiac ICs. Powerful TFs, includingMEF2A, GATA4, 6exerted positive effect on ICs in hiPSC-CM and the adult LV. In the hiPSC-CMs,HDAC1, HDAC10andSIRT6were found to be the strongest predictors of the expression of individual cardiac ICs, as revealed by permutation importance. Further studies will involve determination of the role of different cell types using single-cell sequencing data from the adult LV. Our analysis offers new insights about the role of epigenetic modifiers on cardiac electrophysiology and informs the utility of hiPSC-CM as a scalable, experimental model for cardiotoxicity testing of HDAC inhibitors.

Circulation, Volume 146, Issue Suppl_1, Page A13473-A13473, November 8, 2022. Introduction:Atrial natriuretic peptide (ANP) is the most potent endogenous activator of the guanylyl cyclase A receptor (GC-A) mediating cardiorenal protective actions by increasing cGMP production. To test the hypothesis that acute decompensated heart failure (ADHF) is associated with increased circulating ANP but with reduced GC-A potency, we developed an ex-vivo precision medicine assay to quantify the potency of circulating ANP in plasma from healthy and ADHF subjects. Further, we assessed the ex-vivo potency of MANP, a novel ANP analog with greater cGMP activation than ANP, also using plasma of healthy and ADHF subjects.Methods:For the potency assay, we engineered HEK293 cells to overexpress human GC-A. Plasma from individual healthy (n=4) and ADHF (n=4) subjects was incubated in the assay and cGMP was assessed. The endogenous ANP derived cGMP was evaluated against the cGMP response of equimolar synthetic ANP (SANP) to assess potency of ANP from each cohort. MANP mediated cGMP generation was also assessed in the assay using healthy and ADHF plasma.Results:Healthy plasma, in which plasma ANP was 26±5 pg/mL, generated 10.3±0.7 pmol/mL of cGMP. SANPadded to the assay to mimic endogenous ANP levels in healthy subjects produced 7.8±0.5 pmol/mL of cGMP. ADHF plasma with markedly elevated ANP (350±57pg/mL) generated 23.5±3.1 pmol/mL of cGMP. SANPmimicking plasma ANP in ADHF produced 59.7±13.8 pmol/mL of cGMP, which was greater than cGMP generated from ADHF patient-derived plasma (p

Circulation, Volume 146, Issue Suppl_1, Page A9624-A9624, November 8, 2022. Introduction:Incliciran, a small interfering ribonucleic acid (siRNA), is the new modulator of the proprotein convertase subtilisin kexin 9 (PCSK9) that has proven safe and effective in reducing LDL-C and ASCVD. However, it is unclear if incliciran is superior to the already established PCSK9 monoclonal antibodies (mAb) in reducing major adverse cardiac events (MACE).Methods:We performed a meta-analysis of randomized controlled trials of patients with high cardiovascular risk who underwent PCSK9 inhibition by either mAb or siRNA. The primary outcome was the reduction of MACE.Results:Thirty RCTs with a total of 92,775 patients were included. For patients with a high cardiovascular risk profile, the modulation of PCSK9 by mAb decreases the risk of MACE (HR 0.84, 95% CI 0.8 – 0.89, p

Circulation, Volume 146, Issue Suppl_1, Page A14818-A14818, November 8, 2022. Introduction:Endothelial mechano-transduction mechanisms are instrumental to vascular health and disease. Novel strategies targeting disease-causing mechano-sensitive pathways in dysfunctional endothelial cells could revolutionize future cardiovascular therapeutics. Vascular complications such as atherosclerosis and stenosis preferentially develop at arterial curvatures and bifurcations where endothelial cells are activated by local disturbed blood flow, leading to peripheral artery disease, carotid artery disease and ischemic stroke.Hypothesis:Current vascular therapies mainly target systematic risk factors (e.g. hypercholesterolemia and hypertension) but not the diseased vasculature, distinct molecular/cellular signatures of which can be targeted by innovated precision nanomedicine approaches.Methods:We first elucidated novel mechano-sensitive molecular mechanisms in endothelium activated by disturbed flow (DF) and then engineered rationally-designed nano-materials with purposed-constructed functionalities to deliver therapeutic nucleotides to DF-activated endothelial cells.Results:Our results elucidated previously unrecognized endothelial mechano-sensitive pathways in endothelial activation, with emphasis upon cellular metabolism (DF-induced glycolysis), human genetic variants (DF-induced suppression of PLPP3, a CAD GWAS gene), miRNA, protein stability (DF-induced NOS3 protein degradation via TXNDC5) and mRNA chemical modification/epi-transcriptome (DF-induced suppression of m7G). VCAM1-targeting nanoparticles were engineered to deliver therapeutic nucleotides such as mRNA, miRNA inhibitor, or CRISPR/Cas9 constructs specifically to inflamed endothelial cells to intervene aforementioned mechano-sensitive pathways, effectively reducing atherosclerosis and stenosis in mice. Similar approaches were very effective to promote endothelial health and lessen acute respiratory distress syndrome (ARDS) in mice induced by influenza or SARS-CoV-2 viruses.Conclusions:These results elucidate novel endothelial mechano-sensing mechanisms and provide a proof of concept of innovative targeted nanomedicine approaches, addressing an unmet medical need in vascular therapies.

Circulation, Volume 146, Issue Suppl_1, Page A15042-A15042, November 8, 2022. Introduction:Use of gene therapy for cardiac transplantation is limited by the lack of an effective recombinant Adeno-associated viral vector (rAAV) gene delivery strategy.Ex vivonormothermic perfusion has been increasingly used in clinical practice and is an effective platform for achieving robust global gene delivery to a donor heart. Here we utilized a rationally designed cardiac-enhanced AAV3b variant, SASTG, to deliver the gene firefly luciferase to porcine hearts and evaluated transgene expression 30-days post-transplantation.Methods:Heterotopic heart transplantation was performed using blood-type and MHC matched Yucatan pigs (n=4). Donor hearts underwentex vivoperfusion for 2-hours with perfusate containing the rAAV SASTG-Luciferase. Three different doses of rAAV were used: 2×1013, 8×1013, 1x1014viral genome copies (VGC). Recipients were survived for 30 days at which time they were euthanized and donor and recipient hearts were examined for overall levels of luciferase activity, luciferase DNA copy number, and histology for luciferase expression on immunostaining and evidence of inflammation.Results:There was a dose-dependent response in luciferase activity observed in the donor heart ventricles with the lowest activity detected at the 2x1013VGC dose (1.5-9.9x103RLU/mg) mid-level activity at the 8x1013VGC dose (32.7-71.2x103RLU/mg) and highest activity at the 1x1014VGC dose (27.6-278.1x103RLU/mg). Luciferase DNA was detectable in a dose-dependent manner in the donor heart ventricles (0.02-131.1x103VGC/pg of DNA), but undetectable in all native hearts. Finally, immunostaining demonstrated diffuse luciferase expression in the donor heart but none in the recipient hearts. There was also no evidence of inflammation in any of the donor hearts.Conclusions:The cardiac-enhanced AAV3b variant, SASTG, is associated with robust, global transgene expression in porcine cardiac allografts when administered duringex vivoperfusion.

Circulation, Volume 146, Issue Suppl_1, Page A12802-A12802, November 8, 2022. Introduction:Limited access is a key challenge in the management of chronic diseases, like heart failure (HF), in rural areas. Telehealth (TH), which is the delivery of care via remote technologies, and remote monitoring (RM), which allows a patient’s implanted device to be interrogated at a distance, are potential ways to improve access. However, many older adults live in rural areas and age is negatively associated with the adoption of novel technologies. The aim of this study is to understand the association between age and use of TH and RM in urban vs. rural areas.Methods:We created annual, cross-sectional cohorts of patients with HF in each year from 2013 to 2018 with approximately 5 million patients/year by requiring ≥1 inpatient or ≥2 outpatient ICD9/10 codes for HF in the 2 years prior. TH and RM use were determined using validated ICD-9/10 and CPT codes. Rurality was determined using the Rural/Urban Commuting Areas (RUCA) classification. Multivariable, logistic regression models were used to determine the association between age and use of TH and RM in urban vs. rural areas.Results:Increasing age was associated with a decreased odds of TH use in both urban and rural areas. However, the decline in use with age was greater in urban (OR=0.41 for 85+ vs 65-69) than rural areas (OR=0.65). In contrast, increasing age was associated with an increased odds of RM in both urban and rural areas, with a larger increase with increase age in rural (OR=1.47 for 85+ vs 65-69) compared to urban areas (OR=1.35).Conclusion:TH and RM are viable ways to improve access for rural patients with HFrEF across the age spectrum. With increasing age, TH use decreases but RM increases, suggesting that patients’ ability to navigate technology plays a significant role – TH technology requires more active engagement by the patient compared to RM technology. Efforts to simplify TH technology would likely mitigate the drop in TH use seen with increasing age and improve access to care for older adults with HFrEF.

Circulation, Volume 146, Issue Suppl_1, Page A13129-A13129, November 8, 2022. Introduction:Catheter cardiac ablation is a standard treatment for atrial fibrillation for patients that do not respond to drugs. Pulsed electric field (PEF) cardiac ablation has been proposed as a nonthermal technology to cause cell death by irreversible electroporation (IRE). IRE has shown to minimize off target damage. The lack of systematic studies to inform optimal PEF ablation parameters has significantly slowed down device development and regulatory process.Hypothesis:As part of a larger effort to develop a standard laboratory assay for PEF parameter optimization, in this study we investigate tissue specificity of IRE cardiac ablation treatments.Methods:Human induced pluripotent stem cell cardiomyocytes derived (hiPSC-CMs, Fujifilm Cellular Dynamic, Inc.), human esophageal muscle cells (HESMC, ScienCell Research Laboratories), and human bronchial epithelial cells (HBEC3-KT, ATCC) were cultured in monolayer formats on 96-well NanoFiber plates. PEF were delivered by an FID pulse generator connected to two needle electrodes (1.7 mm center-to-center distance). PEF parameters investigated in this study are: phase duration tp=1, 5, 10 μs; phase intensity Vp=284, 372, 568V; pulse repetition frequency PRF=10, 100, 1000 Hz, pulse number p#=100. We quantified IRE areas 4 hours post-treatment by propidium iodide (PI).Results:Preliminary results show that increasing the tp, Vp and decreasing the PRF resulted in a larger IRE area in all the cell types studied. For the range of PEF parameters investigated, a treatment with same tp, p# and PRF required higher Vp to induce IRE in HESMC and HBEC3-KT than hiPSC-CMs, i.e., 100 pulses with tp=5 μs and PRF=1000 Hz, to achieve the same IRE area, hiPSC-CMs needed half Vp than HESMC and HBEC3-KT.Conclusions:Our data suggests that same PEF treatments induced larger IRE areas in hiPSC-CM than in HESMC and HBEC3-KT. Future work will focus on validating these results in tissues to assess safety of IRE cardiac ablation devices.