A5032725801- Cardiomyopathy and Myosin Studies
- Congenital heart defects research
- Pluripotent Stem Cells Research
- Cardiac electrophysiology and arrhythmias
- CRISPR and Genetic Engineering
- Tissue Engineering and Regenerative Medicine
- Neuroscience and Neural Engineering
- Cardiovascular Effects of Exercise
- 3D Printing in Biomedical Research
- Cardiac Fibrosis and Remodeling
- Viral Infections and Immunology Research
- Signaling Pathways in Disease
- Ion channel regulation and function
- Muscle Physiology and Disorders
- Peptidase Inhibition and Analysis
- RNA modifications and cancer
- Cancer-related gene regulation
- Genomic variations and chromosomal abnormalities
- Neurogenetic and Muscular Disorders Research
- Neuroscience and Neuropharmacology Research
- Autophagy in Disease and Therapy
- Single-cell and spatial transcriptomics
- Genomics and Chromatin Dynamics
- Protein Kinase Regulation and GTPase Signaling
- Trauma, Hemostasis, Coagulopathy, Resuscitation
Boston Children's Hospital
2019-2025
Harvard University
2019-2025
University Medical Center Hamburg-Eppendorf
2017-2024
Universität Hamburg
2017-2024
Boston Children's Museum
2023-2024
German Centre for Cardiovascular Research
2017-2024
Institute of Experimental Pharmacology and Toxicology of the Slovak Academy of Sciences
2018-2023
Boston University
2019-2021
Netherlands Heart Institute
2017
University of Oxford
2017
Sarcomeric gene mutations frequently underlie hypertrophic cardiomyopathy (HCM), a prevalent and complex condition leading to left ventricle thickening heart dysfunction. We evaluated isogenic genome-edited human pluripotent stem cell-cardiomyocytes (hPSC-CM) for their validity model, add clarity to, HCM. CRISPR/Cas9 editing produced 11 variants of the HCM-causing mutation c.C9123T-MYH7 [(p.R453C-β-myosin heavy chain (MHC)] in 3 independent hPSC lines. Isogenic sets were differentiated...
Abstract Human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CM) are a promising tool for drug testing and modelling genetic disorders. Abnormally low upstroke velocity is current limitation. Here we investigated the use of 3D engineered heart tissue (EHT) as culture method with greater resemblance to human in comparison standard technique 2D monolayer (ML) format. I Na was measured ML or EHT using patch-clamp technique. density ~1.8 fold larger (−18.5 ± 1.9 pA/pF; n = 17) than...
Hypertrophic cardiomyopathy (HCM) is a cardiac genetic disease accompanied by structural and contractile alterations. We identified rare c.740C>T (p.T247M) mutation in ACTN2, encoding α-actinin 2 HCM patient, who presented with left ventricular hypertrophy, outflow tract obstruction, atrial fibrillation. generated patient-derived human-induced pluripotent stem cells (hiPSCs) show that hiPSC-derived cardiomyocytes engineered heart tissues recapitulated several hallmarks of HCM, such as...
Gene therapy is a promising option for severe forms of genetic diseases. We previously provided evidence the feasibility trans-splicing, exon skipping, and gene replacement in mouse model hypertrophic cardiomyopathy (HCM) carrying mutation MYBPC3, encoding cardiac myosin-binding protein C (cMyBP-C). Here we used human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) from an HCM patient heterozygous c.1358-1359insC MYBPC3 healthy donor. hiPSC-CMs exhibited ∼50% lower mRNA...
Background: Cardiac chamber-selective transcriptional programs underpin the structural and functional differences between atrial ventricular cardiomyocytes (aCMs vCMs). The mechanisms responsible for these remain largely undefined. Methods: We nominated candidate enhancers (CSEs) by determining genome-wide occupancy of 7 key cardiac transcription factors (GATA4, MEF2A, MEF2C, NKX2-5, SRF, TBX5, TEAD1) coactivator P300 in atria ventricles. Candidate were tested using an adeno-associated...
Human iPSC-derived cardiomyocytes (hiPSC-CMs) have proven invaluable for cardiac disease modeling and regeneration. Challenges with quality, inter-batch consistency, cryopreservation scale remain, reducing experimental reproducibility clinical translation. Here, we report a robust stirred suspension differentiation protocol, perform extensive morphological functional characterization of the resulting bioreactor-differentiated iPSC-CMs (bCMs). Across multiple different iPSC lines, protocol...
Background: Cardiac repolarization abnormalities in drug-induced and genetic long-QT syndrome may lead to afterdepolarizations life-threatening ventricular arrhythmias. Human induced pluripotent stem cell–derived cardiomyocytes (hiPSC-CMs) should help overcome the limitations of animal models based on species differences reserve. Here, we compared head-to-head contribution I Ks (long QT1) Kr QT2) action potentials (APs) human left (LV) tissue hiPSC-CM–derived engineered heart (EHT). Methods:...
Human-induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CM) may serve as a new assay for drug testing in human context, but their validity particularly the evaluation of inotropic effects remains unclear. In this blinded analysis, we compared 10 indicator compounds with known electrically stimulated (1.5 Hz) hiPSC-CM-derived 3-dimensional engineered heart tissue (EHT) and atrial trabeculae (hAT). Human EHTs were prepared from iCell hiPSC-CM, hAT obtained at routine surgery. Mean...
Mutations in tafazzin (TAZ), a gene required for biogenesis of cardiolipin, the signature phospholipid inner mitochondrial membrane, causes Barth syndrome (BTHS). Cardiomyopathy and risk sudden cardiac death are prominent features BTHS, but mechanisms by which impaired cardiolipin muscle weakness arrhythmia poorly understood.We performed vivo electrophysiology to define vulnerability cardiac-specific TAZ knockout mice. Using cardiomyocytes derived from human induced pluripotent stem cells...
Alterations in autophagy have been reported hypertrophic cardiomyopathy (HCM) caused by Danon disease, Vici syndrome, or LEOPARD but not HCM mutations genes encoding sarcomeric proteins, which account for most of cases. MYBPC3, cMyBP-C (cardiac myosin-binding protein C), is the frequently mutated gene.We evaluated patients with carrying MYBPC3 and a Mybpc3-targeted knockin mouse model, as well effect modulators on development mice. Microtubule-associated 1 light chain 3 (LC3)-II levels were...
Chronic tachypacing is commonly used in animals to induce cardiac dysfunction and study mechanisms of heart failure arrhythmogenesis. Human induced pluripotent stem cells (hiPSC) may replace animal models overcome species differences ethical problems. Here, 3D engineered tissue (EHT) was investigate the effect chronic on hiPSC-cardiomyocytes (hiPSC-CMs).To avoid cell toxicity by electrical pacing, we developed an optogenetic approach. EHTs were transduced with lentivirus expressing...
Genetic variants in α-actinin-2 (ACTN2) are associated with several forms of (cardio)myopathy. We previously reported a heterozygous missense (c.740C>T) ACTN2 gene variant, hypertrophic cardiomyopathy, and characterized by an electro-mechanical phenotype human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs). Here, we created CRISPR/Cas9 genetic tools two functional knock-out hiPSC lines second wild-type (ACTN2wt) (ACTN2mut) allele, respectively. evaluated their impact on...
BACKGROUND: How the sarcomeric complex is continuously turned over in long-living cardiomyocytes unclear. According to prevailing model of sarcomere maintenance, sarcomeres are maintained by cytoplasmic soluble protein pools with free recycling between and sarcomeres. METHODS: We imaged quantified turnover expressed endogenous proteins, including giant titin, culture vivo, at single cell level using pulse-chase labeling Halo-tagged proteins covalent ligands. RESULTS: disprove pool instead...
<title>Abstract</title> Heart failure is a multifaceted syndrome contributing significantly to mortality and hospitalization rates among the global population1. One of prevalent causes heart ischemic disease (IHD), often caused by blockage in coronary artery, ultimately leading loss myocardial tissue contractile force2. The impact this ambiance on cardiomyocyte genome transcriptome has not been thoroughly studied. During normal aging, cardiomyocytes progressively accumulate somatic mutations...
N-terminal acetyltransferases including NAA10 catalyze acetylation, an evolutionarily conserved co- and post-translational modification. However, little is known about the role of acetylation in cardiac homeostasis. To gain insight into cardiac-dependent function, we studied a previously unidentified variant p.(Arg4Ser) segregating with QT-prolongation, cardiomyopathy, developmental delay large kindred. Here, show that NAA10R4S reduced enzymatic activity, decreased NAA10-NAA15 complex...
Polysialic acid (PSA) is a homopolymeric glycan that plays crucial roles in the developing and adult nervous system. So far only few PSA-binding proteins have been identified. Here, we identify myristoylated alanine-rich C kinase substrate (MARCKS) as novel PSA binding partner. Binding assays showed direct interaction between peptide comprising effector domain of MARCKS (MARCKS-ED). Co-immunoprecipitation PSA-carrying neural cell adhesion molecule (PSA-NCAM) with co-immunostaining at...
BACKGROUND: Calmodulinopathies are rare inherited arrhythmia syndromes caused by dominant heterozygous variants in CALM1 , CALM2 or CALM3 which each encode the identical CaM (calmodulin) protein. We hypothesized that antisense oligonucleotide (ASO)–mediated depletion of an affected calmodulin gene would ameliorate disease manifestations, whereas other 2 genes preserve level and function. METHODS: tested this hypothesis using human induced pluripotent stem cell–derived cardiomyocyte mouse...
Colloids, known as volume expanders, have been used resuscitation fluids for hypovolemic shock decades, they increase plasma oncotic pressure and expand intravascular volume. However, recent studies show that commonly synthetic colloids adverse interactions with human biological systems. In this work, a low-fouling amine(N)-oxide-based zwitterionic polymer an alternative expander improved biocompatibility efficacy is designed. It demonstrated the possesses antifouling ability, resisting cell...
Human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) represent an unlimited source of human CMs that could be a standard tool in drug research. However, there is concern whether hiPSC-CMs express all cardiac ion channels at physiological level and they might non-cardiac channels. In control hiPSC line, we found large, “noisy” outward K+ currents, when measured potassium currents isolated hiPSC-CMs. Currents were sensitive to iberiotoxin, the selective blocker big...
In the last decade human iPSC-derived cardiomyocytes (hiPSC-CMs) proved to be valuable for cardiac disease modeling and regeneration, yet challenges with scale, quality, inter-batch consistency, cryopreservation remain, reducing experimental reproducibility limiting clinical translation. Here, we report a robust differentiation protocol that uses Wnt modulation stirred suspension bioreactor produce on average 124 million hiPSC-CMs >90% purity using variety of hiPSC lines (19...
<title>Abstract</title> N-terminal-acetyltransferases including NAA10 catalyze N-terminal acetylation (Nt-acetylation), an evolutionarily conserved co-translational modification. Little is known about the role of Nt-acetylation in cardiac homeostasis. To gain insights, we studied a novel variant (p.R4S) segregating with QT-prolongation, cardiomyopathy and developmental delay large kindred. Here show that NAA10-R4S mutation reduced enzymatic activity, decreased expression levels NAA10/NAA15...