Michael Gotthardt

Max-Delbrück-Centrum für Molekulare Medizin, Berlín, Berlin, Germany

Are you Michael Gotthardt?

Claim your profile

Publications (66)519.48 Total impact

  • Heart (British Cardiac Society) 06/2015; 101(Suppl 4):A126-A126. DOI:10.1136/heartjnl-2015-308066.232 · 6.02 Impact Factor
  • [Show abstract] [Hide abstract]
    ABSTRACT: sec> Introduction TTN truncating variants (TTNtv) cause severe dilated cardiomyopathy (DCM), but sometimes occur in healthy individuals, posing significant challenges for the interpretation of these variants in an era of accessible genome sequencing. The mechanism by which TTNtv impact clinical outcomes is poorly understood. Methods Here, we integrated the power of quantitative cardiac MRI and capacity of next generation sequencing to assess the relationship between TTN genotype and cardiac phenotype. We sequenced TTN in 4,440 subjects including 308 healthy volunteers, 3,603 Framingham Heart Study (FHS) and Jackson Heart Study (JHS) participants, 374 prospective, unselected DCM cases and 155 end-stage retrospective DCM cases including 84 for whom left ventricular (LV) tissue was available for RNA and protein studies. Results TTNtv were identified in 1.4% of controls (healthy volunteers, FHS and JHS participants), in 13% of unselected and 22% of end-stage DCM cases (OR 16.6, P = 4.8 × 10–45, DCM vs controls). More than 45% of controls have at least one rare TTN non-synonymous SNP (nsSNP). Rare and novel TTN nsSNPs were not enriched in DCM, either alone or in combination with a TTNtv ( P = 0 .8 (38.85% in DCM vs 38.24% in controls) suggesting that TTN nsSNPs are not an important cause of DCM. To improve TTN transcript annotations, we determined average cardiac TTN exon usage de novo from RNA-sequencing. TTNtv in DCM cases were enriched in highly utilised exons and isoforms (P = 2.5 × 10–4) compared to controls. We estimate that TTNtv in highly utilised exons have >93% probability of pathogenicity (likelihood ratio 14) in DCM cases. TTNtv-positive DCM patients had more depressed LV ejection fraction (LVEF: P = 0.02), thinner LV walls (P < 0.02), and a higher incidence of sustained ventricular tachycardia (P = 0.001). C-terminus TTNtv were associated with lower LVEF vs N-terminus (β=–18 ± 7%, p = 0.006) and were more common in end-stage disease. No change was detected in total TTN mRNA or protein levels in TTNtv-positive hearts. Conclusion TTNtv are the most common cause of DCM. TTN nsSNPs are not an important cause of DCM in the absence of other discriminating features. Incorporation of variant position and exon-specific expression improves interpretation of TTNtv. Most individuals with TTNtv do not develop DCM, but TTNtv in highly utilised, particularly distal exons commonly cause DCM with severely impaired LV function and life-threatening ventricular arrhythmias, likely through dominant-negative mechanisms. In DCM patients, presence and position of TTNtv may aid prognostication and management. Abstract 163 Figure 1 </sec
    Heart (British Cardiac Society) 06/2015; 101(Suppl 4). DOI:10.1136/heartjnl-2015-308066.163 · 6.02 Impact Factor
  • Science translational medicine 01/2015; 7(270). · 14.41 Impact Factor
  • [Show abstract] [Hide abstract]
    ABSTRACT: The recent discovery of heterozygous human mutations that truncate full-length titin (TTN, an abundant structural, sensory, and signaling filament in muscle) as a common cause of end-stage dilated cardiomyopathy (DCM) promises new prospects for improving heart failure management. However, realization of this opportunity has been hindered by the burden of TTN-truncating variants (TTNtv) in the general population and uncertainty about their consequences in health or disease. To elucidate the effects of TTNtv, we coupled TTN gene sequencing with cardiac phenotyping in 5267 individuals across the spectrum of cardiac physiology and integrated these data with RNA and protein analyses of human heart tissues. We report diversity of TTN isoform expression in the heart, define the relative inclusion of TTN exons in different isoforms (using the TTN transcript annotations available at http://cardiodb.org/titin), and demonstrate that these data, coupled with the position of the TTNtv, provide a robust strategy to discriminate pathogenic from benign TTNtv. We show that TTNtv is the most common genetic cause of DCM in ambulant patients in the community, identify clinically important manifestations of TTNtv-positive DCM, and define the penetrance and outcomes of TTNtv in the general population. By integrating genetic, transcriptome, and protein analyses, we provide evidence for a length-dependent mechanism of disease. These data inform diagnostic criteria and management strategies for TTNtv-positive DCM patients and for TTNtv that are identified as incidental findings. Copyright © 2015, American Association for the Advancement of Science.
    Science translational medicine 01/2015; 7(270-270):270ra6. DOI:10.1126/scitranslmed.3010134 · 14.41 Impact Factor
  • Source
    [Show abstract] [Hide abstract]
    ABSTRACT: ShcA (Src Homology and Collagen A) is an adaptor protein that binds to tyrosine kinase receptors. Its germ line deletion is embryonic lethal with abnormal cardiovascular system formation, and its role in cardiovascular development is unknown. To investigate its functional role in cardiovascular development in mice, ShcA was deleted in cardiomyocytes and vascular smooth muscle cells by crossing ShcA flox mice with SM22a-Cre transgenic mice. Conditional mutant mice developed signs of severe dilated cardiomyopathy, myocardial infarctions, and premature death. No evidence of a vascular contribution to the phenotype was observed. Histological analysis of the heart revealed aberrant sarcomeric Z-disk and M-band structures, and misalignments of T-tubules with Z-disks. We find that not only the ErbB3/Neuregulin signaling pathway but also the baroreceptor reflex response, which have been functionally associated, are altered in the mutant mice. We further demonstrate that ShcA interacts with Caveolin-1 and the costameric protein plasma membrane Ca2+/calmodulin-dependent ATPase (PMCA), and that its deletion leads to abnormal dystrophin signaling. Collectively, these results demonstrate that ShcA interacts with crucial proteins and pathways that link Z-disk and costamere. Copyright © 2014, The American Society for Biochemistry and Molecular Biology.
    Journal of Biological Chemistry 12/2014; 290(4). DOI:10.1074/jbc.M114.597377 · 4.60 Impact Factor
  • [Show abstract] [Hide abstract]
    ABSTRACT: Congenital core myopathies (CM) remain genetically unexplained in many cases. Heart disease is not considered part of the typical CM spectrum. Childhood-onset dilated cardiomyopathy (DCM) was reported in two families with minicores and homozygous TTN mutations. TTN encodes the giant sarcomere protein titin. Relatively few titin mutations (mostly heterozygous and associated to adult-onset cardiac or skeletal muscle disease) have been reported, TTN size and complexity precluding full screening before NGS. Thus, the prevalence and spectrum of titinopathies is probably underestimated. Here we report the analysis of 23 families with CM and primary heart disease using TTN M-line targeted sequencing followed in selected patients by whole-exome sequencing and functional studies. We found 7 novel homozygous or compound heterozygous TTN mutations (5 in the M-line, 5 truncating) in 5 patients. All heterozygous parents were healthy. We identified 4 novel phenotypes previously unreported or non-associated with TTN, including several congenital cardiopathies. The first antenatal-onset titinopathy presented with multi-minicore disease (MmD), arthrogryposis, left ventricular non-compaction and ventricular septal defect, and was associated with the first-reported absence of a functional titin kinase domain in humans. Other novel phenotypes associate MmD with auricular and/or ventricular septal defects, Rigid Spine, Emery-Dreifuss phenotype and/or adult-onset DCM. The histological pattern consistently showed minicores, central nuclei and structural lesions. Conversely, the spectrum of severity and heart disease was large. Thus, we demonstrate that TTN M-line truncating mutations are typically recessive and represent a significant cause of MmD with heart disease. Our results represent the first series of recessive titinopathies, expand TTN mutational and phenotypic spectrum, and identify TTN as a candidate gene in arthrogryposis, EDMD and pediatric heart defects.
    Neuromuscular Disorders 10/2014; 24(s 9–10):804–805. DOI:10.1016/j.nmd.2014.06.048 · 3.13 Impact Factor
  • [Show abstract] [Hide abstract]
    ABSTRACT: Cardiac titin is the main determinant of sarcomere stiffness during diastolic relaxation. To explore whether titin stiffness affects the kinetics of cardiac myofibrillar contraction and relaxation, we used subcellular myofibrils from left ventricles of homozygous (KO) and heterozygous (HET) N2B-KO mice which express truncated cardiac titins lacking the unique elastic N2B region. Compared to myofibrils from wildtype (WT) mice, myofibrils from KO and HET mice exhibit increased passive myofibrillar stiffness. To determine the kinetics of Ca(2+)-induced force development (rate constant kACT), myofibrils from KO, HET and WT mice were stretched to the same sarcomere length (2.3 µm) and rapidly Ca(2+) activated. Additionally, mechanically-induced force redevelopment kinetics (rate constant kTR) was determined by slackening and re-stretching myofibrils during Ca(2+) activation. Myofibrils from KO mice exhibited significant higher kACT, kTR and maximum Ca(2+) activated tension than myofibrils from WT. In contrast, the kinetic parameters of biphasic force relaxation induced by rapidly reducing [Ca(2+)] were not significantly different among the three genotypes. These results indicate that increased titin stiffness promotes myocardial contraction by accelerating the formation of force-generating cross-bridges without decelerating relaxation.
    Journal of Cell Science 06/2014; 127(17). DOI:10.1242/jcs.141796 · 5.33 Impact Factor
  • [Show abstract] [Hide abstract]
    ABSTRACT: Mutations in the gene encoding the RNA-binding protein RBM20 have been implicated in dilated cardiomyopathy (DCM), a major cause of chronic heart failure, presumably through altering cardiac RNA splicing. Here, we combined transcriptome-wide crosslinking immunoprecipitation (CLIP-seq), RNA-seq, and quantitative proteomics in cell culture and rat and human hearts to examine how RBM20 regulates alternative splicing in the heart. Our analyses revealed the presence of a distinct RBM20 RNA-recognition element that is predominantly found within intronic binding sites and linked to repression of exon splicing with RBM20 binding near 3' and 5' splice sites. Proteomic analysis determined that RBM20 interacts with both U1 and U2 small nuclear ribonucleic particles (snRNPs) and suggested that RBM20-dependent splicing repression occurs through spliceosome stalling at complex A. Direct RBM20 targets included several genes previously shown to be involved in DCM as well as genes not typically associated with this disease. In failing human hearts, reduced expression of RBM20 affected alternative splicing of several direct targets, indicating that differences in RBM20 expression may affect cardiac function. Together, these findings identify RBM20-regulated targets and provide insight into the pathogenesis of human heart failure.
    Journal of Clinical Investigation 06/2014; 124(8). DOI:10.1172/JCI74523 · 13.77 Impact Factor
  • Source
    [Show abstract] [Hide abstract]
    ABSTRACT: The coxsackievirus and adenovirus receptor (CAR) is a cell contact protein with an important role in virus uptake. Its extracellular immunoglobulin domains mediate the binding to coxsackievirus and adenovirus as well as homophilic and heterophilic interactions between cells. The cytoplasmic tail links CAR to the cytoskeleton and intracellular signaling cascades. In the heart, CAR is crucial for embryonic development, electrophysiology, and coxsackievirus B infection. Noncardiac functions are less well understood, in part due to the lack of suitable animal models. Here, we generated a transgenic mouse that rescued the otherwise embryonic-lethal CAR knockout (KO) phenotype by expressing chicken CAR exclusively in the heart. Using this rescue model, we addressed interspecies differences in coxsackievirus uptake and noncardiac functions of CAR. Survival of the noncardiac CAR KO (ncKO) mouse indicates an essential role for CAR in the developing heart but not in other tissues. In adult animals, cardiac activity was normal, suggesting that chicken CAR can replace the physiological functions of mouse CAR in the cardiomyocyte. However, chicken CAR did not mediate virus entry in vivo, so that hearts expressing chicken instead of mouse CAR were protected from infection and myocarditis. Comparison of sequence homology and modeling of the D1 domain indicate differences between mammalian and chicken CAR that relate to the sites important for virus binding but not those involved in homodimerization. Thus, CAR-directed anticoxsackievirus therapy with only minor adverse effects in noncardiac tissue could be further improved by selectively targeting the virus-host interaction while maintaining cardiac function.
    Journal of Virology 04/2014; 88(13). DOI:10.1128/JVI.00104-14 · 4.65 Impact Factor
  • Acta Physiologica 03/2014; 210:42-42. · 4.25 Impact Factor
  • Source
    [Show abstract] [Hide abstract]
    ABSTRACT: Non-physiological activation of the mineralocorticoid receptor (MR), e.g. by aldosterone under conditions of high salt intake, contributes to the pathogenesis of cardiovascular diseases, although beneficial effects of aldosterone also have been described. The epidermal growth factor receptor (EGFR) contributes to cardiovascular alterations and mediates part of the MR effects. Recently, we showed that EGFR is required for physiological homeostasis and function of heart and arteries in adult animals. We hypothesize that moderate high aldosterone/NaCl, at normal blood pressure, affects the cardiovascular system depending on cardiovascular EGFR. Therefore we performed an experimental series in male and female animals each, using a recently established mouse model with EGFR knockout in vascular smooth muscle cells and cardiomyocytes and determined the effects of a mild-high aldosterone-to-NaCl constellation on a.o. marker gene expression, heart size, systolic blood pressure, impulse conduction and heart rate. Our data show that (i) cardiac tissue of male but not of female mice is sensitive to mild aldosterone/NaCl treatment, (ii) EGFR knockout induces stronger cardiac disturbances in male as compared to female animals and (iii) mild aldosterone/NaCl treatment requires the EGFR in order to disturb cardiac tissue homeostasis whereas beneficial effects of aldosterone seem to be independent of EGFR.
  • [Show abstract] [Hide abstract]
    ABSTRACT: Core myopathies (CM), the main non-dystrophic myopathy in childhood, remain genetically unexplained in many cases. Heart disease is not considered part of the typical CM spectrum. No congenital heart defect has been reported, and childhood-onset cardiomyopathy has been documented in only two CM families with homozygous mutations of the TTN gene. TTN encodes titin, a giant protein of striated muscles. Recently, heterozygous TTN truncating mutations have also been reported as a major cause of dominant dilated cardiomyopathy. However, relatively few TTN mutations and phenotypes are known, and titin pathophysiological role in cardiac and skeletal muscle conditions is incompletely understood.We analyzed a series of 23 families with congenital CM and primary heart disease using TTN M-line targeted sequencing followed in selected patients by whole-exome sequencing and functional studies. We identified 7 novel homozygous or compound-heterozygous TTN mutations (5 in the M-line, 5 truncating) in 17% patients. Heterozygous parents were healthy. Phenotype analysis identified four novel titinopathies, including cardiac septal defects, left ventricular non-compaction, Emery-Dreifuss muscular dystrophy or arthrogryposis. Additionally, in vitro studies documented the first-reported absence of the titin kinase domain in humans, leading to a severe antenatal phenotype.We establish that CM are associated with a large range of heart conditions of which TTN mutations are a major cause, thereby expanding TTN mutational and phenotypic spectrum. Additionally, our results suggest titin kinase implication in cardiac morphogenesis and demonstrate that heterozygous TTN truncating mutations may not manifest unless associated with a second mutation, reassessing the paradigm of their dominant expression.
    Human Molecular Genetics 10/2013; DOI:10.1093/hmg/ddt494 · 6.68 Impact Factor
  • Biophysical Journal 01/2013; 104(2):310-. DOI:10.1016/j.bpj.2012.11.1724 · 3.83 Impact Factor
  • Source
    [Show abstract] [Hide abstract]
    ABSTRACT: Objectives To investigate the modulatory effect of the Coxsackie and adenovirus receptor (CAR) on ventricular conduction and arrhythmia vulnerability in the setting of myocardial ischemia. Background A heritable component in risk for ventricular fibrillation (VF) during myocardial infarction (MI) has been well established. A recent genome-wide association study (GWAS) for VF during acute MI has led to the identification of a locus on chromosome 21q21 (rs2824292) in the vicinity of the CXADR gene. CXADR encodes the coxsackie and adenovirus receptor (CAR), a cell adhesion molecule predominantly located at intercalated discs of the cardiomyocyte. Methods The correlation between CAR transcript levels and rs2824292 genotype was investigated in human left ventricular samples. Electrophysiological studies and molecular analyses were performed CAR haploinsufficient mice (CAR+/-). Results In human left ventricular samples, the risk allele at the chr21q21 GWAS locus was associated with lower CXADR mRNA levels, suggesting that decreased cardiac levels of CAR predispose to ischemia-induced VF. Hearts from CAR+/- mice displayed ventricular conduction slowing in addition to an earlier onset of ventricular arrhythmias during the early phase of acute myocardial ischemia following LAD ligation. Connexin43 expression and distribution was unaffected, but CAR+/- hearts displayed increased arrhythmia susceptibility upon pharmacological electrical uncoupling. Patch-clamp analysis of isolated CAR+/- myocytes showed reduced sodium current magnitude specifically at the intercalated disc. Moreover, CAR co-precipitated with NaV1.5 in vitro, suggesting that CAR affects sodium channel function through a physical interaction with NaV1.5. Conclusion We identify CAR as a novel modifier of ventricular conduction and arrhythmia vulnerability in the setting of myocardial ischemia. Genetic determinants of arrhythmia susceptibility (such as CAR) may constitute future targets for risk stratification of potentially lethal ventricular arrhythmias in patients with coronary artery disease
    Journal of the American College of Cardiology 01/2013; 63(6). DOI:10.1016/j.jacc.2013.10.062 · 15.34 Impact Factor
  • [Show abstract] [Hide abstract]
    ABSTRACT: The epidermal growth factor receptor (EGFR), a receptor tyrosine kinase, contributes to parainflammatory dysregulation, possibly causing cardiovascular dysfunction and remodeling. The physiological role of cardiovascular EGFR is not completely understood. To investigate the physiological importance of EGFR in vascular smooth muscle cells and cardiomyocytes, we generated a mouse model with targeted deletion of the EGFR using the SM22 (smooth muscle-specific protein 22) promoter. While the reproduction of knockout animals was not impaired, life span was significantly reduced. Systolic blood pressure was not different between the 2 genotypes-neither in tail cuff nor in intravascular measurements-whereas total peripheral vascular resistance, diastolic blood pressure, and mean blood pressure were reduced. Loss of vascular smooth muscle cell-EGFR results in a dilated vascular phenotype with minor signs of fibrosis and inflammation. Echocardiography, necropsy, and histology revealed a dramatic eccentric cardiac hypertrophy in knockout mice (2.5-fold increase in heart weight), with increased stroke volume and cardiac output as well as left ventricular wall thickness and lumen. Cardiac hypertrophy is accompanied by an increase in cardiomyocyte volume, a strong tendency to cardiac fibrosis and inflammation, as well as enhanced NADPH-oxidase 4 and hypertrophy marker expression. Thus, in cardiomyocytes, EGFR prevents excessive hypertrophic growth through its impact on reactive oxygen species balance, whereas in vascular smooth muscle cells EGFR contributes to the appropriate vascular wall architecture and vessel reactivity, thereby supporting a physiological vascular tone.
    Hypertension 12/2012; 61(2). DOI:10.1161/HYPERTENSIONAHA.112.196543 · 7.63 Impact Factor
  • [Show abstract] [Hide abstract]
    ABSTRACT: The cellular basis of the Frank-Starling "Law of the Heart" is the length-dependence of activation, but the mechanisms by which the sarcomere detects length changes and converts this information to altered calcium sensitivity has remained elusive. Here the effect of titin-based passive tension on the length-dependence of activation (LDA) was studied by measuring the tension-pCa relation in skinned mouse LV muscle at two sarcomere lengths (SLs). N2B KO myocardium, where the N2B spring element in titin is deleted and passive tension is elevated, was compared to WT myocardium. Myofilament lattice structure was studied with low-angle X-ray diffraction; the myofilament lattice spacing (d(10)) was measured as well as the ratio of the intensities of the 1,1 and 1,0 diffraction peaks (I(11)/I(10)) as an estimate of the degree of association of myosin heads with the thin filaments. Experiments were carried out in skinned muscle in which the lattice spacing was reduced with Dextran-T500. Experiments with and without lattice compression were also carried out following PKA phosphorylation of the skinned muscle. Under all conditions that were tested, LDA was significantly larger in N2B KO myocardium compared to WT myocardium, with the largest differences following PKA phosphorylation. A positive correlation between passive tension and LDA was found that persisted when the myofilament lattice was compressed with Dextran and that was enhanced following PKA phosphorylation. Low-angle X-ray diffraction revealed a shift in mass from thin filaments to thick filaments as sarcomere length was increased. Furthermore, a positive correlation was obtained between myofilament lattice spacing and passive tension and the change in I(11)/I(10) and passive tension and these provide possible explanations for how titin-based passive tension might regulate calcium sensitivity.
    Archives of Biochemistry and Biophysics 12/2012; DOI:10.1016/j.abb.2012.12.004 · 3.04 Impact Factor
  • Source
    [Show abstract] [Hide abstract]
    ABSTRACT: Alternative splicing has a major role in cardiac adaptive responses, as exemplified by the isoform switch of the sarcomeric protein titin, which adjusts ventricular filling. By positional cloning using a previously characterized rat strain with altered titin mRNA splicing, we identified a loss-of-function mutation in the gene encoding RNA binding motif protein 20 (Rbm20) as the underlying cause of pathological titin isoform expression. The phenotype of Rbm20-deficient rats resembled the pathology seen in individuals with dilated cardiomyopathy caused by RBM20 mutations. Deep sequencing of the human and rat cardiac transcriptome revealed an RBM20-dependent regulation of alternative splicing. In addition to titin (TTN), we identified a set of 30 genes with conserved splicing regulation between humans and rats. This network is enriched for genes that have previously been linked to cardiomyopathy, ion homeostasis and sarcomere biology. Our studies emphasize the key role of post-transcriptional regulation in cardiac function and provide mechanistic insights into the pathogenesis of human heart failure.
    Nature medicine 04/2012; 18(5):766-73. DOI:10.1038/nm.2693 · 28.05 Impact Factor
  • Source
    [Show abstract] [Hide abstract]
    ABSTRACT: Titin (also known as connectin) is an intrasarcomeric muscle protein that functions as a molecular spring and generates passive tension upon muscle stretch. The N2B element is a cardiac-specific spring element within titin's extensible region. Our goal was to study the contribution of the N2B element to the mechanical properties of titin, particularly its hypothesized role in limiting energy loss during repeated stretch (diastole)-shortening (systole) cycles of the heart. We studied energy loss by measuring hysteresis from the area between the stretch and release passive force-sarcomere length curves and used both wild-type (WT) mice and N2B knockout (KO) mice in which the N2B element has been deleted. A range of protocols was used, including those that mimic physiological loading conditions. KO mice showed significant increases in hysteresis. Most prominently, in tissue that had been preconditioned with a physiological stretch-release protocol, hysteresis increased significantly from 320 ± 46 pJ/mm(2)/sarcomere in WT to 650 ± 94 pJ/mm(2)/sarcomere in N2B KO myocardium. These results are supported by experiments in which oxidative stress was used to mechanically inactivate portions of the N2B-Us of WT titin through cysteine cross-linking. Studies on muscle from which the thin filaments had been extracted (using the actin severing protein gelsolin) showed that the difference in hysteresis between WT and KO tissue cannot be explained by filament sliding-based viscosity. Instead the results suggest that hysteresis arises from within titin and most likely involves unfolding of immunoglobulin-like domains. These studies support that the mechanical function of the N2B element of titin includes reducing hysteresis and increasing the efficiency of the heart.
    Biophysical Journal 09/2011; 101(6):1385-92. DOI:10.1016/j.bpj.2011.06.054 · 3.83 Impact Factor
  • [Show abstract] [Hide abstract]
    ABSTRACT: Pathophysiological effects of the epidermal growth factor receptor (EGFR or ErbB1) include vascular remodeling. EGFR transactivation is proposed to contribute significantly to heterologous signaling and remodeling in vascular smooth muscle cells (VSMC). We investigated the importance of EGFR in primary VSMC from aorta of mice with targeted deletion of the EGFR (EGFR(Δ/Δ VSMC)→VSMC(EGFR-/-) and EGFR(Δ/+ VSMC)→VSMC(EGFR+/-)) and the respective littermate controls (EGFR(+/+ VSMC)→VSMC(EGFR+/+)) with respect to survival, pentose phosphate pathway activity, matrix homeostasis, extracellular signal-regulated kinase 1/2 (ERK1/2) phosphorylation, and Ca(2+) homeostasis. In VSMC(EGFR-/-), epidermal growth factor-induced signaling was abolished; VSMC(EGFR+/-) showed an intermediate phenotype. EGFR deletion enhanced spontaneous cell death, reduced pentose phosphate pathway activity, disturbed cellular matrix homeostasis (collagen III and fibronectin), and abolished epidermal growth factor sensitivity. In VSMC(EGFR-/-) endothelin-1- or α(1)-adrenoceptor-induced ERK1/2 phosphorylation and the fraction of Ca(2+) responders were significantly reduced, whereas responsive cells showed a significantly stronger Ca(2+) signal. Oxidative stress (H(2)O(2)) induced ERK1/2 activation in VSMC(EGFR+/+) and VSMC(EGFR+/-) but not in VSMC(EGFR-/-). The Ca(2+) signal was enhanced in VSMC(EGFR-/-), similar to purinergic stimulation by ATP. In conclusion, EGFR was found to be important for basal VSMC homeostasis and ERK1/2 activation by the tested G-protein-coupled receptors or radical stress. Ca(2+) signaling was modulated by EGFR differentially with respect to the fraction of responders and magnitude of the signal. Thus, EGFR seems to be Janus-faced for VSMC biology.
    Arteriosclerosis Thrombosis and Vascular Biology 07/2011; 31(7):1643-52. DOI:10.1161/ATVBAHA.111.223537 · 5.53 Impact Factor
  • Source
    [Show abstract] [Hide abstract]
    ABSTRACT: Viscosity is proposed to modulate diastolic function, but only limited understanding of the source(s) of viscosity exists. In vitro experiments have shown that the proline-glutamic acid-valine-lysine (PEVK) rich element of titin interacts with actin, causing a viscous force in the sarcomere. It is unknown whether this mechanism contributes to viscosity in vivo. We tested the hypothesis that PEVK-actin interaction causes cardiac viscosity and is important in vivo via an integrative physiological study on a unique PEVK knockout (KO) model. Both skinned cardiomyocytes and papillary muscle fibers were isolated from wildtype (WT) and PEVK KO mice and passive viscosity was examined using stretch-hold-release and sinusoidal analysis. Viscosity was reduced by ~60% in KO myocytes and ~50% in muscle fibers at room temperature. The PEVK-actin interaction was not modulated by temperature or diastolic calcium, but was increased by lattice compression. Stretch-hold and sinusoidal frequency protocols on intact isolated mouse hearts showed a smaller, 30-40% reduction in viscosity, possibly due to actomyosin interactions, and showed that microtubules did not contribute to viscosity. Transmitral Doppler echocardiography similarly revealed a 40% decrease in LV chamber viscosity in the PEVK KO in vivo. This integrative study is the first to quantify the influence of a specific molecular (PEVK-actin) viscosity in vivo and shows that PEVK-actin interactions are an important physiological source of viscosity.
    Journal of Molecular and Cellular Cardiology 06/2011; 51(3):428-34. DOI:10.1016/j.yjmcc.2011.06.006 · 5.22 Impact Factor

Publication Stats

4k Citations
519.48 Total Impact Points

Institutions

  • 2005–2015
    • Max-Delbrück-Centrum für Molekulare Medizin
      • Research Team Neuromuscular and Cardiovascular Cell Biology
      Berlín, Berlin, Germany
  • 2014
    • Universitätsklinikum Tübingen
      • Division of Molecular Pathology
      Tübingen, Baden-Württemberg, Germany
  • 2009–2013
    • Max Planck Institute of Molecular Cell Biology and Genetics
      Dresden, Saxony, Germany
    • The University of Arizona
      • Department of Physiology
      Tucson, AZ, United States
  • 2012
    • University of Wisconsin, Madison
      • Muscle Biology Laboratory
      Madison, MS, United States
  • 2003–2011
    • Washington State University
      • Department of Veterinary and Comparative Anatomy, Pharmacology and Physiology (VCAPP)
      پولمن، واشینگتن, Washington, United States
    • University of Texas at Dallas
      • Biochemistry
      Richardson, Texas, United States
  • 2007
    • Universität Mannheim
      Mannheim, Baden-Württemberg, Germany
  • 2004
    • University of Strasbourg
      Strasburg, Alsace, France
  • 1996–2002
    • University of Texas Southwestern Medical Center
      • Department of Molecular Genetics
      Dallas, Texas, United States
  • 1998
    • Howard Hughes Medical Institute
      Ashburn, Virginia, United States