The tight junction protein CAR regulates cardiac conduction and cell–cell communication

The Coxsackievirus-adenovirus receptor (CAR) is known for its role in virus uptake and as a protein of the tight junction. It is predominantly expressed in the developing brain and heart and reinduced upon cardiac remodeling in heart disease. So far, the physiological functions of CAR in the adult heart are largely unknown. We have generated a heart-specific inducible CAR knockout (KO) and found impaired electrical conduction between atrium and ventricle that increased with progressive loss of CAR. The underlying mechanism relates to the cross talk of tight and gap junctions with altered expression and localization of connexins that affect communication between CAR KO cardiomyocytes. Our results indicate that CAR is not only relevant for virus uptake and cardiac remodeling but also has a previously unknown function in the propagation of excitation from the atrium to the ventricle that could explain the association of arrhythmia and Coxsackievirus infection of the heart.

[1]  Jamie I Vandenberg,et al.  Slowed conduction and ventricular tachycardia after targeted disruption of the cardiac sodium channel gene Scn5a , 2002, Proceedings of the National Academy of Sciences of the United States of America.

[2]  S. Butz,et al.  Coxsackievirus-adenovirus receptor (CAR) is essential for early embryonic cardiac development , 2005, Journal of Cell Science.

[3]  J E Saffitz,et al.  Connexin expression and turnover : implications for cardiac excitability. , 2000, Circulation research.

[4]  J. Bergelson,et al.  Isolation of a Common Receptor for Coxsackie B Viruses and Adenoviruses 2 and 5 , 1997, Science.

[5]  K. Otsu,et al.  Direct Association of the Gap Junction Protein Connexin-43 with ZO-1 in Cardiac Myocytes* , 1998, The Journal of Biological Chemistry.

[6]  J. Seidman,et al.  The T-Box transcription factor Tbx5 is required for the patterning and maturation of the murine cardiac conduction system , 2004, Development.

[7]  Yiming Wu,et al.  Targeted deletion of titin N2B region leads to diastolic dysfunction and cardiac atrophy , 2007, Proceedings of the National Academy of Sciences.

[8]  Yuan-Tsong Chen,et al.  ENU mutagenesis identifies mice with cardiac fibrosis and hepatic steatosis caused by a mutation in the mitochondrial trifunctional protein beta-subunit. , 2006, Human molecular genetics.

[9]  D. Roden,et al.  Nkx2-5 mutation causes anatomic hypoplasia of the cardiac conduction system. , 2004, The Journal of clinical investigation.

[10]  D. Gros,et al.  Mouse Model of SCN5A-Linked Hereditary Lenègre’s Disease: Age-Related Conduction Slowing and Myocardial Fibrosis , 2005, Circulation.

[11]  K. Willecke,et al.  Connexin-mediated cardiac impulse propagation: connexin 30.2 slows atrioventricular conduction in mouse heart. , 2006, Trends in cardiovascular medicine.

[12]  Colin McKerlie,et al.  A Gja1 missense mutation in a mouse model of oculodentodigital dysplasia , 2005, Development.

[13]  G. Lemke,et al.  A critical role for the EphA3 receptor tyrosine kinase in heart development. , 2007, Developmental biology.

[14]  L. Philipson,et al.  The Coxsackievirus and Adenovirus Receptor (CAR) Forms a Complex with the PDZ Domain-containing Protein Ligand-of-Numb Protein-X (LNX)* , 2003, The Journal of Biological Chemistry.

[15]  A. Matsumori,et al.  Electrocardiographic findings in experimental myocarditis in DBA/2 mice: complete atrioventricular block in the acute stage, low voltage of the QRS complex in the subacute stage and arrhythmias in the chronic stage. , 1984, Journal of the American College of Cardiology.

[16]  B. Erdmann,et al.  M line–deficient titin causes cardiac lethality through impaired maturation of the sarcomere , 2006, The Journal of cell biology.

[17]  G. Fishman,et al.  Electrical remodeling contributes to complex tachyarrhythmias in connexin43‐deficient mouse hearts , 2008, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[18]  P. Carmeliet,et al.  Overlap Syndrome of Cardiac Sodium Channel Disease in Mice Carrying the Equivalent Mutation of Human SCN5A-1795insD , 2006, Circulation.

[19]  B L Langille,et al.  Cardiac malformation in neonatal mice lacking connexin43. , 1995, Science.

[20]  L. Philipson,et al.  HCAR and MCAR: the human and mouse cellular receptors for subgroup C adenoviruses and group B coxsackieviruses. , 1997, Proceedings of the National Academy of Sciences of the United States of America.

[21]  K. Nishii,et al.  Mice Lacking Connexin45 Conditionally in Cardiac Myocytes Display Embryonic Lethality Similar to That of Germline Knockout Mice Without Endocardial Cushion Defect , 2003, Cell communication & adhesion.

[22]  Christian C Witt,et al.  Conditional Expression of Mutant M-line Titins Results in Cardiomyopathy with Altered Sarcomere Structure* , 2003, The Journal of Biological Chemistry.

[23]  J. Seidman,et al.  Electrophysiological abnormalities and arrhythmias in alpha MHC mutant familial hypertrophic cardiomyopathy mice. , 1997, The Journal of clinical investigation.

[24]  H. Musa,et al.  Connexin43 Remodeling Caused by Inhibition of Plakophilin-2 Expression in Cardiac Cells , 2007, Circulation research.

[25]  I. Larre,et al.  New diseases derived or associated with the tight junction. , 2007, Archives of medical research.

[26]  M. Franz,et al.  Targeted disruption of the Kcnq1 gene produces a mouse model of Jervell and Lange– Nielsen Syndrome , 2001, Proceedings of the National Academy of Sciences of the United States of America.

[27]  W. Edwards,et al.  Pathology of the cardiac conduction system in myotonic dystrophy: a study of 12 cases. , 1988, Journal of the American College of Cardiology.

[28]  Yiming Wu,et al.  Cardiac Hypertrophy and Reduced Contractility in Hearts Deficient in the Titin Kinase Region , 2007, Circulation.

[29]  D. Kass,et al.  Sudden cardiac death in heart failure. The role of abnormal repolarization. , 1994, Circulation.

[30]  Stephen N. Jones,et al.  Coxsackievirus and adenovirus receptor is essential for cardiomyocyte development , 2005, Genesis.

[31]  M. Welsh,et al.  Adenovirus Fiber Disrupts CAR-Mediated Intercellular Adhesion Allowing Virus Escape , 2002, Cell.

[32]  T. Magnuson,et al.  Mice mutant for Egfr and Shp2 have defective cardiac semilunar valvulogenesis , 2000, Nature Genetics.

[33]  E. Stanley,et al.  Cardiac Septal and Valvular Dysmorphogenesis in Mice Heterozygous for Mutations in the Homeobox Gene Nkx2-5 , 2000, Circulation research.

[34]  K. Willecke,et al.  Reduced cardiac conduction velocity and predisposition to arrhythmias in connexin40-deficient mice , 1998, Current Biology.

[35]  S. Varnous,et al.  Mouse model carrying H222P-Lmna mutation develops muscular dystrophy and dilated cardiomyopathy similar to human striated muscle laminopathies. , 2005, Human molecular genetics.

[36]  Misao Suzuki,et al.  Loss of connexin45 causes a cushion defect in early cardiogenesis. , 2000, Development.

[37]  Michael D. Schneider,et al.  Cardiomyocyte-Specific Deletion of the Coxsackievirus and Adenovirus Receptor Results in Hyperplasia of the Embryonic Left Ventricle and Abnormalities of Sinuatrial Valves , 2006, Circulation research.

[38]  E. Schenk,et al.  HISTOCHEMICAL METHODS FOR SEPARATE, CONSECUTIVE AND SIMULTANEOUS DEMONSTRATION OF ACETYLCHOLINESTERASE AND NOREPINEPHRINE IN CRYOSTAT SECTIONS , 1967, The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society.

[39]  B. Persson,et al.  Dye‐coupling between term pregnant human myometrial cells before labor: Carboxyfluorescein versus lucifer yellow , 1995, Cell biology international.

[40]  Tsuyoshi Saito,et al.  Possible involvement of gap junctions in the barrier function of tight junctions of brain and lung endothelial cells , 2006, Journal of cellular physiology.

[41]  H. Yagi,et al.  Complete atrioventricular block associated with regional myocardial scarring in a patient with Coxsackie B2 myocarditis. , 1989, Japanese heart journal.

[42]  K. Alitalo,et al.  Vascular Endothelial Growth Factor-B–Deficient Mice Display an Atrial Conduction Defect , 2001, Circulation.

[43]  N. Peters.  Gap junctions: clarifying the complexities of connexins and conduction. , 2006, Circulation research.

[44]  J. Bergelson,et al.  The Coxsackievirus and Adenovirus Receptor Interacts with the Multi-PDZ Domain Protein-1 (MUPP-1) within the Tight Junction* , 2004, Journal of Biological Chemistry.

[45]  Kazuko Koshiba-Takeuchi,et al.  Tbx5-dependent rheostatic control of cardiac gene expression and morphogenesis. , 2006, Developmental biology.

[46]  Paul J. Wang,et al.  Assessment of Atrioventricular Nodal Physiology in the Mouse , 1999, Journal of Interventional Cardiac Electrophysiology.

[47]  G. Morley,et al.  Cardiac-Specific Loss of N-Cadherin Leads to Alteration in Connexins With Conduction Slowing and Arrhythmogenesis , 2005, Circulation research.

[48]  J. Bergelson,et al.  The coxsackievirus and adenovirus receptor is a transmembrane component of the tight junction , 2001, Proceedings of the National Academy of Sciences of the United States of America.

[49]  J. Nam,et al.  Internalization and trafficking mechanisms of coxsackievirus B3 in HeLa cells. , 2005, Virology.

[50]  G. Barsh,et al.  Overexpression of angiotensin AT1 receptor transgene in the mouse myocardium produces a lethal phenotype associated with myocyte hyperplasia and heart block. , 1997, Proceedings of the National Academy of Sciences of the United States of America.

[51]  M. Van Ranst,et al.  Identification of alternative splice products encoded by the human coxsackie-adenovirus receptor gene. , 2001, Biochemical and biophysical research communications.

[52]  E. Herlenius,et al.  Ro/SSA autoantibodies directly bind cardiomyocytes, disturb calcium homeostasis, and mediate congenital heart block , 2005, The Journal of experimental medicine.

[53]  G. Tomaselli,et al.  Sudden Cardiac Death in Heart Failure , 1998 .

[54]  David L. Paul,et al.  Mice lacking connexin40 have cardiac conduction abnormalities characteristic of atrioventricular block and bundle branch block , 1998, Current Biology.

[55]  J. Engel,et al.  Congenital Deafness and Sinoatrial Node Dysfunction in Mice Lacking Class D L-Type Ca2+ Channels , 2000, Cell.

[56]  G. Salama,et al.  Transcription Enhancer Factor-1–Related Factor–Transgenic Mice Develop Cardiac Conduction Defects Associated With Altered Connexin Phosphorylation , 2004, Circulation.

[57]  A. Dörner,et al.  Alternatively Spliced Soluble Coxsackie-adenovirus Receptors Inhibit Coxsackievirus Infection* , 2004, Journal of Biological Chemistry.

[58]  D. Roden,et al.  Nkx 2-5 mutation causes anatomic hypoplasia of the cardiac conduction system , 2022 .

[59]  B. A. French,et al.  Gene recombination in postmitotic cells. Targeted expression of Cre recombinase provokes cardiac-restricted, site-specific rearrangement in adult ventricular muscle in vivo. , 1997, The Journal of clinical investigation.

[60]  O. Berenfeld,et al.  Altered Right Atrial Excitation and Propagation in Connexin40 Knockout Mice , 2005, Circulation.

[61]  I. Efimov,et al.  Cx43 and Dual-Pathway Electrophysiology of the Atrioventricular Node and Atrioventricular Nodal Reentry , 2003, Circulation research.