Component Of Intercalated Disc Research Articles

Animal Models and Molecular Pathogenesis of Arrhythmogenic Cardiomyopathy Associated with Pathogenic Variants in Intercalated Disc Genes.

Arrhythmogenic cardiomyopathy (ACM) is a rare genetic cardiac disease characterized by the progressive substitution of myocardium with fibro-fatty tissue. Clinically, ACM shows wide variability among patients; symptoms can include syncope and ventricular tachycardia but also sudden death, with the latter often being its sole manifestation. Approximately half of ACM patients have been found with variations in one or more genes encoding cardiac intercalated discs proteins; the most involved genes are plakophilin 2 (PKP2), desmoglein 2 (DSG2), and desmoplakin (DSP). Cardiac intercalated discs provide mechanical and electro-metabolic coupling among cardiomyocytes. Mechanical communication is guaranteed by the interaction of proteins of desmosomes and adheren junctions in the so-called area composita, whereas electro-metabolic coupling between adjacent cardiac cells depends on gap junctions. Although ACM has been first described almost thirty years ago, the pathogenic mechanism(s) leading to its development are still only partially known. Several studies with different animal models point to the involvement of the Wnt/β-catenin signaling in combination with the Hippo pathway. Here, we present an overview about the existing murine models of ACM harboring variants in intercalated disc components with a particular focus on the underlying pathogenic mechanisms. Prospectively, mechanistic insights into the disease pathogenesis will lead to the development of effective targeted therapies for ACM.

Retrofitting the Heart: Explaining the Enigmatic Septal Thickening in Hypertrophic Cardiomyopathy.

Hypertrophic cardiomyopathy is the most common genetic cardiac disease and is characterized by left ventricular hypertrophy. Although this hypertrophy often associates with sarcomeric gene mutations, nongenetic factors also contribute to the disease, leading to diastolic dysfunction. Notably, this dysfunction manifests before hypertrophy and is linked to hypercontractility, as well as nonuniform contraction and relaxation (myofibril asynchrony) of the myocardium. Although the distribution of hypertrophy in hypertrophic cardiomyopathy can vary both between and within individuals, in most cases, it is primarily confined to the interventricular septum. The reasons for septal thickening remain largely unknown. In this article, we propose that alterations in muscle fiber geometry, present from birth, dictate the septal shape. When combined with hypercontractility and exacerbated by left ventricular outflow tract obstruction, these factors predispose the septum to an isometric type of contraction during systole, consequently constraining its mobility. This contraction, or more accurately, this focal increase in biomechanical stress, prompts the septum to adapt and undergo remodeling. Drawing a parallel, this is reminiscent of how earthquake-resistant buildings are retrofitted with vibration dampers to absorb the majority of the shock motion and load. Similarly, the heart adapts by synthesizing viscoelastic elements such as microtubules, titin, desmin, collagen, and intercalated disc components. This pronounced remodeling in the cytoskeletal structure leads to noticeable septal hypertrophy. This structural adaptation acts as a protective measure against damage by attenuating myofibril shortening while reducing cavity tension according to Laplace Law. By examining these events, we provide a coherent explanation for the septum's predisposition toward hypertrophy.

The intercalated disc: a unique organelle for electromechanical synchrony in cardiomyocytes.

The intercalated disc (ID) is a highly specialized structure that connects cardiomyocytes via mechanical and electrical junctions. Although described in some detail by light microscopy in the 19th century, it was in 1966 that electron microscopy images showed that the ID represented apposing cell borders and provided detailed insight into the complex ID nanostructure. Since then, much has been learned about the ID and its molecular composition, and it has become evident that a large number of proteins, not all of them involved in direct cell-to-cell coupling via mechanical or gap junctions, reside at the ID. Furthermore, an increasing number of functional interactions between ID components are emerging, leading to the concept that the ID is not the sum of isolated molecular silos but an interacting molecular complex, an "organelle" where components work in concert to bring about electrical and mechanical synchrony. The aim of the present review is to give a short historical account of the ID's discovery and an updated overview of its composition and organization, followed by a discussion of the physiological implications of the ID architecture and the local intermolecular interactions. The latter will focus on both the importance of normal conduction of cardiac action potentials as well as the impact on the pathophysiology of arrhythmias.

Intercalated disc protein Xin\u03b2 is required for Hippo-YAP signaling in the heart

Intercalated discs (ICD), specific cell-to-cell contacts that connect adjacent cardiomyocytes, ensure mechanical and electrochemical coupling during contraction of the heart. Mutations in genes encoding ICD components are linked to cardiovascular diseases. Here, we show that loss of Xinβ, a newly-identified component of ICDs, results in cardiomyocyte proliferation defects and cardiomyopathy. We uncovered a role for Xinβ in signaling via the Hippo-YAP pathway by recruiting NF2 to the ICD to modulate cardiac function. In Xinβ mutant hearts levels of phosphorylated NF2 are substantially reduced, suggesting an impairment of Hippo-YAP signaling. Cardiac-specific overexpression of YAP rescues cardiac defects in Xinβ knock-out mice—indicating a functional and genetic interaction between Xinβ and YAP. Our study reveals a molecular mechanism by which cardiac-expressed intercalated disc protein Xinβ modulates Hippo-YAP signaling to control heart development and cardiac function in a tissue specific manner. Consequently, this pathway may represent a therapeutic target for the treatment of cardiovascular diseases.

Critical Roles of Xirp Proteins in Cardiac Conduction and Their Rare Variants Identified in Sudden Unexplained Nocturnal Death Syndrome and Brugada Syndrome in Chinese Han Population.

BackgroundSudden unexplained nocturnal death syndrome (SUNDS) remains an autopsy negative entity with unclear etiology. Arrhythmia has been implicated in SUNDS. Mutations/deficiencies in intercalated disc components have been shown to cause arrhythmias. Human cardiomyopathy‐associated 1 (XIRP1) and 3 (XIRP2) are intercalated disc–associated, Xin repeats‐containing proteins. Mouse Xirp1 is necessary for the integrity of intercalated disc and for the surface expression of transient outward and delayed rectifier K+ channels, whereas mouse Xirp2 is required for Xirp1 intercalated disc localization. Thus, XIRP1 and XIRP2 may be potentially causal genes for SUNDS.Methods and ResultsWe genetically screened XIRP genes in 134 sporadic SUNDS victims and 22 Brugada syndrome (BrS) cases in a Chinese Han population. We identified 16 rare variants (6 were in silico predicted as deleterious) in SUNDS victims, including a novel variant, XIRP2‐E215K. There were also four rare variants (2 were in silico predicted as deleterious) detected in BrS cases, including a novel variant, XIRP2‐L2718P. Interestingly, among these 20 variants, we detected 2 likely pathogenic variants: a nonsense variant (XIRP2‐Q2875*) and a frameshift variant (XIRP2‐T2238QfsX7). Analyzing available Xirp2 knockout mice, we further found that mouse hearts without Xirp2 exhibited prolonged PR and QT intervals, slow conduction velocity, atrioventricular conduction block, and an abnormal infranodal ventricular conduction system. Whole‐cell patch‐clamp detected altered ionic currents in Xirp2 −/− cardiomyocytes, consistent with the observed association between Xirp2 and Nav1.5/Kv1.5 in co‐immunoprecipitation.ConclusionsThis is the first report identifying likely pathogenic XIRP rare variants in arrhythmogenic disorders such as SUNDS and Brugada syndrome, and showing critical roles of Xirp2 in cardiac conduction.

Intercalated disc in failing hearts from patients with dilated cardiomyopathy: Its role in the depressed left ventricular function.

Alterations in myocardial structure and reduced cardiomyocyte adhesions have been previously described in dilated cardiomyopathy (DCM). We studied the transcriptome of cell adhesion molecules in these patients and their relationships with left ventricular (LV) function decay. We also visualized the intercalated disc (ID) structure and organization. The transcriptomic profile of 23 explanted LV samples was analyzed using RNA-sequencing (13 DCM, 10 control [CNT]), focusing on cell adhesion genes. Electron microscopy analysis to visualize ID structural differences and immunohistochemistry experiments of ID proteins was also performed. RT-qPCR and western blot experiments were carried out on ID components. We found 29 differentially expressed genes, most of all, constituents of the ID structure. We found that the expression of GJA3, DSP and CTNNA3 was directly associated with LV ejection fraction (r = 0.741, P = 0.004; r = 0.674, P = 0.011 and r = 0.565, P = 0.044, respectively), LV systolic (P = 0.003, P = 0.003, P = 0.028, respectively) and diastolic dimensions (P = 0.006, P = 0.001, P = 0.025, respectively). Electron microscopy micrographs showed a reduced ID convolution index and immunogold labeling of connexin 46 (GJA gene), desmoplakin (DSP gene) and catenin α-3 (CTNNA3 gene) proteins in DCM patients. Moreover, we observed that protein and mRNA levels analyzed by RT-qPCR of these ID components were diminished in DCM group. In conclusion, we report significant gene and protein expression changes and found that the ID components GJA3, DSP and CTNNA3 were highly related to LV function. Microscopic observations indicated that ID is structurally compromised in these patients. These findings give new data for understanding the ventricular depression that characterizes DCM, opening new therapeutic perspectives for these critically diseased patients.

Refining the molecular organization of the cardiac intercalated disc.

This review presents an extensively integrated model of the cardiac intercalated disc (ID), a highly orchestrated structure that connects adjacent cardiomyocytes. Classically, three main structures are distinguished: gap junctions (GJs) metabolically and electrically connect cytoplasm of adjacent cardiomyocytes; adherens junctions (AJs) connect the actin cytoskeleton of adjacent cells; and desmosomes function as cell anchors and connect intermediate filaments. Furthermore, ion channels reside in the ID. Mutations in ID proteins have been associated with cardiac arrhythmias such as Brugada syndrome and arrhythmogenic cardiomyopathy. However, rather than being independent, all ID components work together intensively by multifunctional proteins such as ZO-1, Ankyrin G, and β-catenin, integrating mechanical and electrical functions. GJs form a plaque surrounded by the perinexus in which free connexons reside; the connexome integrates NaV channels, the desmosome and GJs; and the area composita hosts AJs and desmosomes, also integrated as adhering junctions. Furthermore, the transitional junction connects sarcomeres to the plasma membrane. Lastly, this review integrates all these findings in comprehensible figures, illustrating the interdependencies of ID proteins.

Intercalated discs and arrhythmogenic cardiomyopathy.

Heart tissue is subjected to high mechanical stress. Different junctional complexes exist within the intercalated disc (ID) at the site of end-to-end contacts between cardiomyocytes. These junctions are essential for adhesive integrity, morphogenesis, differentiation, and maintenance of cardiac tissue. Recent findings of molecular interactions among intercellular adhesion molecules, gap junctions, and the voltage-gated sodium channel complex suggest that IDs should be considered an organelle in which macromolecular complexes interact specifically to maintain cardiac structure and cardiomyocyte synchrony. It is within this organelle that most of the mutated proteins involved in arrhythmogenic right ventricular cardiomyopathy (ARVC) reside. This inherited cardiomyopathy is characterized by both structural and electric abnormalities of the heart, particularly in young people and athletes. This review highlights recent advances in understanding the link between ID alterations and the molecular genetics and pathogenesis of ARVC. Cardiomyocytes are extensively interconnected at their ends through their IDs, a complex region composed of different kinds of intercellular junctions essential for electric, mechanical, and signaling communication between adjacent cells and, hence, for maintaining correct heart function and growth. Although traditionally depicted as a composition of different separate units, recent data indicate that the ID of cardiomyocytes should be considered a single functional unit in which macromolecular complexes interact mechanically and electrically to maintain cardiomyocyte rigidity and synchrony. The ID in vertebrates was originally described as consisting of 3 main junctional complexes: desmosomes, adherens junctions (AJ, also called fascia adherens in cardiac muscle), and gap junctions. It has been proposed that while gap junctions, being essential for chemical and electric coupling of neighboring cells, represent the electric component of ID, desmosomes together with AJ form the mechanical intercellular junctions in cardiomyocytes.1 Desmosomes and AJ are highly specialized anchoring junctions. They are particularly important for maintenance of adhesion and integrity of tissues exposed to …

Abstract 421: Alterations in Adherens Junction and Gap Junction Precede Desmosomes Remodeling During the Transition from Experimental Compensated Cardiac Hypertrophy to Decompensation

Hypertension causes cardiac hypertrophy, cardiac dysfunction, and heart failure (HF). The mechanisms implicated in the transition from physiological to pathological hypertrophy are not fully understood. Growing evidence points out to the role of inappropriate intercellular mechanical and electrical coupling in the pathogenesis of HF. We investigated adherens junction, desmosome and gap junction proteins and their potential role in the pathogenesis of HF in rats submitted to chronic abdominal aorta stenosis. Wistar rats were submitted to abdominal aorta stenosis and killed at 90 days post surgery: 60% presented hypertrophied hearts (HH) and 40% hypertrophied and dilated hearts (DH), as evaluated by echocardiography. Blood pressure was evaluated. The hearts were collected, Western blot (WB) and immunofluorescence (IF) were performed for cadherin, plakoglobin, B-catenin, desmoglein, desmocollin and connexin-43. Cardiac systolic function was evaluated using Vevo 2100 ultrasound system. Data were considered significant when p<0.05. Cadherin expression increased 55% in HH and decreased 33% in DH; plakoglobin increased 67% in HH and decreased 29% in DH; B-catenin increased 73% in HH and decreased 32% in DH; connexin-43 decreased 17% in HH and 32% in DH; desmocolin increased 57% only in HH group and no alteration in desmoglein expression was observed in both groups. Mean blood pressure increased 61% in HH and 59% in DH. Ejection fraction and fractional shortening decreased 45% and 41%, respectively, only in DH. These results suggest different sensitivity of the intercalated disc components: proteins of the adherens junction and gap junction are more sensitive than desmosomes proteins and present significant reduced expression. This reduced expression could contribute to the transition of the compensated cardiac hypertrophy to decompensation. These abnormal parameters may emerge as therapeutic targets to modulate the evolution of HF.

Heart rhythm genomic fabric in hypoxia

The molecular mechanisms by which chronic hypoxia, whether constant (CCH) or intermittent (CIH), alters the heart rhythm are still under debate. Expression level, control, maturational profile and intercoordination of 54 genes encoding heart rhythm determinants (HRDs) were analyzed in 36 mice subjected for 1, 2 or 4 weeks of their early life to normal atmospheric conditions or to CCH or CIH. Our analysis revealed a complex network of genes encoding various heart rate, inotropy and development controllers, receptors, ion channels and transporters, ankyrins, epigenetic modulators and intercalated disc components (adherens, cadherins, catenins, desmosomal, gap and tight junction proteins). The network is remodeled during maturation and substantially and differently altered by CIH and CCH. Gene Prominence Analysis that ranks the genes according to their expression stability and networking within functional gene webs, confirmed the HRD status of certain epigenetic modulators and components of the intercalated discs not yet associated with arrhythmia.

Emerin and the Nuclear Lamina in Muscle and Cardiac Disease

The human genome is contained within the nucleus and is separated from the cytoplasm by the nuclear envelope. Mutations in the nuclear envelope proteins emerin and lamin A cause a number of diseases including premature aging syndromes, muscular dystrophy, and cardiomyopathy. Emerin and lamin A are implicated in regulating muscle- and heart-specific gene expression and nuclear architecture. For example, lamin A regulates the expression and localization of gap junction and intercalated disc components. Additionally, emerin and lamin A are also required to maintain nuclear envelope integrity. Demonstrating the importance of maintaining nuclear integrity in heart disease, atrioventricular node cells lacking lamin A exhibit increased nuclear deformation and apoptosis. This review highlights the present understanding of lamin A and emerin function in regulating nuclear architecture, gene expression, and cell signaling and discusses putative mechanisms for how specific mutations in lamin A and emerin cause cardiac- or muscle-specific disease.

Desmin tail mutation linked to human dilated cardiomyopathy promotes cleavage of the N-terminus and abolishes z-disc localization

A missense mutation (Ile 451 to Met) at the tail domain of the muscle specific intermediate filament protein desmin has been suggested to be a genetic cause of dilated cardiomyopathy. Most of the desmin mutations (usually at the central rod domain) have as result the intracytoplasmic accumulation of desmin and other proteins aggregates in muscle cells. These aggregates are a major histological hallmark of desminopathies. Surprisingly most of the desmin tail domain mutants have the ability to form proper filaments on their own, so we hypothesize that filament-forming mutations will critically interfere with the binding of the filaments to z-disc or intercalated disc components, or proteins that are part of muscle-specific signaling cascades.

Reduction of gap and adherens junction proteins and intercalated disc structural remodeling in the hearts of mice submitted to severe cecal ligation and puncture sepsis*

The present study describes intercalated disc remodeling under both protein expression and structural features in experimental severe sepsis induced by cecal ligation and puncture in mice. Controlled animal study. University research laboratory. Male C57BL/6 mice. Mice were submitted to moderate and severe septic injury by cecal ligation and puncture. Severe septic injury was accompanied by a large number of bacteria in the peritoneal cavity and blood, high levels of tumor necrosis factor-alpha, and monocyte inflammatory protein-1alpha in the septic focus and serum, marked hypotension, and a high mortality rate. Western blot analysis and immunofluorescence showed a marked decrease of key gap and adherens junction proteins (connexin43 and N-cadherin, respectively) in mice submitted to severe septic injury. These changes may result in the loss of intercalated disc structural integrity, characterized in the electron microscopic study by partial separation or dehiscence of gap junctions and adherens junctions. Our data provide important insight regarding the alterations in intercalated disc components resulting from severe septic injury. The intercalated disc remodeling under both protein expression and structural features in experimental severe sepsis induced by cecal ligation and puncture may be partly responsible for myocardial depression in sepsis/septic shock. Although further electrophysiological studies in animals and humans are needed to determine the effect of these alterations on myocardial conduction velocity, the abnormal variables may emerge as therapeutic targets, and their modulation might provide beneficial effects on future cardiovascular outcomes and mortality in sepsis.

Ultrastructure of cultured atrial cardiac muscle cells from adult rats.

Atrial cardiac muscle cells enzymatically isolated from adult rats were maintained in culture for 0-17 days and examined by transmission electron microscopy (TEM). Cells were stained with conventional TEM stains as well as with osmium ferrocyanide and tannic acid. Our results show that cultured adult atrial cells are capable of in vitro ultrastructural reorganization and possess differentiated ultrastructural characteristics including specific atrial granules, sarcomerically arranged myofilaments, appropriately organized sarcoplasmic reticulum (both junctional and nonjunctional), and intercalated disc components. In addition, the cultured atrial cells also possess rare, but ultrastructurally typical, elements of the transverse tubular system. These can be identified on the basis of size, location, association with internal junctional sarcoplasmic reticulum, and accumulation of extracellular tracer. Atrial muscle cells are capable of reestablishing a myotypic ultrastructure, although they have a considerably less complex and organized in vitro ultrastructure than similarly cultured adult ventricular myocytes. This lessened in vitro ultrastructural specialization is in accord with the in vivo comparative ultrastructure of atrial vs. ventricular myocytes.