Mechanical Forces Regulate Cardiomyocyte Myofilament Maturation via the VCL-SSH1-CFL Axis

Fukuda, Ryūji; Gunawan, Felix; Ramadass, Radhan; Beisaw, Arica; Konzer, Anne; Mullapudi, Sri Teja; Gentile, Alessandra; Maischein, Hans-Martin; Graumann, Johannes; Stainier, Didier Y.R.

doi:10.1016/j.devcel.2019.08.006

Cited by 40 publications

(53 citation statements)

References 98 publications

(138 reference statements)

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“…Directed hypertrophy determines the overall shape of a chamber while cell division is more evident during ballooning stages in mammalian hearts that undergo rapid increases in size [ 6 ]. Outgrowth occurs as the linear heart has already commenced peristaltic beating and zebrafish studies have shown that blood fluid forces can influence final chamber morphology by affecting cytoskeletal protein localization, cardiomyocyte maturation and also endocardial proliferation [ 7 , 8 , Reviewed 9 ]. Underpinning the directed growth is myofibrillogenesis, the process of assembling the contractile protein machinery within the muscle cells.…”

Section: Introductionmentioning

confidence: 99%

Defective heart chamber growth and myofibrillogenesis after knockout of adprhl1 gene function by targeted disruption of the ancestral catalytic active site

et al. 2020

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ADP-ribosylhydrolase-like 1 (Adprhl1) is a pseudoenzyme expressed in the developing heart myocardium of all vertebrates. In the amphibian Xenopus laevis, knockdown of the two cardiac Adprhl1 protein species (40 and 23 kDa) causes failure of chamber outgrowth but this has only been demonstrated using antisense morpholinos that interfere with RNAsplicing. Transgenic production of 40 kDa Adprhl1 provides only part rescue of these defects. CRISPR/Cas9 technology now enables targeted mutation of the adprhl1 gene in G0-generation embryos with routine cleavage of all alleles. Testing multiple gRNAs distributed across the locus reveals exonic locations that encode critical amino acids for Adprhl1 function. The gRNA recording the highest frequency of a specific ventricle outgrowth phenotype directs Cas9 cleavage of an exon 6 sequence, where microhomology mediated endjoining biases subsequent DNA repairs towards three small in-frame deletions. Mutant alleles encode discrete loss of 1, 3 or 4 amino acids from a di-arginine (Arg271-Arg272) containing peptide loop at the centre of the ancestral ADP-ribosylhydrolase site. Thus despite lacking catalytic activity, it is the modified (adenosine-ribose) substrate binding cleft of Adprhl1 that fulfils an essential role during heart formation. Mutation results in striking loss of myofibril assembly in ventricle cardiomyocytes. The defects suggest Adprhl1 participation from the earliest stage of cardiac myofibrillogenesis and are consistent with previous MO results and Adprhl1 protein localization to actin filament Z-disc boundaries. A single nucleotide change to the gRNA sequence renders it inactive. Mice lacking Adprhl1 exons 3-4 are normal but production of the smaller ADPRHL1 species is unaffected, providing further evidence that cardiac activity is concentrated at the C-terminal protein portion.

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Section: Introductionmentioning

confidence: 99%

Defective heart chamber growth and myofibrillogenesis after knockout of adprhl1 gene function by targeted disruption of the ancestral catalytic active site

et al. 2020

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“…As such, it is interesting to note that focal adhesion proteins such as FAK and vinculin also localize to intercalated discs once they form ( Koteliansky and Gneushev, 1983 ; Yi et al , 2003 ). Indeed, a recent study found that vinculin localizes to cell–cell-adhesion sites under increased load in zebrafish hearts in vivo , driving the thickening of myofibrils ( Fukuda et al , 2019 ). Determining the mechanisms driving the localization of such proteins to sites of increased load should be the focus of future studies.…”

Section: Discussionmentioning

confidence: 99%

Coupling to substrate adhesions drives the maturation of muscle stress fibers into myofibrils within cardiomyocytes

Taneja

Neininger

Burnette

2020

MBoC

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“…A proteomic screen in non-contracting and contracting rat cardiomyocytes identified the slingshot protein phosphatase (SSH1) as a vinculin-binding partner in contracting cells (Fukuda et al 2019 ). Indeed, the vinculin-SSH1 interaction was increased by cyclical stretch and abolished when cardiac contractility was inhibited, suggesting that the interaction is force-dependent (Fukuda et al 2019 ). SSH1 can dephosphorylate and activate the actin regulator cofilin (CFL), which severs F-actin filaments to supply monomeric G-actin and promote filament reassembly (Ohashi 2015 ).…”

Section: Mechanosensing and Transduction At The Idmentioning

confidence: 99%

“…Interestingly, neonatal rat cardiomyocytes infected with a phosphomimetic CFL2 adenovirus displayed ‘stress-like’ fibre structures and reduced contractility compared with cells infected with wild-type adenovirus, suggesting that excess actin filament formation is detrimental for contraction (Subramanian et al 2015 ). It may be that CFL recruitment to the vinculin-SSH1 complex is a protective mechanism in response to increased mechanical load since decreased CFL activity is associated with DCM (Fukuda et al 2019 ). It will be interesting to study the precise localisation of CFL in cardiomyocytes, but immunofluorescence microscopy of human hearts revealed that CFL2 is present predominantly at the intercalated disc (Subramanian et al 2015 ), further supporting its role in mechanotransduction at this specialised site.…”

Section: Mechanosensing and Transduction At The Idmentioning

confidence: 99%

The intercalated disc: a mechanosensing signalling node in cardiomyopathy

Pruna

Ehler

2020

Biophys Rev

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Cardiomyocytes, the cells generating contractile force in the heart, are connected to each other through a highly specialised structure, the intercalated disc (ID), which ensures force transmission and transduction between neighbouring cells and allows the myocardium to function in synchrony. In addition, cardiomyocytes possess an intrinsic ability to sense mechanical changes and to regulate their own contractile output accordingly. To achieve this, some of the components responsible for force transmission have evolved to sense changes in tension and to trigger a biochemical response that results in molecular and cellular changes in cardiomyocytes. This becomes of particular importance in cardiomyopathies, where the heart is exposed to increased mechanical load and needs to adapt to sustain its contractile function. In this review, we will discuss key mechanosensing elements present at the intercalated disc and provide an overview of the signalling molecules involved in mediating the responses to changes in mechanical force. Keywords Cardiomyopathy. Mechanobiology. Cell-cell contact. Intercalated disc. Cytoskeleton Mechanical cues in the heart Mechanical stimuli play a key role in both heart morphogenesis and in the mature heart. During heart development, mechanical forces orchestrate the molecular and cellular changes that transform the linear tubular heart into a multichambered machine with four valves (Lindsey et al. 2014). In the chick embryo, primordial heart contraction and the resulting pulsatile blood flow occurs before active oxygen transport is required (Burggren 2004), suggesting that contractile force is required not only for blood pumping but also for morphogenesis. Internal forces from cardiac contraction exert strain on the cell-cell junctions, whereas blood flow exerts both perpendicular (cyclic strain) and parallel forces (shear stress) to the vessel wall (Granados-Riveron and Brook 2012). In the developing heart, these mechanical forces are essential for shaping the chambered structure as well as for myofibrillogenesis (Geach et al. 2015), whereas in the fully formed heart, these cues are important in maintaining the structural and functional integrity of the myocardium. In cardiomyopathies, increased mechanical load triggers compensatory molecular and cellular changes temporarily allowing the myocardium to sustain pump function, but with time, these adaptive responses fail to meet the increased demand, resulting in cardiac dysfunction and heart failure (reviewed in Harvey and Leinwand 2011; McNally et al. 2013). Cardiomyocyte cytoarchitecture Cells that make up the contractile tissue of the heart, the cardiomyocytes, are characterised by a highly regular architecture of cytoskeletal elements to ensure force generation and transduction with each heartbeat (reviewed in Ehler 2016). Cytoskeletal elements are organised into two major multiprotein complexes: the myofibrils and the intercalated disc (Fig. 1). Myofibrils, consisting of thin, thick and elastic filaments, contain the contractile machine...

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Mechanical Forces Regulate Cardiomyocyte Myofilament Maturation via the VCL-SSH1-CFL Axis

Cited by 40 publications

References 98 publications

Defective heart chamber growth and myofibrillogenesis after knockout of adprhl1 gene function by targeted disruption of the ancestral catalytic active site

Defective heart chamber growth and myofibrillogenesis after knockout of adprhl1 gene function by targeted disruption of the ancestral catalytic active site

Coupling to substrate adhesions drives the maturation of muscle stress fibers into myofibrils within cardiomyocytes

The intercalated disc: a mechanosensing signalling node in cardiomyopathy

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