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The heart’s response to varying demands of the body is regulated by signaling pathways that activate protein kinases which phosphorylate sarcomeric proteins. Although phosphorylation of cardiac myosin binding protein-C (cMyBP-C) has been recognized as a key regulator of myocardial contractility, little is known about its mechanism of action. Here, we used protein kinase A (PKA) and Cε (PKCε), as well as ribosomal S6 kinase II (RSK2), which have different specificities for cMyBP-C’s multiple phosphorylation sites, to show that individual sites are not independent, and that phosphorylation of cMyBP-C is controlled by positive and negative regulatory coupling between those sites. PKA phosphorylation of cMyBP-C’s N terminus on 3 conserved serine residues is hierarchical and antagonizes phosphorylation by PKCε, and vice versa. In contrast, RSK2 phosphorylation of cMyBP-C accelerates PKA phosphorylation. We used cMyBP-C’s regulatory N-terminal domains in defined phosphorylation states for protein–protein interaction studies with isolated cardiac native thin filaments and the S2 domain of cardiac myosin to show that site-specific phosphorylation of this region of cMyBP-C controls its interaction with both the actin-containing thin and myosin-containing thick filaments. We also used fluorescence probes on the myosin-associated regulatory light chain in the thick filaments and on troponin C in the thin filaments to monitor structural changes in the myofilaments of intact heart muscle cells associated with activation of myocardial contraction by the N-terminal region of cMyBP-C in its different phosphorylation states. Our results suggest that cMyBP-C acts as a sarcomeric integrator of multiple signaling pathways that determines downstream physiological function.

The auto-inhibited, super-relaxed (SRX) state of cardiac myosin is thought to be crucial for regulating contraction, relaxation, and energy conservation in the heart. We used single ATP turnover experiments to demonstrate that a dilated cardiomyopathy (DCM) mutation (E525K) in human beta-cardiac myosin increases the fraction of myosin heads in the SRX state (with slow ATP turnover), especially in physiological ionic strength conditions. We also utilized FRET between a C-terminal GFP tag on the myosin tail and Cy3ATP bound to the active site of the motor domain to estimate the fraction of heads in the closed, interacting-heads motif (IHM); we found a strong correlation between the IHM and SRX state. Negative stain electron microscopy and 2D class averaging of the construct demonstrated that the E525K mutation increased the fraction of molecules adopting the IHM. Overall, our results demonstrate that the E525K DCM mutation may reduce muscle force and power by stabilizing the auto-inhibited SRX state. Our studies also provide direct evidence for a correlation between the SRX biochemical state and the IHM structural state in cardiac muscle myosin. Furthermore, the E525 residue may be implicated in crucial electrostatic interactions that modulate this conserved, auto-inhibited conformation of myosin.

Skeletal myopathies and ataxias with secondary cardiac involvement are complex, progressive and debilitating conditions. As life expectancy increases across these conditions, cardiac involvement often becomes more prominent. This highlights the need for targeted therapies that address these evolving cardiac pathologies. Musculopathies by and large lack cures that directly target the genetic basis of the diseases; however, as our understanding of the genetic causes of these conditions has evolved, it has become tractable to develop targeted therapies using biologics, to design precision approaches to target the primary genetic causes of these varied diseases. Using the examples of Duchenne muscular dystrophy, Friedreich ataxia and Pompe disease, we discuss how the genetic causes of such diseases derail diverse homeostatic, energetic and signalling pathways, which span multiple cellular systems in varied tissues across the body. We outline existing therapeutics and treatments in the context of emerging novel genetic approaches. We discuss the hurdles that the field must overcome to deliver targeted therapies across the many tissue types affected in primary myopathies. What is the topic of this review? Overlapping disease pathomechanisms and therapeutic opportunities in neuromuscular, skeletal and cardiac muscle diseases in the context of novel genetic therapies. What advances does it highlight? This review outlines the diverse genetic changes that drive pathomechanism across a set of neuromuscular conditions and highlight the emerging targeted biological therapies that are being developed to treat these conditions, with additional discussion of the hurdles to actualising genetically targeted precision medicine.

Omecamtiv mecarbil (OM) is a small molecule that has been shown to improve the function of the slow human ventricular myosin (MyHC) motor through a complex perturbation of the thin/thick filament regulatory state of the sarcomere mediated by binding to myosin allosteric sites coupled to inorganic phosphate (Pi) release. Here, myofibrils from samples of human left ventricle (β-slow MyHC-7) and left atrium (α-fast MyHC-6) from healthy donors were used to study the differential effects of μmolar [OM] on isometric force in relaxing conditions (pCa 9.0) and at maximal (pCa 4.5) or half-maximal (pCa 5.75) calcium activation, both under control conditions (15 °C; equimolar DMSO; contaminant inorganic phosphate [Pi] ~170 μM) and in the presence of 5 mM [Pi]. The activation state and OM concentration within the contractile lattice were rapidly altered by fast solution switching, demonstrating that the effect of OM was rapid and fully reversible with dose-dependent and myosin isoform-dependent features. In MyHC-7 ventricular myofibrils, OM increased submaximal and maximal Ca2+-activated isometric force with a complex dose-dependent effect peaking (40% increase) at 0.5 μM, whereas in MyHC-6 atrial myofibrils, it had no effect or—at concentrations above 5 µM—decreased the maximum Ca2+-activated force. In both ventricular and atrial myofibrils, OM strongly depressed the kinetics of force development and relaxation up to 90% at 10 μM [OM] and reduced the inhibition of force by inorganic phosphate. Interestingly, in the ventricle, but not in the atrium, OM induced a large dose-dependent Ca2+-independent force development and an increase in basal ATPase that were abolished by the presence of millimolar inorganic phosphate, consistent with the hypothesis that the widely reported Ca2+-sensitising effect of OM may be coupled to a change in the state of the thick filaments that resembles the on–off regulation of thin filaments by Ca2+. The complexity of this scenario may help to understand the disappointing results of clinical trials testing OM as inotropic support in systolic heart failure compared with currently available inotropic drugs that alter the calcium signalling cascade.

Cachexia is a multifactorial syndrome that presents with, among other characteristics, progressive loss of muscle mass and anti-cardiac remodeling effect that may lead to heart failure. This condition affects about 80% of patients with advanced cancer and contributes to worsening patients’ tolerance to anticancer treatments and to their premature death. Its pathogenesis involves an imbalance in metabolic homeostasis, with increased catabolism and inflammatory cytokines levels, leading to proteolysis and lipolysis, with insufficient food intake. A multimodal approach is indicated for patients with cachexia, with the aim of reducing the speed of muscle wasting and improving their quality of life, which may include nutritional, physical, pharmacologic, and psychological support. This review aims to outline the mechanisms of muscle loss, as well as to evaluate the current clinical evidence of the use of physical exercise in patients with cachexia.

The Frank-Starling relation is a fundamental auto-regulatory property of the heart that ensures the volume of blood ejected in each heartbeat is matched to the extent of venous filling. At the cellular level, heart muscle cells generate higher force when stretched, but despite intense efforts the underlying molecular mechanism remains unknown. We applied a fluorescence-based method, which reports structural changes separately in the thick and thin filaments of rat cardiac muscle, to elucidate that mechanism. The distinct structural changes of troponin C in the thin filaments and myosin regulatory light chain in the thick filaments allowed us to identify two aspects of the Frank-Starling relation. Our results show that the enhanced force observed when heart muscle cells are maximally activated by calcium is due to a change in thick filament structure, but the increase in calcium sensitivity at lower calcium levels is due to a change in thin filament structure. The heart needs to pump out the same volume of blood that enters it. This is not as simple as it sounds, as changes in heart rate – for example, in response to exercise – alter how hard the heart must pump. When blood flows into the heart it stretches the heart muscle, which consists of units called sarcomeres. Sarcomeres contain two types of protein filament, known as thick filaments and thin filaments. When a heartbeat is triggered by calcium ions flowing into the heart muscle cells, the thick filaments slide over the thin filaments. This causes the heart muscle cell to contract. The Frank–Starling mechanism helps to regulate the contraction of the heart. This mechanism has two aspects. Firstly, as the sarcomere lengthens, its protein filaments are able to contract with more force for a given high level of calcium ions. Secondly, the lengthening of the sarcomere makes the filaments more sensitive to calcium ions, which again causes the heart to contract more forcefully. However, the molecular mechanisms that underlie these effects were not clear. Zhang et al. have now studied rat heart muscle cells using a new fluorescence-based method that can detect structural changes in the thick and thin filaments. The results show that the increased force that is generated when sarcomeres are stretched can be accounted for by changes in the structure of the thick filament. In contrast, the increase in calcium sensitivity that occurs as the sarcomere lengthens is largely due to structural alterations in the thin filament. These two processes can be controlled independently, but work together in the Frank–Starling mechanism. Now that we better understand the molecular basis of the Frank–Starling mechanism, further work could investigate new strategies for designing and testing treatments for heart disease.

The Cardiomyopathy–associated gene 5 ( Cmya5 ) encodes myospryn, a large tripartite motif (TRIM)-related protein found predominantly in cardiac and skeletal muscle. Cmya5 is an expression biomarker for a number of diseases affecting striated muscle and may also be a schizophrenia risk gene. To further understand the function of myospryn in striated muscle, we searched for additional myospryn paralogs. Here we identify a novel muscle-expressed TRIM-related protein minispryn, encoded by Fsd2 , that has extensive sequence similarity with the C-terminus of myospryn. Cmya5 and Fsd2 appear to have originated by a chromosomal duplication and are found within evolutionarily-conserved gene clusters on different chromosomes. Using immunoaffinity purification and mass spectrometry we show that minispryn co-purifies with myospryn and the major cardiac ryanodine receptor (RyR2) from heart. Accordingly, myospryn, minispryn and RyR2 co-localise at the junctional sarcoplasmic reticulum of isolated cardiomyocytes. Myospryn redistributes RyR2 into clusters when co-expressed in heterologous cells whereas minispryn lacks this activity. Together these data suggest a novel role for the myospryn complex in the assembly of ryanodine receptor clusters in striated muscle.

An overview of noteworthy new methods of biomarker determination based on surface-enhanced Raman scattering (SERS) is presented. Biomarkers can be used to identify the occurrence and development of diseases, which furthers the understanding of biological processes in the body. Accurate detection of a disease-specific biomarker is helpful for the identification, early diagnosis and prevention of a disease and for monitoring during treatment. The search for and discovery of valuable biomarkers have become important research hotspots. Different diseases have different biomarkers, some of which are involved in metabolic processes. Therefore, the fingerprint characteristics and band intensities in SERS spectra have been used to identify metabolites and analyze markers. As a promising technique, SERS has been widely used for the quantitative and qualitative determination of different types of biomarkers for different diseases. SERS techniques provide new technologies for the diagnosis of disease-related markers and determining the basis for clinical treatment. Herein, several SERS-based methods with excellent sensitivity and selectivity for the determination of biomarkers for tumors, viruses, Alzheimer’s disease, cardiac muscle tissue injury, and cell activity are highlighted.

In non-diabetic patients with severe disease, such as acute myocardial infarction or acute heart failure, admission blood glucose level is associated with their short-term and long-term mortality. We examined whether transient elevation of glucose affects contractile properties in non-diabetic hearts. Force, intracellular Ca 2+ ([Ca 2+ ] i ), and sarcomere length were measured in trabeculae from rat hearts. To assess contractile properties, maximum velocity of contraction (Max dF / dt ) and minimum velocity of relaxation (Min dF / dt ) were calculated. The ratio of phosphorylated troponin I (P-TnI) to troponin I (TnI) was measured. One hour after elevation of glucose from 150 to 400 mg/dL, developed force, Max dF / dt , and Min dF / dt were reduced without changes in [Ca 2+ ] i transients at 2.5 Hz stimulation and 2.0 mM [Ca 2+ ] o , while developed force and [Ca 2+ ] i transients showed no changes at 0.5 Hz stimulation and 0.7 mM [Ca 2+ ] o . In the presence of 1 μM KN-93, a Ca 2+ /calmodulin-dependent protein kinaseII (CaMKII) inhibitor, or 50 μM diazo-5-oxonorleucine, a l -glutamine- d -fructose-6-phosphate amidotransferase inhibitor, the reduction of contractile properties after elevation of glucose was suppressed. Furthermore, 1 h after elevation of glucose to 400 mg/dL at 2.0 mM [Ca 2+ ] o , the ratio of P-TnI to TnI was increased. These results suggest that in non-diabetic hearts under higher Ca 2+ -load, transient elevation of glucose for 1 h reduces contractile properties probably by activating CaMKII through O-GlcNAcylation. Thus, in the patients with severe disease, transient elevation of blood glucose, such as due to stress, may worsen cardiac function and thereby affect their mortality without known diabetes.

The article is devoted to the assessment of force and energy characteristics of the heart muscle in microgravity using the method of spatial ballistocardiography. The studies were conducted with the participation of Russian cosmonauts who performed space flights of various durations. Physiological signals were recorded using the Cardiovector device. The experiment was carried out twice before the flight, monthly during the flight, and also on days +3–4 and +7–8 after return to Earth. It has been shown that the force of cardiac contraction does not change significantly, but the energy decreases, which can be explained by the principle of economizing the work of the heart under space flight conditions. Mission extension does not lead to significant energy reduction to levels that could indicate possible signs and risks of energy deficiency.