Gene therapies for RyR1-related myopathies Marty, Isabelle; Beaufils, Mathilde; Fauré, Julien ...
Current opinion in pharmacology,
February 2023, 2023-02-00, 20230201, Letnik:
68
Journal Article
Recenzirano
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Myopathies related to variations in the RYR1 gene are genetic diseases for which the therapeutic options are sparse, in part because of the very large size of the gene and protein, and of the ...distribution of variations all along the sequence. Taking advantage of the progress made in the gene therapy field, different approaches can be applied to the different genetic variations, either at the mRNA level or directly at the DNA level, specifically with the new gene editing tools. Some of those have already been tested in cellulo and/or in vivo, and for the development of the most innovative gene editing technology, inspiration can be sought in other genetic diseases.
Spermatogenesis defects concern millions of men worldwide, yet the vast majority remains undiagnosed. Here we report men with primary infertility due to multiple morphological abnormalities of the ...sperm flagella with severe disorganization of the sperm axoneme, a microtubule-based structure highly conserved throughout evolution. Whole-exome sequencing was performed on 78 patients allowing the identification of 22 men with bi-allelic mutations in DNAH1 (n = 6), CFAP43 (n = 10), and CFAP44 (n = 6). CRISPR/Cas9 created homozygous CFAP43/44 male mice that were infertile and presented severe flagellar defects confirming the human genetic results. Immunoelectron and stimulated-emission-depletion microscopy performed on CFAP43 and CFAP44 orthologs in Trypanosoma brucei evidenced that both proteins are located between the doublet microtubules 5 and 6 and the paraflagellar rod. Overall, we demonstrate that CFAP43 and CFAP44 have a similar structure with a unique axonemal localization and are necessary to produce functional flagella in species ranging from Trypanosoma to human.
Most patients with idiopathic membranous nephropathy (IMN) have IgG4 autoantibodies against phospholipase A2 receptor (PLA2R). C3 and C5b-9 are found in immune deposits of IMN kidney biopsy ...specimens, but the pathway of complement activation in IMN remains elusive. We report the case of a patient who developed IMN with intense staining for PLA2R, IgG4, C3, C5b-9, factor B, and properdin and very weak staining for C1q, C4d, and IgG1. Measurement of mannan binding lectin (MBL) antigenic level and activity revealed MBL deficiency. Genotyping revealed a heterozygous (A/C) polymorphism in codon 57 of MBL2 exon 1 associated with homozygous and heterozygous variations in the promoter region at -550 (L/L) and -221 (X/Y), respectively, suggesting that the patient harbored the LXA/LYC haplotypes linked to MBL deficiency. Genetic sequencing in 77 consecutive patients with IMN identified four patients with MBL2 promoter and coding region variations associated with MBL deficiency and the same complement pattern in immune deposits as the index patient. In contrast, patients with wild-type MBL2 had immune deposits with intense Cd4 staining. Thus, IMN can develop in patients with complete MBL deficiency, with complement activated mainly by the alternative pathway, whereas the lectin pathway is also activated in those with wild-type MBL2.
Airborne dust is the main climatic agent in the Martian environment. Local dust storms play a key role in the dust cycle; yet their life cycle is poorly known. Here we use mesoscale modeling that ...includes the transport of radiatively active dust to predict the evolution of a local dust storm monitored by OMEGA on board Mars Express. We show that the evolution of this dust storm is governed by deep convective motions. The supply of convective energy is provided by the absorption of incoming sunlight by dust particles, rather than by latent heating as in moist convection on Earth. We propose to use the terminology “rocket dust storm,” or conio‐cumulonimbus, to describe those storms in which rapid and efficient vertical transport takes place, injecting dust particles at high altitudes in the Martian troposphere (30–50 km). Combined to horizontal transport by large‐scale winds, rocket dust storms produce detached layers of dust reminiscent of those observed with Mars Global Surveyor and Mars Reconnaissance Orbiter. Since nighttime sedimentation is less efficient than daytime convective transport, and the detached dust layers can convect during the daytime, these layers can be stable for several days. The peak activity of rocket dust storms is expected in low‐latitude regions at clear seasons (late northern winter to late northern summer), which accounts for the high‐altitude tropical dust maxima unveiled by Mars Climate Sounder. Dust‐driven deep convection has strong implications for the Martian dust cycle, thermal structure, atmospheric dynamics, cloud microphysics, chemistry, and robotic and human exploration.
Key Points
Rocket dust storms: radiatively‐induced deep convection in Mars' dust storms
Rocket dust storms are the best scenario to account for detached layers of dust
Dusty deep convection has strong implications for Mars climate and exploration
In skeletal muscle, proteins of the calcium release complex responsible for the excitation-contraction (EC) coupling are exclusively localized in specific reticulum-plasma membrane (ER-PM) contact ...points named triads. The CRC protein triadin (T95) is localized in the sarcoplasmic reticulum (SR) subdomain of triads where it forms large multimers. However, the mechanisms leading to the steady-state accumulation of T95 in these specific areas of SR are largely unknown. To visualize T95 dynamics, fluorescent chimeras were expressed in triadin knockout myotubes, and their mobility was compared with the mobility of Sec61β, a membrane protein of the SR unrelated to the EC coupling process. At all stages of skeletal muscle cells differentiation, we show a permanent flux of T95 diffusing in the SR membrane. Moreover, we find evidence that a longer residence time in the ER-PM contact point is due to the transmembrane domain of T95 resulting in an overall triad localization.
Context. In protoplanetary disks, the inner boundary between the turbulent and laminar regions could be a promising site for planet formation, thanks to the trapping of solids at the boundary itself ...or in vortices generated by the Rossby wave instability. At the interface, the disk thermodynamics and the turbulent dynamics are entwined because of the importance of turbulent dissipation and thermal ionization. Numerical models of the boundary, however, have neglected the thermodynamics, and thus miss a part of the physics. Aims. The aim of this paper is to numerically investigate the interplay between thermodynamics and dynamics in the inner regions of protoplanetary disks by properly accounting for turbulent heating and the dependence of the resistivity on the local temperature. Methods. Using the Godunov code RAMSES, we performed a series of 3D global numerical simulations of protoplanetary disks in the cylindrical limit, including turbulent heating and a simple prescription for radiative cooling. Results. We find that waves excited by the turbulence significantly heat the dead zone, and we subsequently provide a simple theoretical framework for estimating the wave heating and consequent temperature profile. In addition, our simulations reveal that the dead-zone inner edge can propagate outward into the dead zone, before stalling at a critical radius that can be estimated from a mean-field model. The engine driving the propagation is in fact density wave heating close to the interface. A pressure maximum appears at the interface in all simulations, and we note the emergence of the Rossby wave instability in simulations with extended azimuth. Conclusions. Our simulations illustrate the complex interplay between thermodynamics and turbulent dynamics in the inner regions of protoplanetary disks. They also reveal how important activity at the dead-zone interface can be for the dead-zone thermodynamic structure.
Catecholaminergic polymorphic ventricular tachycardia (CPVT) is an inherited arrhythmogenic disease so far related to mutations in the cardiac ryanodine receptor (RYR2) or the cardiac calsequestrin ...(CASQ2) genes. Because mutations in RYR2 or in CASQ2 are not retrieved in all CPVT cases, we searched for mutations in the physiological protein partners of RyR2 and CSQ2 in a large cohort of CPVT patients with no detected mutation in these two genes. Based on a candidate gene approach, we focused our investigations on triadin and junctin, two proteins that link RyR2 and CSQ2. Mutations in the triadin (TRDN) and in the junctin (ASPH) genes were searched in a cohort of 97 CPVT patients. We identified three mutations in triadin which cosegregated with the disease on a recessive mode of transmission in two families, but no mutation was found in junctin. Two TRDN mutations, a 4 bp deletion and a nonsense mutation, resulted in premature stop codons; the third mutation, a p.T59R missense mutation, was further studied. Expression of the p.T59R mutant in COS-7 cells resulted in intracellular retention and degradation of the mutant protein. This was confirmed after in vivo expression of the mutant triadin in triadin knock-out mice by viral transduction. In this work, we identified TRDN as a new gene responsible for an autosomal recessive form of CPVT. The mutations identified in the two families lead to the absence of the protein, thereby demonstrating the importance of triadin for the normal function of the cardiac calcium release complex in humans.
Arthrogryposis multiplex congenita (AMC) refers to a clinical presentation of congenital contractures involving two or more body areas. More than 400 distinct conditions may lead to AMC, making the ...aetiological diagnosis challenging. The objective of this work was to set up evidence-based recommendations for the diagnosis of AMC by taking advantage of both data from our nation-wide cohort of children with AMC and from the literature.
We conducted a retrospective single-centre observational study. Patients had been evaluated at least once at a paediatric age in the AMC clinic of Grenoble University Hospital between 2007 and 2019. After gathering data about their diagnostic procedure, a literature review was performed for each paraclinical investigation to discuss their relevance.
One hundred and twenty-five patients were included, 43% had Amyoplasia, 27% had distal arthrogryposis and 30% had other forms. A definitive aetiological diagnosis was available for 66% of cases. We recommend a two-time diagnostic process: first, non-invasive investigations that aim at classifying patients into one of the three groups, and second, selected investigations targeting a subset of patients.
The aetiological management for patients with AMC remains arduous. This process will be facilitated by the increasing use of next-generation sequencing combined with detailed phenotyping. Invasive investigations should be avoided because of their limited yield.
Recessive forms of catecholaminergic polymorphic ventricular tachycardia (CPVT) are induced by mutations in genes encoding triadin or calsequestrin, two proteins that belong to the Ca2+ release ...complex, responsible for intracellular Ca2+ release triggering cardiac contractions. To better understand the mechanisms of triadin-induced CPVT and to assay multiple therapeutic interventions, we used a triadin knockout mouse model presenting a CPVT-like phenotype associated with a decrease in calsequestrin protein level. We assessed different approaches to rescue protein expression and to correct intracellular Ca2+ release and cardiac function: pharmacological treatment with kifunensine or a viral gene transfer-based approach, using adeno-associated virus serotype 2/9 (AAV2/9) encoding the triadin or calsequestrin. We observed that the levels of triadin and calsequestrin are intimately linked, and that reduction of both proteins contributes to the CPVT phenotype. Different combinations of triadin and calsequestrin expression level were obtained using these therapeutic approaches. A full expression of each is not necessary to correct the phenotype; a fine-tuning of the relative re-expression of both triadin and calsequestrin is required to correct the CPVT phenotype and rescue the cardiac function. AAV-mediated gene delivery of calsequestrin or triadin and treatment with kifunensine are potential treatments for recessive forms of CPVT due to triadin mutations.
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Catecholaminergic polymorphic ventricular tachycardia (CPVT) is induced by mutations in triadin or calsequestrin. Deletion/mutation of triadin is associated with reduction in calsequestrin and leads to CPVT. Cacheux and colleagues show that partial re-expression of triadin and calsequestrin through gene or pharmacological therapy reverses this cardiac arrhythmia.
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