Distal Myopathy Research Articles

Extra-muscular manifestations in GNE myopathy patients: A nationwide repository questionnaire survey in Japan

ObjectiveGNE myopathy is a rare autosomal recessive adult-onset distal myopathy caused by biallelic pathogenic variants in GNE. Although some extra-muscular manifestations associated with GNE myopathy have been reported, little is known about whether they are disease-specific and how often they present. This study aimed to characterize extra-muscular manifestations of GNE myopathy. MethodsWe conducted a questionnaire survey of GNE myopathy patients registered in a national registry in Japan. The questionnaire requested information regarding idiopathic thrombocytopenia, cardiac involvement, respiratory involvement, sleep apnea syndrome (SAS), and psychiatric diseases. ResultsThe response rate was 62.4% (126/198), yielding a total of 51 male and 75 female participants. Of the participants, 4.1% (5/123) had a diagnosis of idiopathic thrombocytopenia, and 16.3% (8/49) of males and 6.6% of females (5/76) had a diagnosis of SAS. In total, 0.8% (1/126) of participants had pervasive developmental disabilities and 14.7% (16/109) had a psychiatric disease. ConclusionThe frequencies of idiopathic thrombocytopenia and SAS among Japanese GNE myopathy patients were higher than those observed in the general Japanese population. Routine blood tests and evaluation of sleep-disordered breathing should be considered in order to better manage GNE myopathy patients.

Muscle “islands”: An MRI signature distinguishing neurogenic from myopathic causes of early onset distal weakness

Muscle MRI has an increasing role in diagnosis of inherited neuromuscular diseases, but no features are known which reliably differentiate myopathic and neurogenic conditions. Using patients presenting with early onset distal weakness, we aimed to identify an MRI signature to distinguish myopathic and neurogenic conditions. We identified lower limb MRI scans from patients with either genetically (n = 24) or clinically (n = 13) confirmed diagnoses of childhood onset distal myopathy or distal spinal muscular atrophy. An initial exploratory phase reviewed 11 scans from genetically confirmed patients identifying a single potential discriminatory marker concerning the pattern of fat replacement within muscle, coined “islands”. This pattern comprised small areas of muscle tissue with normal signal intensity completely surrounded by areas with similar intensity to subcutaneous fat. In the subsequent validation phase, islands correctly classified scans from all 12 remaining genetically confirmed patients, and 12/13 clinically classified patients. In the genetically confirmed patients MRI classification of neurogenic/myopathic aetiology had 100% accuracy (24/24) compared with 65% accuracy (15/23) for EMG, and 79% accuracy (15/19) for muscle biopsy. Future studies are needed in other clinical contexts, however the presence of islands appears to highly suggestive of a neurogenic aetiology in patients presenting with early onset distal motor weakness.

A case report of deglutition triggered atrial fibrillation in a patient with Laing distal myopathy

BackgroundDeglutition-induced atrial fibrillation is a rare clinical entity with a reported prevalence of 0.6%. Laing distal myopathy is a rare autosomal dominant muscular dystrophy that is the result of mutations within the slow skeletal muscle fibre myosin heavy chain gene (MYH7). Atrial fibrillation has not been previously reported in patients with Laing distal myopathy. We describe the first reported case of deglutition triggered atrial fibrillation in a female with a history of Laing distal myopathy.Case summaryA 44-year-old female with a history of Laing distal myopathy diagnosed at age 32, began experiencing intermittent episodes of pre-syncope and palpitations which occurred after deglutition with food. An ambulatory 30-day patient triggered event monitor recorded episodes of atrial fibrillation with rapid ventricular response. Family history was significant for Laing distal myopathy, atrial fibrillation, as well as sudden cardiac death. Laboratory data, transthoracic echocardiogram, cardiac magnetic resonance imaging, and an exercise treadmill SPECT Imaging stress test were normal. An oesophagram revealed a mild oesophageal dysmotility with no other abnormalities. She was started on flecainide 50 mg p.o. every 8 h and verapamil 40 mg p.o. every 8 h with no further episodes of atrial fibrillation.DiscussionGiven the strong genetic component of this myopathy, one could postulate as to a possible genetic component in the development of atrial fibrillation in our patient. Although we cannot make definite correlation between deglutition-induced atrial fibrillation and Laing myopathy, it is important to report this unusual association which has not been described before.

Dominant Distal Myopathy 3 (MPD3) Caused by a Deletion in the HNRNPA1 Gene.

Background and ObjectivesTo determine the genetic cause of the disease in the previously reported family with adult-onset autosomal dominant distal myopathy (myopathy, distal, 3; MPD3).MethodsContinued clinical evaluation including muscle MRI and muscle pathology. A linkage analysis with single nucleotide polymorphism arrays and genome sequencing were used to identify the genetic defect, which was verified by Sanger sequencing. RNA sequencing was used to investigate the transcriptional effects of the identified genetic defect.ResultsSmall hand muscles (intrinsic, thenar, and hypothenar) were first involved with spread to the lower legs and later proximal muscles. Dystrophic changes with rimmed vacuoles and cytoplasmic inclusions were observed in muscle biopsies at advanced stage. A single nucleotide polymorphism array confirmed the previous microsatellite-based linkage to 8p22-q11 and 12q13-q22. Genome sequencing of three affected family members combined with structural variant calling revealed a small heterozygous deletion of 160 base pairs spanning the second last exon 10 of the heterogeneous nuclear ribonucleoprotein A1 (HNRNPA1) gene, which is in the linked region on chromosome 12. Segregation of the mutation with the disease was confirmed by Sanger sequencing. RNA sequencing showed that the mutant allele produces a shorter mutant mRNA transcript compared with the wild-type allele. Immunofluorescence studies on muscle biopsies revealed small p62 and larger TDP-43 inclusions.DiscussionA small exon 10 deletion in the gene HNRNPA1 was identified as the cause of MPD3 in this family. The new HNRNPA1-related phenotype, upper limb presenting distal myopathy, was thus confirmed, and the family displays the complexities of gene identification.

First Family of MATR3-Related Distal Myopathy From Italy: The Role of Muscle Biopsy in the Diagnosis and Characterization of a Still Poorly Understood Disease.

Mutations in the MATR3 gene are associated to distal myopathy with vocal cord and pharyngeal weakness (VCPDM), as well as familiar and sporadic motor neuron disease. To date, 12 VCPDM families from the United States, Germany, Japan, Bulgary, and France have been described in the literature. Here we report an Italian family with a propositus of a 40-year-old woman presenting progressive bilateral foot drop, rhinolalia, and distal muscular atrophy, without clinical signs of motor neuron affection. Her father, deceased some years before, presented a similar distal myopathy phenotype, while her 20-year-old son is asymptomatic. Myopathic changes with vacuolization were observed in muscle biopsy from the propositus. These results, together with the peculiar clinical picture, lead to MATR3 gene sequencing, which revealed a heterozygous p.S85C mutation in the propositus. The same mutation was found in her son. Over a 5-year follow-up, progression is mild in the propositus, while her son remains asymptomatic. Clinical, radiological, and pathological data of our propositus are presented and compared to previously reported cases of VCPDM. VCPDM turns out to be a quite homogenous phenotype of late-onset myopathy associated to p.S85C mutation in MATR3 gene. MATR3-related pathology, encompassing myopathy and motor neuron disease, represents an illustrative example of multisystem proteinopathy (MSP), such as other diseases associated to mutations in VCP, HNRNPA2B1, HNRNPA1, and SQSTM1 genes. The present report contributes to a further characterization of this still poorly understood pathology and points out the diagnostic utility of muscle biopsy in challenging cases.

Asymmetrical scapular winging, bilateral facial muscle involvement and distal myopathy with normal CPK; unusual features in anti JO-I positive polymyositis

Introduction Polymyositis is an idiopathic inflammatory myopathy which present with typical symmetrical proximal muscle weakness with the evidence of muscle inflammation. The acute inflammation of muscle results in high levels of CPK and has a relapsing remitting course with a good response to anti-inflammatory treatment. We present here a patient presented with a chronic myopathy with relapsing remitting fashion, with asymmetrical scapula winging, and bilateral facial weakness, with predominant distal muscle weakness which was supported by EMG.

Clinicopathological features of titinopathy from a Chinese neuromuscular center.

Titin, one of the largest proteins in humans, is a major component of muscle sarcomeres. Pathogenic variants in the titin gene (TTN) have been reported to cause a range of skeletal muscle diseases, collectively known as titinopathy. Titinopathy is a heterogeneous group of disabling diseases characterized by muscle weakness. In our study, we aimed to establish the clinicopathological-genetic spectrum of titinopathy from a single neuromuscular center. Three patients were diagnosed as having definite titinopathy, and additional three patients were diagnosed as having possible titinopathy according to the diagnostic criteria. All the patients showed initial symptoms from age one to 40 years. Physical examination revealed that five patients had muscle weakness, and that one patient experienced behavioral changes. Muscle biopsy specimens obtained from all six patients demonstrated multiple myopathological changes, including increased fiber size variation, muscle fiber hypertrophy or atrophy, formation of centralized cell nuclei, necklace cytoplasmic bodies, and formation of rimmed vacuoles and cores. Genetic testing revealed 11 different TTN alterations, including missense (6/11), nonsense (2/11), frameshift (2/11), and splicing (1/11) mutations. Our study provides further evidence that TTN mutations are more likely to be responsible for an increasing proportion of various myopathies, such as hereditary myopathy with early respiratory failure (HMERF), core myopathy, and distal myopathy with rimmed vacuoles, than currently recognized mutations. Our findings expand the clinical, pathohistological and genetic spectrum of titinopathy.

DISTAL MYOPATHIES: EP.75 Functional validation of a novel variant of the SPTAN1 gene identified in a family with distal motor myopathy with nerve involvement

Hereditary distal motor neuropathies are a heterogeneous group of inherited neuromuscular disorders characterized by weakness mostly predominantly in lower limbs. We describe a family with two affected members, presenting a distal motor weakness. The propositus, a 33-year-old man, still fully ambulatory, presents with motor weakness and wasting of legs, forearms and hands. At the age of 10, the first symptoms encompass i.e., tiptoe walking, pes cavovarus and claw toes, which required multiple surgeries.

DISTAL MYOPATHIES: EP.76 Mutations in the prion-like domain of heterogenous nuclear ribonuclearprotein A1 (HNRNPA1) cause proximal and distal myopathies

DISTAL MYOPATHIES: EP.76 Mutations in the prion-like domain of heterogenous nuclear ribonuclearprotein A1 (HNRNPA1) cause proximal and distal myopathies

DISTAL MYOPATHIES: EP.74 GNE pathogenic variant p.D207V rarely leads to myopathy in homozygotes; GNE might not be the only pathogenic determinant of GNE myopathy

DISTAL MYOPATHIES: EP.74 GNE pathogenic variant p.D207V rarely leads to myopathy in homozygotes; GNE might not be the only pathogenic determinant of GNE myopathy

DISTAL MYOPATHIES: EP.71 Long-term evaluation parameters and complications in GNE myopathy: a five-year observational follow-up natural history study

DISTAL MYOPATHIES: EP.71 Long-term evaluation parameters and complications in GNE myopathy: a five-year observational follow-up natural history study

DISTAL MYOPATHIES: EP.73 Effects of SMPX on stress granule dynamics

DISTAL MYOPATHIES: EP.73 Effects of SMPX on stress granule dynamics

DISTAL MYOPATHIES: EP.72 Isoform specific variant in MLIP as a potential cause of adult-onset distal myopathy

Despite recent advancements in next-generation DNA-sequencing technologies, at least 25% of myopathy patients remain without a genetic diagnosis. This represents a major burden to both the patient and the healthcare system. In order to increase diagnostic yield, we combined omic technologies to identify novel gene mutations for functional studies in a cohort of 17 patients with rare undiagnosed myopathies. Among those, we have identified a homozygous nonsense variant in the MLIP gene encoding muscular LMNA-interacting protein (MLIP) in a patient affected with an adult-onset distal myopathy.

Prospects for the Etiotropic Treatment of Dysferlinopathy

Dysferlinopathies belong to a phenotypically heterogeneous group of neuromuscular diseases caused by mutations in the DYSF gene, which disrupt the expression of dysferlin protein in human skeletal muscle cells. These pathologies are of an autosomal recessive inheritance pattern, their prevalence is 1: 200000. Dysferlinopathies include diseases such as Miyoshi myopathy with primary lesion of the distal fragments of the lower extremities and limb-gridle muscular dystrophy type 2B with primary lesion of the proximal fragments of both the lower and upper limbs, also distal myopathy with anterior tibial onset (DMAT). Nowdays, there are various pathogenetic and symptomatic treatments for hereditary muscular dystrophies but there are very few registered drugs for the etiological treatment of these diseases. This review discusses the main modern methods of gene therapy that can be used to treat dysferlinopathies, such as stop-codon passing, exon skipping, overexpression of other genes, gene transfer, splicosome-mediated trans-splicing, and also describes the latest experimental studies using these methods. In conclusion, exon-skipping and trans-splicing have been identified as the most optimal approaches in the treatment of muscular dystrophies, in particular dysferlinopathies.

Out-of-Frame Mutations in ACTN2 Last Exon Cause a Dominant Distal Myopathy With Facial Weakness.

Background and ObjectivesTo clinically, genetically, and histopathologically characterize patients presenting with an unusual combination of distal myopathy and facial weakness, without involvement of upper limb or shoulder girdle muscles.MethodsTwo families with a novel form of actininopathy were identified. Patients had been followed up over 10 years. Their molecular genetic diagnosis was not clear after extensive investigations, including analysis of candidate genes and FSHD1-related D4Z4 repeats.ResultsPatients shared a similar clinical phenotype and a common pattern of muscle involvement. They presented with a very slowly progressive myopathy involving anterior lower leg and facial muscles. Muscle MRI finding showed complete fat replacement of anterolateral compartment muscles of the lower legs with variable involvement of soleus and gastrocnemius but sparing thigh muscles. Muscle biopsy showed internalized nuclei, myofibrillar disorganization, and rimmed vacuoles. High-throughput sequencing identified in each proband a heterozygous single nucleotide deletion (c.2558del and c.2567del) in the last exon of the ACTN2 gene. The deletions are predicted to lead to a novel but unstructured slightly extended C-terminal amino acid sequence.DiscussionOur findings indicate an unusual form of actininopathy with specific molecular and clinical features. Actininopathy should be considered in the differential diagnosis of distal myopathy combined with facial weakness.

An Integrated Clinical-Biological Approach to Identify Interindividual Variability and Atypical Phenotype-Genotype Correlations in Myopathies: Experience on A Cohort of 156 Families.

Diagnosis of myopathies is challenged by the high genetic heterogeneity and clinical overlap of the various etiologies. We previously reported a Next-Generation Sequencing strategy to identify genetic etiology in patients with undiagnosed Limb-Girdle Muscular Dystrophies, Congenital Myopathies, Congenital Muscular Dystrophies, Distal Myopathies, Myofibrillar Myopathies, and hyperCKemia or effort intolerance, using a large gene panel including genes classically associated with other entry diagnostic categories. In this study, we report the comprehensive clinical-biological strategy used to interpret NGS data in a cohort of 156 pediatric and adult patients, that included Copy Number Variants search, variants filtering and interpretation according to ACMG guidelines, segregation studies, deep phenotyping of patients and relatives, transcripts and protein studies, and multidisciplinary meetings. Genetic etiology was identified in 74 patients, a diagnostic yield (47.4%) similar to previous studies. We identified 18 patients (10%) with causative variants in different genes (ACTA1, RYR1, NEB, TTN, TRIP4, CACNA1S, FLNC, TNNT1, and PAPBN1) that resulted in milder and/or atypical phenotypes, with high intrafamilial variability in some cases. Mild phenotypes could mostly be explained by a less deleterious effect of variants on the protein. Detection of inter-individual variability and atypical phenotype-genotype associations is essential for precision medicine, patient care, and to progress in the understanding of the molecular mechanisms of myopathies.

P 45. Different K+-release in distal myopathy and motor neuropathy during non-ischemic exercise

Background. In myotonic dystrophy, a significantly increased potassium release upon exercise was previously described. However, it remained unclear so far whether this is a specific feature of myotonic dystrophies or a consequence of distal muscular weakness in general. Methods. We performed non-ischemic forearm exercise test (NIFET) and measured venous K + concentration at rest and at three different force levels (20–30%, 50–60%, 70–80%) related to maximal contraction force (MCF) in patients with different myopathies ( n = 7), motor neuropathies ( n = 8) with distal phenotype and healthy volunteers ( n = 12). The specific K + production was defined as venous K+ increase related to workload. Results. Maximal contraction force, workload and venous K + increase were lower at all force levels in both myopathy and neuropathy groups compared to the control group. With increasing workload, the control group showed a quasi-linear increase in K + release, resulting in a nearly constant specific K + production within the three force levels ( p = 0.752, Kruskal-Wallis-Test). In contrast, in myopathies a higher specific K + production was measured at low force levels (20-30% MCF) in comparison to higher force levels ( p = 0.015, Kruskal-Wallis test). In neuropathies, the lowest specific K + production was obtained at 20–30% MCF. At this force level, the specific K + production was significantly higher in myopathies compared to neuropathies (Fig. 1; p = 0.005, Mann-Whitney U Test). At 50–60% and 70–80% MCF, the specific K + values increasingly converged and did not significantly differ between the three groups (Fig. 1; p = 0.09 and p = 0.37, Kruskal-Wallis test). Conclusion. In our study, increased K + efflux related to workload occurred in patients with myogenic in comparison to neurogenic distal muscular weakness at low force levels. High specific K + production does not seem to be an exclusive feature of myotonic dystrophies but of myogenic muscular weakness in general. Our results point to a different regulation of K+ balance in neurogenic and myogenic muscular weakness possibly due to a different recruitment behaviour of motor units and the action potential frequency of motor neurons in neurogenic and myogenic processes. Fig. 1.

Cybernic treatment with wearable cyborg Hybrid Assistive Limb (HAL) improves ambulatory function in patients with slowly progressive rare neuromuscular diseases: a multicentre, randomised, controlled crossover trial for efficacy and safety (NCY-3001)

BackgroundRare neuromuscular diseases such as spinal muscular atrophy, spinal bulbar muscular atrophy, muscular dystrophy, Charcot-Marie-Tooth disease, distal myopathy, sporadic inclusion body myositis, congenital myopathy, and amyotrophic lateral sclerosis lead to incurable amyotrophy and consequent loss of ambulation. Thus far, no therapeutic approaches have been successful in recovering the ambulatory ability. Thus, the aim of this trial was to evaluate the efficacy and safety of cybernic treatment with a wearable cyborg Hybrid Assistive Limb (HAL, Lower Limb Type) in improving the ambulatory function in those patients.ResultsWe conducted an open-label, randomised, controlled crossover trial to test HAL at nine hospitals between March 6, 2013 and August 8, 2014. Eligible patients were older than 18 years and had a diagnosis of neuromuscular disease as specified above. They were unable to walk for 10 m independently and had neither respiratory failure nor rapid deterioration in gait. The primary endpoint was the distance passed during a two-minute walk test (2MWT). The secondary endpoints were walking speed, cadence, and step length during the 10-m walk test (10MWT), muscle strength by manual muscle testing (MMT), and a series of functional measures. Adverse events and failures/problems/errors with HAL were also evaluated. Thirty patients were randomly assigned to groups A or B, with each group of 15 receiving both treatments in a crossover design. The efficacy of a 40-min walking program performed nine times was compared between HAL plus a hoist and a hoist only. The final analysis included 13 and 11 patients in groups A and B, respectively. Cybernic treatment with HAL resulted in a 10.066% significantly improved distance in 2MWT (95% confidence interval, 0.667–19.464; p = 0.0369) compared with the hoist only treatment. Among the secondary endpoints, the total scores of MMT and cadence at 10MWT were the only ones that showed significant improvement. The only adverse effects were slight to mild myalgia, back pain, and contact skin troubles, which were easily remedied.ConclusionsHAL is a new treatment device for walking exercise, proven to be more effective than the conventional method in patients with incurable neuromuscular diseases.Trial registration: JMACTR, JMA-IIA00156

THE CHALLENGING DIAGNOSIS OF DYSFERLINOPATHY – A CASE REPORT

Objectives. Dysferlinopathies are a group of rare genetic myopathies characterized by muscle weakness and atrophy with four distinct clinical phenotypes: Miyoshi myopathy, limb girdle muscular dystrophy type 2B, distal myopathy with anterior tibial onset and an intermediate proximo-distal phenotype. We report a case of dysferlinopathy and discuss relevant clinical, pathological and genetic data. Material and methods. We present the case of a 36 years old man with more than ten years history of progressive muscle weakness and atrophies in the distal lower limbs. He had a first clinical evaluation at the age of 24 years and a muscle biopsy was performed but it was inconclusive. On current admission, he presented with clinical suspicion of Charcot-Marie-Tooth disease, but because of increased serum creatine kinase levels and EMG (electromyography) findings of myopathy, another muscle biopsy was recommended. The muscle tissue obtained was analysed by histopathology, immunohistochemistry, and Western blot techniques, and also a genetic confirmation was recommended. Results. Based on morphological findings, electrodiagnostic study results and clinical context, the diagnosis of primary dysferlinopathy was established, supported by genetic data. Conclusions. The diagnosis may be challenging in these rare genetic myopathies, due to the high variability of phenotypes, ranging from asymptomatic hyperCKemia (high serum creatine kinase level) to a severe clinical picture with loss of ambulation. Repeated evaluation, increasing access to genetic testing and a multidisciplinary approach made an accurate diagnosis possible in our case.

Generation and Analysis of Genetically Modified CD14+ Monocytes with Dysferlin Overexpression

IntroductionDysferlinopathies is a group of orphan neuromuscular diseases associated with impairment of mRNA and defect in dysferlin protein expression in skeletal muscle tissue due to mutations in DYSF (Dystrophy-associated fer-1-like) gene. This group of diseases is having autosomal recessive type of inheritance and the frequency of occurrence is 1/1,300-1/200,000 in different populations. There are three main clinical phenotypes associated with dysferlin gene mutations: Limb girdle muscular dystrophy type 2B (LGMD2B), Miyoshi myopathy (MM) and distal myopathy with anterior tibial onset (DMAT) (Yakovlev et. al. Human Gene Therapy 2016). Currently there is no treatment for these diseases. Hence, the development of new methods of treating this group of diseases remains an urgent task, in particular, with the use of gene therapy. According to the literature, CD14+ cells play a key role in the pathogenesis of human dysferlinopathies and are involved in inflammatory reactions and pathological angiogenesis.MethodsIn the present work, by using previously created recombinant adenovirus Ad5-Dysf (Starostina et. al. Medicine 2016) that encodes human cDNA dysferlin gene and enables the expression of the recombinant dysferlin protein, genetic modification of CD14 + monocytes was carried out. Therefore, the following aims were set:1. Isolation of CD14+ cells from a patient with dysferlinopathy and a healthy donor.2. Genetic modification of CD14+ monocytes.3. Conduct western blot analysis of mRNA expression of recombinant dysferlin protein in modified cells after viral transduction. Evaluate the effectiveness of viral transduction by analyzing the expression of EGFP protein (adenovirus Ad5-EGFP) which was taken as a positive control by fluorescence microscopy.ResultsCD14+ cells were isolated from the freshly prepared peripheral blood leukocyte buffy coat layer of the patient with dysferlinopathy and a healthy donor by use of EasySep ™ Human Buffy Coat CD14 Positive Selection Kit. Since the correct assembly and effective DYSF gene delivery of the adenovirus Ad5-Dysf was confirmed in previous work, the viral transduction of human monocytes was carried out successfully. Ad5-EGFP adenovirus was used as a positive control to evaluate the effectiveness of viral transduction. EGFP protein expression analysis was conducted by fluorescence microscopy on an inverted fluorescent microscope of the research class AxioObserver.Z1 (Carl Zeiss) and demonstrated 100% of EGFP protein expression (Fig. 1). After genetic modification of the cells with resulting Ad5-Dysf adenovirus the evaluation of the recombinant dysferlin protein expression in cells was carried out by western blot analysis. Positive reaction was revealed with rabbit polyclonal antibodies to dysferlin (Fig. 2).ConclusionsIn this work we demonstrated that genetically modified CD14+ cells with DYSF overexpression can be successfully geberated and considered as one of the approaches for the therapy of human dysferlinopathies. Future studies are needed to analyze the functionality of the obtained genetic construct in animal models. SVV was supported by RFBR grant 16-34-00657.Fig. 1. CD14+ monocytes genetically modified with Ad5-EGFP. A - phase-contrast microscopy. B - fluorescence microscopy. 48 hours after genetic modification. Magnification 200X.Fig. 2. Western blot analysis of DYSF protein expression in CD14+ monocytes isolated from the peripheral blood of a patient with dysferlinopathy. Electrophoresis in 12% SDS-PAGE gel according to the Laemmli system. Antibodies to dysferlin (Abcam, ab15108) were used in a dilution of 1:250. The expected size of DYSF protein is 231 kDa. Well 1 - transduced CD14+ monocytes with Ad5-Dysf. Well 2 - native CD14+ monocytes. M - molecular weight marker. [Display omitted] DisclosuresNo relevant conflicts of interest to declare.