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1.
Brain ; 144(2): 574-583, 2021 03 03.
Artigo em Inglês | MEDLINE | ID: mdl-33459760

RESUMO

The von Willebrand Factor A domain containing 1 protein, encoded by VWA1, is an extracellular matrix protein expressed in muscle and peripheral nerve. It interacts with collagen VI and perlecan, two proteins that are affected in hereditary neuromuscular disorders. Lack of VWA1 is known to compromise peripheral nerves in a Vwa1 knock-out mouse model. Exome sequencing led us to identify bi-allelic loss of function variants in VWA1 as the molecular cause underlying a so far genetically undefined neuromuscular disorder. We detected six different truncating variants in 15 affected individuals from six families of German, Arabic, and Roma descent. Disease manifested in childhood or adulthood with proximal and distal muscle weakness predominantly of the lower limbs. Myopathological and neurophysiological findings were indicative of combined neurogenic and myopathic pathology. Early childhood foot deformity was frequent, but no sensory signs were observed. Our findings establish VWA1 as a new disease gene confidently implicated in this autosomal recessive neuromyopathic condition presenting with child-/adult-onset muscle weakness as a key clinical feature.


Assuntos
Proteínas da Matriz Extracelular/genética , Doenças Neuromusculares/genética , Adolescente , Adulto , Criança , Feminino , Predisposição Genética para Doença , Humanos , Masculino , Pessoa de Meia-Idade , Músculo Esquelético/patologia , Mutação , Doenças Neuromusculares/patologia , Linhagem , Sequenciamento Completo do Exoma
2.
Nat Commun ; 11(1): 4625, 2020 09 15.
Artigo em Inglês | MEDLINE | ID: mdl-32934225

RESUMO

A hallmark of neurodegeneration is defective protein quality control. The E3 ligase Listerin (LTN1/Ltn1) acts in a specialized protein quality control pathway-Ribosome-associated Quality Control (RQC)-by mediating proteolytic targeting of incomplete polypeptides produced by ribosome stalling, and Ltn1 mutation leads to neurodegeneration in mice. Whether neurodegeneration results from defective RQC and whether defective RQC contributes to human disease have remained unknown. Here we show that three independently-generated mouse models with mutations in a different component of the RQC complex, NEMF/Rqc2, develop progressive motor neuron degeneration. Equivalent mutations in yeast Rqc2 selectively interfere with its ability to modify aberrant translation products with C-terminal tails which assist with RQC-mediated protein degradation, suggesting a pathomechanism. Finally, we identify NEMF mutations expected to interfere with function in patients from seven families presenting juvenile neuromuscular disease. These uncover NEMF's role in translational homeostasis in the nervous system and implicate RQC dysfunction in causing neurodegeneration.


Assuntos
Doenças Neuromusculares/metabolismo , Ribossomos/metabolismo , Sequência de Aminoácidos , Animais , Feminino , Humanos , Masculino , Camundongos , Camundongos Knockout , Mutação , Doenças Neuromusculares/genética , Doenças Neuromusculares/patologia , Proteólise , Proteínas de Ligação a RNA/química , Proteínas de Ligação a RNA/genética , Proteínas de Ligação a RNA/metabolismo , Ribossomos/genética , Saccharomyces cerevisiae/genética , Saccharomyces cerevisiae/metabolismo , Proteínas de Saccharomyces cerevisiae/química , Proteínas de Saccharomyces cerevisiae/genética , Proteínas de Saccharomyces cerevisiae/metabolismo , Alinhamento de Sequência
3.
J Neuropathol Exp Neurol ; 79(7): 719-733, 2020 07 01.
Artigo em Inglês | MEDLINE | ID: mdl-32529201

RESUMO

Competence in muscle biopsy evaluation is a core component of neuropathology practice. The practicing neuropathologist should be able to prepare frozen sections of muscle biopsies with minimal artifacts and identify key histopathologic features of neuromuscular disease in hematoxylin and eosin-stained sections as well as implement and interpret a basic panel of additional histochemical, enzyme histochemical, and immunohistochemical stains. Important to everyday practice is a working knowledge of normal muscle histology at different ages, muscle motor units, pitfalls of myotendinous junctions, nonpathologic variations encountered at traditional and nontraditional muscle sites, the pathophysiology of myonecrosis and regeneration, and approaches to distinguish muscular dystrophies from inflammatory myopathies and other necrotizing myopathies. Here, we provide a brief overview of what every neuropathologist needs to know concerning the muscle biopsy.


Assuntos
Músculo Esquelético/patologia , Músculo Liso/patologia , Doenças Neuromusculares/patologia , Neuropatologia/normas , Biópsia/métodos , Biópsia/normas , Humanos , Neuropatologia/educação , Neuropatologia/métodos
4.
J Clin Neurophysiol ; 37(3): 197-199, 2020 May.
Artigo em Inglês | MEDLINE | ID: mdl-32358244

RESUMO

There is extensive evidence in the literature that both peripheral nerve fibers and muscle fibers are affected in the course of intensive care unit-acquired weakness. Peripheral nerve lesion is characterized by axonal degeneration, without inflammatory changes. Muscle fiber involvement is characterized by muscle fiber atrophy and loss of thick filaments, predominantly involving type 2 fibers, but myonecrosis ("acute necrotizing myopathy of intensive care") has also been reported. Steroids can precipitate thick myofilament damage, probably to some extent also triggered by immobilization and neuromuscular junction blockade. Sepsis and a systemic inflammatory response cause muscle fiber injury because of the release of cytokines and chemokines that modulate enzymatic reactions related to proteolysis. Regarding axonal injury, hyperglycemia, hypoalbuminemia, inflammatory response, and hypoperfusion are accepted risk factors. Nerve and muscle biopsy are the best methods for detection of structural abnormalities, but these are invasive investigations; although not suitable for repeated studies, in selected cases, biopsies may have a role in diagnosis.


Assuntos
Cuidados Críticos , Debilidade Muscular/etiologia , Debilidade Muscular/patologia , Doenças do Sistema Nervoso Periférico/etiologia , Doenças do Sistema Nervoso Periférico/patologia , Cuidados Críticos/métodos , Humanos , Unidades de Terapia Intensiva , Fibras Musculares Esqueléticas/patologia , Doenças Neuromusculares/etiologia , Doenças Neuromusculares/patologia , Neuropatologia , Nervos Periféricos/patologia , Fatores de Risco
5.
Nat Protoc ; 15(2): 421-449, 2020 02.
Artigo em Inglês | MEDLINE | ID: mdl-31932771

RESUMO

This protocol describes the design, fabrication and use of a 3D physiological and pathophysiological motor unit model consisting of motor neurons coupled to skeletal muscles interacting via the neuromuscular junction (NMJ) within a microfluidic device. This model facilitates imaging and quantitative functional assessment. The 'NMJ chip' enables real-time, live imaging of axonal outgrowth, NMJ formation and muscle maturation, as well as synchronization of motor neuron activity and muscle contraction under optogenetic control for the study of normal physiological events. The proposed protocol takes ~2-3 months to be implemented. Pathological behaviors associated with various neuromuscular diseases, such as regression of motor neuron axons, motor neuron death, and muscle degradation and atrophy can also be recapitulated in this system. Disease models can be created by the use of patient-derived induced pluripotent stem cells to generate both the motor neurons and skeletal muscle cells used. This is demonstrated by the use of cells from a patient with sporadic amyotrophic lateral sclerosis but can be applied more generally to models of neuromuscular disease, such as spinal muscular atrophy, NMJ disorder and muscular dystrophy. Models such as this hold considerable potential for applications in precision medicine, drug screening and disease risk assessment.


Assuntos
Avaliação Pré-Clínica de Medicamentos/instrumentação , Procedimentos Analíticos em Microchip/métodos , Doenças Neuromusculares/tratamento farmacológico , Medicina de Precisão/instrumentação , Humanos , Contração Muscular/efeitos dos fármacos , Músculo Esquelético/efeitos dos fármacos , Músculo Esquelético/patologia , Músculo Esquelético/fisiopatologia , Doenças Neuromusculares/patologia , Doenças Neuromusculares/fisiopatologia , Junção Neuromuscular/efeitos dos fármacos , Junção Neuromuscular/patologia , Neurônios/efeitos dos fármacos , Neurônios/patologia , Medição de Risco
7.
Muscle Nerve ; 60(6): 687-692, 2019 12.
Artigo em Inglês | MEDLINE | ID: mdl-31478199

RESUMO

INTRODUCTION: Nerve imaging has a limited role in axonal and muscle fiber loss. In this study, we sought to explore the utility of standardized muscle ultrasound (US) assessment in these clinical scenarios. METHODS: We performed a prospective study from March to August 2018 of patients attending the neuromuscular clinic. All patients underwent clinical evaluation and standardized muscle thickness measurement by US in seven muscles. RESULTS: The study cohort consisted of 114 participants, including patients with polyneuropathy, motor neuron disease, and myopathy. The smallest distal muscle thickness was found in patients with polyneuropathy, while the smallest proximal muscle thickness was found in patients with myopathy. Muscle thickness was strongly correlated with muscle strength (r 2 = 0.62), electrophysiological findings (r 2 : 0.44-0.55), and disability score (r 2 = 0.53). DISCUSSION: Standardized muscle thickness measured by US shows diagnostic usefulness in a spectrum of neuromuscular disorders and correlates with clinical and electrophysiological findings.


Assuntos
Músculo Esquelético/diagnóstico por imagem , Doenças Neuromusculares/diagnóstico por imagem , Potenciais de Ação/fisiologia , Adulto , Idoso , Esclerose Amiotrófica Lateral/diagnóstico por imagem , Esclerose Amiotrófica Lateral/patologia , Esclerose Amiotrófica Lateral/fisiopatologia , Eletromiografia , Feminino , Humanos , Masculino , Sintomas Inexplicáveis , Pessoa de Meia-Idade , Força Muscular , Músculo Esquelético/patologia , Músculo Esquelético/fisiopatologia , Atrofia Muscular Espinal/diagnóstico por imagem , Atrofia Muscular Espinal/patologia , Atrofia Muscular Espinal/fisiopatologia , Doenças Musculares/diagnóstico por imagem , Doenças Musculares/patologia , Doenças Musculares/fisiopatologia , Condução Nervosa/fisiologia , Doenças Neuromusculares/patologia , Doenças Neuromusculares/fisiopatologia , Tamanho do Órgão , Polineuropatias/diagnóstico por imagem , Polineuropatias/patologia , Polineuropatias/fisiopatologia , Estudos Prospectivos , Radiculopatia/diagnóstico por imagem , Radiculopatia/patologia , Radiculopatia/fisiopatologia , Ultrassonografia
8.
Nutrients ; 11(8)2019 Aug 13.
Artigo em Inglês | MEDLINE | ID: mdl-31412596

RESUMO

Autophagy is the major intracellular machinery for degrading proteins, lipids, polysaccharides, and organelles. This cellular process is essential for the maintenance of the correct cellular balance in both physiological and stress conditions. Because of its role in maintaining cellular homeostasis, dysregulation of autophagy leads to various disease manifestations, such as inflammation, metabolic alterations, aging, and neurodegeneration. A common feature of many neurologic and neuromuscular diseases is the alteration of the autophagy-lysosomal pathways. For this reason, autophagy is considered a target for the prevention and/or cure of these diseases. Dietary intake of polyphenols has been demonstrated to prevent/ameliorate several of these diseases. Thus, natural products that can modulate the autophagy machinery are considered a promising therapeutic strategy. In particular, curcumin, a phenolic compound widely used as a dietary supplement, exerts an important effect in modulating autophagy. Herein, we report on the current knowledge concerning the role of curcumin in modulating the autophagy machinery in various neurological and neuromuscular diseases as well as its role in restoring the autophagy molecular mechanism in several cell types that have different effects on the progression of neurological and neuromuscular disorders.


Assuntos
Autofagia/efeitos dos fármacos , Curcumina/uso terapêutico , Suplementos Nutricionais , Músculo Esquelético/efeitos dos fármacos , Doenças do Sistema Nervoso/tratamento farmacológico , Sistema Nervoso/efeitos dos fármacos , Doenças Neuromusculares/tratamento farmacológico , Animais , Proteínas Relacionadas à Autofagia/metabolismo , Curcumina/efeitos adversos , Suplementos Nutricionais/efeitos adversos , Humanos , Músculo Esquelético/metabolismo , Músculo Esquelético/patologia , Sistema Nervoso/metabolismo , Sistema Nervoso/patologia , Doenças do Sistema Nervoso/metabolismo , Doenças do Sistema Nervoso/patologia , Doenças Neuromusculares/metabolismo , Doenças Neuromusculares/patologia , Transdução de Sinais
9.
J R Soc Interface ; 16(157): 20190402, 2019 08 30.
Artigo em Inglês | MEDLINE | ID: mdl-31431186

RESUMO

Physics-based predictive simulations of human movement have the potential to support personalized medicine, but large computational costs and difficulties to model control strategies have limited their use. We have developed a computationally efficient optimal control framework to predict human gaits based on optimization of a performance criterion without relying on experimental data. The framework generates three-dimensional muscle-driven simulations in 36 min on average-more than 20 times faster than existing simulations-by using direct collocation, implicit differential equations and algorithmic differentiation. Using this framework, we identified a multi-objective performance criterion combining energy and effort considerations that produces physiologically realistic walking gaits. The same criterion also predicted the walk-to-run transition and clinical gait deficiencies caused by muscle weakness and prosthesis use, suggesting that diverse healthy and pathological gaits can emerge from the same control strategy. The ability to predict the mechanics and energetics of a broad range of gaits with complex three-dimensional musculoskeletal models will allow testing novel hypotheses about gait control and hasten the development of optimal treatments for neuro-musculoskeletal disorders.


Assuntos
Simulação por Computador , Marcha/fisiologia , Modelos Biológicos , Fenômenos Biomecânicos , Humanos , Doenças Neuromusculares/patologia
10.
Elife ; 82019 07 31.
Artigo em Inglês | MEDLINE | ID: mdl-31364990

RESUMO

Cytoplasmic dynein plays critical roles within the developing and mature nervous systems, including effecting nuclear migration, and retrograde transport of various cargos. Unsurprisingly, mutations in dynein are causative of various developmental neuropathies and motor neuron diseases. These 'dyneinopathies' define a broad spectrum of diseases with no known correlation between mutation identity and disease state. To circumvent complications associated with dynein studies in human cells, we employed budding yeast as a screening platform to characterize the motility properties of seventeen disease-correlated dynein mutants. Using this system, we determined the molecular basis for several classes of etiologically related diseases. Moreover, by engineering compensatory mutations, we alleviated the mutant phenotypes in two of these cases, one of which we confirmed with recombinant human dynein. In addition to revealing molecular insight into dynein regulation, our data provide additional evidence that the type of disease may in fact be dictated by the degree of dynein dysfunction.


Assuntos
Dineínas/genética , Dineínas/metabolismo , Regulação da Expressão Gênica , Proteínas Mutantes/genética , Proteínas Mutantes/metabolismo , Doenças Neuromusculares/patologia , Doenças Neuromusculares/fisiopatologia , Humanos , Modelos Teóricos , Fenótipo , Saccharomyces cerevisiae/genética , Saccharomyces cerevisiae/metabolismo , Supressão Genética
11.
NMR Biomed ; 32(8): e4111, 2019 08.
Artigo em Inglês | MEDLINE | ID: mdl-31180167

RESUMO

Quantitative imaging techniques are emerging in the field of magnetic resonance imaging of neuromuscular diseases (NMD). T2 of water (T2w ) is considered an important imaging marker to assess acute and chronic alterations of the muscle fibers, being generally interpreted as an indicator for "disease activity" in the muscle tissue. To validate the accuracy and robustness of quantitative imaging methods, 1 H magnetic resonance spectroscopy (MRS) can be used as a gold standard. The purpose of the present work was to investigate T2w of remaining muscle tissue in regions of higher proton density fat fraction (PDFF) in 40 patients with defined NMD using multi-TE single-voxel 1 H MRS. Patients underwent MR measurements on a 3 T system to perform a multi-TE single-voxel stimulated echo acquisition method (STEAM) MRS (TE = 11/15/20/25(/35) ms) in regions of healthy, edematous and fatty thigh muscle tissue. Muscle regions for MRS were selected based on T2 -weighted water and fat images of a two-echo 2D Dixon TSE. MRS results were confined to regions with qualitatively defined remaining muscle tissue without edema and high fat content, based on visual grading of the imaging data. The results showed decreased T2w values with increasing PDFF with R2  = 0.45 (p < 10-3 ) (linear fit) and with R2  = 0.51 (exponential fit). The observed dependence of T2w on PDFF should be considered when using T2w as a marker in NMD imaging and when performing single-voxel MRS for T2w in regions enclosing edematous, nonedematous and fatty infiltrated muscle tissue.


Assuntos
Tecido Adiposo/patologia , Músculo Esquelético/patologia , Doenças Neuromusculares/patologia , Água/química , Adulto , Idoso , Idoso de 80 Anos ou mais , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Prótons
12.
J Surg Res ; 243: 27-32, 2019 11.
Artigo em Inglês | MEDLINE | ID: mdl-31151034

RESUMO

BACKGROUND: An operative biopsy is an important component in the diagnosis and treatment of neuromuscular disorders (NMDs). However, recent advances in molecular genetics suggest less invasive genetic testing should be the initial approach. The purpose of our study was to demonstrate the diagnostic value of muscle or nerve biopsy within the pediatric population at a pediatric academic center and offer recommendations for genetic testing in relation to biopsy to achieve the highest diagnostic yield. METHODS: Following institutional review board approval, we retrospectively reviewed the electronic medical record of 221 pediatric patients who underwent muscle and/or nerve biopsy for suspicion of NMD from January 2007 to March 2018. Demographics, family history, clinical presentations, genetic testing results, pathology results, anesthesia complications, clinical diagnoses, and clinic follow-up data were collected. Chi-square analysis was done for statistical significance. RESULTS: A total of 220 underwent muscle biopsy, and 15 underwent nerve biopsy. Not all patients received genetic testing. The average age at biopsy was 7.7 y. Biopsy revealed significant histologic abnormalities in 62.9% (139), directly leading to a specific clinical diagnosis in 33.9% (75). When genetic testing was done before biopsy, definite pathogenic variants were found in 7.6% (9). When genetic testing was done after biopsy, definite pathogenic variants were found in 45.0% (27). Genetic testing yield for pathogenic variants was higher when done after biopsy (P value < 0.00001). CONCLUSIONS: Muscle and nerve biopsies may provide significant diagnostic value. Biopsy helped to rule in or out NMD and guide genetic testing. Our data suggest NMD genetic testing yield was higher when done after biopsy.


Assuntos
Testes Genéticos , Músculo Esquelético/patologia , Doenças Neuromusculares/diagnóstico , Nervos Periféricos/patologia , Biópsia , Criança , Feminino , Seguimentos , Humanos , Masculino , Doenças Neuromusculares/genética , Doenças Neuromusculares/patologia , Estudos Retrospectivos
13.
Am J Hum Genet ; 104(5): 847-860, 2019 05 02.
Artigo em Inglês | MEDLINE | ID: mdl-31051113

RESUMO

Collagen type IV alpha 1 and alpha 2 chains form heterotrimers ([α1(IV)]2α2(IV)) that represent a fundamental basement membrane constituent. Dominant COL4A1 and COL4A2 mutations cause a multisystem disorder that is marked by clinical heterogeneity and variable expressivity and that is generally characterized by the presence of cerebrovascular disease with ocular, renal, and muscular involvement. Despite the fact that muscle pathology is reported in up to one-third of individuals with COL4A1 and COL4A2 mutations and in animal models with mutations in COL4A1 and COL4A2 orthologs, the pathophysiological mechanisms underlying COL4A1-related myopathy are unknown. In general, mutations are thought to impair [α1(IV)]2α2(IV) secretion. Whether pathogenesis results from intracellular retention, extracellular deficiency, or the presence of mutant proteins in basement membranes represents an important gap in knowledge and a major obstacle for developing targeted interventions. We report that Col4a1 mutant mice develop progressive neuromuscular pathology that models human disease. We demonstrate that independent muscular, neural, and vascular insults contribute to neuromyopathy and that there is mechanistic heterogeneity among tissues. Importantly, we provide evidence of a COL4A1 functional subdomain with disproportionate significance for tissue-specific pathology and demonstrate that a potential therapeutic strategy aimed at promoting [α1(IV)]2α2(IV) secretion can ameliorate or exacerbate myopathy in a mutation-dependent manner. These data have important translational implications for prediction of clinical outcomes based on genotype, development of mechanism-based interventions, and genetic stratification for clinical trials. Collectively, our data underscore the importance of the [α1(IV)]2α2(IV) network as a multifunctional signaling platform and show that allelic and tissue-specific mechanistic heterogeneities contribute to the variable expressivity of COL4A1 and COL4A2 mutations.


Assuntos
Colágeno Tipo IV/genética , Doenças Musculares/etiologia , Mutação , Doenças Neuromusculares/etiologia , Animais , Feminino , Humanos , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Doenças Musculares/patologia , Doenças Neuromusculares/patologia , Especificidade de Órgãos , Fenótipo
16.
Pediatr Infect Dis J ; 38(7): 667-672, 2019 07.
Artigo em Inglês | MEDLINE | ID: mdl-30985511

RESUMO

BACKGROUND: Acute flaccid myelitis (AFM) is defined as an acute onset of limb weakness with longitudinal spinal gray matter lesions. Reporting bias and misdiagnosis confound epidemiologic studies of AFM. We mitigated these confounders by using a large data set to assess AFM incidence, risk factors and outcomes in a fixed population. METHODS: A retrospective cohort study was conducted within Kaiser Permanente Northern California population among children 1-18 years. Cases met radiographic and clinical criteria for AFM and were confirmed by two clinicians. Clinical and demographic data were assessed. RESULTS: A total of 28 patients met study criteria for AFM between January 1, 2011 and December 31, 2016, an overall rate of 1.46 per 100,000 person-years. Incidence increased from 0.30 to 1.43 cases/per 100,000 person-years between 2011 and 2016, respectively. Median age was 9 years. Risk factors included male sex, Asian ancestry and history of asthma, atopic dermatitis or head injury. Risk factors associated with poliomyelitis were absent. Prodromal illness was common; enterovirus was the most common pathogen detected (n = 5). Among the 27 patients with 12-month follow-up, most demonstrated some improvement, 11 (41.0%) had full recovery, but several had significant deficits with one death reported after the study period. CONCLUSIONS: We employed a closed-population study to generate AFM incidence, risk and outcome data. Our findings support previous reports of male sex and atopy as risk factors. Interval increase in incidence, predisposing Asian ancestry and history of head injury were unique findings to this study. Overall prognosis was better than prior reports, but recovery was incomplete in several patients.


Assuntos
Viroses do Sistema Nervoso Central/epidemiologia , Viroses do Sistema Nervoso Central/patologia , Infecções por Enterovirus/epidemiologia , Infecções por Enterovirus/patologia , Enterovirus/isolamento & purificação , Mielite/epidemiologia , Mielite/patologia , Doenças Neuromusculares/epidemiologia , Doenças Neuromusculares/patologia , Adolescente , California/epidemiologia , Criança , Pré-Escolar , Enterovirus/classificação , Feminino , Humanos , Incidência , Lactente , Masculino , Estudos Retrospectivos , Fatores de Risco , Resultado do Tratamento
18.
J Hum Genet ; 64(6): 551-559, 2019 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-30867548

RESUMO

Genetic diagnoses are becoming a routine in the medical practice of neuromuscular diseases. Many diagnoses, however, can have an influence on relatives and family members and thus must be handled carefully by genetic counseling (GC). Here, we aimed to assess the purpose of undergoing GC to verify the utility of collaborations between clinical and genetic divisions. We investigated consecutive GC cases of neuromuscular disease and examined the role of GC. Our study included 102 cases who underwent GC in our hospital from July 2005 to March 2018: 86.3% were women and 45.1% were in their 30's. Disease explanation was the most common reason for attending GC (29.4%), followed by prenatal diagnosis (25.5%), pre-symptomatic diagnosis (17.6%), and carrier diagnosis (14.7%). Clients typically visited the hospital for GC when some kind of life event occurred, such as marriage, had a desire to bear a child, or a change in the condition of the proband. Clinicians should be conscious of such life events from the perspective of both the client and their relatives, and guide the GC at an appropriate time. Overall, the degree of recognition of genetic risk by clients differed; thus, it is important for GC to determine the status of each unique situation and respond individually.


Assuntos
Família , Aconselhamento Genético , Doenças Neuromusculares/diagnóstico , Adulto , Feminino , Triagem de Portadores Genéticos , Testes Genéticos , Humanos , Masculino , Pessoa de Meia-Idade , Doenças Neuromusculares/genética , Doenças Neuromusculares/patologia , Linhagem , Fatores de Risco , Adulto Jovem
19.
Theranostics ; 9(5): 1232-1246, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-30867827

RESUMO

The study of human neuromuscular diseases has traditionally been performed in animal models, due to the difficulty of performing studies in human subjects. Despite the unquestioned value of animal models, inter-species differences hamper the translation of these findings to clinical trials. Tissue-engineered models of the neuromuscular junction (NMJ) allow for the recapitulation of the human physiology in tightly controlled in vitro settings. Methods: Here we report the first human patient-specific tissue-engineered model of the neuromuscular junction (NMJ) that combines stem cell technology with tissue engineering, optogenetics, microfabrication and image processing. The combination of custom-made hardware and software allows for repeated, quantitative measurements of NMJ function in a user-independent manner. Results: We demonstrate the utility of this model for basic and translational research by characterizing in real time the functional changes during physiological and pathological processes. Principal Conclusions: This system holds great potential for the study of neuromuscular diseases and drug screening, allowing for the extraction of quantitative functional data from a human, patient-specific system.


Assuntos
Modelos Teóricos , Doenças Neuromusculares/patologia , Doenças Neuromusculares/fisiopatologia , Optogenética/métodos , Engenharia Tecidual/métodos , Humanos , Células-Tronco Pluripotentes Induzidas/fisiologia , Junção Neuromuscular/patologia , Junção Neuromuscular/fisiologia , Junção Neuromuscular/fisiopatologia
20.
PLoS One ; 14(2): e0212198, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-30794581

RESUMO

INTRODUCTION: The system of clustered regularly interspaced short palindromic repeats (CRISPR) and CRISPR-associated proteins (cas) is a new technology that allows easier manipulation of the genome. Its potential to edit genes opened a new door in treatment development for incurable neurological monogenic diseases (NMGDs). The aim of this systematic review was to summarise the findings on the current development of CRISPR-cas for therapeutic purposes in the most frequent NMGDs and provide critical assessment. METHODS AND DATA ACQUISITION: We searched the MEDLINE and EMBASE databases, looking for original studies on the use of CRISPR-cas to edit pathogenic variants in models of the most frequent NMGDs, until end of 2017. We included all the studies that met the following criteria: 1. Peer-reviewed study report with explicitly described experimental designs; 2. In vitro, ex vivo, or in vivo study using human or other animal biological systems (including cells, tissues, organs, organisms); 3. focusing on CRISPR as the gene-editing method of choice; and 5. featured at least one NMGD. RESULTS: We obtained 404 papers from MEDLINE and 513 from EMBASE. After removing the duplicates, we screened 490 papers by title and abstract and assessed them for eligibility. After reading 50 full-text papers, we finally selected 42 for the review. DISCUSSION: Here we give a systematic summary on the preclinical development of CRISPR-cas for therapeutic purposes in NMGDs. Furthermore, we address the clinical interpretability of the findings, giving a comprehensive overview of the current state of the art. Duchenne's muscular dystrophy (DMD) paves the way forward, with 26 out of 42 studies reporting different strategies on DMD gene editing in different models of the disease. Most of the strategies aimed for permanent exon skipping by deletion with CRISPR-cas. Successful silencing of the mHTT gene with CRISPR-cas led to successful reversal of the neurotoxic effects in the striatum of mouse models of Huntington's disease. Many other strategies have been explored, including epigenetic regulation of gene expression, in cellular and animal models of: myotonic dystrophy, Fraxile X syndrome, ataxias, and other less frequent dystrophies. Still, before even considering the clinical application of CRISPR-cas, three major bottlenecks need to be addressed: efficacy, safety, and delivery of the systems. This requires a collaborative approach in the research community, while having ethical considerations in mind.


Assuntos
Sistemas CRISPR-Cas , Expansão das Repetições de DNA , Edição de Genes/métodos , Técnicas de Transferência de Genes , Doenças Genéticas Inatas , Doenças Neuromusculares , Animais , Modelos Animais de Doenças , Edição de Genes/tendências , Inativação Gênica , Doenças Genéticas Inatas/genética , Doenças Genéticas Inatas/patologia , Doenças Genéticas Inatas/terapia , Humanos , MEDLINE , Camundongos , Doenças Neuromusculares/genética , Doenças Neuromusculares/patologia , Doenças Neuromusculares/terapia
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