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Amyotrophic lateral sclerosis (ALS) is a disease that progressively annihilates spinal cord motor neurons, causing severe motor decline and death. The disease is divided into familial and sporadic ALS. Mutations in the TAR DNA binding protein 43 (TDP-43) have been involved in the pathological emergence and progression of ALS, although the molecular mechanisms eliciting the disease are unknown. Transposable elements (TEs) and DNA sequences capable of transposing within the genome become dysregulated and transcribed in the presence of TDP-43 mutations. We performed RNA-Seq in human motor neurons (iMNs) derived from induced pluripotent stem cells (iPSCs) from TDP-43 wild-type-iMNs-TDP-43WT-and mutant-iMNs-TDP-43M337V-genotypes at 7 and 14 DIV, and, with state-of-the-art bioinformatic tools, analyzed whether TDP-43M337V alters both gene expression and TE activity. Our results show that TDP-43M337V induced global changes in the gene expression and TEs levels at all in vitro stages studied. Interestingly, many genetic pathways overlapped with that of the TEs activity, suggesting that TEs control the expression of several genes. TEs correlated with genes that played key roles in the extracellular matrix and RNA processing: all the regulatory pathways affected in ALS. Thus, the loss of TE regulation is present in TDP-43 mutations and is a critical determinant of the disease in human motor neurons. Overall, our results support the evidence that indicates TEs are critical regulatory sequences contributing to ALS neurodegeneration.
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Esclerose Lateral Amiotrófica , Humanos , Esclerose Lateral Amiotrófica/metabolismo , Elementos de DNA Transponíveis/genética , Neurônios Motores/metabolismo , Mutação , Proteínas de Ligação a DNA/genética , Proteínas de Ligação a DNA/metabolismoRESUMO
The development of cell culture models that recapitulate the etiology and features of nervous system diseases is central to the discovery of new drugs and their translation onto therapies. Neuronal tissues are inaccessible due to skeletal constraints and the invasiveness of the procedure to obtain them. Thus, the emergence of induced pluripotent stem cell (iPSC) technology offers the opportunity to model different neuronal pathologies. Our focus centers on iPSCs derived from amyotrophic lateral sclerosis (ALS) patients, whose pathology remains in urgent need of new drugs and treatment. In this sense, we aim to revise the process to obtain motor neurons derived iPSCs (iPSC-MNs) from patients with ALS as a drug screening model, review current 3D-models and offer a perspective on bioinformatics as a powerful tool that can aid in the progress of finding new pharmacological treatments.
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BACKGROUND: Glutathione S-transferase Pi (GSTP1) enzyme has a major antioxidant effect on the central nervous system (CNS), where it acts against oxidative damage, an established risk factor for amyotrophic lateral sclerosis (ALS). Hence, the purpose of this study was to evaluate a possible relationship between GSTP1 rs1695 polymorphism and the survival rate of male ALS patients, which is the gender more affected by the disease. METHODS AND RESULTS: A case-control study was performed with 56 male ALS patients and 70 healthy male individuals from Midwestern Brazil, which were age-adjusted. GSTP1 rs1695 polymorphism molecular analysis was carried out with restriction fragment length polymorphism. The relationship between ALS patients and GSTP1 rs1695 polymorphism was analyzed using cumulative survival rate as the major outcome, where differences in survival were evaluated through the log-rank test. Our results revealed that mutant genotype (G/G) did not influence the cumulative survival rate of male ALS patients regarding the age of diagnosis (p = 0.5) and time from symptom to diagnosis (p = 0.3). On the other hand, mutant carriers exhibited a significant survival of fewer than 25 months compared to A/A and A/G genotypes that survive more than 100 months (p = 7-E10) in comparison with symptom onset to outcome (p = 0.00006). CONCLUSIONS: In summary, our findings revealed that mutant genotype carriers' male patients had a reduced lifetime, which probably may be resulted from oxidative stress exposure in CNS.
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Esclerose Lateral Amiotrófica/genética , Glutationa S-Transferase pi/metabolismo , Adulto , Esclerose Lateral Amiotrófica/metabolismo , Brasil/epidemiologia , Estudos de Casos e Controles , Estudos de Associação Genética/métodos , Predisposição Genética para Doença , Genótipo , Glutationa S-Transferase pi/genética , Humanos , Masculino , Pessoa de Meia-Idade , Polimorfismo de Fragmento de Restrição , Polimorfismo de Nucleotídeo Único/genética , Fatores de RiscoRESUMO
INTRODUCTION: The use of machine learning (ML) techniques in healthcare encompasses an emerging concept that envisages vast contributions to the tackling of rare diseases. In this scenario, amyotrophic lateral sclerosis (ALS) involves complexities that are yet not demystified. In ALS, the biomedical signals present themselves as potential biomarkers that, when used in tandem with smart algorithms, can be useful to applications within the context of the disease. METHODS: This Systematic Literature Review (SLR) consists of searching for and investigating primary studies that use ML techniques and biomedical signals related to ALS. Following the definition and execution of the SLR protocol, 18 articles met the inclusion, exclusion, and quality assessment criteria, and answered the SLR research questions. DISCUSSIONS: Based on the results, we identified three classes of ML applications combined with biomedical signals in the context of ALS: diagnosis (72.22%), communication (22.22%), and survival prediction (5.56%). CONCLUSIONS: Distinct algorithmic models and biomedical signals have been reported and present promising approaches, regardless of their classes. In summary, this SLR provides an overview of the primary studies analyzed as well as directions for the construction and evolution of technology-based research within the scope of ALS.
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Esclerose Lateral Amiotrófica , Biomarcadores , Progressão da Doença , Humanos , Aprendizado de MáquinaRESUMO
Patients suffering of amyotrophic lateral sclerosis (ALS) present motoneuron degeneration leading to muscle atrophy, dysphagia, and dysarthria. The Wobbler mouse, an animal model of ALS, shows a selective loss of motoneurons, astrocytosis, and microgliosis in the spinal cord. The incidence of ALS is greater in men; however, it increases in women after menopause, suggesting a role of sex steroids in ALS. Testosterone is a complex steroid that exerts its effects directly via androgen (AR) or Sigma-1 receptors and indirectly via estrogen receptors (ER) after aromatization into estradiol. Its reduced-metabolite 5α-dihydrotestosterone acts via AR. This study analyzed the effects of testosterone in male symptomatic Wobblers. Controls or Wobblers received empty or testosterone-filled silastic tubes for 2 months. The cervical spinal cord from testosterone-treated Wobblers showed (1) similar androgen levels to untreated control and (2) increased levels of testosterone, and its 5α-reduced metabolites, 5α- dihydrotestosterone, and 3ß-androstanediol, but (3) undetectable levels of estradiol compared to untreated Wobblers. Testosterone-treated controls showed comparable steroid concentrations to its untreated counterpart. In testosterone- treated Wobblers a reduction of AR, ERα, and aromatase and high levels of Sigma-1 receptor mRNAs was demonstrated. Testosterone treatment increased ChAT immunoreactivity and the antiinflammatory mediator TGFß, while it lessened vacuolated motoneurons, GFAP+ astrogliosis, the density of IBA1+ microgliosis, proinflammatory mediators, and oxidative/nitrosative stress. Clinically, testosterone treatment in Wobblers slowed the progression of paw atrophy and improved rotarod performance. Collectively, our findings indicate an antiinflammatory and protective effect of testosterone in the degenerating spinal cord. These results coincided with a high concentration of androgen-reduced derivatives after testosterone treatment suggesting that the steroid profile may have a beneficial role on disease progression.
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Esclerose Lateral Amiotrófica/tratamento farmacológico , Neurônios Motores/efeitos dos fármacos , Fármacos Neuroprotetores/uso terapêutico , Testosterona/uso terapêutico , Esclerose Lateral Amiotrófica/metabolismo , Esclerose Lateral Amiotrófica/patologia , Animais , Aromatase/metabolismo , Astrócitos/efeitos dos fármacos , Astrócitos/metabolismo , Astrócitos/patologia , Modelos Animais de Doenças , Receptor alfa de Estrogênio/metabolismo , Masculino , Camundongos , Neurônios Motores/metabolismo , Neurônios Motores/patologia , Fármacos Neuroprotetores/farmacologia , Receptores Androgênicos/metabolismo , Medula Espinal/efeitos dos fármacos , Medula Espinal/metabolismo , Medula Espinal/patologia , Testosterona/metabolismo , Testosterona/farmacologia , Resultado do TratamentoRESUMO
TDP-43 is a major component of cytoplasmic inclusions observed in neurodegenerative diseases like frontotemporal dementia (FTD) and amyotrophic lateral sclerosis (ALS). To further understand the role of TDP-43 in mRNA/protein metabolism and proteostasis, we used a combined approach with cellular and animal models overexpressing a cytoplasmic form of human TDP-43 (TDP-43-ΔNLS), recapitulating ALS/FTD features. We applied in HEK293 cells a method for labeling de novo translation, surface sensing of translation (SUnSET), based on puromycin (PURO) incorporation. While control cells displayed robust puromycilation, TDP-43-ΔNLS transfected cells exhibited reduced ongoing protein synthesis. Next, by using a transgenic mouse overexpressing cytoplasmic TDP-43 in the forebrain (TDP-43-ΔNLS mice) we assessed whether cytoplasmic TDP-43 regulates global translation in vivo. Polysome profiling of brain cortices from transgenic mice showed a shift toward non-polysomal fractions as compared to wild-type littermates, indicating a decrease in global translation. Lastly, cellular level translational assessment by SUNSET was performed in TDP-43-ΔNLS mice brain slices. Control mice slices incubated with PURO exhibited robust cytoplasmic PURO signal in layer 5 neurons from motor cortex, and normal nuclear TDP-43 staining. Neurons in TDP-43-ΔNLS mice slices incubated with PURO exhibited high cytoplasmic expression of TDP-43 and reduced puromycilation respect to control mice. These in vitro and in vivo results indicate that cytoplasmic TDP-43 decreases global translation and potentially cause functional/cytotoxic effects as observed in ALS/FTD. Our study provide in vivo evidence (by two independent and complementary methods) for a role of mislocalized TDP-43 in the regulation of global mRNA translation, with implications for TDP-43 proteinopathies.
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Motor neuron disease (MND) have an incidence of 2 in 100 000 persons, resulting in the death of 1 in every 500 people affected. The most common disease in MND spectrum is amyotrophic lateral sclerosis (ALS). We describe the case of an ALS-like syndrome in a HIV patient. This case report presents a 38 years old male from Peru with HIV who after 2 months of combined antiretroviral treatment (cART) initiation was admitted to the hospital for spastic paraplegia. On his first admission, rapid plasma reagent (RPR) was positive and he was treated for neurosyphilis and discharged. Nevertheless, one month after, he was admitted for the second time because paraplegia persisted. Laboratory tests, electromyography and imaging were performed, and ALS was diagnosed. Normally, HIV treated patient with ALS tend to have a better prognosis, however this was not the case. In this case report, we discuss possible association between ALS and immune reconstitution inflammatory syndrome in HIV patients.
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In vitro modeling of neurodegenerative diseases is now possible by using patient-derived induced pluripotent stem cells (iPS). Through them, it is nowadays conceivable to obtain human neurons and glia, and study diseases cellular and molecular mechanisms, an attribute that was previously unavailable to any human condition. Amyotrophic lateral sclerosis (ALS) is one of the diseases that has gained a rapid advance with iPS technology. By differentiating motor neurons from iPS cells of ALS- patients, we are studying the mechanisms underlying ALS- disease onset and progression. Here, we introduce a cellular platform to help maintain longevity of ALS iPS-motor neurons, a cellular feature relevant for most late-onset human diseases. Long term cultures of patient-derived iPS cells might prove to be critical for the development of personalized-drugs.
Actualmente es posible modelar in vitro enfermedades neurodegenerativas humanas mediante el uso de células madre pluripotentes inducidas (iPS) derivadas del paciente. A través de ellas, es hoy concebible obtener neuronas y glía humanas, y estudiar mecanismos celulares y moleculares de enfermedades, un atributo que anteriormente no era posible para ninguna condición humana. La esclerosis lateral amiotrófica (ELA) es una de las enfermedades que se ha beneficiado con la tecnología de iPS. Al diferenciar neuronas motoras de células iPS obtenidas de pacientes con ELA, hemos iniciado estudios sobre los mecanismos que subyacen a la aparición y progresión de la enfermedad. Aquí, presentamos el desarrollo de una plataforma celular que permite extender la longevidad de las neuronas motoras derivadas de iPS, una característica relevante para la mayoría de las enfermedades humanas de inicio tardío. Los cultivos a largo plazo de células iPS provenientes de pacientes pueden ser determinantes en el desarrollo de terapias asociadas a la medicina de precisión.
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Humanos , Animais , Camundongos , Células-Tronco Pluripotentes Induzidas/citologia , Esclerose Lateral Amiotrófica/metabolismo , Imuno-Histoquímica , Linhagem Celular , Técnicas de Cocultura , Esclerose Lateral Amiotrófica/patologia , Esclerose Lateral Amiotrófica/terapiaRESUMO
Genetics has led to a new focus regarding approaches to the most prevalent diseases today. Ascertaining the molecular secrets of neurodegenerative diseases will lead to developing drugs that will change natural history, thereby affecting the quality of life and mortality of patients. The sequencing of candidate genes in patients suffering neurodegenerative pathologies is faster, more accurate, and has a lower cost, thereby enabling algorithms to be proposed regarding the risk of neurodegeneration onset in healthy persons including the year of onset and neurodegeneration severity. Next generation sequencing has resulted in an explosion of articles regarding the diagnosis of neurodegenerative diseases involving exome sequencing or sequencing a whole gene for correlating phenotypical expression with genetic mutations in proteins having key functions. Many of them occur in neuronal glia, which can trigger a proinflammatory effect leading to defective proteins causing sporadic or familial mutations. This article reviews the genetic diagnosis techniques and the importance of bioinformatics in interpreting results from neurodegenerative diseases. Risk scores must be established in the near future regarding diseases with a high incidence in healthy people for defining prevention strategies or an early start for giving drugs in the absence of symptoms.
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Since proving adenosine triphosphate (ATP) functions as a neurotransmitter in neuron/glia interactions, the purinergic system has been more intensely studied within the scope of the central nervous system. In neurological disorders with associated motor symptoms, including Parkinson's disease (PD), motor neuron diseases (MND), multiple sclerosis (MS), amyotrophic lateral sclerosis (ALS), Huntington's Disease (HD), restless leg syndrome (RLS), and ataxias, alterations in purinergic receptor expression and activity have been noted, indicating a potential role for this system in disease etiology and progression. In neurodegenerative conditions, neural cell death provokes extensive ATP release and alters calcium signaling through purinergic receptor modulation. Consequently, neuroinflammatory responses, excitotoxicity and apoptosis are directly or indirectly induced. This review analyzes currently available data, which suggests involvement of the purinergic system in neuro-associated motor dysfunctions and underlying mechanisms. Possible targets for pharmacological interventions are also discussed.
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Introducción: La esclerosis lateral amiotrófica (ELA) es una enfermedad neurodegenerativa devastadora que se manifiesta por debilidad muscular y produce dificultades progresivas de movilización, comunicación, alimentación y, en última instancia, respiración, creando una dependencia creciente de familiares y de otros cuidadores La experiencia de cuidado demanda esfuerzo físico, emocional y social requiriendo todo tipo de apoyos, en especial aquellos que brinda la tecnología para proveer soporte social ya que algunas experiencias con TIC's han demostrado eliminar barreras en el acceso al cuidado, disminuir los costos que implican en los cuidadores el desplazamiento y el tiempo, garantizando un servicio disponible para la persona enferma en su ausencia. Objetivo: Describir cómo se relacionan la apropiación de las tecnologías de la información y comunicación y la percepción de soporte social en cuidadores familiares de personas con Esclerosis Lateral Amiotrófica que pertenecen a la Asociación Colombiana de Esclerosis Lateral Amiotrófica. Método: Estudio descriptivo, de relación, abordaje cuantitativo, de corte trasversal. Participaron 76 cuidadores familiares de personas con ELA pertenecientes a ACELA. Instrumentos utilizados: GCPC-UN-D Este instrumento se define como una encuesta de caracterización para el cuidado de una "Diada persona con enfermedad crónica- cuidador familiar" se identifican las necesidades básicas de información para cuidar a una Diada a través de 42 ítems y tres dimensiones: 1) Las condiciones y perfil socio demográfico de la Diada. 2) La percepción de carga y apoyo. 3) Los medios de información y comunicación " y el Cuestionario estudio de desenlaces médicos de apoyo social MOS (Sherbourne & Stewart, 1991) el cual contiene 4 dimensiones de apoyo social funcional o cualitativo: a) apoyo emocional/informacional; b) interacción social positiva; c) apoyo afectivo y d) apoyo instrumental de ayuda material o tangible. El análisis estadístico de relación se realizó mediante la determinación de los coeficientes de correlación de Spearman. Resultados: Las características socio demográficas de los cuidadores de personas con ELA son: mayoría mujeres, edades entre los 30 a 49 años, estado civil casados, estrato socioeconómico 5 y 6, nivel educativo bachillerato, ocupación empleados. Cuando se analiza la apropiación de todas las TIC´s surge de revisar el acceso, el conocimiento y el uso de estas TIC's. Los hallazgos señalan que la apropiación de las tecnologías es heterogénea dentro del grupo de cuidadores además que tienen una elevada apropiación de las mismas. Se evidencia que predomina una percepción alta del soporte social en cada una de las dimensiones. A pesar de no encontrar relación estadísticamente significativa entre la percepción de soporte social y la apropiación de las TIC´s, si se presenta relación fuerte y estadísticamente significativa entre las variables Uso de las TIC´s para el cuidado de la persona con ELA y el apoyo percibido para el cuidado por parte del cuidador familiar de estas personas. Conclusiones: Al establecer la relación entre el soporte social percibido y la apropiación de las TIC's en cuidadores de personas con ELA, se acepta la hipótesis nula de que no existe relación estadísticamente significativa entre las dos variables. Se requiere, a partir de estudios posteriores, qué otras variables inciden en el soporte social percibido ya que hay una relación estadísticamente significativa y fuerte entre el uso de las TIC´s para el cuidado y la percepción de soporte a través de ellas genera alternativas para mayores exploraciones del fenómeno abordado.
Introduction: Amyotrophic lateral sclerosis (ALS) is a devastating neurodegenerative disease manifested by progressive muscle weakness and produces difficulties of mobilization, communication, food and, ultimately, breathing, creating a growing dependence on family and other caregivers Experience care demands physical, emotional and social effort requiring all kinds of support, especially those offered by technology to provide social support as some experiences with ICT have proven to eliminate barriers to access to care, reduce costs involved in caregivers the displacement and time, ensuring a service available to the sick person in his absence. To describe how the appropriation of information technologies and communication and perception of social support family caregivers of people with ALS who belong to the Colombian Association of Amyotrophic Lateral Sclerosis relate. Method: Descriptive, relationship, quantitative approach to cross-cutting. They involved 76 family caregivers of people with ALS belonging to ACELA. Instruments used: GCPC-A-D This instrument is defined as a characterization survey for the care of a "National Day person familiar chronicle disease caregiver" basic information needs are identified to care for a National Day through 42 items and three dimensions: 1) The conditions and sociodemographic profile of the Day. 2) The perception of load and support. 3) The means of information and communication "and the Medical Outcomes Study Questionnaire of social support MOS (Sherbourne & Stewart, 1991) which contains 4 functional or qualitative dimensions of social support: a) emotional / informational support; b) positive social interaction; c) emotional support d) instrumental support material or tangible help. Statistical analysis was performed relationship by determining the Spearman correlation coefficients. Results: Demographic partner caregivers of people with ALS features are mostly women aged 30-49 years married marital status, socioeconomic stratum 5 and 6, high school education, occupation employees. When all the appropriation of ICTs arises to review the access, knowledge and use of these ICTs it is analyzed. The findings indicate that the appropriation of technologies is heterogeneous within the group of caregivers also having a high ownership of them. It is evident that dominates a high perception of social support in each of the dimensions. Despite not find statistically significant relationship between perceived social support and appropriation of ICTs, if strong and statistically significant relationship occurs between the variables use of ICT for the care of the person with ALS and perceived support for care of the family caregiver of these people. Conclusions: To establish the relationship between perceived social support and appropriation of ICTs in carers of people with ALS, the null hypothesis that there is no statistically significant relationship between the two variables is accepted. It requires from later studies, what other variables affect the perceived social support as there is a statistically significant and strong relationship between the use of ICT for the care and support perception through them generates alternatives further exploration of the phenomenon addressed.
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Humanos , Masculino , Feminino , Apoio Social , Tecnologia da Informação , Esclerose Lateral Amiotrófica , Estudos Transversais , CuidadoresRESUMO
The role of oxidative post-translational modifications of human superoxide dismutase 1 (hSOD1) in the amyotrophic lateral sclerosis (ALS) pathology is an attractive hypothesis to explore based on several lines of evidence. Among them, the remarkable stability of hSOD1(WT) and several of its ALS-associated mutants suggests that hSOD1 oxidation may precede its conversion to the unfolded and aggregated forms found in ALS patients. The bicarbonate-dependent peroxidase activity of hSOD1 causes oxidation of its own solvent-exposed Trp(32) residue. The resulting products are apparently different from those produced in the absence of bicarbonate and are most likely specific for simian SOD1s, which contain the Trp(32) residue. The aims of this work were to examine whether the bicarbonate-dependent peroxidase activity of hSOD1 (hSOD1(WT) and hSOD1(G93A) mutant) triggers aggregation of the enzyme and to comprehend the role of the Trp(32) residue in the process. The results showed that Trp(32) residues of both enzymes are oxidized to a similar extent to hSOD1-derived tryptophanyl radicals. These radicals decayed to hSOD1-N-formylkynurenine and hSOD1-kynurenine or to a hSOD1 covalent dimer cross-linked by a ditryptophan bond, causing hSOD1 unfolding, oligomerization, and non-amyloid aggregation. The latter process was inhibited by tempol, which recombines with the hSOD1-derived tryptophanyl radical, and did not occur in the absence of bicarbonate or with enzymes that lack the Trp(32) residue (bovine SOD1 and hSOD1(W32F) mutant). The results support a role for the oxidation products of the hSOD1-Trp(32) residue, particularly the covalent dimer, in triggering the non-amyloid aggregation of hSOD1.