Your browser doesn't support javascript.
Mostrar: 20 | 50 | 100
Resultados 1 - 20 de 663
Filtrar
1.
Cell Mol Life Sci ; 76(18): 3497-3514, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31089747

RESUMO

Accurate determination of microRNA expression levels is a prerequisite in using these small non-coding RNA molecules as novel biomarkers in disease diagnosis and prognosis. Quantitative PCR is the method of choice for measuring the expression levels of microRNAs. However, a major obstacle that affects the reliability of results is the lack of validated reference controls for data normalization. Various non-coding RNAs have previously been used as reference controls, but their use may lead to variations and lack of comparability of microRNA data among the studies. Despite the growing number of studies investigating microRNA profiles to discriminate between healthy and disease stages, robust reference controls for data normalization have so far not been established. In the present article, we provide an overview of different reference controls used in various diseases, and highlight the urgent need for the identification of suitable reference controls to produce reliable data. Our analysis shows, among others, that RNU6 is not an ideal normalizer in studies using patient material from different diseases. Finally, our article tries to disclose the challenges to find a reference control which is uniformly and stably expressed across all body tissues, fluids, and diseases.


Assuntos
MicroRNAs/metabolismo , Biomarcadores/metabolismo , Doenças Cardiovasculares/genética , Doenças Cardiovasculares/patologia , Doenças do Sistema Nervoso Central/genética , Doenças do Sistema Nervoso Central/patologia , Hepatite B/genética , Hepatite B/patologia , Humanos , MicroRNAs/sangue , Neoplasias/genética , Neoplasias/patologia , Prognóstico , RNA Nuclear Pequeno/sangue , RNA Nuclear Pequeno/metabolismo , Tuberculose/genética , Tuberculose/patologia
2.
Nat Methods ; 16(5): 381-386, 2019 05.
Artigo em Inglês | MEDLINE | ID: mdl-30962620

RESUMO

Single-cell transcriptomics provides an opportunity to characterize cell-type-specific transcriptional networks, intercellular signaling pathways and cellular diversity with unprecedented resolution by profiling thousands of cells in a single experiment. However, owing to the unique statistical properties of scRNA-seq data, the optimal measures of association for identifying gene-gene and cell-cell relationships from single-cell transcriptomics remain unclear. Here, we conducted a large-scale evaluation of 17 measures of association for their ability to reconstruct cellular networks, cluster cells of the same type and link cell-type-specific transcriptional programs to disease. Measures of proportionality were consistently among the best-performing methods across datasets and tasks. Our analysis provides data-driven guidance for gene and cell network analysis in single-cell transcriptomics.


Assuntos
Biologia Computacional/métodos , Perfilação da Expressão Gênica/métodos , Redes Reguladoras de Genes/genética , Análise de Célula Única/métodos , Animais , Doenças Cardiovasculares/genética , Linhagem Celular , Doenças do Sistema Nervoso Central/genética , Análise por Conglomerados , Humanos , Camundongos , Reprodutibilidade dos Testes , Análise de Sequência de RNA/métodos , Software
3.
Prog Retin Eye Res ; 69: 137-158, 2019 03.
Artigo em Inglês | MEDLINE | ID: mdl-30982505

RESUMO

ELOngation of Very Long chain fatty acids-4 (ELOVL4) is an elongase responsible for the biosynthesis of very long chain (VLC, ≥C28) saturated (VLC-SFA) and polyunsaturated (VLC-PUFA) fatty acids in brain, retina, skin, Meibomian glands, and testes. Fascinatingly, different mutations in this gene have been reported to cause vastly different phenotypes in humans. Heterozygous inheritance of seven different mutations in the coding sequence and 5' untranslated region of ELOVL4 causes autosomal dominant Stargardt-like macular dystrophy (STGD3), while homozygous inheritance of three more mutant variants causes severe seizures with ichthyosis, hypertonia, and even death. Some recent studies have described heterozygous inheritance in yet another three mutant ELOVL4 variants, two that cause spinocerebellar ataxia-34 (SCA34) with erythrokeratodermia (EKV) and one that causes SCA34 without EKV. We identified the specific enzymatic reactions catalyzed by ELOVL4 and, using a variety of genetically engineered mouse models, have actively searched for the mechanisms by which ELOVL4 impacts neural function and health. In this review, we critically compare and contrast the various animal model and case studies involving ELOVL4 deficiency via either mutation or deletion, and the resulting consequences on neuronal health and function in both the retina and central nervous system.


Assuntos
Fenômenos Fisiológicos Celulares/fisiologia , Doenças do Sistema Nervoso Central/fisiopatologia , Proteínas do Olho/fisiologia , Mamíferos/fisiologia , Proteínas de Membrana/fisiologia , Células Fotorreceptoras de Vertebrados/fisiologia , Doenças Retinianas/fisiopatologia , Animais , Doenças do Sistema Nervoso Central/genética , Proteínas do Olho/genética , Proteínas do Olho/metabolismo , Humanos , Proteínas de Membrana/genética , Proteínas de Membrana/metabolismo , Mutação , Doenças Retinianas/genética , Doenças Retinianas/metabolismo
4.
Front Biosci (Landmark Ed) ; 24: 870-889, 2019 03 01.
Artigo em Inglês | MEDLINE | ID: mdl-30844719

RESUMO

Congenital and genetic disorders cause many diseases in Arab countries due to large family sizes and high levels of inbreeding. Saudi Arabia (SA) has the highest consanguinity rates among Middle Eastern countries (~60% of all marriages) and is burdened by the highest number of genetic diseases. Genetic diseases can be life-threatening, often manifesting early in life. Approximately 8% of births in SA are affected, and more common genetic diseases, such as metabolic disease and cancer, manifest later in life in up to 20% of the population. This represents a massive healthcare burden to SA hospitals. The number of genetic disorders in the human population ranges from 7000 to 8000, over 3000 of which are caused by unknown mutations. In 2013, SA initiated the Saudi Human Genome Program (SHGP), which aims to sequence over 100,000 human genomes, with the goal of identifying strategies to discover, prevent, diagnose and treat genetic disorders through precision therapy. High-technology genomics and informatic-based centers that exploit next-generation sequencing (NGS) have now identified mutations underlying many unexplained diseases.


Assuntos
Genoma Humano , Medicina de Precisão/métodos , Análise de Sequência de DNA/métodos , Doenças Autoimunes/genética , Doenças do Sistema Nervoso Central/genética , Anormalidades Congênitas/genética , Consanguinidade , Feminino , Genômica , Humanos , Deficiência Intelectual/genética , Nefropatias/genética , Deformidades Congênitas dos Membros/genética , Masculino , Mutação , Síndromes Neoplásicas Hereditárias/genética , Arábia Saudita
5.
Pharmacol Res ; 141: 602-608, 2019 03.
Artigo em Inglês | MEDLINE | ID: mdl-30708051

RESUMO

The zebrafish (Danio rerio) is increasingly utilized as a powerful new model organism in neurobehavioral research. Aggression is a common symptom of many CNS disorders, has some genetic determinants and can be modulated pharmacologically in humans and animal model species. Mounting evidence suggests zebrafish as a useful tool to study neurobiology of aggression, and its pharmacological and genetic regulation. Here, we discuss mechanisms of zebrafish aggression and their pharmacological, pharmacogenetic and pharmacogenomic models, as well as recent developments and existing challenges in this field. We also emphasize the growing utility of zebrafish models in translational neuropharmacological research of aggression, fostering future discoveries of potential therapeutic agents for aggressive behavior.


Assuntos
Agressão , Doenças do Sistema Nervoso Central/genética , Modelos Animais de Doenças , Peixe-Zebra/genética , Agressão/efeitos dos fármacos , Animais , Fármacos do Sistema Nervoso Central/farmacologia , Doenças do Sistema Nervoso Central/tratamento farmacológico , Doenças do Sistema Nervoso Central/patologia , Neurofarmacologia , Farmacogenética , Pesquisa Médica Translacional , Peixe-Zebra/fisiologia
6.
Glia ; 67(2): 277-290, 2019 02.
Artigo em Inglês | MEDLINE | ID: mdl-30565754

RESUMO

Genetically caused neurological disorders of the central nervous system (CNS) usually result in poor or even fatal clinical outcome and few or no causative treatments are available. Often, these disorders are associated with disease-amplifying neuroinflammation, a feature shared by progressive forms of multiple sclerosis (PMS), another poorly treatable disorder of the CNS. We have previously generated two mouse lines carrying distinct mutations in the oligodendrocytic PLP1 gene that have initially been identified in patients fulfilling clinical criteria for multiple sclerosis (MS). These mutations cause a loss of function of the gene product resulting in a histopathological and clinical phenotype common to both PMS and genetic CNS disorders, like hereditary spastic paraplegias. Importantly, neuroinflammation comprising adaptive immune reactions promotes disease progression in these PLP1 mutant models, opening the possibility to improve disease outcome of the respective disorders by targeting/modulating inflammation. We here show that PLX3397, a potent inhibitor of the CSF-1R and targeting innate immune cells, attenuates neuroinflammation in our models by reducing numbers of resident microglia and attenuating T-lymphocyte recruitment in the CNS. This leads to an amelioration of demyelination, axonopathic features and neuron loss in the retinotectal system, also reflected by reduced thinning of the inner retinal composite layer in longitudinal studies using noninvasive optical coherence tomography. Our findings identify microglia as important promoters of neuroinflammation-related neural damage and CSF-1R inhibition as a possible therapeutic strategy not only for PMS but also for inflammation-related genetic diseases of the nervous system for which causal treatment options are presently lacking.


Assuntos
Doenças do Sistema Nervoso Central/complicações , Doenças do Sistema Nervoso Central/genética , Inflamação , Microglia/metabolismo , Mutação/genética , Proteína Proteolipídica de Mielina/genética , Aminopiridinas/uso terapêutico , Animais , Anti-Inflamatórios/uso terapêutico , Citocinas/metabolismo , Modelos Animais de Doenças , Relação Dose-Resposta a Droga , Citometria de Fluxo , Humanos , Inflamação/tratamento farmacológico , Inflamação/etiologia , Inflamação/genética , Inflamação/patologia , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Transgênicos , Microglia/efeitos dos fármacos , Microglia/patologia , Microglia/ultraestrutura , Microscopia Eletrônica de Transmissão , Proteína Proteolipídica de Mielina/metabolismo , Neurônios/efeitos dos fármacos , Neurônios/patologia , Neurônios/ultraestrutura , Pirróis/uso terapêutico , Linfócitos T/efeitos dos fármacos , Linfócitos T/patologia , Tomografia de Coerência Óptica
8.
PLoS Negl Trop Dis ; 12(7): e0006648, 2018 07.
Artigo em Inglês | MEDLINE | ID: mdl-30001342

RESUMO

Chandipura Virus (CHPV), a negative-stranded RNA virus belonging to the Rhabdoviridae family, has been previously reported to bring neuronal apoptosis by activating several factors leading to neurodegeneration. Following virus infection of the central nervous system, microglia, the ontogenetic and functional equivalents of macrophages in somatic tissues gets activated and starts secreting chemokines, thereby recruiting peripheral leukocytes into the brain parenchyma. In the present study, we have systemically examined the effect of CHPV on microglia and the activation of cellular signalling pathways leading to chemokine expression upon CHPV infection. Protein and mRNA expression profiles of chemokine genes revealed that CHPV infection strongly induces the expression of CXC chemokine ligand 10 (CXCL10) and CC chemokine ligand 5 (CCL5) in microglia. CHPV infection triggered the activation of signalling pathways mediated by mitogen-activated protein kinases, including p38, JNK 1 and 2, and nuclear factor κB (NF-kappaB). CHPV-induced expression of CXCL10 and CCL5 was achieved by the activation of p38 and NF-kappaB pathways. Considering the important role of inflammation in neurodegeneration, we have targeted NF-kappaB using a newly synthesised natural product nitrosporeusine analogue and showed incapability of microglial supernatant of inducing apoptosis in neurons after treatment.


Assuntos
Alcaloides/administração & dosagem , Antivirais/administração & dosagem , Doenças do Sistema Nervoso Central/tratamento farmacológico , Microglia/imunologia , NF-kappa B/imunologia , Infecções por Rhabdoviridae/imunologia , Vesiculovirus/fisiologia , Animais , Linhagem Celular , Sistema Nervoso Central/imunologia , Sistema Nervoso Central/virologia , Doenças do Sistema Nervoso Central/genética , Doenças do Sistema Nervoso Central/imunologia , Doenças do Sistema Nervoso Central/virologia , Quimiocina CCL5/genética , Quimiocina CCL5/imunologia , Quimiocina CXCL10/genética , Quimiocina CXCL10/imunologia , Humanos , Camundongos , Microglia/efeitos dos fármacos , Microglia/virologia , NF-kappa B/genética , Infecções por Rhabdoviridae/tratamento farmacológico , Infecções por Rhabdoviridae/genética , Infecções por Rhabdoviridae/virologia , Transdução de Sinais/efeitos dos fármacos , Vesiculovirus/efeitos dos fármacos , Vesiculovirus/genética
9.
Wiad Lek ; 71(2 pt 2): 413-416, 2018.
Artigo em Polonês | MEDLINE | ID: mdl-29786595

RESUMO

Mutations leading to disorders within ion (mainly potassium and sodium) channels, have different degrees of expression in the brain and in the heart, which can cause simultaneous occurrence of disorders in both organs. This is manifested by the occurrence of epileptic seizures and cardiac electrical disturbances, further exacerbated by stimulation of autonomic structures within the central nervous system. In all patients with unclear paroxysmal disorders, and in those with unexplained sudden cardiac death, consideration should be given to the possibility of occurrence of genetically determined disorders in the ion channels. This article concerns the most common genetically determined epilepsy syndromes and genetically determined cardiac diseases.


Assuntos
Arritmias Cardíacas/genética , Encéfalo/fisiopatologia , Doenças do Sistema Nervoso Central/genética , Canais Iônicos/genética , Humanos , Mutação
10.
Ticks Tick Borne Dis ; 9(4): 763-767, 2018 05.
Artigo em Inglês | MEDLINE | ID: mdl-29496490

RESUMO

The progression of infectious diseases depends on causative agents, the environment and the host's genetic susceptibility. To date, human genetic susceptibility to tick-borne encephalitis (TBE) virus-induced disease has not been sufficiently studied. We have combined whole-exome sequencing with a candidate gene approach to identify genes that are involved in the development of predisposition to TBE in a Russian population. Initially, six exomes from TBE patients with severe central nervous system (CNS) disease and seven exomes from control individuals were sequenced. Despite the small sample size, two nonsynonymous single nucleotide polymorphisms (SNPs) were significantly associated with TBE virus-induced severe CNS disease. One of these SNPs is rs6558394 (G/A, Pro422Leu) in the scribbled planar cell polarity protein (SCRIB) gene and the other SNP is rs17576 (A/G, Gln279Arg) in the matrix metalloproteinase 9 (MMP9) gene. Subsequently, these SNPs were genotyped in DNA samples of 150 non-immunized TBE patients with different clinical forms of the disease from two cities and 228 control randomly selected samples from the same populations. There were no statistically significant differences in genotype and allele frequencies between the case and control groups for rs6558394. However, the frequency of the rs17576 G allele was significantly higher in TBE patients with severe CNS diseases such as meningo-encephalitis (43.5%) when compared with TBE patients with milder meningitis (26.3%; P = 0.01), as well as with the population control group (32.5%; P = 0.042). The results suggest that the MMP9 gene may affect genetic predisposition to TBE in a Russian population.


Assuntos
Doenças do Sistema Nervoso Central/genética , Doenças do Sistema Nervoso Central/virologia , Encefalite Transmitida por Carrapatos/genética , Predisposição Genética para Doença , Metaloproteinase 9 da Matriz/genética , Polimorfismo de Nucleotídeo Único , Alelos , Animais , Doenças do Sistema Nervoso Central/epidemiologia , Doenças do Sistema Nervoso Central/etiologia , Vírus da Encefalite Transmitidos por Carrapatos/genética , Vírus da Encefalite Transmitidos por Carrapatos/isolamento & purificação , Encefalite Transmitida por Carrapatos/sangue , Encefalite Transmitida por Carrapatos/epidemiologia , Encefalite Transmitida por Carrapatos/virologia , Frequência do Gene , Genótipo , Humanos , Camundongos , Camundongos Knockout , Federação Russa/epidemiologia , Sequenciamento Completo do Genoma
11.
Handb Clin Neurol ; 147: 23-36, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-29325614

RESUMO

Many neurogenetic conditions are inherited and therefore diagnosis of a patient will have implications for the patient's relatives and can raise ethical issues. Predictive genetic testing offers asymptomatic relatives the opportunity to determine their risk status for a neurogenetic condition, and professional guidelines emphasize patients' autonomy and informed, voluntary decision making. Beneficence and nonmaleficence both need to be considered when making decisions about disclosure and nondisclosure of genetic information and test results. There can be disclosure concerns and challenges in determining whose autonomy to prioritize when a patient makes a genetic testing decision that can reveal the genetic status of a relative (e.g., testing an adult child when the at-risk parent has not been tested). Ethical issues are prominent when genetic testing for neurogenetic conditions is requested prenatally, on minors, adoptees, adult children at 25% risk, and for individuals with psychiatric issues or cognitive impairment. Neurogenetic conditions can result in cognitive decline which can affect decisional capacity and lead to ethical challenges with decision making, informed consent, and determining the patient's ability to comprehend test results. The ethical implications of genetic testing and emerging issues, including direct-to-consumer genetic testing, disclosure of secondary findings from genomic sequencing, and use of apolipoprotein E testing in clinical and research settings, are also discussed. Resources for information about genetic testing practice guidelines, insurance laws, and directories of genetics clinics are included.


Assuntos
Doenças do Sistema Nervoso Central , Transtornos Cognitivos/etiologia , Ética Médica , Testes Genéticos/ética , Testes Genéticos/métodos , Doenças do Sistema Nervoso Central/complicações , Doenças do Sistema Nervoso Central/diagnóstico , Doenças do Sistema Nervoso Central/genética , Aconselhamento Genético/ética , Aconselhamento Genético/legislação & jurisprudência , Humanos
12.
Biochim Biophys Acta Biomembr ; 1860(1): 65-71, 2018 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-28735901

RESUMO

Pannexins are a family of integral membrane proteins with distinct post-translational modifications, sub-cellular localization and tissue distribution. Panx1 is the most studied and best-characterized isoform of this gene family. The ubiquitous expression, as well as its function as a major ATP release and nucleotide permeation channel, makes Panx1 a primary candidate for participating in the pathophysiology of CNS disorders. While many investigations revolve around Panx1 functions in health and disease, more recently, details started emerging about mechanisms that control Panx1 channel activity. These advancements in Panx1 biology have revealed that beyond its classical role as an unopposed plasma membrane channel, it participates in alternative pathways involving multiple intracellular compartments, protein complexes and a myriad of extracellular participants. Here, we review recent progress in our understanding of Panx1 at the center of these pathways, highlighting its modulation in a context specific manner. This article is part of a Special Issue entitled: Gap Junction Proteins edited by Jean Claude Herve.


Assuntos
Membrana Celular/metabolismo , Doenças do Sistema Nervoso Central/metabolismo , Conexinas/metabolismo , Ativação do Canal Iônico , Canais Iônicos/metabolismo , Proteínas do Tecido Nervoso/metabolismo , Animais , Membrana Celular/genética , Doenças do Sistema Nervoso Central/genética , Doenças do Sistema Nervoso Central/fisiopatologia , Conexinas/genética , Humanos , Canais Iônicos/genética , Proteínas do Tecido Nervoso/genética
14.
Expert Opin Ther Targets ; 21(12): 1161-1170, 2017 12.
Artigo em Inglês | MEDLINE | ID: mdl-29072508

RESUMO

INTRODUCTION: Aquaporin-4 (AQP4) is a water transporting protein expressed at the plasma membrane of astrocytes throughout the central nervous system (CNS). Analysis of AQP4 knockout mice has suggested its broad involvement in brain water balance, neuroexcitation, glial scarring, neuroinflammation, and even neurodegenerative and neuropsychiatric disorders. Broad clinical utility of AQP4 modulators has been speculated. Area covered: This review covers the biology of AQP4, evidence for its roles in normal CNS function and neurological disorders, and progress in AQP4 drug discovery. Expert opinion: Critical examination of available data reduces the lengthy potential applications list to AQP4 inhibitors for early therapy of ischemic stroke and perhaps for reduction of glial scarring following CNS injury. Major challenges in identification and clinical development of AQP4 inhibitors include the apparent poor druggability of AQPs, the many homologous AQP isoforms with broad tissue distribution and functions, technical issues with water transport assays, predicted undesired CNS and non-CNS actions, and the need for high blood-brain barrier permeation. To date, despite considerable effort, validated small-molecule AQP4 inhibitors have not been advanced. However, a biologic ('aquaporumab') is in development for neuromyelitis optica, an autoimmune inflammatory demyelinating disease where CNS pathology is initiated by binding of anti-AQP4 autoantibodies to astrocyte AQP4.


Assuntos
Aquaporina 4/antagonistas & inibidores , Doenças do Sistema Nervoso Central/tratamento farmacológico , Desenho de Drogas , Animais , Aquaporina 4/metabolismo , Astrócitos/metabolismo , Barreira Hematoencefálica/metabolismo , Sistema Nervoso Central/fisiologia , Sistema Nervoso Central/fisiopatologia , Fármacos do Sistema Nervoso Central/farmacocinética , Fármacos do Sistema Nervoso Central/farmacologia , Doenças do Sistema Nervoso Central/genética , Doenças do Sistema Nervoso Central/fisiopatologia , Descoberta de Drogas/métodos , Humanos , Camundongos , Camundongos Knockout , Terapia de Alvo Molecular
15.
Carcinogenesis ; 38(8): 812-820, 2017 08 01.
Artigo em Inglês | MEDLINE | ID: mdl-28854563

RESUMO

Central nervous system (CNS) relapse is a devastating complication that occurs in about 5% of diffuse large B-cell lymphoma (DLBCL) patients. Currently, there are no predictive biological markers. We wanted to study potential biomarkers of CNS tropism that play a role in adhesion, migration and/or in the regulation of inflammatory responses. The expression levels of ITGA10, CD44, PTEN, cadherin-11, CDH12, N-cadherin, P-cadherin, lactoferrin and E-cadherin were studied with IHC and IEM. GEP was performed to see whether found expressional changes are regulated at DNA/RNA level. IHC included 96 samples of primary CNS lymphoma (PCNSL), secondary CNS lymphoma (sCNSL) and systemic DLBCL (sDLBCL). IEM included two PCNSL, one sCNSL, one sDLBCL and one reactive lymph node samples. GEP was performed on two DLBCL samples, one with and one without CNS relapse. CNS disease was associated with enhanced expression of cytoplasmic and membranous ITGA10 and nuclear PTEN (P < 0.0005, P = 0.002, P = 0.024, respectively). sCNSL presented decreased membranous CD44 and nuclear and cytoplasmic cadherin-11 expressions (P = 0.001, P = 0.006, P = 0.048, respectively). In PCNSL lactoferrin expression was upregulated (P < 0.0005). IEM results were mainly supportive of the IHC results. In GEP CD44, cadherin-11, lactoferrin and E-cadherin were under-expressed in CNS disease. Our results are in line with previous studies, where gene expressions in extracellular matrix and adhesion-related pathways are altered in CNS lymphoma. This study gives new information on the DLBCL CNS tropism. If further verified, these markers might become useful in predicting CNS relapses.


Assuntos
Caderinas/genética , Doenças do Sistema Nervoso Central/genética , Receptores de Hialuronatos/genética , Cadeias alfa de Integrinas/genética , Lactoferrina/genética , Linfoma Difuso de Grandes Células B/genética , PTEN Fosfo-Hidrolase/genética , Adulto , Idoso , Idoso de 80 Anos ou mais , Biomarcadores Tumorais/genética , Caderinas/biossíntese , Sistema Nervoso Central/patologia , Doenças do Sistema Nervoso Central/etiologia , Doenças do Sistema Nervoso Central/patologia , Feminino , Regulação Neoplásica da Expressão Gênica , Humanos , Receptores de Hialuronatos/biossíntese , Cadeias alfa de Integrinas/biossíntese , Lactoferrina/biossíntese , Linfonodos/metabolismo , Linfonodos/patologia , Linfoma Difuso de Grandes Células B/complicações , Linfoma Difuso de Grandes Células B/patologia , Masculino , Pessoa de Meia-Idade , PTEN Fosfo-Hidrolase/biossíntese
16.
Cell Rep ; 20(12): 2980-2991, 2017 Sep 19.
Artigo em Inglês | MEDLINE | ID: mdl-28930690

RESUMO

Neurodegenerative disorders are a major public health problem because of the high frequency of these diseases. Genome editing with the CRISPR/Cas9 system is making it possible to modify the sequence of genes linked to these disorders. We designed the KamiCas9 self-inactivating editing system to achieve transient expression of the Cas9 protein and high editing efficiency. In the first application, the gene responsible for Huntington's disease (HD) was targeted in adult mouse neuronal and glial cells. Mutant huntingtin (HTT) was efficiently inactivated in mouse models of HD, leading to an improvement in key markers of the disease. Sequencing of potential off-targets with the constitutive Cas9 system in differentiated human iPSC revealed a very low incidence with only one site above background level. This off-target frequency was significantly reduced with the KamiCas9 system. These results demonstrate the potential of the self-inactivating CRISPR/Cas9 editing for applications in the context of neurodegenerative diseases.


Assuntos
Sistemas CRISPR-Cas/genética , Doenças do Sistema Nervoso Central/genética , Edição de Genes , Animais , Astrócitos/citologia , Astrócitos/metabolismo , Sequência de Bases , Células Cultivadas , Córtex Cerebral/citologia , Células HEK293 , Humanos , Proteína Huntingtina/genética , Células-Tronco Pluripotentes Induzidas/citologia , Células-Tronco Pluripotentes Induzidas/metabolismo , Cinética , Camundongos , Neurônios/citologia , Neurônios/metabolismo
17.
Brain Nerve ; 69(8): 901-912, 2017 Aug.
Artigo em Japonês | MEDLINE | ID: mdl-28819074

RESUMO

Polyglutamine diseases result from gain-of-function mutations. The expanded polyglutamine tracts lead to conformational changes in proteins, resulting in their aggregation. The intermediates including monomers or oligomers, are more toxic than the aggregates to neurons. At the molecular level, protein misfolding, transcriptional dysregulation, deranged calcium homeostasis, impaired cytoskeleton/axonal transport, mitochondrial dysfunction, and RNA toxicity contribute to disease progression. Understanding the underlying pathogenesis facilitates development of therapy for polyglutamine diseases.


Assuntos
Doenças do Sistema Nervoso Central/metabolismo , Peptídeos/metabolismo , Animais , Transporte Biológico , Doenças do Sistema Nervoso Central/genética , Homeostase , Humanos , Mitocôndrias/metabolismo , Peptídeos/genética , Agregação Patológica de Proteínas
18.
Hum Genet ; 136(9): 1079-1091, 2017 09.
Artigo em Inglês | MEDLINE | ID: mdl-28664341

RESUMO

The central nervous system-specific serotonin receptor 2C (5HT2C) controls key physiological functions, such as food intake, anxiety, and motoneuron activity. Its deregulation is involved in depression, suicidal behavior, and spasticity, making it the target for antipsychotic drugs, appetite controlling substances, and possibly anti-spasm agents. Through alternative pre-mRNA splicing and RNA editing, the 5HT2C gene generates at least 33 mRNA isoforms encoding 25 proteins. The 5HT2C is a G-protein coupled receptor that signals through phospholipase C, influencing the expression of immediate/early genes like c-fos. Most 5HT2C isoforms show constitutive activity, i.e., signal without ligand binding. The constitutive activity of 5HT2C is decreased by pre-mRNA editing as well as alternative pre-mRNA splicing, which generates a truncated isoform that switches off 5HT2C receptor activity through heterodimerization; showing that RNA processing regulates the constitutive activity of the 5HT2C system. RNA processing events influencing the constitutive activity target exon Vb that forms a stable double stranded RNA structure with its downstream intron. This structure can be targeted by small molecules and oligonucleotides that change exon Vb alternative splicing and influence 5HT2C signaling in mouse models, leading to a reduction in food intake. Thus, the 5HT2C system is a candidate for RNA therapy in multiple models of CNS disorders.


Assuntos
Processamento Alternativo , Éxons , Multimerização Proteica , Precursores de RNA , Receptores de Serotonina , Animais , Doenças do Sistema Nervoso Central/genética , Doenças do Sistema Nervoso Central/metabolismo , Doenças do Sistema Nervoso Central/terapia , Humanos , Camundongos , Isoformas de Proteínas/biossíntese , Isoformas de Proteínas/genética , Precursores de RNA/genética , Precursores de RNA/metabolismo , Receptores de Serotonina/biossíntese , Receptores de Serotonina/genética
19.
Glia ; 65(9): 1407-1422, 2017 09.
Artigo em Inglês | MEDLINE | ID: mdl-28568966

RESUMO

Genetically caused neurological disorders of the central nervous system (CNS) are usually orphan diseases with poor or even fatal clinical outcome and few or no treatments that will improve longevity or at least quality of life. Neuroinflammation is common to many of these disorders, despite the fact that a plethora of distinct mutations and molecular changes underlie the disorders. In this article, data from corresponding animal models are analyzed to define the roles of innate and adaptive inflammation as modifiers and amplifiers of disease. We describe both common and distinct patterns of neuroinflammation in genetically mediated CNS disorders and discuss the contrasting mechanisms that lead to adverse versus neuroprotective effects. Moreover, we identify the juxtaparanode as a neuroanatomical compartment commonly associated with inflammatory cells and ongoing axonopathic changes, in models of diverse diseases. The identification of key immunological effector pathways that amplify neuropathic features should lead to realistic possibilities for translatable therapeutic interventions using existing immunomodulators. Moreover, evidence emerges that neuroinflammation is not only able to modify primary neural damage-related symptoms but also may lead to unexpected clinical outcomes such as neuropsychiatric syndromes.


Assuntos
Doenças do Sistema Nervoso Central/imunologia , Doenças do Sistema Nervoso Central/terapia , Inflamação/fisiopatologia , Inflamação/terapia , Animais , Doenças do Sistema Nervoso Central/genética , Humanos , Inflamação/genética , Neuroimunomodulação/fisiologia
20.
J Diabetes Complications ; 31(7): 1243-1246, 2017 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-28502589

RESUMO

AIMS: Precise diagnosis of maturity-onset diabetes of the young (MODY) has proven valuable for understanding mechanism of diabetes and selecting optimal therapy. A proband and her mother with diabetic kidney disease (DKD) were studied to investigate potential genes responsible for diabetes and different severity of DKD between the parent and offspring. METHODS: The family with suspected MODY underwent mutational analyses by the whole exome sequencing (WES). Candidate pathogenic variants were validated by Sanger sequencing and tested for co-segregation. The clinical parameters of subjects were collected from medical records. RESULTS: A novel missense heterozygous mutation in exon 4 of the hepatocyte nuclear factor 1ß (HNF1ß), c.1007A > G (p.H336R), was identified in both the proband and her mother. Moreover, comparing the family's WES results, we found that the proband had acquired a KCNQ1 gene mutation from her father and acquired ACE and SORBS1 gene mutations from her mother. These three genes are known susceptibility genes of DKD and may impose additional effects contributing to DKD severity. CONCLUSIONS: A novel mutation in HNF1ß-MODY was identified in a Chinese family complicated with DKD, and the additional effect of pathogenic variants in susceptibility genes was speculated to contribute to DKD severity.


Assuntos
Doenças do Sistema Nervoso Central/genética , Esmalte Dentário/anormalidades , Diabetes Mellitus Tipo 2/genética , Fator 1-beta Nuclear de Hepatócito/genética , Doenças Renais Císticas/genética , Mutação de Sentido Incorreto , Substituição de Aminoácidos , Doenças do Sistema Nervoso Central/metabolismo , Doenças do Sistema Nervoso Central/fisiopatologia , China , Análise Mutacional de DNA , Esmalte Dentário/metabolismo , Esmalte Dentário/fisiopatologia , Diabetes Mellitus Tipo 2/metabolismo , Diabetes Mellitus Tipo 2/fisiopatologia , Feminino , Fator 1-beta Nuclear de Hepatócito/metabolismo , Humanos , Canal de Potássio KCNQ1/genética , Canal de Potássio KCNQ1/metabolismo , Doenças Renais Císticas/metabolismo , Doenças Renais Císticas/fisiopatologia , Proteínas dos Microfilamentos/genética , Proteínas dos Microfilamentos/metabolismo , Pessoa de Meia-Idade , Mães , Mutação , Peptidil Dipeptidase A/genética , Peptidil Dipeptidase A/metabolismo , Índice de Gravidade de Doença , Sequenciamento Completo do Exoma
SELEÇÃO DE REFERÊNCIAS
DETALHE DA PESQUISA