Asunto(s)
Enfermedad de Alzheimer/complicaciones , Paraparesia Espástica/complicaciones , Parálisis Supranuclear Progresiva/complicaciones , Anciano , Enfermedad de Alzheimer/genética , Enfermedad de Alzheimer/patología , Péptidos beta-Amiloides/metabolismo , Encéfalo/metabolismo , Encéfalo/patología , Análisis Mutacional de ADN , Humanos , Masculino , Mutación/genética , Paraparesia Espástica/genética , Paraparesia Espástica/patología , Presenilina-1/genética , Parálisis Supranuclear Progresiva/genética , Parálisis Supranuclear Progresiva/patologíaRESUMEN
It has been reported that organic anion-transporting polypeptide (OATP) 1B1, OATP1B3 and multidrug resistance-associated protein 2 are involved in the hepatobiliary transport of olmesartan. We investigated the association of SLCO1B1, SLCO1B3 and ABCC2 polymorphisms with the pharmacokinetics of olmesartan. We sequenced all exons, exon-intron junctions and the 5' and 3' flanking regions of the three genes in 115 individuals from African-American, Hispanic and Caucasian populations who had participated in our clinical studies. A total of 348 single-nucleotide polymorphisms (SNPs) were identified with a minor allele frequency of ≥0.01 in at least one population; 132 SNPs were detected in SLCO1B1, 130 in SLCO1B3 and 86 in ABCC2. We characterized the linkage disequilibrium (LD) and haplotypes shared across the populations and then evaluated the association between the haplotypes and the pharmacokinetics of olmesartan. Seven inter-ethnic LD blocks were observed in SLCO1B1, while three in SLCO1B3 and four in ABCC2. Although extensive variability in the sequences of SLCO1B1, SLCO1B3 and ABCC2 existed across the three populations, there was no remarkable difference in any pharmacokinetic parameters of olmesartan between subjects with and without any major haplotypes in the three transporter genes we tested.
Asunto(s)
Imidazoles , Proteínas Asociadas a Resistencia a Múltiples Medicamentos/genética , Transportadores de Anión Orgánico Sodio-Independiente/genética , Transportadores de Anión Orgánico/genética , Tetrazoles , Adulto , Negro o Afroamericano/genética , Femenino , Frecuencia de los Genes , Estudios de Asociación Genética , Haplotipos , Humanos , Imidazoles/farmacocinética , Imidazoles/uso terapéutico , Desequilibrio de Ligamiento , Transportador 1 de Anión Orgánico Específico del Hígado , Masculino , Persona de Mediana Edad , Proteína 2 Asociada a Resistencia a Múltiples Medicamentos , Polimorfismo de Nucleótido Simple , Miembro 1B3 de la Familia de los Transportadores de Solutos de Aniones Orgánicos , Tetrazoles/farmacocinética , Tetrazoles/uso terapéutico , Población Blanca/genéticaRESUMEN
OBJECTIVE: Sorting mechanisms that cause the amyloid precursor protein (APP) and the ß-secretases and γ-secretases to colocalize in the same compartment play an important role in the regulation of Aß production in Alzheimer's disease (AD). We and others have reported that genetic variants in the Sortilin-related receptor (SORL1) increased the risk of AD, that SORL1 is involved in trafficking of APP, and that underexpression of SORL1 leads to overproduction of Aß. Here we explored the role of one of its homologs, the sortilin-related VPS10 domain containing receptor 1 (SORCS1), in AD. METHODS: We analyzed the genetic associations between AD and 16 SORCS1-single nucleotide polymorphisms (SNPs) in 6 independent data sets (2,809 cases and 3,482 controls). In addition, we compared SorCS1 expression levels of affected and unaffected brain regions in AD and control brains in microarray gene expression and real-time polymerase chain reaction (RT-PCR) sets, explored the effects of significant SORCS1-SNPs on SorCS1 brain expression levels, and explored the effect of suppression and overexpression of the common SorCS1 isoforms on APP processing and Aß generation. RESULTS: Inherited variants in SORCS1 were associated with AD in all datasets (0.001 < p < 0.049). In addition, SorCS1 influenced APP processing. While overexpression of SorCS1 reduced γ-secretase activity and Aß levels, the suppression of SorCS1 increased γ-secretase processing of APP and the levels of Aß. INTERPRETATIONS: These data suggest that inherited or acquired changes in SORCS1 expression or function may play a role in the pathogenesis of AD.
Asunto(s)
Enfermedad de Alzheimer/etiología , Enfermedad de Alzheimer/genética , Variación Genética/genética , Receptores de Superficie Celular/genética , Receptores de Superficie Celular/fisiología , Enfermedad de Alzheimer/fisiopatología , Secretasas de la Proteína Precursora del Amiloide/genética , Secretasas de la Proteína Precursora del Amiloide/metabolismo , Precursor de Proteína beta-Amiloide/metabolismo , Encéfalo/metabolismo , Expresión Génica/genética , Expresión Génica/fisiología , Predisposición Genética a la Enfermedad/genética , Genotipo , Humanos , Proteínas Relacionadas con Receptor de LDL/genética , Proteínas de Transporte de Membrana/genética , Metaanálisis como Asunto , Mutación/genética , Polimorfismo de Nucleótido Simple/genética , Polimorfismo de Nucleótido Simple/fisiología , Factores de Riesgo , Población Blanca/genéticaRESUMEN
OBJECTIVE: To reexamine the association between the neuronal sortilin-related receptor gene (SORL1) and Alzheimer disease (AD). DESIGN: Comprehensive and unbiased meta-analysis of all published and unpublished data from case-control studies for the SORL1 single-nucleotide polymorphisms (SNPs) that had been repeatedly assessed across studies. SETTING: Academic research institutions in the United States, the Netherlands, Canada, Belgium, the United Kingdom, Singapore, Japan, Sweden, Germany, France, and Italy. PARTICIPANTS: All published white and Asian case-control data sets, which included a total of 12,464 cases and 17,929 controls. MAIN OUTCOME MEASURES: Alzheimer disease according to the Diagnostic and Statistical Manual of Mental Disorders (Fourth Edition) and the National Institute of Neurological and Communicative Disorders and Stroke and the Alzheimer's Disease and Related Disorders Association (now known as the Alzheimer's Association). RESULTS: In the white data sets, several markers were associated with AD after correction for multiple testing, including previously reported SNPs 8, 9, and 10 (P < .001). In addition, the C-G-C haplotype at SNPs 8 through 10 was associated with AD risk (P < .001). In the combined Asian data sets, SNPs 19 and 23 through 25 were associated with AD risk (P < .001). The disease-associated alleles at SNPs 8, 9, and 10 (120,873,131-120,886,175 base pairs [bp]; C-G-C alleles), at SNP 19 (120,953,300 bp; G allele), and at SNPs 24 through 25 (120,988,611 bp; T and C alleles) were the same previously reported alleles. The SNPs 4 through 5, 8 through 10, 12, and 19 through 25 belong to distinct linkage disequilibrium blocks. The same alleles at SNPs 8 through 10 (C-G-C), 19 (G), and 24 and 25 (T and C) have also been associated with AD endophenotypes, including white matter hyperintensities and hippocampal atrophy on magnetic resonance imaging, cerebrospinal fluid measures of amyloid ß-peptide 42, and full-length SORL1 expression in the human brain. CONCLUSION: This comprehensive meta-analysis provides confirmatory evidence that multiple SORL1 variants in distinct linkage disequilibrium blocks are associated with AD.
Asunto(s)
Enfermedad de Alzheimer/genética , Variación Genética/genética , Proteínas Relacionadas con Receptor de LDL/genética , Proteínas de Transporte de Membrana/genética , Proteínas del Tejido Nervioso/genética , Alelos , Enfermedad de Alzheimer/epidemiología , Enfermedad de Alzheimer/etiología , Estudios de Casos y Controles , Humanos , Polimorfismo de Nucleótido Simple/genéticaRESUMEN
For gene validation and the development of oligonucleotide agents, 2'-O,4'-C-ethylene-bridged nucleic acid (ENA) antisense gapmers are widely available. An in vitro Escherichia coli RNase H reaction analysis using ENA gapmers and an RNA oligonucleotide with mouse peptidylarginine deiminase 4 (PADI4) gene sequences revealed that the RNA oligonucleotide was specifically cleaved in the only reported case of the use of an ENA gapmer with an antisense sequence. On the other hand, duplexes of the full-length transcripts of PADI4 mRNA and ENA gapmers with a wide DNA window were cleaved not only at the target site, but also at nontarget sites by RNase H derived from partial base-pairing between the transcript and the ENA gapmer. When the DNA window region of the ENA gapmer was shortened to 5 or 6 nucleotides, the nontarget cleavage was effectively diminished. Moreover, the specific inhibition of PADI4 mRNA expression was observed in the cotransfection of PADI4 cDNA and ENA gapmers containing a short DNA region into NIH3T3 cells. These results demonstrated that ENA gapmers with a short DNA region improved the sequence-specificity of mRNA downregulation. These optimized ENA gapmers could reduce the "off-target" effect and be applicable to gene validation and oligonucleotide therapeutics.
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Hidrolasas/genética , Oligonucleótidos Antisentido/metabolismo , ARN Mensajero/metabolismo , Ribonucleasa H/metabolismo , Animales , Línea Celular , Hidrolasas/metabolismo , Ratones , Oligonucleótidos Antisentido/genética , Arginina Deiminasa Proteína-Tipo 4 , Desiminasas de la Arginina Proteica , ARN Mensajero/genéticaRESUMEN
In order to evaluate the function of the mouse peptidylarginine deiminase 4 (PADI4) gene, gapmer-designed ENA antisense oligonucleotides were utilized. Antisense ENA gapmers were found to inhibit the mRNA expression of the PADI4 gene, but ENA gapmers with sense sequences as controls only partially inhibited this mRNA expression. An in vitro E. coli RNase H reaction analysis using transcripts of the PADI4 gene and their ENA gapmers revealed that the mixtures of the transcripts and ENA gapmers with sense sequences were cleaved at non-target sites by RNase H derived from partial base-pairing between the transcript and the ENA gapmer. In an in vitro E. coli RNase H reaction analysis, when the DNA region of the ENA gapmer was shortened to 5 or 6 nucleotides, the non-target cleavage disappeared and the specific inhibition of PADI4 mRNA expression was observed. These results demonstrated that ENA gapmers with a short DNA region improved the sequence-specificity of mRNA down-regulation. As well, they suggest that we should be alert to the utility of antisense oligonucleotides with a wide DNA region in terms of sequence specificity, and additionally, that optimized ENA gapmers could be useful for application to gene validation.
Asunto(s)
Hidrolasas/genética , Oligonucleótidos Antisentido/química , Animales , Secuencia de Bases , Regulación hacia Abajo , Hidrolasas/metabolismo , Ratones , Arginina Deiminasa Proteína-Tipo 4 , Desiminasas de la Arginina Proteica , ARN Mensajero/química , ARN Mensajero/metabolismoRESUMEN
Rheumatoid arthritis is a common autoimmune disease with a complex genetic etiology. Here we identify a SNP in the promoter region of FCRL3, a member of the Fc receptor-like family, that is associated with susceptibility to rheumatoid arthritis (odds ratio = 2.15, P = 0.00000085). This polymorphism alters the binding affinity of nuclear factor-kappaB and regulates FCRL3 expression. We observed high FCRL3 expression on B cells and augmented autoantibody production in individuals with the disease-susceptible genotype. We also found associations between the SNP and susceptibility to autoimmune thyroid disease and systemic lupus erythematosus. FCRL3 may therefore have a pivotal role in autoimmunity.
Asunto(s)
Artritis Reumatoide/genética , Autoinmunidad/genética , Receptores Inmunológicos/genética , Artritis Reumatoide/inmunología , Artritis Reumatoide/metabolismo , Autoinmunidad/inmunología , Autoinmunidad/fisiología , Estudios de Casos y Controles , Cromosomas Humanos Par 1 , Regulación de la Expresión Génica/fisiología , Antígenos HLA-DR/genética , Antígenos HLA-DR/inmunología , Cadenas HLA-DRB1 , Humanos , Desequilibrio de Ligamiento , Datos de Secuencia Molecular , Familia de Multigenes , Mutación , Polimorfismo de Nucleótido Simple , Receptores Inmunológicos/metabolismoRESUMEN
Recently we reported that SLC22A4 and RUNX1 are associated with rheumatoid arthritis (RA). SLC22A4 is an organic cation transporter with unknown physiological function, and RUNX1 is a hematological transcriptional regulator that has been shown to be responsible for acute myelogenic leukemia. It is suggested that the association of RUNX1 with RA is due to its regulation of expression of SLC22A4. Because the physiological function of SLC22A4 is still unclear, further investigation is needed into how SLC22A4 affects RA susceptibility. Although the association of RUNX1 with RA was identified as a regulatory factor of SLC22A4, it is possible that RUNX1 is a key molecule in autoimmunity, as it has been reported to be associated with systemic lupus erythematosus and psoriasis, two other autoimmune diseases.
Asunto(s)
Artritis Reumatoide/genética , Enfermedades Autoinmunes/genética , Proteínas de Unión al ADN/genética , Proteínas de Transporte de Membrana/genética , Polimorfismo de Nucleótido Simple , Proteínas Proto-Oncogénicas/genética , Factores de Transcripción/genética , Animales , Antígenos de Superficie/genética , Antígenos de Superficie/fisiología , Proteínas Reguladoras de la Apoptosis , Artritis Reumatoide/epidemiología , Pueblo Asiatico/genética , Enfermedades Autoinmunes/epidemiología , Estudios de Casos y Controles , Cromosomas Humanos Par 5/genética , Subunidad alfa 2 del Factor de Unión al Sitio Principal , Proteínas de Unión al ADN/biosíntesis , Proteínas de Unión al ADN/fisiología , Modelos Animales de Enfermedad , Regulación de la Expresión Génica , Predisposición Genética a la Enfermedad , Humanos , Japón/epidemiología , Lupus Eritematoso Sistémico/epidemiología , Lupus Eritematoso Sistémico/genética , Proteínas de Transporte de Membrana/biosíntesis , Proteínas de Transporte de Membrana/fisiología , Ratones , Modelos Genéticos , Herencia Multifactorial , Proteínas de Transporte de Catión Orgánico , Receptor de Muerte Celular Programada 1 , Regiones Promotoras Genéticas/genética , Unión Proteica , Proteínas Proto-Oncogénicas/biosíntesis , Proteínas Proto-Oncogénicas/fisiología , Psoriasis/epidemiología , Psoriasis/genética , Simportadores , Factores de Transcripción/biosíntesis , Factores de Transcripción/fisiologíaRESUMEN
Rheumatoid arthritis is a common inflammatory disease with complex genetic components. We investigated the genetic contribution of the cytokine gene cluster in chromosome 5q31 to susceptibility to rheumatoid arthritis in the Japanese population by case-control linkage disequilibrium (LD) mapping using single nucleotide polymorphisms (SNPs). Here we report that there is significant association between rheumatoid arthritis and the organic cation transporter gene SLC22A4 (P = 0.000034). We show that expression of SLC22A4 is specific to hematological and immunological tissues and that SLC22A4 is also highly expressed in the inflammatory joints of mice with collagen-induced arthritis. A SNP affects the transcriptional efficiency of SLC22A4 in vitro, owing to an allelic difference in affinity to Runt-related transcription factor 1 (RUNX1), a transcriptional regulator in the hematopoietic system. A SNP in RUNX1 is also strongly associated with rheumatoid arthritis (P = 0.00035). Our data indicate that the regulation of SLC22A4 expression by RUNX1 is associated with susceptibility to rheumatoid arthritis, which may represent an example of an epistatic effect of two genes on this disorder.
Asunto(s)
Artritis Reumatoide/genética , Proteínas de Unión al ADN/genética , Intrones/genética , Desequilibrio de Ligamiento , Transportadores de Anión Orgánico/genética , Proteínas de Transporte de Catión Orgánico , Polimorfismo de Nucleótido Simple/genética , Proteínas Proto-Oncogénicas , Factores de Transcripción/genética , Animales , Artritis Reumatoide/inducido químicamente , Proteínas Portadoras/genética , Estudios de Casos y Controles , Cromosomas Humanos Par 5/genética , Colágeno/farmacología , Subunidad alfa 2 del Factor de Unión al Sitio Principal , Citocinas/genética , Ensayo de Cambio de Movilidad Electroforética , Femenino , Frecuencia de los Genes , Predisposición Genética a la Enfermedad , Humanos , Células Jurkat , Luciferasas , Masculino , Proteínas de la Membrana/genética , Ratones , Persona de Mediana Edad , Regiones Promotoras Genéticas , Miembro 5 de la Familia 22 de Transportadores de SolutosRESUMEN
Individuals with rheumatoid arthritis frequently have autoantibodies to citrullinated peptides, suggesting the involvement of the peptidylarginine deiminases citrullinating enzymes (encoded by PADI genes) in rheumatoid arthritis. Previous linkage studies have shown that a susceptibility locus for rheumatoid arthritis includes four PADI genes but did not establish which PADI gene confers susceptibility to rheumatoid arthritis. We used a case-control linkage disequilibrium study to show that PADI type 4 is a susceptibility locus for rheumatoid arthritis (P = 0.000008). PADI4 was expressed in hematological and rheumatoid arthritis synovial tissues. We also identified a haplotype of PADI4 associated with susceptibility to rheumatoid arthritis that affected stability of transcripts and was associated with levels of antibody to citrullinated peptide in sera from individuals with rheumatoid arthritis. Our results imply that the PADI4 haplotype associated with susceptibility to rheumatoid arthritis increases production of citrullinated peptides acting as autoantigens, resulting in heightened risk of developing the disease.
Asunto(s)
Artritis Reumatoide/enzimología , Artritis Reumatoide/genética , Hidrolasas/genética , Adulto , Anciano , Anciano de 80 o más Años , Artritis Reumatoide/inmunología , Autoanticuerpos/sangre , Autoantígenos/química , Autoantígenos/metabolismo , Estudios de Casos y Controles , Cromosomas Humanos Par 1/genética , Citrulina/química , Citrulina/metabolismo , Femenino , Proteínas Filagrina , Haplotipos , Humanos , Hidrolasas/metabolismo , Proteínas de Filamentos Intermediarios/química , Proteínas de Filamentos Intermediarios/inmunología , Proteínas de Filamentos Intermediarios/metabolismo , Desequilibrio de Ligamiento , Masculino , Persona de Mediana Edad , Péptidos/química , Péptidos/inmunología , Péptidos/metabolismo , Polimorfismo de Nucleótido Simple , Arginina Deiminasa Proteína-Tipo 4 , Desiminasas de la Arginina Proteica , Estabilidad del ARN , ARN Mensajero/genética , ARN Mensajero/metabolismoRESUMEN
beta-Amyloid peptide (A beta), a major component of senile plaques, the formation of which is characteristic of Alzheimer's disease (AD), is believed to induce inflammation of the brain mediated by microglia, leading to neuronal cell loss. In this study, we performed an oligonucleotide microarray analysis to investigate the molecular events underlying the A beta-induced activation of macrophages and its specific suppression by the A beta-specific-macrophage-activation inhibitor, RS-1178. Of the approximately 36,000 genes and expressed sequence tags analyzed, eight genes were specifically and significantly upregulated by a treatment with interferon gamma (IFN gamma) and A beta compared to a treatment with IFN gamma alone (p<0.002). We found that the gene for a well-characterized lipogenetic enzyme, stearoyl coenzyme A desaturase-1 (SCD-1), was specifically upregulated by A beta treatment and was suppressed to basal levels by RS-1178. Although the underlying mechanisms remain unknown, our results suggest the presence of a link between AD and SCD-1.
Asunto(s)
Péptidos beta-Amiloides/metabolismo , Macrófagos/enzimología , Macrófagos/metabolismo , Estearoil-CoA Desaturasa/química , Estearoil-CoA Desaturasa/genética , Regulación hacia Arriba , Enfermedad de Alzheimer/metabolismo , Animales , Humanos , Interferón gamma/metabolismo , Macrófagos Peritoneales/metabolismo , Ratones , Análisis de Secuencia por Matrices de Oligonucleótidos , Pirimidinas/farmacología , ARN/metabolismo , ARN Mensajero/metabolismo , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Factores de TiempoRESUMEN
beta-Amyloid peptide (Abeta), a major component of senile plaques, the formation of which is characteristic of Alzheimer's disease (AD), is believed to induce inflammation in the brain leading to cell loss and cognitive decline. Accumulating evidence shows Abeta activates microglia, which play the role of the brain's immune system, and mediates inflammatory responses in the brain. Thus, a compound inhibiting Abeta-induced activation of microglia may lead to a novel therapy for AD. However, the compound should not inhibit natural immune responses during events such as bacterial infections. We investigated the effect of a synthesized compound, 7,8-dihydro-5-methyl-8-(1-phenylethyl)-6H-pyrrolo [3,2-e] [1,2,4] triazolo [1,5-a] pyrimidine (RS-1178) on macrophage activation induced by various stimulants. The activation of macrophages was determined by nitric oxide or tumor necrosis factor alpha production. RS-1178 inhibited Abeta-induced macrophage activation but did not inhibit zymosan A- nor lipopolysaccharide (LPS)-induced macrophage activation. Moreover, RS-1178 attenuated neurotoxicity due to Abeta-induced macrophage activation in neuron-macrophage co-cultures but not neurotoxicity due to zymosan A- or LPS-induced macrophage activation. In conclusion, RS-1178 showed a specific inhibitory effect on Abeta-induced macrophage activation. Although the exact mechanisms of this effect remain unknown, RS-1178 may provide a novel therapy for AD.