RESUMEN
RATIONALE: Patterns of longitudinal lung function growth and decline in childhood asthma have been shown to be important in determining risk for future respiratory ailments including chronic airway obstruction and chronic obstructive pulmonary disease. OBJECTIVES: To determine the genetic underpinnings of lung function patterns in subjects with childhood asthma. METHODS: We performed a genome-wide association study of 581 non-Hispanic white individuals with asthma that were previously classified by patterns of lung function growth and decline (normal growth, normal growth with early decline, reduced growth, and reduced growth with early decline). The strongest association was also measured in two additional cohorts: a small asthma cohort and a large chronic obstructive pulmonary disease metaanalysis cohort. Interaction between the genomic region encompassing the most strongly associated single-nucleotide polymorphism and nearby genes was assessed by two chromosome conformation capture assays. MEASUREMENTS AND MAIN RESULTS: An intergenic single-nucleotide polymorphism (rs4445257) on chromosome 8 was strongly associated with the normal growth with early decline pattern compared with all other pattern groups (P = 6.7 × 10-9; odds ratio, 2.8; 95% confidence interval, 2.0-4.0); replication analysis suggested this variant had opposite effects in normal growth with early decline and reduced growth with early decline pattern groups. Chromosome conformation capture experiments indicated a chromatin interaction between rs4445257 and the promoter of the distal CSMD3 gene. CONCLUSIONS: Early decline in lung function after normal growth is associated with a genetic polymorphism that may also protect against early decline in reduced growth groups. Clinical trial registered with www.clinicaltrials.gov (NCT00000575).
Asunto(s)
Asma/genética , Asma/fisiopatología , Predisposición Genética a la Enfermedad/genética , Genómica/métodos , Pulmón/fisiopatología , Niño , Preescolar , Femenino , Volumen Espiratorio Forzado , Estudio de Asociación del Genoma Completo , Humanos , Estudios Longitudinales , Masculino , Países Bajos , Polimorfismo de Nucleótido Simple/genética , Polimorfismo de Nucleótido Simple/fisiologíaRESUMEN
Many neurological and psychiatric disorders exhibit gender disparities, and sex differences in the brain likely explain some of these effects. Recent work in rodents points to a role for epigenetics in the development or maintenance of neural sex differences, although genome-wide studies have so far been lacking. Here we review the existing literature on epigenetics and brain sexual differentiation and present preliminary analyses on the genome-wide distribution of histone-3 lysine-4 trimethylation in a sexually dimorphic brain region in male and female mice. H3K4me3 is a histone mark primarily organized as 'peaks' surrounding the transcription start site of active genes. We microdissected the bed nucleus of the stria terminalis and preoptic area (BNST/POA) in adult male and female mice and used ChIP-Seq to compare the distribution of H3K4me3 throughout the genome. We found 248 genes and loci with a significant sex difference in H3K4me3. Of these, the majority (71%) had larger H3K4me3 peaks in females. Comparisons with existing databases indicate that genes and loci with increased H3K4me3 in females are associated with synaptic function and with expression atlases from related brain areas. Based on RT-PCR, only a minority of genes with a sex difference in H3K4me3 has detectable sex differences in expression at baseline conditions. Together with previous findings, our data suggest that there may be sex biases in the use of epigenetic marks. Such biases could underlie sex differences in vulnerabilities to drugs or diseases that disrupt specific epigenetic processes.
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Encéfalo/fisiología , Epigenómica , Histonas/genética , Caracteres Sexuales , Animales , Femenino , Lisina/genética , Masculino , RatonesRESUMEN
Many cellular constituents in the human brain permanently exit from the cell cycle during pre- or early postnatal development, but little is known about epigenetic regulation of neuronal and glial epigenomes during maturation and aging, including changes in mood and psychosis spectrum disorders and other cognitive or emotional disease. Here, we summarize the current knowledge base as it pertains to genome organization in the human brain, including the regulation of DNA cytosine methylation and hydroxymethylation, and a subset of (altogether >100) residue-specific histone modifications associated with gene expression, and silencing and various other functional chromatin states. We propose that high-resolution mapping of epigenetic markings in postmortem brain tissue or neural cultures derived from induced pluripotent cells (iPS), in conjunction with transcriptome profiling and whole-genome sequencing, will increasingly be used to define the molecular pathology of specific cases diagnosed with depression, schizophrenia, autism, or other major psychiatric disease. We predict that these highly integrative explorations of genome organization and function will provide an important alternative to conventional approaches in human brain studies, which mainly are aimed at uncovering group effects by diagnosis but generally face limitations because of cohort size.
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Química Encefálica/genética , Epigénesis Genética/fisiología , Metilación de ADN/fisiología , Humanos , Trastornos Mentales/genética , Trastornos Mentales/metabolismoRESUMEN
Cognitive abilities and disorders unique to humans are thought to result from adaptively driven changes in brain transcriptomes, but little is known about the role of cis-regulatory changes affecting transcription start sites (TSS). Here, we mapped in human, chimpanzee, and macaque prefrontal cortex the genome-wide distribution of histone H3 trimethylated at lysine 4 (H3K4me3), an epigenetic mark sharply regulated at TSS, and identified 471 sequences with human-specific enrichment or depletion. Among these were 33 loci selectively methylated in neuronal but not non-neuronal chromatin from children and adults, including TSS at DPP10 (2q14.1), CNTN4 and CHL1 (3p26.3), and other neuropsychiatric susceptibility genes. Regulatory sequences at DPP10 and additional loci carried a strong footprint of hominid adaptation, including elevated nucleotide substitution rates and regulatory motifs absent in other primates (including archaic hominins), with evidence for selective pressures during more recent evolution and adaptive fixations in modern populations. Chromosome conformation capture at two neurodevelopmental disease loci, 2q14.1 and 16p11.2, revealed higher order chromatin structures resulting in physical contact of multiple human-specific H3K4me3 peaks spaced 0.5-1 Mb apart, in conjunction with a novel cis-bound antisense RNA linked to Polycomb repressor proteins and downregulated DPP10 expression. Therefore, coordinated epigenetic regulation via newly derived TSS chromatin could play an important role in the emergence of human-specific gene expression networks in brain that contribute to cognitive functions and neurological disease susceptibility in modern day humans.
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Metilación de ADN , Histonas/metabolismo , Neuronas/metabolismo , Corteza Prefrontal/citología , Sitio de Iniciación de la Transcripción , Adulto , Animales , Secuencia de Bases , Niño , Cromatina/metabolismo , Ensamble y Desensamble de Cromatina , Mapeo Cromosómico , Cognición , Contactinas/genética , Dipeptidil-Peptidasas y Tripeptidil-Peptidasas/genética , Epigénesis Genética , Evolución Molecular , Redes Reguladoras de Genes , Sitios Genéticos , Histonas/genética , Humanos , Lisina/metabolismo , Macaca/genética , Trastornos Mentales/genética , Neuronas/citología , Pan troglodytes/genética , Filogenia , Proteínas del Grupo Polycomb/metabolismo , Corteza Prefrontal/metabolismo , Secuencias Reguladoras de Ácidos Nucleicos , Especificidad de la Especie , Transcripción GenéticaRESUMEN
Maternal immune activation during prenatal development, including treatment with the viral RNA mimic, polyriboinosinic-polyribocytidilic acid (poly IC), serves as a widely used animal model to induce behavioral deficits reminiscent of schizophrenia and related disease. Here, we report that massive cytokine activation after a single dose of poly IC in the prenatal period is associated with lasting working memory deficits in adult offspring. To explore whether dysregulated gene expression in cerebral cortex, contributes to cognitive dysfunction, we profiled the cortical transcriptome, and in addition, mapped the genome-wide distribution of trimethylated histone H3-lysine 4 (H3K4me3), an epigenetic mark sharply regulated at the 5' end of transcriptional units. However, deep sequencing-based H3K4me3 mapping and mRNA profiling by microarray did not reveal significant alterations in mature cerebral cortex after poly IC exposure at embryonic days E17.5 or E12.5. At a small set of genes (including suppressor of cytokine signaling Socs3), H3K4me3 was sensitive to activation of cytokine signaling in primary cultures from fetal forebrain but adult cortex of saline- and poly IC-exposed mice did not show significant differences. A limited set of transcription start sites (TSS), including Disrupted-in-Schizophrenia 1 (Disc1), a schizophrenia risk gene often implicated in gene-environment interaction models, showed altered H3K4me3 after prenatal poly IC but none of these differences survived after correcting for multiple comparisons. We conclude that prenatal poly IC is associated with cognitive deficits later in life, but without robust alterations in epigenetic regulation of gene expression in the cerebral cortex.
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Corteza Cerebral/efectos de los fármacos , Corteza Cerebral/metabolismo , Epigenómica , Trastornos de la Memoria/inducido químicamente , Efectos Tardíos de la Exposición Prenatal/inmunología , Transcriptoma/efectos de los fármacos , Animales , Células Cultivadas , Corteza Cerebral/citología , Corteza Cerebral/embriología , Inmunoprecipitación de Cromatina , Citocinas/sangre , Embrión de Mamíferos , Ensayo de Inmunoadsorción Enzimática , Femenino , Factor 2 de Crecimiento de Fibroblastos/farmacología , Regulación de la Expresión Génica/efectos de los fármacos , Inductores de Interferón/farmacología , Ratones , Ratones Endogámicos C57BL , Análisis por Micromatrices , Neuronas/efectos de los fármacos , Poli I-C/farmacología , Embarazo , Efectos Tardíos de la Exposición Prenatal/fisiopatología , Ratas , Ratas Sprague-DawleyRESUMEN
The Ets family transcription factor PU.1, encoded by the gene Sfpi1, is essential for normal hematopoiesis. A number of studies have suggested that changes in PU.1 concentration play a role in directing cell fate decisions during hematopoiesis. However, the stages of hematopoietic development at which changes in PU.1 concentration are important have not been defined until recently. Experiments using conditional null alleles, reporter alleles, and hypomorphic alleles of the Sfpi1 gene in mice demonstrate that PU.1 concentration is uniformly high during early stages of hematopoietic development. However, reduction of PU.1 concentration is required for normal development of megakaryocyte-erythroid progenitors, B cell progenitors, and T cell progenitors. PU.1 concentration increases in granulocyte-macrophage progenitors. Furthermore, experimental reduction of PU.1 concentration in the myeloid lineages leads to failed differentiation, abnormal proliferation, and leukemia. In this review, we summarize recent studies to develop a new model of PU.1 function in hematopoiesis.
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Alelos , Hematopoyesis , Leucemia Mieloide Aguda/metabolismo , Proteínas Proto-Oncogénicas/metabolismo , Transactivadores/metabolismo , Animales , Hematopoyesis/genética , Proteínas Proto-Oncogénicas/genética , Transactivadores/genéticaRESUMEN
OBJECTIVE: It has been demonstrated that high concentration of the transcription factor PU.1 (encoded by Sfpi1) promotes macrophage development, whereas low concentration induces B-cell development in vitro. This has led to the hypothesis that lower levels of PU.1 activity are required for B cell than for macrophage development in vivo. We utilized an allele of Sfpi1 (termed BN) with a mutation in the first coding exon, which resulted in a reduction of PU.1 expression in order to test this hypothesis. MATERIALS AND METHODS: Using gene targeting in embryonic stem cells, two ATG-start site codons of PU.1 were mutated, resulting in reduced PU.1 expression originating from a third start codon. Mice were assayed for phenotypic abnormalities using fluorescence-activated cell sorting, microscopy, and colony-forming ability. In addition, isolated cells were tested for their differentiation potential in vitro and in vivo. RESULTS: Lymphoid and myeloid cells derived from cultured Sfpi1(BN/BN) fetal liver cells had reduced levels of PU.1 expression and activity. B-cell development was intrinsically blocked in cells isolated from Sfpi1(BN/BN) mice. In addition, myeloid development was impaired in Sfpi1(BN/BN) fetal liver. However, neonatal Sfpi1(BN/BN) mice had a dramatic expansion and infiltration of immature myeloid cells. CONCLUSION: Contrary to our original hypothesis, high levels of PU.1 activity are required to induce both myeloid and B-cell development. In addition, neonatal mice homozygous for the hypomorphic allele acquire a myeloproliferative disorder and die within 1 month of age.
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Linfocitos B/fisiología , Trastornos Mieloproliferativos/etiología , Proteínas Proto-Oncogénicas/fisiología , Transactivadores/fisiología , Alelos , Animales , Animales Recién Nacidos , Proliferación Celular , Células Cultivadas , Humanos , Ratones , Ratones Endogámicos C57BL , Mielopoyesis , Proteínas Proto-Oncogénicas/genética , Transactivadores/genéticaRESUMEN
OBJECTIVE: The Ets family transcription factor PU.1 is essential for both myeloid and lymphoid development. PU.1 was discovered because of its involvement in murine erythroleukemia. We previously described that infection with a retroviral vector encoding PU.1 immortalizes fetal liver progenitor cells in response to granulocyte-macrophage colony-stimulating factor (GM-CSF) signaling. In this study, we sought to characterize PU.1-immortalized progenitor (PIP) cells. METHODS: PIP cells were characterized using microscopy, reverse-transcriptase polymerase chain reaction analysis, and flow cytometric analysis. In addition, progenitors were immortalized with a retrovirus containing a PU.1 cDNA flanked by loxP sites. The differentiation potential of immortalized progenitors was tested by Cre-mediated excision of the proviral PU.1 cDNA. RESULTS: PIP cells are blastlike in morphology and express cell surface markers indicative of myeloid development. Immortalization of progenitor cells requires both an acidic activation domain and an intact DNA-binding domain of PU.1. Gene expression analysis of PIP cells demonstrated the expression of genes of both myeloid and erythroid lineages. Proliferation of PIP cells was GM-CSF dependent and restricted. Upon Cre-mediated excision of proviral PU.1 cDNA, increased expression of myeloid and erythroid-specific genes was observed; as well as the appearance of both macrophages and erythrocytes in culture. CONCLUSION: We demonstrate that ectopic expression of PU.1 is sufficient to immortalize a hematopoietic progenitor with myeloid and erythroid differentiation potential in response to GM-CSF. These data highlight the importance of the level of PU.1 expression at critical stages of hematopoiesis.