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Controlling large-scale many-body quantum systems at the level of single photons and single atomic systems is a central goal in quantum information science and technology. Intensive research and development has propelled foundry-based silicon-on-insulator photonic integrated circuits to a leading platform for large-scale optical control with individual mode programmability. However, integrating atomic quantum systems with single-emitter tunability remains an open challenge. Here, we overcome this barrier through the hybrid integration of multiple InAs/InP microchiplets containing high-brightness infrared semiconductor quantum dot single photon emitters into advanced silicon-on-insulator photonic integrated circuits fabricated in a 300 mm foundry process. With this platform, we achieve single-photon emission via resonance fluorescence and scalable emission wavelength tunability. The combined control of photonic and quantum systems opens the door to programmable quantum information processors manufactured in leading semiconductor foundries.
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People often modify the shoreline to mitigate erosion and protect property from storm impacts. The 2 main approaches to modification are gray infrastructure (e.g., bulkheads and seawalls) and natural or green infrastructure (NI) (e.g., living shorelines). Gray infrastructure is still more often used for coastal protection than NI, despite having more detrimental effects on ecosystem parameters, such as biodiversity. We assessed the impact of gray infrastructure on biodiversity and whether the adoption of NI can mitigate its loss. We examined the literature to quantify the relationship of gray infrastructure and NI to biodiversity and developed a model with temporal geospatial data on ecosystem distribution and shoreline modification to project future shoreline modification for our study location, coastal Georgia (United States). We applied the literature-derived empirical relationships of infrastructure effects on biodiversity to the shoreline modification projections to predict change in biodiversity under different NI versus gray infrastructure scenarios. For our study area, which is dominated by marshes and use of gray infrastructure, when just under half of all new coastal infrastructure was to be NI, previous losses of biodiversity from gray infrastructure could be mitigated by 2100 (net change of biodiversity of +0.14%, 95% confidence interval -0.10% to +0.39%). As biodiversity continues to decline from human impacts, it is increasingly imperative to minimize negative impacts when possible. We therefore suggest policy and the permitting process be changed to promote the adoption of NI.
Cuantificación del impacto de la futura modificación de la costa sobre la biodiversidad en un estudio de caso de la costa de Georgia, Estados Unidos Resumen Las personas modifican con frecuencia la costa para mitigar la erosión o proteger su propiedad del impacto de las tormentas. Los dos enfoques principales para la modificación son la infraestructura gris (p. ej.: mamparos y malecones) y la infraestructura verde o natural (IN) (p.ej.: costas vivientes). La infraestructura gris es más común que la IN, a pesar de que tiene efectos dañinos sobre los parámetros ambientales, como la biodiversidad. Evaluamos el impacto de la infraestructura gris sobre la biodiversidad y si la adopción de la IN puede mitigar su pérdida. Analizamos la literatura para cuantificar la relación de la infraestructura gris y la IN con la biodiversidad. También desarrollamos un modelo con datos geoespaciales temporales sobre la distribución de los ecosistemas y la modificación de la costa para proyectar la modificación costera en el futuro en nuestra localidad de estudio: la costa de Georgia, Estados Unidos. Aplicamos las relaciones empíricas derivadas de la literatura de los efectos de la infraestructura sobre la biodiversidad a las proyecciones de modificación de la costa para predecir el cambio en la biodiversidad bajo diferentes escenarios de infraestructura gris versus IN. En nuestra área de estudio, que está dominada por marismas y usa infraestructura gris, cuando un poco menos de la mitad de toda la infraestructura costera nueva debería ser IN, las pérdidas previas de biodiversidad a partir de la infraestructura gris podrían mitigarse para 2100 (cambio neto de la biodiversidad de +0.14%, 95% intervalo de confianza 0.10% a +0.39%). Conforme la biodiversidad siga en declive por el impacto humano, cada vez es más imperativo minimizar el impacto negativo cuando sea posible. Por lo tanto, sugerimos que se modifiquen las políticas y el proceso de permisos para promover la adopción de la IN.
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AML is characterized by mutations in genes associated with growth regulation such as internal tandem duplications (ITD) in the receptor kinase FLT3. Inhibitors targeting FLT3 (FLT3i) are being used to treat patients with FLT3-ITD+ but most relapse and become resistant. To elucidate the resistance mechanism, we compared the gene regulatory networks (GRNs) of leukemic cells from patients before and after relapse, which revealed that the GRNs of drug-responsive patients were altered by rewiring their AP-1-RUNX1 axis. Moreover, FLT3i induces the upregulation of signaling genes, and we show that multiple cytokines, including interleukin-3 (IL-3), can overcome FLT3 inhibition and send cells back into cycle. FLT3i leads to loss of AP-1 and RUNX1 chromatin binding, which is counteracted by IL-3. However, cytokine-mediated drug resistance can be overcome by a pan-RAS inhibitor. We show that cytokines instruct AML growth via the transcriptional regulators AP-1 and RUNX1 and that pan-RAS drugs bypass this barrier.
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Acute Myeloid Leukemia (AML) is caused by multiple mutations which dysregulate growth and differentiation of myeloid cells. Cells adopt different gene regulatory networks specific to individual mutations, maintaining a rapidly proliferating blast cell population with fatal consequences for the patient if not treated. The most common treatment option is still chemotherapy which targets such cells. However, patients harbour a population of quiescent leukemic stem cells (LSCs) which can emerge from quiescence to trigger relapse after therapy. The processes that allow such cells to re-grow remain unknown. Here, we examine the well characterised t(8;21) AML sub-type as a model to address this question. Using four primary AML samples and a novel t(8;21) patient-derived xenograft model, we show that t(8;21) LSCs aberrantly activate the VEGF and IL-5 signalling pathways. Both pathways operate within a regulatory circuit consisting of the driver oncoprotein RUNX1::ETO and an AP-1/GATA2 axis allowing LSCs to re-enter the cell cycle while preserving self-renewal capacity.
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Leucemia Mieloide Aguda , Humanos , Leucemia Mieloide Aguda/genética , Leucemia Mieloide Aguda/metabolismo , Mutação , Células-Tronco/metabolismo , Células-Tronco Neoplásicas/metabolismoRESUMO
Acute myeloid leukemia (AML) is a heterogeneous disease caused by different mutations. Previously, we showed that each mutational subtype develops its specific gene regulatory network (GRN) with transcription factors interacting within multiple gene modules, many of which are transcription factor genes themselves. Here, we hypothesize that highly connected nodes within such networks comprise crucial regulators of AML maintenance. We test this hypothesis using FLT3-ITD-mutated AML as a model and conduct an shRNA drop-out screen informed by this analysis. We show that AML-specific GRNs predict crucial regulatory modules required for AML growth. Furthermore, our work shows that all modules are highly connected and regulate each other. The careful multi-omic analysis of the role of one (RUNX1) module by shRNA and chemical inhibition shows that this transcription factor and its target genes stabilize the GRN of FLT3-ITD+ AML and that its removal leads to GRN collapse and cell death.
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Redes Reguladoras de Genes , Leucemia Mieloide Aguda , Humanos , Regulon , Leucemia Mieloide Aguda/genética , Leucemia Mieloide Aguda/metabolismo , Mutação/genética , RNA Interferente Pequeno , Tirosina Quinase 3 Semelhante a fms/genéticaRESUMO
Sea level rise (SLR) is the most significant climate change-related threat to coastal wetlands, driving major transformations in coastal regions through marsh migration. Landscape transformations due to marsh migration are manifested in terms of horizontal and vertical changes in land cover and elevation, respectively. These processes will have an impact on saltmarsh wave attenuation that is yet to be explored. This study stands as a comprehensive analysis of spatially distributed wave attenuation by vegetation in the context of a changing climate. Our results show that: i) changes in saltmarsh cover have little to no effect on the attenuation of floods, while ii) changes in elevation can significantly reduce flood extents and water depths; iii) overland wave heights are directly influenced by marsh migration, although iv) being indirectly attenuated by the water depth limiting effects of water depth attenuation driven by changes in elevation; v) the influence of saltmarsh accretion on wave attenuation is largely evident near the marsh edge, where the increasing elevations can drive major wave energy losses via wave breaking. Lastly, vi) considering the synergy between SLR, marsh migration, and changes in elevation results in significantly more wave attenuation than considering the eustatic effects of SLR and/or horizontal marsh migration alone, and therefore should be adopted in future studies.
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Elevação do Nível do Mar , Áreas Alagadas , Monitoramento Ambiental , Mudança Climática , Água , EcossistemaRESUMO
Coastal communities are vulnerable to wave and storm surges during extreme events, highlighting the need to increase community resilience. The effectiveness of natural wetlands in attenuating waves is vital to designing strategies for protecting public safety. This study aimed to understand how vegetation attenuates waves and determine the best method for modeling vegetation's impact on wave dynamics. The researchers compared two different vegetation representations in numerical models, implicit and explicit, using SWAN and XBeach at varying spatial resolutions. The study focused on two marshes in the Chesapeake Bay, using field measurements to investigate the accuracy of each method in representing wave attenuation by vegetation and the implications of explicitly representing average characteristics of one vegetation species on a regional level. Results showed that explicit modeling using average vegetation characteristics provided more accurate results than the implicit model, which only showed wave attenuation due to topography. The finer scale resolution and site-specific vegetation characteristics further improved the accuracy of wave attenuation observed. Understanding the trade-offs between different vegetation representations in numerical models is essential to accurately represent wave attenuation and design effective protection strategies for coastal communities.
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Baías , Monitoramento Ambiental , Áreas AlagadasRESUMO
AML is a heterogenous disease caused by different mutations. We have previously shown that each mutational sub-type develops its specific gene regulatory network (GRN) with transcription factors interacting with multiple gene modules, many of which are transcription factor genes themselves. Here we hypothesized that highly connected nodes within such networks comprise crucial regulators of AML maintenance. We tested this hypothesis using FLT3-ITD mutated AML as a model and conducted an shRNA drop-out screen informed by this analysis. We show that AML-specific GRNs predict identifying crucial regulatory modules required for AML but not normal cellular growth. Furthermore, our work shows that all modules are highly connected and regulate each other. The careful multi-omic analysis of the role of one (RUNX1) module by shRNA and chemical inhibition shows that this transcription factor and its target genes stabilize the GRN of FLT3-ITD AML and that its removal leads to GRN collapse and cell death.
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River regulation by dams can alter flow regimes and organic matter dynamics, but less is known about how unregulated tributaries regulate organic matter composition and processing in the regulated river below the confluence. This study reports on water chemistry, especially dissolved organic matter (DOM) concentration and composition (dissolved organic carbon (DOC), organic nitrogen (DON), organic phosphorus (DOP) and combined amino acids (DCAA)) along the regulated Tumut and unregulated Goobarragandra (tributary) rivers under different flow conditions (base flow vs storm event) in south-east Australia. The tributary was significantly different from regulated and downstream sites during base flow conditions with higher temperature, pH, buffering capacity, DOC and nutrient concentrations (DON, DOP, DCAA). DOM characterisation by spectrometry and size exclusion chromatography revealed that the tributary contained a higher proportion of terrestrially derived humic-like and fulvic-like DOM. In contrast, regulated and downstream sites contained higher proportion of microbially derived DOM such as low molecular weight neutrals and protein-like components. Storm pulses of tributary flows into the regulated system, influenced both concentration and composition of DOM at the downstream site, which more strongly resembled the tributary site than the regulated site during the storm event. Additionally, we found that the tributary supplied fresh DOM, including small organic molecules to the regulated system during storm events. The presence of these different types of labile DOM can increase primary productivity and ecological functioning within regulated river reaches downstream of tributary junctions. This has important implications for the protection of unregulated tributary inflows within regulated river basins.
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Matéria Orgânica Dissolvida , Rios , Rios/química , Nitrogênio/química , FósforoRESUMO
Mutations in Fms-like tyrosine kinase 3 (FLT3) are common drivers in acute myeloid leukemia (AML) yet FLT3 inhibitors only provide modest clinical benefit. Prior work has shown that inhibitors of lysine-specific demethylase 1 (LSD1) enhance kinase inhibitor activity in AML. Here we show that combined LSD1 and FLT3 inhibition induces synergistic cell death in FLT3-mutant AML. Multi-omic profiling revealed that the drug combination disrupts STAT5, LSD1, and GFI1 binding at the MYC blood superenhancer, suppressing superenhancer accessibility as well as MYC expression and activity. The drug combination simultaneously results in the accumulation of repressive H3K9me1 methylation, an LSD1 substrate, at MYC target genes. We validated these findings in 72 primary AML samples with the nearly every sample demonstrating synergistic responses to the drug combination. Collectively, these studies reveal how epigenetic therapies augment the activity of kinase inhibitors in FLT3-ITD (internal tandem duplication) AML. IMPLICATIONS: This work establishes the synergistic efficacy of combined FLT3 and LSD1 inhibition in FLT3-ITD AML by disrupting STAT5 and GFI1 binding at the MYC blood-specific superenhancer complex.
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Leucemia Mieloide Aguda , Tirosina Quinase 3 Semelhante a fms , Humanos , Apoptose , Tirosina Quinase 3 Semelhante a fms/genética , Tirosina Quinase 3 Semelhante a fms/metabolismo , Histona Desmetilases/genética , Histona Desmetilases/metabolismo , Leucemia Mieloide Aguda/tratamento farmacológico , Leucemia Mieloide Aguda/genética , Leucemia Mieloide Aguda/metabolismo , Mutação , Inibidores de Proteínas Quinases/farmacologia , Inibidores de Proteínas Quinases/uso terapêutico , Fator de Transcrição STAT5/metabolismoRESUMO
Diffuse large B-cell lymphoma (DLBCL) exhibits significant genetic heterogeneity which contributes to drug resistance, necessitating development of novel therapeutic approaches. Pharmacological inhibitors of cyclin-dependent kinases (CDK) demonstrated pre-clinical activity in DLBCL, however many stalled in clinical development. Here we show that AZD4573, a selective inhibitor of CDK9, restricted growth of DLBCL cells. CDK9 inhibition (CDK9i) resulted in rapid changes in the transcriptome and proteome, with downmodulation of multiple oncoproteins (eg, MYC, Mcl-1, JunB, PIM3) and deregulation of phosphoinotiside-3 kinase (PI3K) and senescence pathways. Following initial transcriptional repression due to RNAPII pausing, we observed transcriptional recovery of several oncogenes, including MYC and PIM3. ATAC-Seq and ChIP-Seq experiments revealed that CDK9i induced epigenetic remodeling with bi-directional changes in chromatin accessibility, suppressed promoter activation and led to sustained reprograming of the super-enhancer landscape. A CRISPR library screen suggested that SE-associated genes in the Mediator complex, as well as AKT1, confer resistance to CDK9i. Consistent with this, sgRNA-mediated knockout of MED12 sensitized cells to CDK9i. Informed by our mechanistic findings, we combined AZD4573 with either PIM kinase or PI3K inhibitors. Both combinations decreased proliferation and induced apoptosis in DLBCL and primary lymphoma cells in vitro as well as resulted in delayed tumor progression and extended survival of mice xenografted with DLBCL in vivo. Thus, CDK9i induces reprogramming of the epigenetic landscape, and super-enhancer driven recovery of select oncogenes may contribute to resistance to CDK9i. PIM and PI3K represent potential targets to circumvent resistance to CDK9i in the heterogeneous landscape of DLBCL.
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Quinase 9 Dependente de Ciclina , Epigênese Genética , Linfoma Difuso de Grandes Células B , Animais , Camundongos , Apoptose , Linhagem Celular Tumoral , Linfoma Difuso de Grandes Células B/tratamento farmacológico , Linfoma Difuso de Grandes Células B/genética , Fosfatidilinositol 3-Quinases/metabolismo , Fatores de Transcrição/genética , Quinase 9 Dependente de Ciclina/antagonistas & inibidores , Resistencia a Medicamentos AntineoplásicosRESUMO
Mutations in the gene Additional Sex-Combs Like 1 (ASXL1) are recurrent in myeloid malignancies as well as the pre-malignant condition clonal hematopoiesis, where they are universally associated with poor prognosis. However, the role of ASXL1 in myeloid lineage maturation is incompletely described. To define the role of ASXL1 in myelopoiesis, we employed single cell RNA sequencing and a murine model of hematopoietic-specific Asxl1 deletion. In granulocyte progenitors, Asxl1 deletion leads to hyperactivation of MYC and a quantitative decrease in neutrophil production. This loss of granulocyte production was not accompanied by significant changes in the landscape of covalent histone modifications. However, Asxl1 deletion results in a decrease in RNAPII promoter-proximal pausing in granulocyte progenitors, indicative of a global increase in productive transcription. These results suggest that ASXL1 inhibits productive transcription in granulocyte progenitors, identifying a new role for this epigenetic regulator in myeloid development.
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Síndromes Mielodisplásicas , RNA Polimerase II , Proteínas Repressoras , Animais , Humanos , Camundongos , Células Precursoras de Granulócitos/patologia , Mutação , Síndromes Mielodisplásicas/genética , Proteínas Repressoras/genética , RNA Polimerase II/genética , Fatores de Transcrição/genéticaRESUMO
This article considers designed experiments for stability, comparability, and formulation testing that are analyzed with regression models in which the degradation rate is a fixed effect. In this setting, we investigate how the number of lots, the number of time points and their locations affect the precision of the entities of interest, leverages of the time points, detection of non-linearity and interim analyses. This investigation shows that modifying time point locations suggested by ICH for stability studies can significantly improve these objectives. In addition, we show that estimates of precision can be biased when a regression model that assumes independent measurements is used in the presence of within-assay session correlation. This bias can lead to longer shelf life estimates in stability studies and loss of power in comparability studies. Mixed-effect models that take into account within-assay session correlation are shown to reduce this bias. The findings in this article are obtained from well known statistical theory but provide valuable practical advice to scientists and statisticians designing and interpreting these types of experiments.
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Bioensaio , Modelos Estatísticos , Humanos , Viés , Fatores de TempoRESUMO
Acute myeloid leukemia (AML) is a heterogeneous hematological malignancy caused by mutations in genes encoding transcriptional and epigenetic regulators together with signaling genes. It is characterized by a disturbance of differentiation and abnormal proliferation of hematopoietic progenitors. We have previously shown that each AML subtype establishes its own core gene regulatory network (GRN), consisting of transcription factors binding to their target genes and imposing a specific gene expression pattern that is required for AML maintenance. In this study, we integrate gene expression, open chromatin and ChIP data with promoter-capture Hi-C data to define a refined core GRN common to all patients with CEBPA-double mutant (CEBPAN/C) AML. These mutations disrupt the structure of a major regulator of myelopoiesis. We identify the binding sites of mutated C/EBPα proteins in primary cells, we show that C/EBPα, AP-1 factors and RUNX1 colocalize and are required for AML maintenance, and we employ single cell experiments to link important network nodes to the specific differentiation trajectory from leukemic stem to blast cells. Taken together, our study provides an important resource which predicts the specific therapeutic vulnerabilities of this AML subtype in human cells.
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Redes Reguladoras de Genes , Leucemia Mieloide Aguda , Humanos , Proteínas Estimuladoras de Ligação a CCAAT/genética , Proteínas Estimuladoras de Ligação a CCAAT/metabolismo , Proteína alfa Estimuladora de Ligação a CCAAT/genética , Proteína alfa Estimuladora de Ligação a CCAAT/metabolismo , Mutação , Diferenciação Celular/genética , Leucemia Mieloide Aguda/patologiaRESUMO
The identification of critical process parameters in biologics and small molecule process development is a key element of quality by design. The objectivity and consistency of procedures to identify critical process parameters can be improved with the use of impact ratios. Impact ratios quantify a process parameter's practical effect on a critical quality attribute relative to the critical quality attribute's acceptance limits. If the impact ratio is large, i.e., exceeds a predefined impact ratio threshold, the recommendation is to classify the process parameter as a critical process parameter. This article introduces an improved and mathematically well-defined impact ratio. Benefits of this impact ratio are a consistent interpretation for many scenarios commonly encountered in practice, high suitability to automation, and the possibility of standardizing on a single impact ratio definition for pharmaceutical manufacturing.
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Produtos Biológicos , Tecnologia Farmacêutica , Tecnologia Farmacêutica/métodos , Preparações FarmacêuticasRESUMO
Genome-wide mapping of histone modifications is critical to understanding transcriptional regulation. CUT&Tag is a new method for profiling histone modifications, offering improved sensitivity and decreased cost compared with ChIP-seq. Here, we present GoPeaks, a peak calling method specifically designed for histone modification CUT&Tag data. We compare the performance of GoPeaks against commonly used peak calling algorithms to detect histone modifications that display a range of peak profiles and are frequently used in epigenetic studies. We find that GoPeaks robustly detects genome-wide histone modifications and, notably, identifies a substantial number of H3K27ac peaks with improved sensitivity compared to other standard algorithms.
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Código das Histonas , Processamento de Proteína Pós-Traducional , Imunoprecipitação da Cromatina/métodos , Genoma , Análise de Sequência de DNA/métodosRESUMO
Responses to kinase-inhibitor therapy in AML are frequently short-lived due to the rapid development of resistance, limiting the clinical efficacy. Combination therapy may improve initial therapeutic responses by targeting pathways used by leukemia cells to escape monotherapy. Here we report that combined inhibition of KIT and lysine-specific demethylase 1 (LSD1) produces synergistic cell death in KIT-mutant AML cell lines and primary patient samples. This drug combination evicts both MYC and PU.1 from chromatin driving cell cycle exit. Using a live cell biosensor for AKT activity, we identify early adaptive changes in kinase signaling following KIT inhibition that are reversed with the addition of LSD1 inhibitor via modulation of the GSK3a/b axis. Multi-omic analyses, including scRNA-seq, ATAC-seq and CUT&Tag, confirm these mechanisms in primary KIT-mutant AML. Collectively, this work provides rational for a clinical trial to assess the efficacy of KIT and LSD1 inhibition in patients with KIT-mutant AML.
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Histona Desmetilases , Leucemia Mieloide Aguda , Ciclo Celular , Linhagem Celular Tumoral , Redes Reguladoras de Genes , Humanos , Leucemia Mieloide Aguda/tratamento farmacológico , Leucemia Mieloide Aguda/genética , Leucemia Mieloide Aguda/metabolismoRESUMO
The transcription factor RUNX1 is essential for correct hematopoietic development; in its absence in the germ line, blood stem cells are not formed. RUNX1 orchestrates dramatic changes in the chromatin landscape at the onset of stem cell formation, which set the stage for both stem self-renewal and further differentiation. However, once blood stem cells are formed, the mutation of the RUNX1 gene is not lethal but can lead to various hematopoietic defects and a predisposition to cancer. Here we summarize the current literature on inherited and acquired RUNX1 mutations, with a particular emphasis on mutations that alter the structure of the RUNX1 protein itself, and place these changes in the context of what is known about RUNX1 function. We also summarize which mutant RUNX1 proteins are actually expressed in cells and discuss the molecular mechanism underlying how such variants reprogram the epigenome setting stem cells on the path to malignancy.
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Subunidade alfa 2 de Fator de Ligação ao Core , Doenças Hematológicas , Cromatina/genética , Subunidade alfa 2 de Fator de Ligação ao Core/genética , Subunidade alfa 2 de Fator de Ligação ao Core/metabolismo , Doenças Hematológicas/genética , Hematopoese/genética , Humanos , MutaçãoRESUMO
T cell immunological memory is established within days of an infection, but little is known about the in vivo changes in gene regulatory networks accounting for their ability to respond more efficiently to secondary infections. To decipher the timing and nature of immunological memory we performed genome-wide analyses of epigenetic and transcriptional changes in a mouse model generating antigen-specific T cells. Epigenetic reprogramming for Th differentiation and memory T cell formation was already established by the peak of the T cell response after 7 days. The Th memory T cell program was associated with a gain of open chromatin regions, enriched for RUNX, ETS and T-bet motifs, which remained stable for 56 days. The epigenetic programs for both effector memory, associated with T-bet, and central memory, associated with TCF-1, were established in parallel. Memory T cell-specific regulatory elements were associated with greatly enhanced inducible Th1-biased responses during secondary exposures to antigen. Furthermore, memory T cells responded in vivo to re-exposure to antigen by rapidly reprograming the entire ETS factor gene regulatory network, by suppressing Ets1 and activating Etv6 expression. These data show that gene regulatory networks are epigenetically reprogrammed towards memory during infection, and undergo substantial changes upon re-stimulation.