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1.
Mol Cell Proteomics ; 23(6): 100770, 2024 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-38641226

RESUMO

Inhalation of crystalline silica dust induces incurable lung damage, silicosis, and pulmonary fibrosis. However, the mechanisms of the lung injury remain poorly understood, with limited therapeutic options aside from lung transplantation. Posttranslational modifications can regulate the function of proteins and play an important role in studying disease mechanisms. To investigate changes in posttranslational modifications of proteins in silicosis, combined quantitative proteome, acetylome, and succinylome analyses were performed with lung tissues from silica-injured and healthy mice using liquid chromatography-mass spectrometry. Combined analysis was applied to the three omics datasets to construct a protein landscape. The acetylation and succinylation of the key transcription factor STAT1 were found to play important roles in the silica-induced pathophysiological changes. Modulating the acetylation level of STAT1 with geranylgeranylacetone effectively inhibited the progression of silicosis. This report revealed a comprehensive landscape of posttranslational modifications in silica-injured mouse and presented a novel therapeutic strategy targeting the posttranslational level for silica-induced lung diseases.


Assuntos
Lisina , Processamento de Proteína Pós-Traducional , Proteoma , Fator de Transcrição STAT1 , Silicose , Animais , Silicose/metabolismo , Silicose/tratamento farmacológico , Silicose/patologia , Fator de Transcrição STAT1/metabolismo , Proteoma/metabolismo , Lisina/metabolismo , Acetilação/efeitos dos fármacos , Camundongos , Dióxido de Silício , Pulmão/metabolismo , Pulmão/efeitos dos fármacos , Pulmão/patologia , Camundongos Endogâmicos C57BL , Proteômica/métodos , Masculino , Ácido Succínico/metabolismo
2.
Circulation ; 149(17): 1354-1371, 2024 Apr 23.
Artigo em Inglês | MEDLINE | ID: mdl-38314588

RESUMO

BACKGROUND: Pulmonary hypertension (PH) is a progressive cardiopulmonary disease with a high mortality rate. Although growing evidence has revealed the importance of dysregulated energetic metabolism in the pathogenesis of PH, the underlying cellular and molecular mechanisms are not fully understood. In this study, we focused on ME1 (malic enzyme 1), a key enzyme linking glycolysis to the tricarboxylic acid cycle. We aimed to determine the role and mechanistic action of ME1 in PH. METHODS: Global and endothelial-specific ME1 knockout mice were used to investigate the role of ME1 in hypoxia- and SU5416/hypoxia (SuHx)-induced PH. Small hairpin RNA and ME1 enzymatic inhibitor (ME1*) were used to study the mechanism of ME1 in pulmonary artery endothelial cells. Downstream key metabolic pathways and mediators of ME1 were identified by metabolomics analysis in vivo and ME1-mediated energetic alterations were examined by Seahorse metabolic analysis in vitro. The pharmacological effect of ME1* on PH treatment was evaluated in PH animal models induced by SuHx. RESULTS: We found that ME1 protein level and enzymatic activity were highly elevated in lung tissues of patients and mice with PH, primarily in vascular endothelial cells. Global knockout of ME1 protected mice from developing hypoxia- or SuHx-induced PH. Endothelial-specific ME1 deletion similarly attenuated pulmonary vascular remodeling and PH development in mice, suggesting a critical role of endothelial ME1 in PH. Mechanistic studies revealed that ME1 inhibition promoted downstream adenosine production and activated A2AR-mediated adenosine signaling, which leads to an increase in nitric oxide generation and a decrease in proinflammatory molecule expression in endothelial cells. ME1 inhibition activated adenosine production in an ATP-dependent manner through regulating malate-aspartate NADH (nicotinamide adenine dinucleotide plus hydrogen) shuttle and thereby balancing oxidative phosphorylation and glycolysis. Pharmacological inactivation of ME1 attenuated the progression of PH in both preventive and therapeutic settings by promoting adenosine production in vivo. CONCLUSIONS: Our findings indicate that ME1 upregulation in endothelial cells plays a causative role in PH development by negatively regulating adenosine production and subsequently dysregulating endothelial functions. Our findings also suggest that ME1 may represent as a novel pharmacological target for upregulating protective adenosine signaling in PH therapy.

3.
Am J Respir Cell Mol Biol ; 68(2): 213-227, 2023 02.
Artigo em Inglês | MEDLINE | ID: mdl-36227848

RESUMO

Progressive fibrosing interstitial lung diseases (PF-ILDs) result in high mortality and lack effective therapies. The pathogenesis of PF-ILDs involves macrophages driving inflammation and irreversible fibrosis. Fc-γ receptors (FcγRs) regulate macrophages and inflammation, but their roles in PF-ILDs remain unclear. We characterized the expression of FcγRs and found upregulated FcγRIIB in human and mouse lungs after exposure to silica. FcγRIIB deficiency aggravated lung dysfunction, inflammation, and fibrosis in silica-exposed mice. Using single-cell transcriptomics and in vitro experiments, FcγRIIB was found in alveolar macrophages, where it regulated the expression of fibrosis-related genes Spp1 and Ctss. In mice with macrophage-specific overexpression of FcγRIIB and in mice treated with adenovirus by intratracheal instillation to upregulate FcγRIIB, silica-induced functional and histological changes were ameliorated. Our data from three genetic models and a therapeutic model suggest that FcγRIIB plays a protective role that can be enhanced by adenoviral overexpression, representing a potential therapeutic strategy for PF-ILDs.


Assuntos
Doenças Pulmonares Intersticiais , Pneumonia , Humanos , Animais , Camundongos , Adenoviridae/genética , Adenoviridae/metabolismo , Pneumonia/genética , Inflamação/genética , Inflamação/metabolismo , Receptores de IgG/genética , Receptores de IgG/metabolismo , Fibrose , Dióxido de Silício
4.
Circulation ; 143(10): 1014-1030, 2021 03 09.
Artigo em Inglês | MEDLINE | ID: mdl-33305586

RESUMO

BACKGROUND: Immunoglobulin E (IgE) belongs to a class of immunoglobulins involved in immune response to specific allergens. However, the roles of IgE and IgE receptor (FcεR1) in pathological cardiac remodeling and heart failure are unknown. METHODS: Serum IgE levels and cardiac FcεR1 expression were assessed in diseased hearts from human and mouse. The role of FcεR1 signaling in pathological cardiac remodeling was explored in vivo by FcεR1 genetic depletion, anti-IgE antibodies, and bone marrow transplantation. The roles of the IgE-FcεR1 pathway were further evaluated in vitro in primary cultured rat cardiomyocytes and cardiac fibroblasts (CFs). RNA sequencing and bioinformatic analyses were used to identify biochemical changes and signaling pathways that are regulated by IgE/FcεR1. RESULTS: Serum IgE levels were significantly elevated in patients with heart failure as well as in 2 mouse cardiac disease models induced by chronic pressure overload via transverse aortic constriction and chronic angiotensin II infusion. Interestingly, FcεR1 expression levels were also significantly upregulated in failing hearts from human and mouse. Blockade of the IgE-FcεR1 pathway by FcεR1 knockout alleviated transverse aortic constriction- or angiotensin II-induced pathological cardiac remodeling or dysfunction. Anti-IgE antibodies (including the clinical drug omalizumab) also significantly alleviated angiotensin II-induced cardiac remodeling. Bone marrow transplantation experiments indicated that IgE-induced cardiac remodeling was mediated through non-bone marrow-derived cells. FcεR1 was found to be expressed in both cardiomyocytes and CFs. In cultured rat cardiomyocytes, IgE-induced cardiomyocyte hypertrophy and hypertrophic marker expression were abolished by depleting FcεR1. In cultured rat CFs, IgE-induced CF activation and matrix protein production were also blocked by FcεR1 deficiency. RNA sequencing and signaling pathway analyses revealed that transforming growth factor-ß may be a critical mediator, and blocking transforming growth factor-ß indeed alleviated IgE-induced cardiomyocyte hypertrophy and cardiac fibroblast activation in vitro. CONCLUSIONS: Our findings suggest that IgE induction plays a causative role in pathological cardiac remodeling, at least partially via the activation of IgE-FcεR1 signaling in cardiomyocytes and CFs. Therapeutic strategies targeting the IgE-FcεR1 axis may be effective for managing IgE-mediated cardiac remodeling.


Assuntos
Imunoglobulina E/metabolismo , Miócitos Cardíacos/metabolismo , Remodelação Ventricular/genética , Animais , Humanos , Masculino , Camundongos , Camundongos Knockout
5.
Acta Pharmacol Sin ; 43(4): 908-918, 2022 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-34316030

RESUMO

Silicosis is a global occupational disease characterized by lung dysfunction, pulmonary inflammation, and fibrosis, for which there is a lack of effective drugs. Pirfenidone has been shown to exert anti-inflammatory and anti-fibrotic properties in the lung. However, whether and how pirfenidone is effective against silicosis remains unknown. Here, we evaluated the efficacy of pirfenidone in the treatment of early and advanced silicosis in an experimental mouse model and explored its potential pharmacological mechanisms. We found that pirfenidone alleviated silica-induced lung dysfunction, secretion of inflammatory cytokines (TNF-α, IL-1ß, IL-6) and deposition of fibrotic proteins (collagen I and fibronectin) in both early and advanced silicosis models. Moreover, we observed that both 100 and 200 mg/kg pirfenidone can effectively treat early-stage silicosis, while 400 mg/kg was recommended for advanced silicosis. Mechanistically, antibody array and bioinformatic analysis showed that the pathways related to IL-17 secretion, including JAK-STAT pathway, Th17 differentiation, and IL-17 pathway, might be involved in the treatment of silicosis by pirfenidone. Further in vivo experiments confirmed that pirfenidone reduced the production of IL-17A induced by silica exposure via inhibiting STAT3 phosphorylation. Neutralizing IL-17A by anti-IL-17A antibody improved lung function and reduced pulmonary inflammation and fibrosis in silicosis animals. Collectively, our study has demonstrated that pirfenidone effectively ameliorated silica-induced lung dysfunction, pulmonary inflammation and fibrosis in mouse models by inhibiting the secretion of IL-17A.


Assuntos
Interleucina-17 , Pneumonia , Animais , Modelos Animais de Doenças , Fibrose , Inflamação/induzido quimicamente , Inflamação/tratamento farmacológico , Inflamação/metabolismo , Interleucina-17/metabolismo , Janus Quinases/metabolismo , Janus Quinases/uso terapêutico , Pulmão/metabolismo , Camundongos , Camundongos Endogâmicos C57BL , Pneumonia/induzido quimicamente , Pneumonia/tratamento farmacológico , Pneumonia/metabolismo , Piridonas , Fatores de Transcrição STAT/metabolismo , Fatores de Transcrição STAT/uso terapêutico , Transdução de Sinais , Dióxido de Silício/toxicidade
6.
Ecotoxicol Environ Saf ; 244: 114043, 2022 Oct 01.
Artigo em Inglês | MEDLINE | ID: mdl-36087468

RESUMO

Silicosis is one of the most important occupational diseases worldwide, caused by inhalation of silica particles or free crystalline silicon dioxide. As a disease with high mortality, it has no effective treatment and new therapeutic targets are urgently needed. Recent studies have identified FCER1A, encoding α-subunit of the immunoglobulin E (IgE) receptor FcεRI, as a candidate gene involved in the biological pathways leading to respiratory symptoms. FcεRI is known to be important in allergic asthma, but its role in silicosis remains unclear. In this study, serum IgE concentrations and FcεRI expression were assessed in pneumoconiosis patients and silica-exposed mice. The role of FcεRI was explored in a silica-induced mouse model using wild-type and FcεRI-deficient mice. The results showed that serum IgE concentrations were significantly elevated in both pneumoconiosis patients and mice exposed to silica compared with controls. The mRNA and protein expression of FcεRI were also significantly increased in the lung tissue of patients and silica-exposed mice. FcεRI deficiency significantly attenuated the changes in lung function caused by silica exposure. Silica-induced elevations of IL-1ß, IL-6, and TNF-α were significantly attenuated in the lung tissue and bronchoalveolar lavage fluid (BALF) of FcεRI-deficient mice compared with wild-type controls. Additionally, FcεRI-deficient mice showed a significantly lower score of pulmonary fibrosis than wild-type mice following exposure to silica, with significantly lower hydroxyproline content and expression of fibrotic genes Col1a1 and Fn1. Immunofluorescent staining suggested FcεRI mainly on mast cells. Mast cell degranulation took place after silica exposure, as shown by increased serum histamine levels and ß-hexosaminidase activity, which were significantly reduced in FcεRI-deficient mice compared with wild-type controls. Together, these data showed that FcεRI deficiency had a significant protective effect against silica-induced pulmonary inflammation and fibrosis. Our findings provide new insights into the pathophysiological mechanisms of silica-induced pulmonary fibrosis and a potential target for the treatment of silicosis.


Assuntos
Pneumonia , Fibrose Pulmonar , Silicose , Animais , Fibrose , Histamina/metabolismo , Histamina/toxicidade , Hidroxiprolina/metabolismo , Hidroxiprolina/farmacologia , Hidroxiprolina/uso terapêutico , Imunoglobulina E , Interleucina-6/metabolismo , Pulmão , Camundongos , Camundongos Endogâmicos C57BL , Pneumonia/patologia , Fibrose Pulmonar/induzido quimicamente , Fibrose Pulmonar/genética , RNA Mensageiro/metabolismo , Receptores de IgE/genética , Receptores de IgE/metabolismo , Receptores de IgE/uso terapêutico , Dióxido de Silício/toxicidade , Silicose/genética , Silicose/metabolismo , Fator de Necrose Tumoral alfa/metabolismo , beta-N-Acetil-Hexosaminidases/metabolismo , beta-N-Acetil-Hexosaminidases/farmacologia , beta-N-Acetil-Hexosaminidases/uso terapêutico
7.
Plant J ; 98(1): 19-32, 2019 04.
Artigo em Inglês | MEDLINE | ID: mdl-30548709

RESUMO

In maize, kernel traits strongly impact overall grain yields, and it is known that sophisticated spatiotemporal programs of gene expression coordinate kernel development, so advancing our knowledge of kernel development can help efforts to improve grain yields. Here, using phenotype, genotype and transcriptomics data of maize kernels at 5 and 15 days after pollination (DAP) for a large association mapping panel, we employed multiple quantitative genetics approaches-genome-wide association studies (GWAS) as well as expression quantitative trait loci (eQTL) and quantitative trait transcript (QTT) analyses-to gain insights about molecular genetic basis of kernel development in maize. This resulted in the identification of 137 putative kernel length-related genes at 5 DAP, of which 43 are located in previously reported QTL regions. Strikingly, we identified an eQTL that overlaps the locus encoding a maize homolog of the recently described m6 A methylation reader protein ECT2 from Arabidopsis; this putative epi eQTL is associated with 53 genes and may represent a master epi-transcriptomic regulator of kernel development. Notably, among the genes associated with this epi eQTL, 10 are for the main storage proteins in the maize endosperm (zeins) and two are known regulators of zein expression or endosperm development (Opaque2 and ZmICE1). Collectively, beyond cataloging and characterizing genomic attributes of a large number of eQTL associated with kernel development in maize, our study highlights how an eQTL approach can bolster the impact of both GWAS and QTT studies and can drive insights about the basic biology of plants.


Assuntos
Estudo de Associação Genômica Ampla , Polimorfismo de Nucleotídeo Único/genética , Locos de Características Quantitativas/genética , Zea mays/genética , Grão Comestível/genética , Grão Comestível/crescimento & desenvolvimento , Endosperma/genética , Endosperma/crescimento & desenvolvimento , Ontologia Genética , Genótipo , Modelos Lineares , Fenótipo , Filogenia , Zea mays/crescimento & desenvolvimento
8.
Plant Mol Biol ; 102(3): 339-357, 2020 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-31894455

RESUMO

KEY MESSAGE: NCP1, a NINJA family protein lacking EAR motif, acts as a negative regulator of ABA signaling by interacting with and inhibiting the activity of transcriptional activator ABP9. The phytohormone abscisic acid plays a pivotal role in regulating plant responses to a variety of abiotic stresses including drought and salinity. Maize ABP9 is an ABRE-binding bZIP transcription activator that enhances plant tolerance to multiple stresses by positively regulating ABA signaling, but the molecular mechanism by which ABP9 is regulated in mediating ABA responses remains unknown. Here, we report the identification of an ABP9-interacting protein, named ABP Nine Complex Protein 1 (NCP1) and its functional characterization. NCP1 belongs to the recently identified NINJA family proteins, but lacks the conserved EAR motif, which is a hallmark of this class of transcriptional repressors. In vitro and in vivo assays confirmed that NCP1 physically interacts with ABP9 and that they are co-localized in the nucleus. In addition, NCP1 and ABP9 are similarly induced with similar patterns by ABA treatment and osmotic stress. Interestingly, NCP1 over-expressing Arabidopsis plants exhibited a reduced sensitivity to ABA and decreased drought tolerance. Transient assay in maize protoplasts showed that NCP1 inhibits the activity of ABP9 in activating ABRE-mediated reporter gene expression, a notion further supported by genetic analysis of drought and ABA responses in the transgenic plants over-expressing both ABP9 and NCP1. These data together suggest that NCP1 is a novel negative regulator of ABA signaling via interacting with and inhibiting the activity of ABP9.


Assuntos
Secas , Reguladores de Crescimento de Plantas/farmacologia , Proteínas de Plantas/metabolismo , Transdução de Sinais/efeitos dos fármacos , Fatores de Transcrição/metabolismo , Zea mays/metabolismo , Arabidopsis/genética , Arabidopsis/metabolismo , Proteínas de Arabidopsis , Regulação da Expressão Gênica de Plantas/efeitos dos fármacos , Genes de Plantas , Pressão Osmótica , Reguladores de Crescimento de Plantas/genética , Proteínas de Plantas/genética , Plantas Geneticamente Modificadas/genética , Plantas Geneticamente Modificadas/metabolismo , Proteínas Repressoras/metabolismo , Salinidade , Estresse Fisiológico , Fatores de Transcrição/genética , Transcriptoma , Zea mays/genética
9.
Ecotoxicol Environ Saf ; 202: 110834, 2020 Oct 01.
Artigo em Inglês | MEDLINE | ID: mdl-32622305

RESUMO

Silicosis is caused by massive inhalation of silica-based particles, which leads to pulmonary inflammation, pulmonary fibrosis and lung dysfunction. Currently, the pathophysiological process of silicosis has not been well studied. Here, we defined the progression of silicosis as four stages by unsupervised clustering analysis: normal stage, inflammatory stage, progressive stage and fibrotic stage. Specifically, in normal stage, the lung function was normal, and no inflammation or fibrosis was detected in the lung tissue. Inflammatory stage showed a remarkable pulmonary inflammation but mild fibrosis and lung dysfunction. In progressive stage, significant lung dysfunction was observed, while pulmonary inflammation and fibrosis continued to deteriorate. Fibrotic stage revealed the most severe pulmonary fibrosis and lung dysfunction but no significant deterioration in inflammation. Since the common features were founded in both silicosis patients and rodents, we speculated that the pathophysiological processes of silicosis in patients might be similar to the rodents. Collectively, our new classification identified the process of silicosis, clarified the pathophysiological features of each stage, and provided some new insights for the progression of the disease.


Assuntos
Silicose/fisiopatologia , Animais , Fibrose , Humanos , Inflamação/patologia , Pulmão/patologia , Pneumonia/fisiopatologia , Fibrose Pulmonar/fisiopatologia , Dióxido de Silício
10.
Water Sci Technol ; 81(4): 668-678, 2020 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-32460271

RESUMO

Wastewater ecological soil infiltration system (WESIS) is a land treatment technology for decentralized wastewater treatment that has been applied all over the world. In this study, the pollutant removal, emission of greenhouse gases (GHGs) and functional gene abundances with different influent C/N ratios were evaluated in WESISs with/without intermittent aeration. Intermittent aeration and influent C/N ratio affect pollutant removal and GHG emission. Increased influent C/N ratio led to high total nitrogen (TN) removal, low CH4 and N2O emission in the aerated WESIS, which was different from the non-aerated WESIS. High average removal efficiencies of chemical oxygen demand (COD) (94.8%), NH4 +-N (95.1%), TN (91.2%), total phosphorus (TP) (91.1%) and low emission rates for CH4 (27.2 mg/(m2 d)) and N2O (10.5 mg/(m2 d)) were achieved with an influent C/N ratio of 12:1 in the aerated WESIS. Intermittent aeration enhanced the abundances of bacterial 16S rRNA, amoA, nxrA, narG, napA, nirK, nirS, qnorB, nosZ genes and decreased the abundances of the mcrA gene, which are involved in pollutant removal and GHG emission. Intermittent aeration would be an effective alternative to achieving high pollutant removal and low CH4 and N2O emission in high influent C/N ratio wastewater treatment.


Assuntos
Poluentes Ambientais , Gases de Efeito Estufa , Análise da Demanda Biológica de Oxigênio , Desnitrificação , Nitrogênio , RNA Ribossômico 16S , Solo , Águas Residuárias
11.
Water Sci Technol ; 80(1): 144-152, 2019 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-31461431

RESUMO

Nitrogen removal is an obstacle for the wide application of wastewater ecological soil infiltration (WESI) system in domestic wastewater treatment. In this study, matrix dissolved oxygen (DO), nitrogen removal and nitrous oxide (N2O) emission in aerated pilot WESI systems were investigated under different aeration times (1, 2, 3, 4 and 6 h/d) and aeration rates (1, 2, 3 and 4 L/min). The results showed that aerobic conditions in upper matrix and anoxic or anaerobic conditions in the subsequent matrix were developed in an aerated/non-aerated cycle at the optimal aeration condition of aeration time of 4 h/d and aeration rate of 3 L/min. Simultaneously, high removal efficiency of chemical oxygen demand (COD) (97.9%), NH4 +-N (98.2%), total nitrogen (TN) (90.7%) and low N2O emission rate (13.2 mg/(m2 d)) were obtained. The results would provide optimal aeration parameters for application of intermittent aerated WESI systems.


Assuntos
Nitrogênio/análise , Óxido Nitroso , Eliminação de Resíduos Líquidos/métodos , Águas Residuárias , Desnitrificação , Solo
12.
Proc Natl Acad Sci U S A ; 112(11): E1263-71, 2015 Mar 17.
Artigo em Inglês | MEDLINE | ID: mdl-25733907

RESUMO

The ability of centromeres to alternate between active and inactive states indicates significant epigenetic aspects controlling centromere assembly and function. In maize (Zea mays), misdivision of the B chromosome centromere on a translocation with the short arm of chromosome 9 (TB-9Sb) can produce many variants with varying centromere sizes and centromeric DNA sequences. In such derivatives of TB-9Sb, we found a de novo centromere on chromosome derivative 3-3, which has no canonical centromeric repeat sequences. This centromere is derived from a 288-kb region on the short arm of chromosome 9, and is 19 megabases (Mb) removed from the translocation breakpoint of chromosome 9 in TB-9Sb. The functional B centromere in progenitor telo2-2 is deleted from derivative 3-3, but some B-repeat sequences remain. The de novo centromere of derivative 3-3 becomes inactive in three further derivatives with new centromeres being formed elsewhere on each chromosome. Our results suggest that de novo centromere initiation is quite common and can persist on chromosomal fragments without a canonical centromere. However, we hypothesize that when de novo centromeres are initiated in opposition to a larger normal centromere, they are cleared from the chromosome by inactivation, thus maintaining karyotype integrity.


Assuntos
Centrômero/genética , Cromossomos de Plantas/genética , Zea mays/genética , Pareamento de Bases/genética , Imunoprecipitação da Cromatina , Hibridização in Situ Fluorescente , Meiose/genética , Zea mays/citologia
13.
Plant J ; 88(5): 854-866, 2016 12.
Artigo em Inglês | MEDLINE | ID: mdl-27531446

RESUMO

The inheritance and function of centromeres are not strictly dependent on any specific DNA sequence, but involve an epigenetic component in most species. CENH3, a centromere histone H3 variant, is one of the best-described epigenetic factors in centromere identity, but the chromatin features required during centromere formation have not yet been revealed. We previously identified two de novo centromeres on Zea mays (maize) minichromosomes derived from euchromatic sites with high-density gene distributions but low-density transposon distributions. The distribution of gene location and gene expression in these sites indicates that transcriptionally active regions can initiate de novo centromere formation, and CENH3 seeding shows a preference for gene-free regions or regions with no gene expression. The locations of the expressed genes detected were at relatively hypomethylated loci, and the altered gene expression resulted from de novo centromere formation, but not from the additional copy of the minichromosome. The initial overall DNA methylation level of the two de novo regions was at a low level, but increased substantially to that of native centromeres after centromere formation. These results illustrate the dynamic chromatin changes during euchromatin-originated de novo centromere formation, which provides insight into the mechanism of de novo centromere formation and regulation of subsequent consequences.


Assuntos
Centrômero/metabolismo , Cromatina/metabolismo , Eucromatina/metabolismo , Zea mays/metabolismo , Metilação de DNA/genética , Eucromatina/genética , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Zea mays/genética
15.
Plant Cell ; 25(6): 1979-89, 2013 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-23771890

RESUMO

The maize (Zea mays) B centromere is composed of B centromere-specific repeats (ZmBs), centromere-specific satellite repeats (CentC), and centromeric retrotransposons of maize (CRM). Here we describe a newly formed B centromere in maize, which has lost CentC sequences and has dramatically reduced CRM and ZmBs sequences, but still retains the molecular features of functional centromeres, such as CENH3, H2A phosphorylation at Thr-133, H3 phosphorylation at Ser-10, and Thr-3 immunostaining signals. This new centromere is stable and can be transmitted to offspring through meiosis. Anti-CENH3 chromatin immunoprecipitation sequencing revealed that a 723-kb region from the short arm of chromosome 9 (9S) was involved in the formation of the new centromere. The 723-kb region, which is gene poor and enriched for transposons, contains two abundant DNA motifs. Genes in the new centromere region are still transcribed. The original 723-kb region showed a higher DNA methylation level compared with native centromeres but was not significantly changed when it was involved in new centromere formation. Our results indicate that functional centromeres may be formed without the known centromere-specific sequences, yet the maintenance of a high DNA methylation level seems to be crucial for the proper function of a new centromere.


Assuntos
Centrômero/genética , DNA de Plantas/genética , Meiose/genética , Zea mays/genética , Imunoprecipitação da Cromatina , Cromossomos de Plantas/genética , Metilação de DNA , Genoma de Planta/genética , Imuno-Histoquímica , Hibridização in Situ Fluorescente , Análise de Sequência com Séries de Oligonucleotídeos , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Sequências Repetitivas de Ácido Nucleico/genética , Retroelementos/genética , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Análise de Sequência de DNA , Deleção de Sequência , Transcriptoma , Zea mays/citologia , Zea mays/metabolismo
16.
Proc Natl Acad Sci U S A ; 110(15): 6033-6, 2013 Apr 09.
Artigo em Inglês | MEDLINE | ID: mdl-23530217

RESUMO

The centromere is the part of the chromosome that organizes the kinetochore, which mediates chromosome movement during mitosis and meiosis. A small fragment from chromosome 3, named Duplication 3a (Dp3a), was described from UV-irradiated materials by Stadler and Roman in the 1940s [Stadler LJ, Roman H (1948) Genetics 33(3):273-303]. The genetic behavior of Dp3a is reminiscent of a ring chromosome, but fluoresecent in situ hybridization detected telomeres at both ends, suggesting a linear structure. This small chromosome has no detectable canonical centromeric sequences, but contains a site with protein features of functional centromeres such as CENH3, the centromere specific H3 histone variant, and CENP-C, a foundational kinetochore protein, suggesting the de novo formation of a centromere on the chromatin fragment. To examine the sequences associated with CENH3, chromatin immunoprecipitation was carried out with anti-CENH3 antibodies using material from young seedlings with and without the Dp3a chromosome. A novel peak was detected from the ChIP-Sequencing reads of the Dp3a sample. The peak spanned 350 kb within the long arm of chromosome 3 covering 22 genes. Collectively, these results define the behavior and molecular features of de novo centromere formation in the Dp3a chromosome, which may shed light on the initiation of new centromere sites during evolution.


Assuntos
Centrômero/metabolismo , Cromossomos de Plantas/ultraestrutura , Zea mays/genética , Ciclo Celular , Cromatina/ultraestrutura , Imunoprecipitação da Cromatina , Epigênese Genética , Evolução Molecular , Genes de Plantas , Genoma de Planta , Genômica , Hibridização in Situ Fluorescente , Cinetocoros/metabolismo , Telômero/ultraestrutura
17.
Acta Pharm Sin B ; 14(4): 1726-1741, 2024 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-38572107

RESUMO

Pulmonary hypertension (PH) is a fatal disorder characterized by pulmonary vascular remodeling and obstruction. The phosphodiesterase 4 (PDE4) family hydrolyzes cyclic AMP (cAMP) and is comprised of four subtypes (PDE4A-D). Previous studies have shown the beneficial effects of pan-PDE4 inhibitors in rodent PH; however, this class of drugs is associated with side effects owing to the broad inhibition of all four PDE4 isozymes. Here, we demonstrate that PDE4B is the predominant PDE isozyme in lungs and that it was upregulated in rodent and human PH lung tissues. We also confirmed that PDE4B is mainly expressed in the lung endothelial cells (ECs). Evaluation of PH in Pde4b wild type and knockout mice confirmed that Pde4b is important for the vascular remodeling associated with PH. In vivo EC lineage tracing demonstrated that Pde4b induces PH development by driving endothelial-to-mesenchymal transition (EndMT), and mechanistic studies showed that Pde4b regulates EndMT by antagonizing the cAMP-dependent PKA-CREB-BMPRII axis. Finally, treating PH rats with a PDE4B-specific inhibitor validated that PDE4B inhibition has a significant pharmacological effect in the alleviation of PH. Collectively, our findings indicate a critical role for PDE4B in EndMT and PH, prompting further studies of PDE4B-specific inhibitors as a therapeutic strategy for PH.

18.
Comput Biol Med ; 151(Pt A): 106187, 2022 12.
Artigo em Inglês | MEDLINE | ID: mdl-36327882

RESUMO

Blood leukocyte counts (e.g., eosinophil count) are important biomarkers for the onset, classification, and exacerbation of chronic obstructive pulmonary disease (COPD). The causal relationships between them are necessary for the development of COPD treatment strategy, but remain unclear. Here, we implement two-sample bi-directional univariable Mendelian Randomization (MR) and multivariable MR to investigate the causal relationships. Univariable MR find that elevated blood eosinophil count significantly increases the risk of COPD (odds ratio (OR) = 1.22, 95% confidence interval (CI): 1.14-1.30, P = 1.54 × 10-09) and COPD-related hospitalization (OR = 1.44, 95% CI: 1.15-1.80, P = 1.36 × 10-03). Besides, it also significantly decreases the ratio of forced expiratory volume in the first second over forced vital capacity (FEV1/FVC ratio) (OR = 0.942, 95% CI: 0.914-0.971, P = 1.02 × 10-04). These findings are fully supported by multivariate MR results. Interestingly, univariable MR reveals a weak causal relationship between elevated blood eosinophil count and COPD risk in younger people (<65 years) (OR = 1.39, 95% CI: 1.10-1.75, P = 5.52 × 10-03), but not older individuals (OR = 1.20, 95% CI: 0.926-1.55, P = 0.17). Finally, reverse univariable MR reveals the onset of COPD and the decreased FEV1/FVC ratio both lead to increased blood neutrophil count (OR = 1.03, 95% CI: 1.01-1.05, P = 3.40 × 10-03 and OR = 0.947, 95% CI: 0.91-0.986, P = 8.75 × 10-03 respectively). In summary, this MR study demonstrates that high blood eosinophil count is an independent causal mediator of COPD risk, FEV1/FVC decline, and COPD-related hospitalization. The increase in neutrophil count is induced by COPD onset or FEV1/FVC decline. This suggests eosinophil, but not neutrophil, may be used as a therapeutic target for preventing the onset and exacerbation of COPD and FEV1/FVC decline. Therefore, a non-neutrophil-targeted therapeutic strategy for neutrophilic COPD is required in the future.


Assuntos
Análise da Randomização Mendeliana , Doença Pulmonar Obstrutiva Crônica , Humanos , Doença Pulmonar Obstrutiva Crônica/genética , Volume Expiratório Forçado , Capacidade Vital , Contagem de Leucócitos
19.
Cardiovasc Res ; 118(14): 2985-2999, 2022 11 10.
Artigo em Inglês | MEDLINE | ID: mdl-35048969

RESUMO

AIMS: Hypertension is the major cause of cardiovascular diseases and global mortality. Immunoglobulin E (IgE), which plays crucial roles in allergic diseases, has been implicated in the pathogenesis of vascular and cardiac remodelling via its receptor (FcεR1). In this study, we aimed to reveal the role of IgE and FcεR1 in hypertension. METHODS AND RESULTS: Herein, we reported that IgE levels were significantly increased in hypertensive patients as well as in hypertensive mice induced by angiotensin II (Ang II). Ang II-induced vascular remodelling and hypertension were significantly alleviated in FcεR1 genetic knockout mice or in mice treated with anti-IgE monoclonal antibody. Similarly, treatment with omalizumab (a clinical IgE antagonist) also markedly inhibited Ang II-induced hypertension. Furthermore, the cellular contribution of IgE-FcεR1 in hypertension was evaluated in mice with FcεR1 conditional knockout in mast cell (MC), smooth muscle cell (SMC), or endothelial cell (EC). Our data revealed that IgE-mediated hypertension is largely dependent on FcεR1 in MCs but not SMCs and ECs. Finally, RNA-seq and signalling pathway analyses of mouse bone marrow-derived MCs suggested that interleukin 6 (IL-6) is one of critical mediators in IgE-mediated hypertension. IL-6 derived from IgE-stimulated MCs promoted reactive oxygen species production and decreased the levels of phosphorylated endothelial nitric oxide synthase in ECs, leading to endothelial dysfunction. CONCLUSION: Our findings reveal that IgE contributes to the pathogenesis of hypertension, at least partially through activating the IgE-FcεR1 signalling in MCs. Thus, IgE may represent a new therapeutic target for IgE-mediated hypertension.


Assuntos
Hipertensão , Mastócitos , Camundongos , Animais , Imunoglobulina E/genética , Imunoglobulina E/metabolismo , Receptores de IgE/genética , Receptores de IgE/metabolismo , Interleucina-6/metabolismo , Camundongos Knockout , Hipertensão/metabolismo
20.
Exp Mol Med ; 54(8): 1201-1213, 2022 08.
Artigo em Inglês | MEDLINE | ID: mdl-35999453

RESUMO

Abdominal aortic aneurysm (AAA) is a permanent expansion of the abdominal aorta that has a high mortality but limited treatment options. Phosphodiesterase (PDE) 4 family members are cAMP-specific hydrolyzing enzymes and have four isoforms (PDE4A-PDE4D). Several pan-PDE4 inhibitors are used clinically. However, the regulation and function of PDE4 in AAA remain largely unknown. Herein, we showed that PDE4D expression is upregulated in human and angiotensin II-induced mouse AAA tissues using RT-PCR, western blotting, and immunohistochemical staining. Furthermore, smooth muscle cell (SMC)-specific Pde4d knockout mice showed significantly reduced vascular destabilization and AAA development in an experimental AAA model. The PDE4 inhibitor rolipram also suppressed vascular pathogenesis and AAA formation in mice. In addition, PDE4D deficiency inhibited caspase 3 cleavage and SMC apoptosis in vivo and in vitro, as shown by bulk RNA-seq, western blotting, flow cytometry and TUNEL staining. Mechanistic studies revealed that PDE4D promotes apoptosis by suppressing the activation of cAMP-activated protein kinase A (PKA) instead of the exchange protein directly activated by cAMP (Epac). Additionally, the phosphorylation of BCL2-antagonist of cell death (Bad) was reversed by PDE4D siRNA in vitro, which indicates that PDE4D regulates SMC apoptosis via the cAMP-PKA-pBad axis. Overall, these findings indicate that PDE4D upregulation in SMCs plays a causative role in AAA development and suggest that pharmacological inhibition of PDE4 may represent a potential therapeutic strategy.


Assuntos
Angiotensina II , Aneurisma da Aorta Abdominal , Angiotensina II/efeitos adversos , Animais , Aneurisma da Aorta Abdominal/induzido quimicamente , Aneurisma da Aorta Abdominal/genética , Aneurisma da Aorta Abdominal/metabolismo , Apoptose , Proteínas Quinases Dependentes de AMP Cíclico , Nucleotídeo Cíclico Fosfodiesterase do Tipo 4/efeitos adversos , Nucleotídeo Cíclico Fosfodiesterase do Tipo 4/genética , Humanos , Camundongos , Camundongos Knockout , Miócitos de Músculo Liso/metabolismo
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