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1.
PLoS Genet ; 18(10): e1010433, 2022 10.
Artigo em Inglês | MEDLINE | ID: mdl-36301831

RESUMO

Circular RNAs (circRNAs) are a new group of noncoding/regulatory RNAs that are particularly abundant in the nervous system, however, their physiological functions are underexplored. Here we report that the brain-enriched circular RNA Edis (Ect4-derived immune suppressor) plays an essential role in neuronal development in Drosophila. We show that depletion of Edis in vivo causes defects in axonal projection patterns of mushroom body (MB) neurons in the brain, as well as impaired locomotor activity and shortened lifespan of adult flies. In addition, we find that the castor gene, which encodes a transcription factor involved in neurodevelopment, is upregulated in Edis knockdown neurons. Notably, castor overexpression phenocopies Edis knockdown, and reducing castor levels suppresses the neurodevelopmental phenotypes in Edis-depleted neurons. Furthermore, chromatin immunoprecipitation analysis reveals that the transcription factor Relish, which plays a key role in regulating innate immunity signaling, occupies a pair of sites at the castor promoter, and that both sites are required for optimal castor gene activation by either immune challenge or Edis depletion. Lastly, Relish mutation and/or depletion can rescue both the castor gene hyperactivation phenotype and neuronal defects in Edis knockdown animals. We conclude that the circular RNA Edis acts through Relish and castor to regulate neuronal development.


Assuntos
Proteínas de Drosophila , Drosophila , Animais , Drosophila/genética , RNA Circular/genética , Proteínas de Drosophila/genética , Fatores de Transcrição/genética , Corpos Pedunculados , Drosophila melanogaster/fisiologia
2.
PLoS Genet ; 18(10): e1010429, 2022 10.
Artigo em Inglês | MEDLINE | ID: mdl-36301822

RESUMO

Circular RNAs (circRNAs) are widely expressed in eukaryotes. However, only a subset has been functionally characterized. We identify and validate a collection of circRNAs in Drosophila, and show that depletion of the brain-enriched circRNA Edis (circ_Ect4) causes hyperactivation of antibacterial innate immunity both in cultured cells and in vivo. Notably, Edis depleted flies display heightened resistance to bacterial infection and enhanced pathogen clearance. Conversely, ectopic Edis expression blocks innate immunity signaling. In addition, inactivation of Edis in vivo leads to impaired locomotor activity and shortened lifespan. Remarkably, these phenotypes can be recapitulated with neuron-specific depletion of Edis, accompanied by defective neurodevelopment. Furthermore, inactivation of Relish suppresses the innate immunity hyperactivation phenotype in the fly brain. Moreover, we provide evidence that Edis encodes a functional protein that associates with and compromises the processing and activation of the immune transcription factor Relish. Importantly, restoring Edis expression or ectopic expression of Edis-encoded protein suppresses both innate immunity and neurodevelopment phenotypes elicited by Edis depletion. Thus, our study establishes Edis as a key regulator of neurodevelopment and innate immunity.


Assuntos
Imunidade Inata , RNA Circular , Animais , RNA Circular/genética , Imunidade Inata/genética , Fatores de Transcrição/genética , Drosophila/genética , Drosophila/metabolismo , Transdução de Sinais , RNA/genética
3.
J Neurosci ; 40(31): 5908-5921, 2020 07 29.
Artigo em Inglês | MEDLINE | ID: mdl-32601248

RESUMO

SORLA is a transmembrane trafficking protein associated with Alzheimer's disease risk. Although SORLA is abundantly expressed in neurons, physiological roles for SORLA remain unclear. Here, we show that cultured transgenic neurons overexpressing SORLA feature longer neurites, and accelerated neurite regeneration with wounding. Enhanced release of a soluble form of SORLA (sSORLA) is observed in transgenic mouse neurons overexpressing human SORLA, while purified sSORLA promotes neurite extension and regeneration. Phosphoproteomic analyses demonstrate enrichment of phosphoproteins related to the epidermal growth factor (EGFR)/ERK pathway in SORLA transgenic mouse hippocampus from both genders. sSORLA coprecipitates with EGFR in vitro, and sSORLA treatment increases EGFR Y1173 phosphorylation, which is involved in ERK activation in cultured neurons. Furthermore, sSORLA triggers ERK activation, whereas pharmacological EGFR or ERK inhibition reverses sSORLA-dependent enhancement of neurite outgrowth. In search for downstream ERK effectors activated by sSORLA, we identified upregulation of Fos expression in hippocampus from male mice overexpressing SORLA by RNAseq analysis. We also found that Fos is upregulated and translocates to the nucleus in an ERK-dependent manner in neurons treated with sSORLA. Together, these results demonstrate that sSORLA is an EGFR-interacting protein that activates EGFR/ERK/Fos signaling to enhance neurite outgrowth and regeneration.SIGNIFICANCE STATEMENT SORLA is a transmembrane trafficking protein previously known to reduce the levels of amyloid-ß, which is critical in the pathogenesis of Alzheimer's disease. In addition, SORLA mutations are a risk factor for Alzheimer's disease. Interestingly, the SORLA ectodomain is cleaved into a soluble form, sSORLA, which has been shown to regulate cytoskeletal signaling pathways and cell motility in cells outside the nervous system. We show here that sSORLA binds and activates the EGF receptor to induce downstream signaling through the ERK serine/threonine kinase and the Fos transcription factor, thereby enhancing neurite outgrowth. These findings reveal a novel role for sSORLA in promoting neurite regeneration through the EGF receptor/ERK/Fos pathway, thereby demonstrating a potential neuroprotective mechanism involving SORLA.


Assuntos
Receptores ErbB/fisiologia , Sistema de Sinalização das MAP Quinases/fisiologia , Proteínas de Membrana Transportadoras/fisiologia , Regeneração Nervosa/fisiologia , Neuritos/fisiologia , Receptores de LDL/fisiologia , Animais , Células Cultivadas , Feminino , Regulação da Expressão Gênica , Genes fos , Hipocampo/fisiologia , Masculino , Proteínas de Membrana Transportadoras/genética , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Transgênicos , Fosforilação , Receptores de LDL/genética
4.
PLoS Pathog ; 12(11): e1006034, 2016 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-27893816

RESUMO

microRNAs are endogenous small regulatory RNAs that modulate myriad biological processes by repressing target gene expression in a sequence-specific manner. Here we show that the conserved miRNA miR-34 regulates innate immunity and ecdysone signaling in Drosophila. miR-34 over-expression activates antibacterial innate immunity signaling both in cultured cells and in vivo, and flies over-expressing miR-34 display improved survival and pathogen clearance upon Gram-negative bacterial infection; whereas miR-34 knockout animals are defective in antibacterial defense. In particular, miR-34 achieves its immune-stimulatory function, at least in part, by repressing the two novel target genes Dlg1 and Eip75B. In addition, our study reveals a mutual repression between miR-34 expression and ecdysone signaling, and identifies miR-34 as a node in the intricate interplay between ecdysone signaling and innate immunity. Lastly, we identify cis-regulatory genomic elements and trans-acting transcription factors required for optimal ecdysone-mediated repression of miR-34. Taken together, our study enriches the repertoire of immune-modulating miRNAs in animals, and provides new insights into the interplay between steroid hormone signaling and innate immunity.


Assuntos
Drosophila melanogaster/imunologia , Ecdisona/imunologia , Imunidade Inata/imunologia , MicroRNAs/imunologia , Transdução de Sinais , Animais , Northern Blotting , Imunoprecipitação da Cromatina , Proteínas de Ligação a DNA/imunologia , Modelos Animais de Doenças , Proteínas de Drosophila/imunologia , Técnicas de Inativação de Genes , Imunoprecipitação , Reação em Cadeia da Polimerase , Transdução de Sinais/imunologia , Fatores de Transcrição/imunologia , Proteínas Supressoras de Tumor/imunologia
5.
PLoS Genet ; 11(8): e1005475, 2015 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-26308709

RESUMO

microRNAs (miRNAs) are a class of endogenous regulatory RNAs that play a key role in myriad biological processes. Upon transcription, primary miRNA transcripts are sequentially processed by Drosha and Dicer ribonucleases into ~22-24 nt miRNAs. Subsequently, miRNAs are incorporated into the RNA-induced silencing complexes (RISCs) that contain Argonaute (AGO) family proteins and guide RISC to target RNAs via complementary base pairing, leading to post-transcriptional gene silencing by a combination of translation inhibition and mRNA destabilization. Select pre-mRNA splicing factors have been implicated in small RNA-mediated gene silencing pathways in fission yeast, worms, flies and mammals, but the underlying molecular mechanisms are not well understood. Here, we show that SmD1, a core component of the Drosophila small nuclear ribonucleoprotein particle (snRNP) implicated in splicing, is required for miRNA biogenesis and function. SmD1 interacts with both the microprocessor component Pasha and pri-miRNAs, and is indispensable for optimal miRNA biogenesis. Depletion of SmD1 impairs the assembly and function of the miRISC without significantly affecting the expression of major canonical miRNA pathway components. Moreover, SmD1 physically and functionally associates with components of the miRISC, including AGO1 and GW182. Notably, miRNA defects resulting from SmD1 silencing can be uncoupled from defects in pre-mRNA splicing, and the miRNA and splicing machineries are physically and functionally distinct entities. Finally, photoactivatable-ribonucleoside-enhanced crosslinking and immunoprecipitation (PAR-CLIP) analysis identifies numerous SmD1-binding events across the transcriptome and reveals direct SmD1-miRNA interactions. Our study suggests that SmD1 plays a direct role in miRNA-mediated gene silencing independently of its pre-mRNA splicing activity and indicates that the dual roles of splicing factors in post-transcriptional gene regulation may be evolutionarily widespread.


Assuntos
Drosophila melanogaster/genética , Proteínas de Insetos/genética , MicroRNAs/genética , Processamento Pós-Transcricional do RNA/genética , Splicing de RNA/genética , Animais , Proteínas Argonautas/genética , Sequência de Bases , Linhagem Celular , Proteínas de Drosophila/genética , Regulação da Expressão Gênica , Inativação Gênica , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Proteínas de Ligação a RNA/genética , Complexo de Inativação Induzido por RNA/genética , Análise de Sequência de RNA
6.
Proc Natl Acad Sci U S A ; 110(41): 16520-5, 2013 Oct 08.
Artigo em Inglês | MEDLINE | ID: mdl-24067655

RESUMO

RNAi is an evolutionarily conserved gene regulatory process that operates in a wide variety of organisms. During RNAi, long double-stranded RNA precursors are processed by Dicer proteins into ∼21-nt siRNAs. Subsequently, siRNAs are incorporated into the RNA-induced silencing complexes (RISCs) that contain Argonaute-family proteins and guide RISC to target RNAs via complementary base pairing, leading to posttranscriptional gene silencing. Select pre-mRNA splicing factors have been implicated in RNAi in fission yeast, worms, and flies, but the underlying molecular mechanisms are not well understood. Here, we show that SmD1, a core component of the Drosophila small nuclear ribonucleoprotein particle implicated in splicing, is required for RNAi and antiviral immunity in cultured cells and in vivo. SmD1 interacts with both Dicer-2 and dsRNA precursors and is indispensable for optimal siRNA biogenesis. Depletion of SmD1 impairs the assembly and function of the small interfering RISC without significantly affecting the expression of major canonical siRNA pathway components. Moreover, SmD1 physically and functionally associates with components of the small interfering RISC, including Argonaute 2, both in flies and in humans. Notably, RNAi defects resulting from SmD1 silencing can be uncoupled from defects in pre-mRNA splicing, and the RNAi and splicing machineries are physically and functionally distinct entities. Our results suggest that Drosophila SmD1 plays a direct role in RNAi-mediated gene silencing independently of its pre-mRNA splicing activity and indicate that the dual roles of splicing factors in posttranscriptional gene regulation may be evolutionarily widespread.


Assuntos
Drosophila/genética , Interferência de RNA/fisiologia , Proteínas Centrais de snRNP/genética , Proteínas Centrais de snRNP/metabolismo , Animais , Northern Blotting , Linhagem Celular , Drosophila/fisiologia , Imunoprecipitação , Precursores de RNA/genética , RNA Interferente Pequeno/genética
7.
RNA Biol ; 11(9): 1171-9, 2014.
Artigo em Inglês | MEDLINE | ID: mdl-25483042

RESUMO

RNA interference (RNAi) is a eukaryotic gene-silencing system. Although the biochemistry of RNAi is relatively well defined, how this pathway is regulated remains incompletely understood. To identify genes involved in regulating the RNAi pathway, we screened for genetic mutations in Drosophila that alter the efficiency of RNAi. We identified the Drosophila homolog of the mammalian CR6-interacting factor 1 (CRIF1), also known as growth arrest and DNA-damage-inducible 45-gamma interacting protein (Gadd45GIP1), as a potential new regulator of the RNAi pathway. Loss-of-function mutants of Drosophila CRIF1 (dCRIF) are deficient in RNAi-mediated target gene knock-down, in the biogenesis of small interfering RNA (siRNA) molecules, and in antiviral immunity. Moreover, we show that dCRIF may function by interacting with, and stabilizing, the RNase III enzyme Dicer-2. Our results suggest that dCRIF may play an important role in regulating the RNAi pathway.


Assuntos
Animais Geneticamente Modificados/genética , Proteínas de Drosophila/metabolismo , Drosophila melanogaster/genética , RNA Helicases/química , Interferência de RNA , RNA Interferente Pequeno/genética , Ribonuclease III/química , Glândulas Salivares/imunologia , Animais , Animais Geneticamente Modificados/crescimento & desenvolvimento , Animais Geneticamente Modificados/metabolismo , Proteínas Argonautas/genética , Proteínas Argonautas/metabolismo , Northern Blotting , Western Blotting , Células Cultivadas , Proteínas de Drosophila/química , Proteínas de Drosophila/genética , Drosophila melanogaster/crescimento & desenvolvimento , Drosophila melanogaster/metabolismo , Imunofluorescência , Imunoprecipitação , RNA Helicases/genética , RNA Helicases/metabolismo , RNA Mensageiro/genética , Reação em Cadeia da Polimerase em Tempo Real , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Ribonuclease III/genética , Ribonuclease III/metabolismo , Glândulas Salivares/metabolismo , Glândulas Salivares/virologia
8.
Cell Rep ; 43(4): 113973, 2024 Apr 23.
Artigo em Inglês | MEDLINE | ID: mdl-38507406

RESUMO

We identified and validated a collection of circular RNAs (circRNAs) in Drosophila melanogaster. We show that depletion of the pro-viral circRNA circATP8B(2), but not its linear siblings, compromises viral infection both in cultured Drosophila cells and in vivo. In addition, circATP8B(2) is enriched in the fly gut, and gut-specific depletion of circATP8B(2) attenuates viral replication in an oral infection model. Furthermore, circATP8B(2) depletion results in increased levels of reactive oxygen species (ROS) and enhanced expression of dual oxidase (Duox), which produces ROS. Genetic and pharmacological manipulations of circATP8B(2)-depleted flies that reduce ROS levels rescue the viral replication defects elicited by circATP8B(2) depletion. Mechanistically, circATP8B(2) associates with Duox, and circATP8B(2)-Duox interaction is crucial for circATP8B(2)-mediated modulation of Duox activity. In addition, Gαq, a G protein subunit required for optimal Duox activity, acts downstream of circATP8B(2). We conclude that circATP8B(2) regulates antiviral defense by modulating Duox expression and Duox-dependent ROS production.


Assuntos
Proteínas de Drosophila , Drosophila melanogaster , RNA Circular , Espécies Reativas de Oxigênio , Animais , Espécies Reativas de Oxigênio/metabolismo , RNA Circular/genética , RNA Circular/metabolismo , Drosophila melanogaster/imunologia , Drosophila melanogaster/genética , Drosophila melanogaster/metabolismo , Proteínas de Drosophila/metabolismo , Proteínas de Drosophila/genética , Replicação Viral , RNA/metabolismo , RNA/genética , NADPH Oxidases/metabolismo , NADPH Oxidases/genética , Oxidases Duais/metabolismo , Oxidases Duais/genética
9.
Eur J Med Chem ; 259: 115684, 2023 Nov 05.
Artigo em Inglês | MEDLINE | ID: mdl-37542989

RESUMO

Recently, histone lysine specific demethylase 1 (LSD1) has become an emerging and promising target for cancer immunotherapy. Herein, based on our previously reported LSD1 inhibitor DXJ-1 (also called 6x), a series of novel acridine-based LSD1 inhibitors were identified via structure optimizations. Among them, compound 5ac demonstrated significantly enhanced inhibitory activity against LSD1 with an IC50 value of 13 nM, about 4.6-fold more potent than DXJ-1 (IC50 = 73 nM). Molecular docking studies revealed that compound 5ac could dock well into the active site of LSD1. Further mechanism studies showed that compound 5ac inhibited the stemness and migration of gastric cancer cells, and reduced the expression of PD-L1 in BGC-823 and MFC cells. More importantly, BGC-823 cells were more sensitive to T cell killing when treated with compound 5ac. Besides, the tumor growth was also suppressed by compound 5ac in mice. Together, 5ac could serve as a promising candidate to enhance immune response in gastric cancer.


Assuntos
Antineoplásicos , Neoplasias Gástricas , Animais , Camundongos , Antineoplásicos/química , Relação Estrutura-Atividade , Neoplasias Gástricas/tratamento farmacológico , Simulação de Acoplamento Molecular , Acridinas/farmacologia , Linhagem Celular Tumoral , Imunidade , Histona Desmetilases , Inibidores Enzimáticos/farmacologia , Proliferação de Células
10.
Eur J Med Chem ; 251: 115255, 2023 May 05.
Artigo em Inglês | MEDLINE | ID: mdl-36913900

RESUMO

LSD1 is overexpressed in various cancers and promotes tumor cell proliferation, tumor expansion, and suppresses immune cells infiltration and is closely associated with immune checkpoint inhibitors therapy. Therefore, the inhibition of LSD1 has been recognized as a promising strategy for cancer therapy. In this study, we screened an in-house small-molecule library targeting LSD1, an FDA-approved drug amsacrine for acute leukemia and malignant lymphomas was found to exhibit moderate anti-LSD1 inhibitory activity (IC50 = 0.88 µM). Through further medicinal chemistry efforts, the most active compound 6x increased anti-LSD1 activity significantly (IC50 = 0.073 µM). Further mechanistic studies demonstrated that compound 6x inhibited the stemness and migration of gastric cancer cell, and decreased the expression of PD-L1 (programmed cell death-ligand 1) in BGC-823 and MFC cells. More importantly, BGC-823 cells are more susceptible to T-cell killing when treated with compound 6x. Moreover, tumor growth was also suppressed by compound 6x in mice. Altogether, our findings demonstrated that acridine-based novel LSD1 inhibitor 6x may be a lead compound for the development of activating T cell immune response in gastric cancer cells.


Assuntos
Antineoplásicos , Neoplasias Gástricas , Animais , Camundongos , Antineoplásicos/química , Inibidores Enzimáticos/farmacologia , Neoplasias Gástricas/tratamento farmacológico , Acridinas/farmacologia , Acridinas/uso terapêutico , Linhagem Celular Tumoral , Histona Desmetilases , Proliferação de Células
11.
J Med Chem ; 66(6): 3896-3916, 2023 03 23.
Artigo em Inglês | MEDLINE | ID: mdl-36856685

RESUMO

Histone lysine specific demethylase 1 (LSD1) has been recognized as an important epigenetic target for cancer treatment. Although several LSD1 inhibitors have entered clinical trials, the discovery of novel potent LSD1 inhibitors remains a challenge. In this study, the antipsychotic drug chlorpromazine was characterized as an LSD1 inhibitor (IC50 = 5.135 µM), and a series of chlorpromazine derivatives were synthesized. Among them, compound 3s (IC50 = 0.247 µM) was the most potent one. More importantly, compound 3s inhibited LSD1 in the cellular level and downregulated the expression of programmed cell death-ligand 1 (PD-L1) in BGC-823 and MFC cells to enhance T-cell killing response. An in vivo study confirmed that compound 3s can inhibit MFC cell proliferation without significant toxicity in immunocompetent mice. Taken together, our findings indicated that the novel LSD1 inhibitor 3s tethering a phenothiazine scaffold may serve as a lead compound for further development to activate T-cell immunity in gastric cancer.


Assuntos
Inibidores Enzimáticos , Neoplasias Gástricas , Animais , Camundongos , Inibidores Enzimáticos/farmacologia , Neoplasias Gástricas/tratamento farmacológico , Linhagem Celular Tumoral , Clorpromazina/uso terapêutico , Linfócitos T/metabolismo , Proliferação de Células , Histona Desmetilases/metabolismo , Morte Celular , Relação Estrutura-Atividade
12.
J Virol ; 85(6): 2869-77, 2011 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-21209107

RESUMO

Infectious spleen and kidney necrosis virus (ISKNV), the type species of the genus Megalocytivirus in the family Iridoviridae, causes severe damage to mandarin fish cultures in China. Little is known about the proteins of ISKNV virions. In this study, a total of 38 ISKNV virion-associated proteins were identified by four different workflows with systematic and comprehensive proteomic approaches. Among the 38 identified proteins, 21 proteins were identified by the gel-based workflows (one-dimensional [1-D] and two-dimensional [2-D] gel electrophoresis). Fifteen proteins were identified by 1-D gel electrophoresis, and 16 proteins were identified by 2-D gel electrophoresis, with 10 proteins identified by both methods. Another 17 proteins were identified only by liquid chromatography (LC)-based workflows (LC-matrix-assisted laser desorption ionization [MALDI] and linear trap quadrupole [LTQ]-Orbitrap). Among these 17 LC-identified proteins, 5 proteins were identified uniquely by the LC-MALDI workflow, whereas another 6 proteins were identified only by the LTQ-Orbitrap workflow. These results underscore the importance of incorporation of multiple approaches in identification of viral proteins. Based on viral genomic sequence, genes encoding these 38 viral proteins were cloned and expressed in vitro. Antibodies were produced against these 38 proteins to confirm the ISKNV structural proteins by Western blotting. Of the newly identified proteins, ORF 056L and ORF 118L were identified and confirmed as two novel viral envelope proteins by Western blotting and immunoelectron microscopy (IEM). The ISKNV proteome reported here is currently the only characterized megalocytivirus proteome. The systematic and comprehensive identification of ISKNV structural proteins and their localizations in this study will facilitate future studies of the ISKNV assembly process and infection mechanism.


Assuntos
Iridoviridae/química , Proteínas Estruturais Virais/análise , Vírion/química , Animais , Linhagem Celular , China , Cromatografia Líquida , Eletroforese em Gel Bidimensional , Eletroforese em Gel de Poliacrilamida , Peixes , Proteoma/análise , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz
13.
Fish Shellfish Immunol ; 31(6): 919-24, 2011 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-21888976

RESUMO

Infectious spleen and kidney necrosis virus (ISKNV), belonging to the genus Megalocytivirus in the family Iridoviridae, is one of the major agents causing mortality and economic losses to the freshwater fish culture industry in Asian countries. Currently, little information regarding the antigenic properties of Megalocytivirus (especially ISKNV) is available. Our previous study using four different workflows with systematic and comprehensive proteomic approaches led to the identification of 38 ISKNV virion-associated proteins (J. Virol. 2869-2877, 2011). Thus, in this report, the antigenicity of 31 structural proteins from ISKNV virion was investigated. A one-dimensional gel electrophoresis immunoblot profile coupled with MALDI-TOF-TOF MS/MS was applied to identify six immunogenic viral proteins, namely, ORFs major capsid protein (006L), 054L, 055L, 101L, 117L, and 125L. Then, the antigenicity of 31 structural proteins was characterized by Western blot by using pooled sera from mandarin fish that survived ISKNV infection. Of the 31 viral proteins, 22 were recognized by the fish ISKNV antiserum. Furthermore, this antiserum neutralizes MFF-1 cells ISKNV infection. To our knowledge, this study is the first report on the immunogenicity of viral proteins and characterization of the proteome of megalocytivirus infective agents. Our findings are expected to promote the development of effective vaccine candidates.


Assuntos
Antígenos Virais/genética , Proteínas do Capsídeo/genética , Iridoviridae/genética , Perciformes/virologia , Vírion/genética , Animais , Ásia , Western Blotting , Soros Imunes/imunologia , Testes de Neutralização , Proteômica/métodos , Espectrometria de Massas em Tandem
14.
J Med Chem ; 64(5): 2466-2488, 2021 03 11.
Artigo em Inglês | MEDLINE | ID: mdl-33619958

RESUMO

As a flavin adenine dinucleotide (FAD)-dependent monoamine oxidase, lysine specific demethylase 1 (LSD1/KDM1A) functions as a transcription coactivator or corepressor to regulate the methylation of histone 3 lysine 4 and 9 (H3K4/9), and it has emerged as a promising epigenetic target for anticancer treatment. To date, numerous inhibitors targeting LSD1 have been developed, some of which are undergoing clinical trials for cancer therapy. Although only two reversible LSD1 inhibitors CC-90011 and SP-2577 are in the clinical stage, the past decade has seen remarkable advances in the development of reversible LSD1 inhibitors. Herein, we provide a comprehensive review about structures, biological evaluation, and structure-activity relationship (SAR) of reversible LSD1 inhibitors.


Assuntos
Inibidores Enzimáticos/uso terapêutico , Histona Desmetilases/antagonistas & inibidores , Compostos Orgânicos/uso terapêutico , Animais , Antineoplásicos/química , Antineoplásicos/farmacologia , Antineoplásicos/uso terapêutico , Linhagem Celular Tumoral , Inibidores Enzimáticos/química , Inibidores Enzimáticos/farmacologia , Humanos , Estrutura Molecular , Neoplasias/tratamento farmacológico , Compostos Orgânicos/química , Compostos Orgânicos/farmacologia , Relação Estrutura-Atividade
15.
J Proteome Res ; 9(5): 2573-83, 2010 May 07.
Artigo em Inglês | MEDLINE | ID: mdl-20148598

RESUMO

Bacterium is still a major cause of many infectious diseases and a global threat to human health, aquaculture, and animal feeding. Prevention by vaccination is the most efficient and economical way of fighting bacterial diseases, but one of the persistent challenges to prevent bacterial infections and disease transmissions is the existence of multiple bacterial species, families, and genera and the lack of efficient polyvalent vaccines against them. The information on candidate immunogens for polyvalent vaccine development is elusive, as well. For the development of broad cross-protective vaccines, we have employed heterogeneous antiserum-based immunoproteomics approaches to identify antigenically similar outer membrane (OM) proteins that could be used as potential polyvalent vaccine candidates against Vibrio parahaemolyticus , V. alginolyticus , V. fluvialis , Aeromonas hydrophila , and A. sobria infections. VPA1435, VP0764, VPA1186, VP1061, and VP2850 could be recognized by at least three antisera and demonstrated significantly passive and active immune protection against V. parahaemolyticus infection in a crucian carp model. VP1061 and VP2850 induced higher immune and protective abilities than the other three OM proteins. Furthermore, the abilities of VP1061 and VP2850 in the generation of broad cross-protective immune reaction against the infections of V. alginolyticus , A. hydrophila , and Pseudomonas fluorescens were also investigated in fish and mouse models. Our results suggested that VP1061 and VP2850 could potentially be used as polyvalent vaccine candidates for the development of novel polyvalent vaccines against V. parahaemolyticus and other Gram-negative pathogens. On the basis of these results, characteristics of OM proteins as polyvalent vaccine candidates have been addressed.


Assuntos
Anticorpos Antibacterianos/imunologia , Proteínas da Membrana Bacteriana Externa/imunologia , Vacinas Bacterianas/imunologia , Proteômica/métodos , Vibrio parahaemolyticus/imunologia , Sequência de Aminoácidos , Animais , Anticorpos Antibacterianos/sangue , Proteínas da Membrana Bacteriana Externa/genética , Vacinas Bacterianas/química , Western Blotting , Carpas , Proteção Cruzada , Eletroforese em Gel Bidimensional , Imunização Passiva , Camundongos , Dados de Sequência Molecular , Filogenia , Alinhamento de Sequência , Vacinação , Vibrio parahaemolyticus/química , Vibrio parahaemolyticus/genética
16.
Fish Shellfish Immunol ; 29(5): 810-6, 2010 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-20659563

RESUMO

Vibrio alginolyticus is the etiological agent that causes great losses in aquacultures and clinical emanating cases in humans. Identification of highly efficient vaccine candidates to control V. alginolyticus infection has been highly concerned since vaccines offer a powerful approach to provide efficient protection from bacterial infections. In the present study, we firstly investigated the altered outer membrane proteins (OM proteins) of V. alginolyticus in response to NaCl concentrations and iron limitation using Western blotting, and then identified the protective activity of these altered OM proteins by bacterial challenge post immunization. Ten OM proteins were differentially expressed in response to the osmolarity changing or/and iron limitation, in which VA2212, OmpV, VPA1186, OmpU, VPA1644, VA1061, VA1631 and VPA0860 were markedly altered in response to osmolarity, and VPA1186, OmpU, OmpV, VA0449, VPA0860, VPA1435 and VA1631 were determined to be iron-limited responsive proteins. Out of the ten OM proteins, VA1061, OmpU, VPA1435 and VPA0860 could be effective vaccine candidates against infection by V. alginolyticus in vivo. Further results indicated that VA1061 and VPA0860 were dominant antigens and could stimulate hosts to produce stronger antibody response than other two in live or inactivated whole-cell vaccines. These results not only expand knowledge on osmolarity-, iron-responsive proteins, but also provide a valuable strategy for identify protective proteins suitable for use in vaccine development.


Assuntos
Proteínas da Membrana Bacteriana Externa/imunologia , Proteínas da Membrana Bacteriana Externa/metabolismo , Vacinas Bacterianas/imunologia , Descoberta de Drogas/métodos , Regulação Bacteriana da Expressão Gênica/fisiologia , Vibrio alginolyticus/imunologia , Animais , Western Blotting , Eletroforese em Gel de Poliacrilamida , Ensaio de Imunoadsorção Enzimática , Soros Imunes/imunologia , Ferro/metabolismo , Deficiências de Ferro , Camundongos , Cloreto de Sódio/metabolismo
17.
J Med Chem ; 63(23): 14197-14215, 2020 12 10.
Artigo em Inglês | MEDLINE | ID: mdl-32931269

RESUMO

Histone lysine-specific demethylase 1 (LSD1/KDM1A) has become an important and promising anticancer target since it was first identified in 2004 and specially demethylates lysine residues of histone H3K4me1/2 and H3K9me1/2. LSD1 is ubiquitously overexpressed in diverse cancers, and abrogation of LSD1 results in inhibition of proliferation, invasion, and migration in cancer cells. Over the past decade, a number of biologically active small-molecule LSD1 inhibitors have been developed. To date, six trans-2-phenylcyclopropylamine (TCP)-based LSD1 inhibitors (including TCP, ORY-1001, GSK-2879552, INCB059872, IMG-7289, and ORY-2001) that covalently bind to the flavin adenine dinucleotide (FAD) within the LSD1 catalytic cavity have already entered into clinical trials. Here, we provide an overview about the structures, activities, and structure-activity relationship (SAR) of TCP-based LSD1 inhibitors that mainly covers the literature from 2008 to date. The opportunities, challenges, and future research directions in this emerging and promising field are also discussed.


Assuntos
Inibidores Enzimáticos/farmacologia , Histona Desmetilases/antagonistas & inibidores , Tranilcipromina/farmacologia , Antineoplásicos/química , Antineoplásicos/farmacologia , Desenho de Fármacos , Inibidores Enzimáticos/química , Humanos , Relação Estrutura-Atividade , Tranilcipromina/química
20.
Virology ; 437(2): 89-99, 2013 Mar 15.
Artigo em Inglês | MEDLINE | ID: mdl-23352451

RESUMO

Megalocytiviruses have three main genotypes, which are represented by ISKNV, RSIV, and TRBIV. To date, the virion-associated proteins of RSIV and TRBIV are still unknown. The spotted knifejaw iridovirus (SKIV) is a newly characterized RSIV-type megalocytivirus. In this study, the virion-associated proteins of SKIV were identified by systemic one-dimensional gel electrophoresis-based proteomic approaches. A total of 49 viral proteins and 33 cellular proteins were associated with the SKIV virions by LC MS/MS, including 18 highly abundant structural proteins that were detected by MALDI TOF/TOF-MS. One highly abundant structural protein of interest was identified as the virus-inducible stress protein (VISP) and further characterized as an envelope protein. However, knockdown of mVISP by siRNA method showed no effect in virion production. The current study is the first to present detailed information on the virion-associated proteins of an RSIV-type megalocytivirus and to identify a novel cellular envelope protein of this virus.


Assuntos
Infecções por Vírus de DNA/veterinária , Doenças dos Peixes/virologia , Iridoviridae/química , Perciformes/virologia , Proteínas Virais/análise , Vírion/química , Animais , Cromatografia Líquida , Infecções por Vírus de DNA/virologia , Eletroforese , Proteoma/análise , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz , Espectrometria de Massas em Tandem
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