Background:Sepsis,a life-threatening systemic inflammatory response to infection,presents significant challenges in the intensive care unit(ICU).The condition is frequently accompanied by persistent neurodegenerative ...Background:Sepsis,a life-threatening systemic inflammatory response to infection,presents significant challenges in the intensive care unit(ICU).The condition is frequently accompanied by persistent neurodegenerative changes and muscle atrophy,especially in patients with burn wounds,severely affecting recuperation and quality of life.Traditional physical therapy and advanced sedative screening represent pivotal approaches for the rehabilitation and management of these cases.Methods:A comprehensive transcriptomic study was conducted using gene expression datasets from sepsis patients and important biological models to identify potential targets.We subsequently employed virtual molecular docking and molecular dynamics simulations to screen the FDA drug library and traditional Chinese medicine databases for compounds exhibiting high binding affinity to isocitrate dehydrogenase 2(IDH2).The functional role of this target was validated in vitro using an IDH2 deficient BV-2 microglial cell model stimulated by lipopolysaccharide.Results:Key features were identified through differential expression analysis across multiple datasets.Gene set enrichment analysis(GSEA)highlighted pathways associated with chronic neurodegenerative diseases.Stable interactions between IDH2 and cobicistat or licorice glycoside E were predicted,supporting the identification of IDH2 as a promising therapeutic target.Knockdown of IDH2 in microglial cells led to markedly suppressed production of pro-inflammatory cytokines,including TNF-αand IL-6,confirming its pivotal role in neuroinflammatory response regulation.Conclusion:These findings underscore the pivotal role of IDH2 in persistent neurodegenerative changes associated with sepsis,including burn-related cases.Targeting IDH2 may reduce long-term neurovascular complications and muscle atrophy in septic ICU patients.Our findings suggest that IDH2 is a potential therapeutic target,and the combination of cobicistat and licorice glycoside E warrants further investigation as a promising therapeutic strategy.The study also emphasizes the significance of novel neuroinflammatory and neurovascular biomarkers in understanding and managing long-term sequelae in septic survivors.展开更多
Isocitrate dehydrogenase 2(IDH2)and glutamate dehydrogenase 1(GLUD1)are key enzymes involved in the production ofα-ketoglutarate(α-KG),a metabolite central to the tricarboxylic acid cycle and glutamine metabolism.In...Isocitrate dehydrogenase 2(IDH2)and glutamate dehydrogenase 1(GLUD1)are key enzymes involved in the production ofα-ketoglutarate(α-KG),a metabolite central to the tricarboxylic acid cycle and glutamine metabolism.In this study,we investigated the impact of IDH2 and GLUD1 on early porcine embryonic development following IDH2 and GLUD1 knockdown(KD)via doublestranded RNA(dsRNA)microinjection.Results showed that KD reducedα-KG levels,leading to delayed embryonic development,decreased blastocyst formation,increased apoptosis,reduced blastomere proliferation,and pluripotency.Additionally,IDH2 and GLUD1 KD induced abnormally high levels of trimethylation of lysine 20 of histone H4(H4K20me3)at the 4-cell stage,likely resulting in transcriptional repression of embryonic genome activation(EGA)-related genes.Notably,KD of lysine methyltransferase 5C(KMT5C)and supplementation with exogenousα-KG reduced H4K20me3 expression and partially rescued these defects,suggesting a critical role of IDH2 and GLUD1 in the epigenetic regulation and proper development of porcine embryos.Overall,this study highlights the significance of IDH2 and GLUD1 in maintaining normal embryonic development through their influence onα-KG production and subsequent epigenetic modifications.展开更多
Aim:Hepatocellular carcinoma(HCC)in patients with Metabolic dysfunction-associated steatotic liver disease(MASLD,formerly NAFLD)is expected to be a significant public health issue in the near future.Therefore,understa...Aim:Hepatocellular carcinoma(HCC)in patients with Metabolic dysfunction-associated steatotic liver disease(MASLD,formerly NAFLD)is expected to be a significant public health issue in the near future.Therefore,understanding the tumor microenvironment interactions in MASLD-induced HCC is crucial,and the development of relevant preclinical models is needed.Hence,we aimed to determine the effects of a MASLD-mimicking microenvironment(ME)on the aggressiveness of HCC cells and identify target genes that drive HCC by developing a 3D-in vitro co-culture system.Methods:A 3D co-culture system mimicking the MASLD-ME was created with LX-2 liver stellate cells embedded in 3D collagen gel in the lower and SNU-449 HCC cells on the upper parts of Boyden chambers,and cells were grown in an optimized metabolic medium(MM).The effects of MASLD-ME on motility,sphere formation,proliferation,and cell cycle of SNU-449 cells were tested by Boyden chamber,3D sphere formation,XTT,and Flow cytometry,respectively.The protein expression/activation profiles of motile SNU-449 cells that passed the membrane toward MASLD-ME or control condition were investigated using a multiplex protein profiling system DigiWest and confirmed with RT-PCR,WB,and Flow cytometry.IDH2 levels were examined in primary human HCC and adjacent liver tissues by IHC and in TCGA and CPTAC cohorts by bioinformatics tools.Results:MM treatment increased fat accumulation,motility,and spheroid formation of both SNU-449 and LX-2 cells.MASLD-ME induced activation of LX2 cells,leading to the formation of bigger colonies with many intrusions compared to related controls.DigiWest analysis showed that metabolism-related proteins such as IDH2 were the most affected molecules in SNU-449 cells that migrated toward the MASLD-ME compared to those that migrated toward the control condition.Downregulation of IDH2 expression was confirmed in SNU-449 cells grown in MASLD-ME,in primary HCC tumor samples by IHC,and in HCC patient cohorts by bioinformatics analysis.Conclusion:This study reports the potential involvement of MASLD-ME in the downregulation of IDH2 expression and promoted motility and colonization capacity of HCC cells.The 3D MASLD model presented in this study may be useful in investigating the mechanistic roles of MASLD-ME in HCC.展开更多
基金support and insightful feedback provided by colleagues and affiliated institutions during the preparation of this manuscript.
文摘Background:Sepsis,a life-threatening systemic inflammatory response to infection,presents significant challenges in the intensive care unit(ICU).The condition is frequently accompanied by persistent neurodegenerative changes and muscle atrophy,especially in patients with burn wounds,severely affecting recuperation and quality of life.Traditional physical therapy and advanced sedative screening represent pivotal approaches for the rehabilitation and management of these cases.Methods:A comprehensive transcriptomic study was conducted using gene expression datasets from sepsis patients and important biological models to identify potential targets.We subsequently employed virtual molecular docking and molecular dynamics simulations to screen the FDA drug library and traditional Chinese medicine databases for compounds exhibiting high binding affinity to isocitrate dehydrogenase 2(IDH2).The functional role of this target was validated in vitro using an IDH2 deficient BV-2 microglial cell model stimulated by lipopolysaccharide.Results:Key features were identified through differential expression analysis across multiple datasets.Gene set enrichment analysis(GSEA)highlighted pathways associated with chronic neurodegenerative diseases.Stable interactions between IDH2 and cobicistat or licorice glycoside E were predicted,supporting the identification of IDH2 as a promising therapeutic target.Knockdown of IDH2 in microglial cells led to markedly suppressed production of pro-inflammatory cytokines,including TNF-αand IL-6,confirming its pivotal role in neuroinflammatory response regulation.Conclusion:These findings underscore the pivotal role of IDH2 in persistent neurodegenerative changes associated with sepsis,including burn-related cases.Targeting IDH2 may reduce long-term neurovascular complications and muscle atrophy in septic ICU patients.Our findings suggest that IDH2 is a potential therapeutic target,and the combination of cobicistat and licorice glycoside E warrants further investigation as a promising therapeutic strategy.The study also emphasizes the significance of novel neuroinflammatory and neurovascular biomarkers in understanding and managing long-term sequelae in septic survivors.
基金supported by the National Research Foundation(NRF)of Korea grant funded by the Korean government(MSIT)(2022R1A2C300769),Republic of Korea。
文摘Isocitrate dehydrogenase 2(IDH2)and glutamate dehydrogenase 1(GLUD1)are key enzymes involved in the production ofα-ketoglutarate(α-KG),a metabolite central to the tricarboxylic acid cycle and glutamine metabolism.In this study,we investigated the impact of IDH2 and GLUD1 on early porcine embryonic development following IDH2 and GLUD1 knockdown(KD)via doublestranded RNA(dsRNA)microinjection.Results showed that KD reducedα-KG levels,leading to delayed embryonic development,decreased blastocyst formation,increased apoptosis,reduced blastomere proliferation,and pluripotency.Additionally,IDH2 and GLUD1 KD induced abnormally high levels of trimethylation of lysine 20 of histone H4(H4K20me3)at the 4-cell stage,likely resulting in transcriptional repression of embryonic genome activation(EGA)-related genes.Notably,KD of lysine methyltransferase 5C(KMT5C)and supplementation with exogenousα-KG reduced H4K20me3 expression and partially rescued these defects,suggesting a critical role of IDH2 and GLUD1 in the epigenetic regulation and proper development of porcine embryos.Overall,this study highlights the significance of IDH2 and GLUD1 in maintaining normal embryonic development through their influence onα-KG production and subsequent epigenetic modifications.
基金supported by“Scientific and Technological Research Council of Turkey(TUBITAK)”with the grant number 119S698.
文摘Aim:Hepatocellular carcinoma(HCC)in patients with Metabolic dysfunction-associated steatotic liver disease(MASLD,formerly NAFLD)is expected to be a significant public health issue in the near future.Therefore,understanding the tumor microenvironment interactions in MASLD-induced HCC is crucial,and the development of relevant preclinical models is needed.Hence,we aimed to determine the effects of a MASLD-mimicking microenvironment(ME)on the aggressiveness of HCC cells and identify target genes that drive HCC by developing a 3D-in vitro co-culture system.Methods:A 3D co-culture system mimicking the MASLD-ME was created with LX-2 liver stellate cells embedded in 3D collagen gel in the lower and SNU-449 HCC cells on the upper parts of Boyden chambers,and cells were grown in an optimized metabolic medium(MM).The effects of MASLD-ME on motility,sphere formation,proliferation,and cell cycle of SNU-449 cells were tested by Boyden chamber,3D sphere formation,XTT,and Flow cytometry,respectively.The protein expression/activation profiles of motile SNU-449 cells that passed the membrane toward MASLD-ME or control condition were investigated using a multiplex protein profiling system DigiWest and confirmed with RT-PCR,WB,and Flow cytometry.IDH2 levels were examined in primary human HCC and adjacent liver tissues by IHC and in TCGA and CPTAC cohorts by bioinformatics tools.Results:MM treatment increased fat accumulation,motility,and spheroid formation of both SNU-449 and LX-2 cells.MASLD-ME induced activation of LX2 cells,leading to the formation of bigger colonies with many intrusions compared to related controls.DigiWest analysis showed that metabolism-related proteins such as IDH2 were the most affected molecules in SNU-449 cells that migrated toward the MASLD-ME compared to those that migrated toward the control condition.Downregulation of IDH2 expression was confirmed in SNU-449 cells grown in MASLD-ME,in primary HCC tumor samples by IHC,and in HCC patient cohorts by bioinformatics analysis.Conclusion:This study reports the potential involvement of MASLD-ME in the downregulation of IDH2 expression and promoted motility and colonization capacity of HCC cells.The 3D MASLD model presented in this study may be useful in investigating the mechanistic roles of MASLD-ME in HCC.
