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Juan J Loor - One of the best experts on this subject based on the ideXlab platform.
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effect of heat shock protein b7 on oxidative stress in adipocytes from preruminant calves
Journal of Dairy Science, 2019Co-Authors: Xudong Sun, Juan J Loor, Hongdou Jia, R Bucktrout, Yazhe Wang, Xin Shu, Jihong Dong, Rankun Zuo, Liangyu Yang, Guowen LiuAbstract:Dairy cows with ketosis display excessive lipolysis in adipose tissue. Heat-shock protein B7 (HSPB7), a small heat-shock protein, plays important roles in mediating cytoprotective responses to oxidative stress in rodent adipose tissue. Accordingly, it is assumed that HSPB7 may also play important roles in the antioxidant response in adipose tissue of ketotic cows. Therefore, the aim of this study is to investigate (1) the redox state of adipose tissue in ketotic cows and (2) the role and mechanism of HSPB7 on the regulation of oxidative stress in adipocytes from preruminant calves. An in vivo study consisting of 15 healthy and 15 clinically ketotic cows was performed to harvest subcutaneous adipose tissue and blood samples. In addition, adipocytes isolated from calves were treated with different concentrations of H2O2 (0, 12.5, 25, 50, 100, or 200 μM) for 2 h, transfected with adenovirus-mediated overexpression of HSPB7 for 48 h, or transfected with small interfering RNA of HSPB7 for 48 h followed by exposure to H2O2 (200 μM) for 2 h. Serum concentrations of nonesterified fatty acids and β-hydroxybutyrate were greater in cows with clinical ketosis, whereas serum concentration of glucose was lower. Compared with healthy cows, the malondialdehyde content was greater but the activity of glutathione peroxidase and superoxide dismutase was lower in adipose tissue of clinically ketotic cows. The abundance of HSPB7 and nuclear factor, erythroid 2 like 2 (NFE2L2) was greater in adipose tissue of clinically ketotic cows. In vitro, H2O2 treatment induced the overproduction of reactive oxygen species and malondialdehyde, and inhibited the activity of antioxidant enzymes glutathione peroxidase and superoxide dismutase in adipocytes from preruminant calves. The low concentration of H2O2 (12.5, 25, and 50 μM) increased the abundance of HSPB7 and NFE2L2, but high concentrations of H2O2 (100 or 200 μM) reduced the abundance of HSPB7 and NFE2L2. The overexpression of HSPB7 improved the H2O2-induced oxidative stress in adipocytes via increasing the abundance of NFE2L2 and its downstream target genes heme oxygenase-1 (HMOX1) and NADH quinone oxidoreductase 1 (NQO1). Knockdown of HSPB7 markedly inhibited the expression of NFE2L2, HMOX1, and NQO1 and further exacerbated H2O2-induced oxidative stress. Overall, these results indicate that activation of the HSPB7-NFE2L2 pathway increases cellular antioxidant capacity, thereby alleviating oxidative stress in bovine adipocytes.
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glutathione metabolism and nuclear factor erythroid 2 like 2 NFE2L2 related proteins in adipose tissue are altered by supply of ethyl cellulose rumen protected methionine in peripartal holstein cows
Journal of Dairy Science, 2019Co-Authors: Yusheng Liang, C Parys, F Batistel, Juan J LoorAbstract:ABSTRACT Enhancing the supply of rumen-protected Met (RPM) during the peripartum period alleviates inflammation and oxidative stress status in dairy cows. We tested the hypothesis that RPM could increase abundance of genes and proteins related to glutathione (GSH) metabolism and the antioxidant transcription factor nuclear factor erythroid 2-like 2 (NFE2L2) in subcutaneous adipose tissue. Multiparous Holstein cows were fed a basal diet [control prepartum diet = 1.47 Mcal/kg of dry matter (DM) and 15.3% crude protein; control postpartum diet = 1.67 Mcal/kg of DM and 17.7% crude protein] or the control plus ethyl-cellulose RPM at a rate of 0.09 and 0.10% of DM intake before expected calving and after calving, respectively. Sixty cows were assigned to treatments based on parity, previous 305-d milk yield, and body condition score at 28 d from parturition. Diets were fed from −28 to 30 d. Biopsies of subcutaneous adipose tissue collected on d −10, 10, and 30 relative to parturition from 7 cows in each group were used for measuring concentrations of GSH, reactive oxygen species, superoxide dismutase, malondialdehyde, and mRNA and protein abundance (Western blotting). A repeated-measures ANOVA was used for statistics. The statistical model included the random effect of block and fixed effects of treatment, time, and its interaction. There was a diet × time effect for reactive oxygen species due to lower concentrations in Met versus control cows specifically at d −10. Cows fed Met also had lower concentrations of malondialdehyde in subcutaneous adipose tissue. Compared with controls, overall mRNA abundance of the GSH metabolism-related genes cystathionine-β-synthase (CBS), glutamate-cysteine ligase modifier subunit (GCLM), glutathione reductase (GSR), and glutathione peroxidase 1 (GPX1) was greater in cows fed Met. Furthermore, supply of Met resulted in an overall upregulation of protein abundance of glutathione peroxidase (GPX) 1, GPX3, glutathione S-transferase mu 1 (GSTM1), and glutathione S-transferase α 4 (GSTA4), all related to GSH metabolism. There was a diet × time effect for protein abundance of NFE2L2 and its repressor Kelch-like ECH associated protein 1 (KEAP1) due to lower values at 30 d in cows fed Met versus controls. The abundance of phosphorylated NFE2L2 was lower at 30 d in response to Met. Overall, the data suggest that exogenous Met may play a role in activating GSH metabolism and the antioxidant NFE2L2 pathways in subcutaneous adipose tissue.
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tea polyphenols protect bovine mammary epithelial cells from hydrogen peroxide induced oxidative damage in vitro by activating NFE2L2 hmox1 pathways
Journal of Dairy Science, 2019Co-Authors: Y F, L Zhao, Danielle N Coleman, M Gao, Juan J LoorAbstract:ABSTRACT Periparturient dairy cows are likely subject to altered intracellular reduction-oxidation (redox) balance due to the high metabolic rates and physiological adaptations occurring around parturition. Such conditions could induce oxidative damage. In nonruminants, it is well established that nuclear factor erythroid 2 like 2 (NFE2L2) is a critical transcription factor for maintaining cellular redox balance by inducing adaptive responses against oxidative stress (OS) that can otherwise lead to uncontrolled inflammation. Tea polyphenols (TP), the major polyphenolic constituents of green tea, are potent antioxidants that could exert protective effects on bovine mammary epithelial cells (BMEC) by scavenging free radicals. We used NFE2L2 short interfering RNA (siRNA) to downregulate NFE2L2 expression in cultured BMEC to investigate whether TP could inhibit H2O2-induced OS by activating the NFE2L2/heme oxygenase-1 (HMOX1) pathway. Isolated BMEC were exposed to H2O2 (600 μM) for 6 h to induce OS. Optimal doses of TP (0, 60, 80, and 100 μg/mL) were evaluated by pretreatment of BMEC for 0, 2, 4, 6, 8, 12, and 24 h, followed by a H2O2 (600 μM) challenge for 6 h. The BMEC were transfected with NFE2L2-siRNA for 48 h, pretreated with 100 µg/mL of TP for 12 h, then challenged by 600 μM H2O2 for 6 h. Results revealed that after H2O2 exposure a concentration of TP of 100 μg/mL during a 12-h incubation led to greater cell viability, protein, and mRNA abundance of NFE2L2, and lower intracellular reactive oxygen species (ROS) accumulation. In addition, transfection with NFE2L2-siRNA decreased abundance of NFE2L2 and HMOX1 in spite of exogenous TP supplementation, whereas ROS production was increased in response to exogenous H2O2 (600 μM). Overall, TP had beneficial effects on redox balance in BMEC, slowing down cellular OS-related injury through decreasing the production of ROS and enhancing mechanisms controlled at least in part by the NFE2L2/HMOX1 pathway.
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tea polyphenols protect bovine mammary epithelial cells from hydrogen peroxide induced oxidative damage in vitro
Journal of Animal Science, 2018Co-Authors: L Zhao, M Gao, Juan J LoorAbstract:Periparturient dairy cows are subjected to altered intracellular reduction-oxidation (redox) balance due to the high metabolic rates and physiological adaptations characteristic of the transition into lactation. Such conditions could alter oxidative stress status. The objective of this study was to investigate the cytoprotective effects of tea polyphenols (TP) in cultured bovine mammary epithelial cells (BMEC) exposed to hydrogen peroxide (H2O2)-induced oxidative stress. To establish oxidative stress, isolated BMEC were exposed to increasing concentrations of H2O2 (0, 100, 200, 400, 600, 800, and 1,000 µM) for 0, 2, 4, 6, 8, 12, and 24 h. Doses of TP (0, 60, 80, and 100 µg/mL) were evaluated by pretreatment of BMEC for 0, 2, 4, 6, 8, 12 and 24 h, followed by an H2O2 (600 µM per culture well) challenge for 6 h. Bovine mammary epithelial cells were preincubated for 30 min with or without 2,4-dinitrochloro-benzene (DNCB), then cultured with or without TP (100 µg/mL) for another 12 h followed by H2O2 (600 µM per culture well) exposure. There were 5 replicate cultures for each treatment in each experiment. Treatment with 600 µM H2O2 per culture well for 6 h induced oxidative damage of BMEC, indicating this system could be used to establish an oxidative stress model. After H2O2 (600 µM per culture well) exposure, a concentration of TP of 100 µg/mL during a 12-h incubation increased cell viability, decreased intracellular reactive oxygen species accumulation, and increased the abundance of nuclear factor-erythroid 2-related factor 2 (NFE2L2). Furthermore, TP upregulated mRNA abundance of genes in the NFE2L2 and mitogen-activated protein kinase (MAPK) pathways of BMEC. The DNCB assay allowed further confirmation that the induction of NFE2L2 and HMOX-1 in response to TP was mediated through the sustained upregulation of the abundance of MAPK3/1 [formerly known as extracellular regulated kinases 1/2] and MAPK11/12/13/14 (formerly known as p38). Overall, results indicate that TP has beneficial effects on BMEC redox balance; it can reduce cellular oxidative stress-related injury and may potentially serve as an antioxidant against oxidative stress in dairy cows.
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methionine supply alters mammary gland antioxidant gene networks via phosphorylation of nuclear factor erythroid 2 like 2 NFE2L2 protein in dairy cows during the periparturient period
Journal of Dairy Science, 2018Co-Authors: Liqiang Han, Fernanda Batistel, A S Alharthi, C Parys, Juan J LoorAbstract:The periparturient period is the most critical period during the lactation cycle of dairy cows and is characterized by increased oxidative stress status. The objective of this experiment was to evaluate the effect of supplementing rumen-protected methionine on nuclear factor erythroid 2-like 2 (NFE2L2, formerly NRF2) protein and target gene expression in the mammary gland during the early postpartal period. Multiparous Holstein cows were used in a block design experiment with 30 cows per treatment. Treatments consisting of a basal control diet (control) or the basal diet plus rumen-protected methionine (methionine) were fed from d -28 to 60 relative to parturition. Mammary tissue biopsies were harvested on d 21 postpartum from 5 cows per treatment. Compared with control, methionine increased dry matter intake, milk yield, and milk protein content. Among plasma parameters measured, methionine led to greater methionine and lower reactive oxygen metabolites. Compared with control, methionine supply resulted in greater mRNA abundance of the NFE2L2 target genes glutamate-cysteine ligase catalytic subunit (GCLC), glutamate-cysteine ligase modifier subunit (GCLM), glutathione reductase (GSR), glutathione peroxidase 1 (GPX1), malic enzyme 1 (ME1), ferrochelatase (FECH), ferritin heavy chain 1 (FTH1), and NAD(P) H quinone dehydrogenase 1 (NQO1) in the mammary tissue. In addition, methionine upregulated the mRNA abundance of NFE2L2, NFKB1, MAPK14 and downregulated KEAP1. The ratio of phosphorylated NFE2L2 to total NFE2L2 protein, and total heme oxygenase 1 (HMOX1) protein were markedly greater in response to methionine supply. In contrast, total protein abundance of Kelch-like ECH-associated protein 1 (KEAP1), which sequesters NFE2L2 in the cytosol and reduces its activity, was lower with methionine. Besides the consistent positive effect of methionine supply on systemic inflammation and oxidative stress status, the present data indicate a positive effect also on antioxidant mechanisms within the mammary gland, which are regulated, at least in part, via phosphorylation of NFE2L2 and its target genes. The exact mechanisms for these responses merit further study.
Antonio Cuadrado - One of the best experts on this subject based on the ideXlab platform.
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therapeutic targeting of the nrf2 and keap1 partnership in chronic diseases
Nature Reviews Drug Discovery, 2019Co-Authors: Antonio Cuadrado, Ana I Rojo, Geoffrey Wells, John D Hayes, Sharon P Cousin, William L Rumsey, Otis C Attucks, Stephen Franklin, Annaliisa Levonen, Thomas W KenslerAbstract:The transcription factor NF-E2 p45-related factor 2 (NRF2; encoded by NFE2L2) and its principal negative regulator, the E3 ligase adaptor Kelch-like ECH-associated protein 1 (KEAP1), are critical in the maintenance of redox, metabolic and protein homeostasis, as well as the regulation of inflammation. Thus, NRF2 activation provides cytoprotection against numerous pathologies including chronic diseases of the lung and liver; autoimmune, neurodegenerative and metabolic disorders; and cancer initiation. One NRF2 activator has received clinical approval and several electrophilic modifiers of the cysteine-based sensor KEAP1 and inhibitors of its interaction with NRF2 are now in clinical development. However, challenges regarding target specificity, pharmacodynamic properties, efficacy and safety remain.
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transcription factor NFE2L2 nrf2 modulates chaperone mediated autophagy through the regulation of lamp2a
Autophagy, 2018Co-Authors: Marta Pajares, Antonio Diazcarretero, Ana I Rojo, Ana Maria Cuervo, Esperanza Arias, Antonio CuadradoAbstract:Chaperone-mediated autophagy (CMA) is a selective degradative process for cytosolic proteins that contributes to the maintenance of proteostasis. The signaling mechanisms that control CMA are not f...
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Transcription factor NFE2L2/NRF2 modulates chaperone-mediated autophagy through the regulation of LAMP2A
2018Co-Authors: Marta Pajares, Ana I Rojo, Ana Maria Cuervo, Esperanza Arias, Antonio Díaz-carretero, Antonio CuadradoAbstract:Chaperone-mediated autophagy (CMA) is a selective degradative process for cytosolic proteins that contributes to the maintenance of proteostasis. The signaling mechanisms that control CMA are not fully understood but might involve response to stress conditions including oxidative stress. Considering the role of CMA in redoxtasis and proteostasis, we sought to determine if the transcription factor NFE2L2/NRF2 (nuclear factor, erythroid derived 2, like 2) has an impact on CMA modulation. In this work, we identified and validated 2 NFE2L2 binding sequences in the LAMP2 gene and demonstrated in several human and mouse cell types that NFE2L2 deficiency and overexpression was linked to reduced and increased LAMP2A levels, respectively. Accordingly, lysosomal LAMP2A levels were drastically reduced in NFE2L2-knockout hepatocytes, which also displayed a marked decrease in CMA activity. Oxidant challenge with paraquat or hydrogen peroxide, or pharmacological activation of NFE2L2 with sulforaphane or dimethyl fumarate also increased LAMP2A levels and CMA activity. Overall, our study identifies for the first time basal and inducible regulation of LAMP2A, and consequently CMA activity, by NFE2L2. Abbreviations: ACTB: actin, beta, ARE: antioxidant response element; ATG5: autophagy related 5; BACH1: BTB domain and CNC homolog 1; ChIP: chromatin immunoprecipitation; CMA: chaperone-mediated autophagy; DHE: dihydroethidium; DMF: dimethyl fumarate; ENCODE: Encyclopedia of DNA elements at the University of California, Santa Cruz; GAPDH: glyceraldehyde-3-phosphate dehydrogenase; GBA: glucosylceramidase beta; GFP: green fluorescent protein; HMOX1: heme oxygenase 1; H2O2: hydrogen peroxide; HSPA8/HSC70: heat shock protein family A (Hsp70) member 8; KEAP1: kelch like ECH associated protein 1; LAMP2A: lysosomal associated membrane protein 2A; LAMP2B: lysosomal associated membrane protein 2B; LAMP2C: lysosomal associated membrane protein 2C; LAMP1: lysosomal associated membrane protein 1; MAFF: MAF bZIP transcription factor F; MAFK: MAF bZIP transcription factor K; NFE2L2/NRF2: nuclear factor, erythroid derived 2, like 2; NQO1: NAD(P)H quinone dehydrogenase 1; PQ: paraquat; PI: protease inhibitors; qRT-PCR: quantitative real-time polymerase chain reaction; RNASE: ribonuclease A family member; SFN: sulforaphane; SQSTM1/p62: sequestosome 1; TBP: TATA-box binding protein
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transcription factor NFE2L2 nrf2 is a regulator of macroautophagy genes
Autophagy, 2016Co-Authors: Marta Pajares, Ana I Rojo, Natalia Jimenezmoreno, Angel Juan Garciayague, Maribel Escoll, Maria L De Ceballos, Fred Van Leuven, Alberto Rabano, Masayuki Yamamoto, Antonio CuadradoAbstract:ABSTRACTAutophagy is a highly coordinated process that is controlled at several levels including transcriptional regulation. Here, we identify the transcription factor NFE2L2/NRF2 (nuclear factor, erythroid 2 like 2) as a regulator of autophagy gene expression and its relevance in a mouse model of Alzheimer disease (AD) that reproduces impaired APP (amyloid β precursor protein) and human (Hs)MAPT/TAU processing, clearance and aggregation. We screened the chromatin immunoprecipitation database ENCODE for 2 proteins, MAFK and BACH1, that bind the NFE2L2-regulated enhancer antioxidant response element (ARE). Using a script generated from the JASPAR's consensus ARE sequence, we identified 27 putative AREs in 16 autophagy-related genes. Twelve of these sequences were validated as NFE2L2 regulated AREs in 9 autophagy genes by additional ChIP assays and quantitative RT-PCR on human and mouse cells after NFE2L2 activation with sulforaphane. Mouse embryo fibroblasts of NFE2L2-knockout mice exhibited reduced expres...
Hagir B. Suliman - One of the best experts on this subject based on the ideXlab platform.
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heme oxygenase 1 couples activation of mitochondrial biogenesis to anti inflammatory cytokine expression
Journal of Biological Chemistry, 2011Co-Authors: Claude A. Piantadosi, Karen E Weltywolf, Crystal M Withers, Raquel R Bartz, Nancy Chou Macgarvey, Ping Fu, Timothy E Sweeney, Hagir B. SulimanAbstract:Abstract The induction of heme oxygenase-1 (HO-1; Hmox1) by inflammation, for instance in sepsis, is associated both with an anti-inflammatory response and with mitochondrial biogenesis. Here, we tested the idea that HO-1, acting through the NFE2L2 (Nrf2) transcription factor, links anti-inflammatory cytokine expression to activation of mitochondrial biogenesis. HO-1 induction after lipopolysaccharide (LPS) stimulated anti-inflammatory interleukin (IL)-10 and IL-1 receptor antagonist (IL-1Ra) expression in mouse liver, human HepG2 cells, and mouse J774.1 macrophages, but blunted tumor necrosis factor-alpha expression. This was accompanied by nuclear NFE2L2 accumulation, and led us to identify abundant NFE2L2 and other mitochondrial biogenesis transcription factor binding sites in the promoter regions of IL10 and IL1Ra compared with pro-inflammatory genes regulated by NF-kB. Mechanistically, HO-1, through its CO product, enabled these transcription factors to bind the core IL10 and IL1Ra promoters, which for IL10 included NFE2L2, nuclear respiratory factor (NRF)-2 (GABPA), and MEF2, and for IL1Ra, NRF-1 and MEF2. In cells, Hmox1 or NFE2L2 RNA silencing prevented IL-10 and IL-1Ra up-regulation, and HO-1 induction failed post-LPS in NFE2L2-silenced cells and post-sepsis in NFE2L2-/- mice. NFE2L2-/- mice compared with WT mice, showed more liver damage, higher mortality, and ineffective CO rescue in sepsis. NFE2L2-/- mice in sepsis also generated higher hepatic TNF-alpha mRNA levels, lower NRF-1 and PGC-1alpha mRNA levels, and no enhancement of anti-inflammatory Il10, Socs3 or BclXL gene expression. These findings disclose a highly-structured transcriptional network that couples mitochondrial biogenesis to counter-inflammation with major implications for immune suppression in sepsis.
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heme oxygenase 1 couples activation of mitochondrial biogenesis to anti inflammatory cytokine expression
Journal of Biological Chemistry, 2011Co-Authors: Claude A. Piantadosi, Karen E Weltywolf, Crystal M Withers, Raquel R Bartz, Nancy Chou Macgarvey, Timothy E Sweeney, Hagir B. SulimanAbstract:The induction of heme oxygenase-1 (HO-1; Hmox1) by inflammation, for instance in sepsis, is associated both with an anti-inflammatory response and with mitochondrial biogenesis. Here, we tested the idea that HO-1, acting through the NFE2L2 (Nrf2) transcription factor, links anti-inflammatory cytokine expression to activation of mitochondrial biogenesis. HO-1 induction after LPS stimulated anti-inflammatory IL-10 and IL-1 receptor antagonist (IL-1Ra) expression in mouse liver, human HepG2 cells, and mouse J774.1 macrophages but blunted tumor necrosis factor-α expression. This was accompanied by nuclear NFE2L2 accumulation and led us to identify abundant NFE2L2 and other mitochondrial biogenesis transcription factor binding sites in the promoter regions of IL10 and IL1Ra compared with pro-inflammatory genes regulated by NF-κΒ. Mechanistically, HO-1, through its CO product, enabled these transcription factors to bind the core IL10 and IL1Ra promoters, which for IL10 included NFE2L2, nuclear respiratory factor (NRF)-2 (Gabpa), and MEF2, and for IL1Ra, included NRF-1 and MEF2. In cells, Hmox1 or NFE2L2 RNA silencing prevented IL-10 and IL-1Ra up-regulation, and HO-1 induction failed post-LPS in NFE2L2-silenced cells and post-sepsis in NFE2L2−/− mice. NFE2L2−/− mice compared with WT mice, showed more liver damage, higher mortality, and ineffective CO rescue in sepsis. NFE2L2−/− mice in sepsis also generated higher hepatic TNF-α mRNA levels, lower NRF-1 and PGC-1α mRNA levels, and no enhancement of anti-inflammatory Il10, Socs3, or bcl-xL gene expression. These findings disclose a highly structured transcriptional network that couples mitochondrial biogenesis to counter-inflammation with major implications for immune suppression in sepsis.
Claude A. Piantadosi - One of the best experts on this subject based on the ideXlab platform.
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regulation of mitochondrial biogenesis and its intersection with inflammatory responses
Antioxidants & Redox Signaling, 2015Co-Authors: Anne D Cherry, Claude A. PiantadosiAbstract:Abstract Significance: Mitochondria play a vital role in cellular homeostasis and are susceptible to damage from inflammatory mediators released by the host defense. Cellular recovery depends, in part, on mitochondrial quality control programs, including mitochondrial biogenesis. Recent Advances: Early-phase inflammatory mediator proteins interact with PRRs to activate NF-κB-, MAPK-, and PKB/Akt-dependent pathways, resulting in increased expression or activity of coactivators and transcription factors (e.g., PGC-1α, NRF-1, NRF-2, and NFE2L2) that regulate mitochondrial biogenesis. Inflammatory upregulation of NOS2-induced NO causes mitochondrial dysfunction, but NO is also a signaling molecule upregulating mitochondrial biogenesis via PGC-1α, participating in NFE2L2-mediated antioxidant gene expression and modulating inflammation. NO and reactive oxygen species generated by the host inflammatory response induce the redox-sensitive HO-1/CO system, causing simultaneous induction of mitochondrial biogenesis ...
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heme oxygenase 1 couples activation of mitochondrial biogenesis to anti inflammatory cytokine expression
Journal of Biological Chemistry, 2011Co-Authors: Claude A. Piantadosi, Karen E Weltywolf, Crystal M Withers, Raquel R Bartz, Nancy Chou Macgarvey, Ping Fu, Timothy E Sweeney, Hagir B. SulimanAbstract:Abstract The induction of heme oxygenase-1 (HO-1; Hmox1) by inflammation, for instance in sepsis, is associated both with an anti-inflammatory response and with mitochondrial biogenesis. Here, we tested the idea that HO-1, acting through the NFE2L2 (Nrf2) transcription factor, links anti-inflammatory cytokine expression to activation of mitochondrial biogenesis. HO-1 induction after lipopolysaccharide (LPS) stimulated anti-inflammatory interleukin (IL)-10 and IL-1 receptor antagonist (IL-1Ra) expression in mouse liver, human HepG2 cells, and mouse J774.1 macrophages, but blunted tumor necrosis factor-alpha expression. This was accompanied by nuclear NFE2L2 accumulation, and led us to identify abundant NFE2L2 and other mitochondrial biogenesis transcription factor binding sites in the promoter regions of IL10 and IL1Ra compared with pro-inflammatory genes regulated by NF-kB. Mechanistically, HO-1, through its CO product, enabled these transcription factors to bind the core IL10 and IL1Ra promoters, which for IL10 included NFE2L2, nuclear respiratory factor (NRF)-2 (GABPA), and MEF2, and for IL1Ra, NRF-1 and MEF2. In cells, Hmox1 or NFE2L2 RNA silencing prevented IL-10 and IL-1Ra up-regulation, and HO-1 induction failed post-LPS in NFE2L2-silenced cells and post-sepsis in NFE2L2-/- mice. NFE2L2-/- mice compared with WT mice, showed more liver damage, higher mortality, and ineffective CO rescue in sepsis. NFE2L2-/- mice in sepsis also generated higher hepatic TNF-alpha mRNA levels, lower NRF-1 and PGC-1alpha mRNA levels, and no enhancement of anti-inflammatory Il10, Socs3 or BclXL gene expression. These findings disclose a highly-structured transcriptional network that couples mitochondrial biogenesis to counter-inflammation with major implications for immune suppression in sepsis.
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heme oxygenase 1 couples activation of mitochondrial biogenesis to anti inflammatory cytokine expression
Journal of Biological Chemistry, 2011Co-Authors: Claude A. Piantadosi, Karen E Weltywolf, Crystal M Withers, Raquel R Bartz, Nancy Chou Macgarvey, Timothy E Sweeney, Hagir B. SulimanAbstract:The induction of heme oxygenase-1 (HO-1; Hmox1) by inflammation, for instance in sepsis, is associated both with an anti-inflammatory response and with mitochondrial biogenesis. Here, we tested the idea that HO-1, acting through the NFE2L2 (Nrf2) transcription factor, links anti-inflammatory cytokine expression to activation of mitochondrial biogenesis. HO-1 induction after LPS stimulated anti-inflammatory IL-10 and IL-1 receptor antagonist (IL-1Ra) expression in mouse liver, human HepG2 cells, and mouse J774.1 macrophages but blunted tumor necrosis factor-α expression. This was accompanied by nuclear NFE2L2 accumulation and led us to identify abundant NFE2L2 and other mitochondrial biogenesis transcription factor binding sites in the promoter regions of IL10 and IL1Ra compared with pro-inflammatory genes regulated by NF-κΒ. Mechanistically, HO-1, through its CO product, enabled these transcription factors to bind the core IL10 and IL1Ra promoters, which for IL10 included NFE2L2, nuclear respiratory factor (NRF)-2 (Gabpa), and MEF2, and for IL1Ra, included NRF-1 and MEF2. In cells, Hmox1 or NFE2L2 RNA silencing prevented IL-10 and IL-1Ra up-regulation, and HO-1 induction failed post-LPS in NFE2L2-silenced cells and post-sepsis in NFE2L2−/− mice. NFE2L2−/− mice compared with WT mice, showed more liver damage, higher mortality, and ineffective CO rescue in sepsis. NFE2L2−/− mice in sepsis also generated higher hepatic TNF-α mRNA levels, lower NRF-1 and PGC-1α mRNA levels, and no enhancement of anti-inflammatory Il10, Socs3, or bcl-xL gene expression. These findings disclose a highly structured transcriptional network that couples mitochondrial biogenesis to counter-inflammation with major implications for immune suppression in sepsis.
Marta Pajares - One of the best experts on this subject based on the ideXlab platform.
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transcription factor NFE2L2 nrf2 modulates chaperone mediated autophagy through the regulation of lamp2a
Autophagy, 2018Co-Authors: Marta Pajares, Antonio Diazcarretero, Ana I Rojo, Ana Maria Cuervo, Esperanza Arias, Antonio CuadradoAbstract:Chaperone-mediated autophagy (CMA) is a selective degradative process for cytosolic proteins that contributes to the maintenance of proteostasis. The signaling mechanisms that control CMA are not f...
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Transcription factor NFE2L2/NRF2 modulates chaperone-mediated autophagy through the regulation of LAMP2A
2018Co-Authors: Marta Pajares, Ana I Rojo, Ana Maria Cuervo, Esperanza Arias, Antonio Díaz-carretero, Antonio CuadradoAbstract:Chaperone-mediated autophagy (CMA) is a selective degradative process for cytosolic proteins that contributes to the maintenance of proteostasis. The signaling mechanisms that control CMA are not fully understood but might involve response to stress conditions including oxidative stress. Considering the role of CMA in redoxtasis and proteostasis, we sought to determine if the transcription factor NFE2L2/NRF2 (nuclear factor, erythroid derived 2, like 2) has an impact on CMA modulation. In this work, we identified and validated 2 NFE2L2 binding sequences in the LAMP2 gene and demonstrated in several human and mouse cell types that NFE2L2 deficiency and overexpression was linked to reduced and increased LAMP2A levels, respectively. Accordingly, lysosomal LAMP2A levels were drastically reduced in NFE2L2-knockout hepatocytes, which also displayed a marked decrease in CMA activity. Oxidant challenge with paraquat or hydrogen peroxide, or pharmacological activation of NFE2L2 with sulforaphane or dimethyl fumarate also increased LAMP2A levels and CMA activity. Overall, our study identifies for the first time basal and inducible regulation of LAMP2A, and consequently CMA activity, by NFE2L2. Abbreviations: ACTB: actin, beta, ARE: antioxidant response element; ATG5: autophagy related 5; BACH1: BTB domain and CNC homolog 1; ChIP: chromatin immunoprecipitation; CMA: chaperone-mediated autophagy; DHE: dihydroethidium; DMF: dimethyl fumarate; ENCODE: Encyclopedia of DNA elements at the University of California, Santa Cruz; GAPDH: glyceraldehyde-3-phosphate dehydrogenase; GBA: glucosylceramidase beta; GFP: green fluorescent protein; HMOX1: heme oxygenase 1; H2O2: hydrogen peroxide; HSPA8/HSC70: heat shock protein family A (Hsp70) member 8; KEAP1: kelch like ECH associated protein 1; LAMP2A: lysosomal associated membrane protein 2A; LAMP2B: lysosomal associated membrane protein 2B; LAMP2C: lysosomal associated membrane protein 2C; LAMP1: lysosomal associated membrane protein 1; MAFF: MAF bZIP transcription factor F; MAFK: MAF bZIP transcription factor K; NFE2L2/NRF2: nuclear factor, erythroid derived 2, like 2; NQO1: NAD(P)H quinone dehydrogenase 1; PQ: paraquat; PI: protease inhibitors; qRT-PCR: quantitative real-time polymerase chain reaction; RNASE: ribonuclease A family member; SFN: sulforaphane; SQSTM1/p62: sequestosome 1; TBP: TATA-box binding protein
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transcription factor NFE2L2 nrf2 is a regulator of macroautophagy genes
Autophagy, 2016Co-Authors: Marta Pajares, Ana I Rojo, Natalia Jimenezmoreno, Angel Juan Garciayague, Maribel Escoll, Maria L De Ceballos, Fred Van Leuven, Alberto Rabano, Masayuki Yamamoto, Antonio CuadradoAbstract:ABSTRACTAutophagy is a highly coordinated process that is controlled at several levels including transcriptional regulation. Here, we identify the transcription factor NFE2L2/NRF2 (nuclear factor, erythroid 2 like 2) as a regulator of autophagy gene expression and its relevance in a mouse model of Alzheimer disease (AD) that reproduces impaired APP (amyloid β precursor protein) and human (Hs)MAPT/TAU processing, clearance and aggregation. We screened the chromatin immunoprecipitation database ENCODE for 2 proteins, MAFK and BACH1, that bind the NFE2L2-regulated enhancer antioxidant response element (ARE). Using a script generated from the JASPAR's consensus ARE sequence, we identified 27 putative AREs in 16 autophagy-related genes. Twelve of these sequences were validated as NFE2L2 regulated AREs in 9 autophagy genes by additional ChIP assays and quantitative RT-PCR on human and mouse cells after NFE2L2 activation with sulforaphane. Mouse embryo fibroblasts of NFE2L2-knockout mice exhibited reduced expres...