cometadata/datacite-titles-descriptions-related-identifiers huggingface dataset:
DOI: 10.6084/m9.figshare.30293191.v1
title: Expression, localization and regulation of NADPH oxidases in pancreatic beta cells

description: Reactive oxygen species (ROS) are short-lived and act in a site-specific manner, underscoring the importance of identifying the subcellular localization of their sources. ROS-generating NADPH oxidases (NOX) regulate pancreatic beta cell (dys)function. However, their subcellular localization and cytokine-mediated regulation in these cells remain largely unknown. We characterized the expression, subcellular localization and time-dependent cytokine-induced regulation of NOX isoforms in beta cells. Isoforms were studied via RT-qPCR, immunoblotting and immunofluorescence in rat islets and beta cell lines. Beta cells express DUOX1 and DUOX2 proteins and <i>Duoxa2</i> transcripts; lacking <i>Duoxa1</i> expression. In INS-1E cells, NOX1 and DUOX1 localize in the endoplasmic reticulum (ER); DUOX2 in insulin vesicles; and NOX2 and NOX4 in vesicles, ER and plasma membrane. In INS-1E, cytokines increased expression of <i>Nox1</i> and <i>Duox1</i> at 4-8 h (returning to baseline at 16 h) and <i>Nox2</i> and <i>p47phox</i> at 8 h (persisting until 24 h). <i>Duox(a)2</i>, <i>p67phox</i> and <i>p40phox</i> were downregulated and DUOX1 upregulated at 16-24 h. The absence of <i>Duoxa1</i> in beta cells might lead to DUOX1 mismatching, impairing its trafficking and activity. NOXs in beta cells are diverse in subcellular localization and cytokine-induced regulation, suggesting their isoform-specific involvement in beta cell function, stress and apoptosis.
DOI: 10.6084/m9.figshare.30719727
title: The novel thioredoxin reductase inhibitor butaselen suppresses lung cancer by inducing oxidative stress

description: Lung cancer is one of the leading causes of cancer-related deaths worldwide with limited treatment options available. The anti-tumor effects of the TrxR inhibitor Butaselen (BS/BS1801) on lung cancer and its underlying mechanisms remain unknown. This study utilized lung cancer cell lines, LLC1-bearing mice models, and organoids to detect the inhibitory effects of BS on lung cancer. The ROS-induction and apoptotic role of BS on lung cancer cells and molecular mechanisms were assessed with flow cytometry, western blot, Co-IP, real-time PCR, ChIP, reporter gene assay, ELISA, and bisulfite pyrosequencing. BS can effectively inhibit lung cancer both in vitro and in vivo, by triggering ROS-induced apoptosis. The inactivation of NF-κB and MAPK signaling pathways, along with the activation of PI3K-Akt and HBP1 signaling pathways, are involved in BS's suppression of lung cancer. HBP1 is a novel downstream target of the Trx system. The activation of HBP1 by BS is dependent on ROS accumulation and further leads to the transcriptional inhibition of DNMT1 and the demethylation of the whole genome, as well as the promoters of p21 and HOXA9. The TrxR/Trx inhibitor butaselen suppresses lung cancer by triggering ROS-induced apoptosis. This study provides a novel and effective regimen for treating lung cancer.
DOI: 10.6084/m9.figshare.30719727.v1
title: The novel thioredoxin reductase inhibitor butaselen suppresses lung cancer by inducing oxidative stress

description: Lung cancer is one of the leading causes of cancer-related deaths worldwide with limited treatment options available. The anti-tumor effects of the TrxR inhibitor Butaselen (BS/BS1801) on lung cancer and its underlying mechanisms remain unknown. This study utilized lung cancer cell lines, LLC1-bearing mice models, and organoids to detect the inhibitory effects of BS on lung cancer. The ROS-induction and apoptotic role of BS on lung cancer cells and molecular mechanisms were assessed with flow cytometry, western blot, Co-IP, real-time PCR, ChIP, reporter gene assay, ELISA, and bisulfite pyrosequencing. BS can effectively inhibit lung cancer both in vitro and in vivo, by triggering ROS-induced apoptosis. The inactivation of NF-κB and MAPK signaling pathways, along with the activation of PI3K-Akt and HBP1 signaling pathways, are involved in BS's suppression of lung cancer. HBP1 is a novel downstream target of the Trx system. The activation of HBP1 by BS is dependent on ROS accumulation and further leads to the transcriptional inhibition of DNMT1 and the demethylation of the whole genome, as well as the promoters of p21 and HOXA9. The TrxR/Trx inhibitor butaselen suppresses lung cancer by triggering ROS-induced apoptosis. This study provides a novel and effective regimen for treating lung cancer.
DOI: 10.6084/m9.figshare.30238674
title: FOXO1-NMNAT3 axis dysregulation promotes doxorubicin cardiotoxicity: NAD<sup>+</sup> replenishment as a redox-targeted antioxidant therapy

description: Doxorubicin (DOX) induces dose-dependent cardiotoxicity, primarily through oxidative stress and metabolic dysregulation. Although NAD<sup>+</sup> deficiency has been implicated in cardiovascular pathology, its role in DOX-induced cardiotoxicity (DIC) remains poorly understood. This study investigated NAD<sup>+</sup> metabolism dysregulation as a redox-sensitive mechanism in DIC pathogenesis. Human cardiomyocytes (AC16), mouse atrial myocytes (HL-1), and C57BL/6 mice were used to establish the DIC model. The role and mechanism of NAD<sup>+</sup> in DIC were investigated using a range of methods. Using integrated <i>in vitro</i> and <i>in vivo</i> models, we demonstrated that DOX induces myocardial oxidative damage accompanied by NAD<sup>+</sup> depletion. Exogenous NAD<sup>+</sup> supplementation mitigated the DOX-induced cardiomyocyte death and redox imbalance. Mechanistically, pharmacological CD38 inhibition with 78C or genetic silencing failed to restore the NAD<sup>+</sup> pool, whereas nicotinamide mononucleotide adenylyltransferase 3 (NMNAT3) overexpression, combined with nicotinamide mononucleotide (NMN) administration, effectively rescued NAD<sup>+</sup> levels and attenuated oxidative stress. Computational and functional analyses identified FOXO1 as a transcriptional repressor of NMNAT3 following DOX exposure. This study establishes the dysregulation of the FOXO1-NMNAT3 axis as a key mechanism underlying NAD<sup>+</sup> depletion in DIC. Targeting this axis through NAD<sup>+</sup> replenishment, particularly by activating NMNAT3, offers a novel redox-based therapeutic strategy against DIC.
DOI: 10.6084/m9.figshare.30238674.v1
title: FOXO1-NMNAT3 axis dysregulation promotes doxorubicin cardiotoxicity: NAD<sup>+</sup> replenishment as a redox-targeted antioxidant therapy

description: Doxorubicin (DOX) induces dose-dependent cardiotoxicity, primarily through oxidative stress and metabolic dysregulation. Although NAD<sup>+</sup> deficiency has been implicated in cardiovascular pathology, its role in DOX-induced cardiotoxicity (DIC) remains poorly understood. This study investigated NAD<sup>+</sup> metabolism dysregulation as a redox-sensitive mechanism in DIC pathogenesis. Human cardiomyocytes (AC16), mouse atrial myocytes (HL-1), and C57BL/6 mice were used to establish the DIC model. The role and mechanism of NAD<sup>+</sup> in DIC were investigated using a range of methods. Using integrated <i>in vitro</i> and <i>in vivo</i> models, we demonstrated that DOX induces myocardial oxidative damage accompanied by NAD<sup>+</sup> depletion. Exogenous NAD<sup>+</sup> supplementation mitigated the DOX-induced cardiomyocyte death and redox imbalance. Mechanistically, pharmacological CD38 inhibition with 78C or genetic silencing failed to restore the NAD<sup>+</sup> pool, whereas nicotinamide mononucleotide adenylyltransferase 3 (NMNAT3) overexpression, combined with nicotinamide mononucleotide (NMN) administration, effectively rescued NAD<sup>+</sup> levels and attenuated oxidative stress. Computational and functional analyses identified FOXO1 as a transcriptional repressor of NMNAT3 following DOX exposure. This study establishes the dysregulation of the FOXO1-NMNAT3 axis as a key mechanism underlying NAD<sup>+</sup> depletion in DIC. Targeting this axis through NAD<sup>+</sup> replenishment, particularly by activating NMNAT3, offers a novel redox-based therapeutic strategy against DIC.
DOI: 10.6084/m9.figshare.29615153
title: ROS-Drp1-mitophagy feedback loop regulates myogenic differentiation via actin cytoskeleton remodeling-mediated MRTF-A/SRF axis

description: Mitochondrial division is one of the main characteristics for the initiation of myogenic differentiation. However, the role and mechanism of Dynamin-related protein 1 (Drp1), the most important protein that regulates mitochondrial fission in mammals, in regulating myogenic differentiation are not well understood. Drp1 siRNAs were transfected to C2C12 cells, or AAV9-shDrp1 were injected to C57BL/6J mice to knockdown Drp1 expression. Then, mitochondrial damage, ROS level, myogenic differentiation, mitophagy and actin/MRTF-A/SRF pathway was detected by quantitative real-time PCR, western blotting, immunofluorescence staining and flow cytometry. The results showed that Drp1 was upregulated after C2C12 differentiation; Drp1 knockdown by siRNA transfection impaired myotube formation. ROS are the upstream activators for Drp1 expression, and Drp1 inversely reduces ROS by facilitating mitophagy to form a ROS-Drp1-mitophagy feedback loop during myogenic differentiation. Knockdown of Drp1 disrupted the ROS-Drp1-mitophagy feedback loop-mediated ROS homeostasis, thereby accelerating F-action depolymerization and blocking MRTF-A nuclear translocation by reducing the phosphorylation of cofilin. A decrease in MRTF-A nuclear translocation impaired SRF activity and hindered myogenic differentiation. In summary, this study revealed the functional mechanism of Drp1 and clarified the interactions among ROS, Drp1-mediated mitophagy and actin cytoskeleton remodeling during myogenic differentiation.
DOI: 10.6084/m9.figshare.29615153.v1
title: ROS-Drp1-mitophagy feedback loop regulates myogenic differentiation via actin cytoskeleton remodeling-mediated MRTF-A/SRF axis

description: Mitochondrial division is one of the main characteristics for the initiation of myogenic differentiation. However, the role and mechanism of Dynamin-related protein 1 (Drp1), the most important protein that regulates mitochondrial fission in mammals, in regulating myogenic differentiation are not well understood. Drp1 siRNAs were transfected to C2C12 cells, or AAV9-shDrp1 were injected to C57BL/6J mice to knockdown Drp1 expression. Then, mitochondrial damage, ROS level, myogenic differentiation, mitophagy and actin/MRTF-A/SRF pathway was detected by quantitative real-time PCR, western blotting, immunofluorescence staining and flow cytometry. The results showed that Drp1 was upregulated after C2C12 differentiation; Drp1 knockdown by siRNA transfection impaired myotube formation. ROS are the upstream activators for Drp1 expression, and Drp1 inversely reduces ROS by facilitating mitophagy to form a ROS-Drp1-mitophagy feedback loop during myogenic differentiation. Knockdown of Drp1 disrupted the ROS-Drp1-mitophagy feedback loop-mediated ROS homeostasis, thereby accelerating F-action depolymerization and blocking MRTF-A nuclear translocation by reducing the phosphorylation of cofilin. A decrease in MRTF-A nuclear translocation impaired SRF activity and hindered myogenic differentiation. In summary, this study revealed the functional mechanism of Drp1 and clarified the interactions among ROS, Drp1-mediated mitophagy and actin cytoskeleton remodeling during myogenic differentiation.
DOI: 10.6084/m9.figshare.30163163
title: Manganese (III) tetrakis (4-benzoic acid) porphyrin (MnTBAP) represses sulfide:quinone oxidoreductase expression and targets the sulfido-redox system in glioblastoma models

description: The adaptation of the redox system and bioenergetics is a major factor contributing to cancer metabolism. Redox therapy is promising but still requires molecular studies that consider the reactive species interactome (RSI) concept, which integrates reactive oxygen, nitrogen, sulfur, carbonyl species, and redox enzymes. Our aim was to decipher the role of the RSI in glioblastoma (GBM), including by challenging the RSI with the MnTBAP redox agent. The effects of MnTBAP on the redox system and bioenergetics were investigated on several GBM models, namely <i>in vitro</i> 2D culture, <i>in vitro</i> 3D culture with two human GBM tumoroids, and <i>in vivo</i> preclinical model, which included male and female comparisons. We show – for the first time – that MnTBAP represses the sulfide:quinone oxidoreductase (SQOR) involved in the sulfur metabolism and bioenergetics, and targets the RSI through the sulfido-redox system. Through <i>in vitro</i> silencing and overexpression approaches, we also demonstrate that SQOR contributed to GBM cell growth and that its decrease is involved in the molecular effect of MnTBAP. Consequently, MnTBAP induces a switch between apoptosis, uncontrolled necrosis, and ferroptosis depending on the glioblastoma models. Our findings represent the next step in establishing a better understanding of redox biology in the context of GBM.
DOI: 10.6084/m9.figshare.30163163.v1
title: Manganese (III) tetrakis (4-benzoic acid) porphyrin (MnTBAP) represses sulfide:quinone oxidoreductase expression and targets the sulfido-redox system in glioblastoma models

description: The adaptation of the redox system and bioenergetics is a major factor contributing to cancer metabolism. Redox therapy is promising but still requires molecular studies that consider the reactive species interactome (RSI) concept, which integrates reactive oxygen, nitrogen, sulfur, carbonyl species, and redox enzymes. Our aim was to decipher the role of the RSI in glioblastoma (GBM), including by challenging the RSI with the MnTBAP redox agent. The effects of MnTBAP on the redox system and bioenergetics were investigated on several GBM models, namely <i>in vitro</i> 2D culture, <i>in vitro</i> 3D culture with two human GBM tumoroids, and <i>in vivo</i> preclinical model, which included male and female comparisons. We show – for the first time – that MnTBAP represses the sulfide:quinone oxidoreductase (SQOR) involved in the sulfur metabolism and bioenergetics, and targets the RSI through the sulfido-redox system. Through <i>in vitro</i> silencing and overexpression approaches, we also demonstrate that SQOR contributed to GBM cell growth and that its decrease is involved in the molecular effect of MnTBAP. Consequently, MnTBAP induces a switch between apoptosis, uncontrolled necrosis, and ferroptosis depending on the glioblastoma models. Our findings represent the next step in establishing a better understanding of redox biology in the context of GBM.
DOI: 10.6084/m9.figshare.30676599
title: Ecm19 coordinates mitochondrial fission and cellular redox homeostasis

description: Mitochondrial dynamics are tightly coupled with cellular redox homeostasis; however, the underlying regulatory mechanisms remain poorly defined. We constructed yeast mutants and evaluated mitochondrial function, morphology, and redox states using growth assays, fluorescence imaging, and flow cytometry. RNA sequencing, RIP assays, and RT-qPCR were applied to identify Ecm19p target genes. Deletion of <i>ECM19</i> alone had no evident impact on mitochondrial morphology or respiratory function. In contrast, double deletion of <i>ECM19</i> with the fusion gene <i>FZO1</i> (<i>ecm19D fzo1D</i>) rescued mitochondrial function and morphology and reduced ROS and malondialdehyde levels relative to <i>fzo1D</i>. Conversely, combining <i>ecm19D</i> with fission genes <i>MDV1</i> or <i>CAF4</i> resulted in hyperfused mitochondria, dependent on the division factor Dnm1p. RNA-seq revealed that <i>ecm19D</i> upregulates redox processes, including catalase (<i>CTA1</i>) and thiol peroxidase (<i>TSA2</i>). RIP-PCR confirmed Ecm19p binds directly to <i>CTA1</i> and <i>TSA2</i> transcripts and reduces their mRNA stability. Under H₂O₂ stress, <i>ecm19D cta1D</i> and <i>ecm19D tsa2D</i> double mutants exhibited improved growth, elevated antioxidant capacity, and lower ROS and malondialdehyde than single mutants. Ecm19 collaborates with Mdv1 and Caf4 to promote mitochondrial fission and post-transcriptionally represses <i>CTA1</i> and <i>TSA2</i> expression to regulate cellular redox, thereby coordinating mitochondrial fission with redox homeostasis.
DOI: 10.6084/m9.figshare.30676599.v1
title: Ecm19 coordinates mitochondrial fission and cellular redox homeostasis

description: Mitochondrial dynamics are tightly coupled with cellular redox homeostasis; however, the underlying regulatory mechanisms remain poorly defined. We constructed yeast mutants and evaluated mitochondrial function, morphology, and redox states using growth assays, fluorescence imaging, and flow cytometry. RNA sequencing, RIP assays, and RT-qPCR were applied to identify Ecm19p target genes. Deletion of <i>ECM19</i> alone had no evident impact on mitochondrial morphology or respiratory function. In contrast, double deletion of <i>ECM19</i> with the fusion gene <i>FZO1</i> (<i>ecm19D fzo1D</i>) rescued mitochondrial function and morphology and reduced ROS and malondialdehyde levels relative to <i>fzo1D</i>. Conversely, combining <i>ecm19D</i> with fission genes <i>MDV1</i> or <i>CAF4</i> resulted in hyperfused mitochondria, dependent on the division factor Dnm1p. RNA-seq revealed that <i>ecm19D</i> upregulates redox processes, including catalase (<i>CTA1</i>) and thiol peroxidase (<i>TSA2</i>). RIP-PCR confirmed Ecm19p binds directly to <i>CTA1</i> and <i>TSA2</i> transcripts and reduces their mRNA stability. Under H₂O₂ stress, <i>ecm19D cta1D</i> and <i>ecm19D tsa2D</i> double mutants exhibited improved growth, elevated antioxidant capacity, and lower ROS and malondialdehyde than single mutants. Ecm19 collaborates with Mdv1 and Caf4 to promote mitochondrial fission and post-transcriptionally represses <i>CTA1</i> and <i>TSA2</i> expression to regulate cellular redox, thereby coordinating mitochondrial fission with redox homeostasis.
DOI: 10.6084/m9.figshare.29552962
title: Sesamin protects against Acetaminophen-induced nephrotoxicity by suppressing HMOX1-mediated apoptosis and ferroptosis

description: Acetaminophen (APAP) is a widely used antipyretic and analgesic agent, and acute exposure can lead to renal injury. Sesamin (Ses) is known for its various health benefits. However, it remains unclear whether Ses exerts a protective effect against APAP-induced kidney injury. In vivo, C57BL/6 mice were pretreated with Ses and injected intraperitoneally with APAP. In vitro, human kidney proximal tubule cells 2 were pretreated with Ses, and then models of kidney injury induced by APAP were established. Kidney damage was evaluated by morphological, inflammation, oxidative stress and protein analyzes. Ses significantly improved APAP-induced nephrotoxicity in vitro and in vivo models. Transcriptomic analysis revealed that the differentially expressed genes were enriched in ferroptosis and apoptosis signaling pathways, identifying heme oxygenase 1 (HMOX1) as a core protein. In the Ses-treated group, ferroptosis and apoptosis were significantly inhibited, while HMOX1 was effectively restored. In cell experiments, both the HMOX1 agonist hemin and Ses attenuated ferroptosis and apoptosis. HMOX1 inhibitor Zinc Protoporphyrin significantly eliminated the protective effect of Ses. Ses alleviates APAP-induced renal injury by mediating the inhibition of ferroptosis and apoptosis via HMOX1. This study provides a new strategy for the prevention and treatment of drug-induced renal injury.
DOI: 10.6084/m9.figshare.29552962.v1
title: Sesamin protects against Acetaminophen-induced nephrotoxicity by suppressing HMOX1-mediated apoptosis and ferroptosis

description: Acetaminophen (APAP) is a widely used antipyretic and analgesic agent, and acute exposure can lead to renal injury. Sesamin (Ses) is known for its various health benefits. However, it remains unclear whether Ses exerts a protective effect against APAP-induced kidney injury. In vivo, C57BL/6 mice were pretreated with Ses and injected intraperitoneally with APAP. In vitro, human kidney proximal tubule cells 2 were pretreated with Ses, and then models of kidney injury induced by APAP were established. Kidney damage was evaluated by morphological, inflammation, oxidative stress and protein analyzes. Ses significantly improved APAP-induced nephrotoxicity in vitro and in vivo models. Transcriptomic analysis revealed that the differentially expressed genes were enriched in ferroptosis and apoptosis signaling pathways, identifying heme oxygenase 1 (HMOX1) as a core protein. In the Ses-treated group, ferroptosis and apoptosis were significantly inhibited, while HMOX1 was effectively restored. In cell experiments, both the HMOX1 agonist hemin and Ses attenuated ferroptosis and apoptosis. HMOX1 inhibitor Zinc Protoporphyrin significantly eliminated the protective effect of Ses. Ses alleviates APAP-induced renal injury by mediating the inhibition of ferroptosis and apoptosis via HMOX1. This study provides a new strategy for the prevention and treatment of drug-induced renal injury.
DOI: 10.6084/m9.figshare.29623565
title: Neutrophil extracellular traps drive osteoporosis via NCF2-dependent signaling: integrated transcriptomics with mechanistic validation

description: Inflammation and immune responses play key roles in osteoporosis (OP) pathogenesis. Osteoimmunology highlights immune dysregulation as a significant contributor to OP, but the specific biological mechanisms linking immune dysfunction to bone loss remain unclear. Understanding these mechanisms is essential for targeted therapies. We established a rat OP model via bilateral ovariectomy. Transcriptomic sequencing (RNA-seq) identified differentially expressed genes (DEGs), and summary data-based Mendelian randomization (SMR) analysis validated their causal associations with OP. Primary neutrophils isolated from bone marrow and differentiated HL-60 neutrophil-like cells were induced to form neutrophil extracellular traps (NETs), and siRNA was employed to knock down the NCF2 gene. Conditioned media from these neutrophils were subsequently applied to primary osteoblasts to evaluate effects on osteogenic differentiation. RNA-seq identified 4,497 DEGs (1,606 upregulated, 2,891 downregulated) in OP rats, significantly enriched in immune response and NETs formation pathways. NETs markers (NE, MPO, CitH3) were markedly elevated in OP bone tissue and stimulated neutrophils. SMR analysis identified VDAC1, PLCG2, and NCF2 as key genes significantly associated with OP risk, experimentally validated at the tissue and cellular levels. Knockdown of NCF2 reduced NETs formation in neutrophil-like cells and alleviated NETs-induced osteoblast differentiation impairment. Drug prediction and molecular docking analyses demonstrated high affinity and pharmacological potential targeting these genes. This study unveils the link between NETs formation and OP, highlighting NCF2 as crucial players. These findings provide new insights into immune inflammation’s role in bone metabolism and pave the way for targeted OP therapies.