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NFE2L2

NFE2L2, also known as NRF2 (nuclear factor erythroid 2-related factor 2), is a protein-coding gene located on human chromosome 2q31.2 that encodes a basic leucine zipper (bZIP) transcription factor essential for regulating cellular responses to oxidative stress and environmental toxins.^[1] This gene produces multiple isoforms and is ubiquitously expressed, with particularly high levels in tissues like the esophagus and thyroid.^[1] The NRF2 protein, the primary product of NFE2L2, functions by binding to antioxidant response elements (AREs) in the promoter regions of target genes, thereby activating the transcription of enzymes and proteins involved in antioxidant defense, detoxification, and inflammation modulation.^[1]^[2] Under normal conditions, NRF2 is sequestered in the cytoplasm by its inhibitor KEAP1 and targeted for proteasomal degradation, but stressors such as reactive oxygen species (ROS) or electrophiles disrupt this interaction, allowing NRF2 to translocate to the nucleus and induce protective gene expression.^[3] This mechanism forms the core of the NRF2/ARE pathway, which coordinates responses to cellular insults including free radical production and xenobiotic exposure.^[1]^[4] Beyond its protective roles, dysregulation of NFE2L2 has been implicated in various pathologies; for instance, hyperactivation of NRF2 can promote cancer progression by enhancing tumor cell survival and resistance to therapies,^[5] while deficiencies contribute to neurodegenerative diseases, chronic inflammation, and accelerated aging due to unchecked oxidative damage.^[6] Research continues to explore NFE2L2 as a therapeutic target, with modulators showing promise in conditions like pulmonary fibrosis,^[7] cardiovascular disease,^[8] and ferroptosis-related disorders.^[5]

Gene Characteristics

Genomic Location and Organization

The NFE2L2 gene is located on the long arm of human chromosome 2 at cytogenetic band 2q31.2, spanning genomic coordinates 177,230,303 to 177,264,727 (approximately 34 kb) on the reverse strand in the GRCh38.p14 assembly.^[9] The orthologous Nfe2l2 gene in mice resides on chromosome 2, from positions 75,505,857 to 75,534,985 (approximately 29 kb) on the reverse strand in the GRCm39 assembly. The gene comprises 5 exons separated by 4 introns, with the first intron exceeding 6 kb in length, contributing to the overall genomic span.^[9] Its promoter region features two ARE-like sequences that enable autoregulation by the encoded NRF2 protein, along with binding sites for transcription factors including NF-κB, which facilitate induction by inflammatory signals, and AP-1 family members responsive to stress stimuli.^[4]^[10] NFE2L2 exhibits high evolutionary conservation across vertebrates, with orthologs identified in 212 species, underscoring its fundamental role in cellular stress responses.^[11] The human and mouse proteins share 88.4% amino acid sequence identity, reflecting strong selective pressure on functional domains.^[12] A notable genetic variant is the promoter polymorphism rs6721961 (C>A), which influences basal NFE2L2 expression levels and has been linked to variability in antioxidant responses.^[13]

Expression Patterns

NFE2L2 exhibits ubiquitous basal expression across human tissues, with the highest levels observed in the skeletal muscle, kidney, lung, and liver.^[9] According to data from the Genotype-Tissue Expression (GTEx) project, median transcripts per million (TPM) values reflect this pattern, with kidney cortex showing approximately 20 TPM, skeletal muscle around 15-18 TPM, and lung about 25 TPM, compared to lower values such as ~5-7 TPM in spleen and testis.^[14] During development, NFE2L2 shows high expression in fetal muscle and liver.^[9] In adult tissues, expression remains relatively stable under basal conditions, supporting constitutive antioxidant maintenance without significant fluctuations.^[15] The expression of NFE2L2 is modulated by external cues, including circadian rhythms, where clock proteins like BMAL1 and CLOCK regulate its rhythmic activation to align antioxidant responses with daily redox cycles.^[16] Hormones such as glucocorticoids also influence NFE2L2 levels, often through glucocorticoid receptor-mediated repression that fine-tunes its transcriptional output in response to stress.^[17] Multiple splice variants of NFE2L2 exist, with at least eight transcripts identified, but the canonical isoform encoding the full-length 605-amino acid protein predominates and is responsible for the primary transcriptional activity.^[11]

Protein Features

Domain Architecture

The NRF2 protein, encoded by the NFE2L2 gene, consists of 605 amino acids with a calculated molecular mass of approximately 68 kDa. It belongs to the cap’n’collar (CNC) subfamily of basic leucine zipper (bZIP) transcription factors and is characterized by seven conserved Nrf2-ECH homology (Neh) domains (Neh1–Neh7), which collectively define its functional architecture. Much of the protein, particularly regions outside the structured bZIP domain, is intrinsically disordered, conferring flexibility that facilitates interactions with binding partners and regulatory modifications.^[18]^[19]^[20] The Neh1 domain, located at the C-terminus, encompasses the CNC-bZIP region responsible for DNA binding to antioxidant response elements (AREs) and dimerization with small Maf proteins via a leucine zipper motif. The Neh2 domain at the N-terminus serves as a redox-sensitive degron, featuring two Keap1-binding motifs: a low-affinity DLG motif (residues 29–31) and a high-affinity ETGE motif (residues 79–82), which enable high-specificity recognition by the Keap1 adaptor protein. Neh3 functions as a transactivation domain, while Neh4 and Neh5, adjacent serine-rich regions, also contribute to transcriptional activation by recruiting co-activators such as CBP and RAC3. The Neh6 domain acts as a redox-insensitive degron, containing phosphorylation sites that promote βTrCP-mediated degradation, and Neh7 represses nuclear receptor activity by binding RXRα.^[21]^[19]^[21] Structurally, the Neh2 domain is highly unstructured and intrinsically disordered, as revealed by nuclear magnetic resonance (NMR) spectroscopy, with transient helical elements forming upon Keap1 binding; the full-length NRF2 lacks a complete crystal structure but AlphaFold models predict extensive flexibility, particularly in the Neh regions, underscoring its adaptability in stress responses. The bZIP domain's leucine zipper enables stable heterodimerization with Maf proteins for DNA binding. Post-translational modifications critically influence domain function: Neh2 contains seven lysine residues targeted for ubiquitination by the Cul3-based E3 ligase complex, facilitating proteasomal degradation, while phosphorylation at Ser40 within Neh1 by protein kinase C (PKC) disrupts inhibitory interactions.^[22]^[20]^[23]

Subcellular Localization and Activation

Under homeostatic conditions, NFE2L2 (NRF2) is primarily localized in the cytoplasm, where it forms a complex with Kelch-like ECH-associated protein 1 (KEAP1) and Cullin 3 (Cul3)-based ubiquitin ligase, leading to its ubiquitination and proteasomal degradation, thereby maintaining low NRF2 protein levels.^[24] This basal state ensures that NRF2 has a short half-life of approximately 10-15 minutes.^[25] Activation of NRF2 occurs in response to oxidative stress or electrophilic compounds, which covalently modify reactive cysteine residues on KEAP1, such as Cys151, thereby disrupting the KEAP1-NRF2 interaction and preventing ubiquitination.^[24] Additionally, phosphorylation of NRF2 at serine 40 (Ser40) by protein kinase C (PKC) promotes its dissociation from KEAP1 and facilitates nuclear import through interaction with importin α/β complexes.^[26] These modifications stabilize NRF2, extending its half-life to over 2 hours, and enable rapid nuclear translocation within 15-30 minutes via a CRM1-independent pathway mediated by nuclear localization signals in the Neh1 domain.^[27] In the nucleus, stabilized NRF2 heterodimerizes with small Maf (sMaf) proteins, such as MAFF, MAFG, or MAFK, to form a transcription factor complex that binds to antioxidant response element (ARE) or electrophile response element (EPRE) sequences, characterized by the consensus motif 5'-TGACnnnGCA-3'.^[24] This binding initiates the transcriptional activation of target genes involved in cytoprotective responses.^[25]

Biological Function

Regulatory Mechanisms

The regulation of NFE2L2, encoding the transcription factor NRF2, occurs primarily at the levels of transcription, translation, and post-translational modification, ensuring tight control over cellular responses to oxidative stress. In the canonical pathway, Kelch-like ECH-associated protein 1 (KEAP1) acts as a negative regulator by binding NRF2 through its Neh2 domain motifs (ETGE and DLG), facilitating ubiquitination via the Cullin 3 (CUL3)-RING ubiquitin ligase complex and subsequent proteasomal degradation under homeostatic conditions.^[28] Upon exposure to reactive oxygen species (ROS) or electrophiles, modification of over 20 cysteine residues in KEAP1—particularly the highly reactive C151, C273, and C288—induces conformational changes that disrupt the KEAP1-NRF2 interaction, stabilizing NRF2 and allowing its nuclear translocation to drive antioxidant response element (ARE)-mediated transcription.^[29]^[28] Non-canonical pathways provide additional layers of control independent of KEAP1. Under conditions of sustained high NRF2 levels, glycogen synthase kinase-3β (GSK-3β) phosphorylates serine residues (S344 and S347) in the Neh6 degron domain of NRF2, recruiting the β-TrCP-CUL1 ubiquitin ligase for proteasomal degradation and preventing excessive NRF2 accumulation. Epigenetic modifications, such as histone acetylation at the NFE2L2 promoter facilitated by the coactivator p300, enhance transcriptional activation by opening chromatin structure and promoting RNA polymerase II recruitment. Conversely, microRNAs like miR-28 suppress NFE2L2 expression by binding its 3' untranslated region (UTR), leading to mRNA degradation and reduced NRF2 protein levels.^[30] Feedback loops and inter-pathway cross-talk further refine NRF2 activity. NRF2 auto-regulates its own expression through binding to ARE sequences in the NFE2L2 promoter, creating a positive feedback mechanism that amplifies the oxidative stress response.^[31] Additionally, NRF2 engages in cross-talk with p53, where p53 can transcriptionally repress NFE2L2 under genotoxic stress, and with NF-κB, where NRF2 activation often antagonizes NF-κB-driven inflammation to maintain redox balance.^[32]^[33] Recent advances from 2023 to 2025 highlight emerging regulatory nuances, particularly in pathological contexts. Epigenetic silencing of NFE2L2 in cancers via histone deacetylases (HDACs), such as class I HDACs, promotes tumor progression by repressing NRF2-mediated antioxidant defenses, with HDAC inhibitors showing potential to reverse this silencing and restore NRF2 activity.^[34] Post-translational SUMOylation of NRF2, mediated by SUMO enzymes at specific lysine residues, enhances its protein stability and transcriptional potency by inhibiting ubiquitination.^[34] Furthermore, 2025 studies have elucidated cysteine-mediated activation mechanisms, including direct cysteinylation of KEAP1 cysteines by exogenous cysteine uptake, which stabilizes NRF2 in cancer cells as a survival adaptation to oxidative stress.^[35]

Target Genes and Pathways

NRF2, encoded by NFE2L2, transcriptionally activates a diverse set of target genes primarily through binding to antioxidant response elements (AREs), which are cis-regulatory sequences typically consisting of a core motif (RTGACnnnGCRC) in gene promoters or enhancers. These targets form a cytoprotective network that mitigates oxidative and electrophilic stresses by enhancing antioxidant defenses, detoxification, and cellular homeostasis. Seminal studies have established that NRF2 heterodimerizes with small Maf proteins to bind AREs, driving the expression of over 200-500 genes genome-wide, as revealed by chromatin immunoprecipitation followed by sequencing (ChIP-seq) analyses in various cell types. For instance, ChIP-seq in human lung adenocarcinoma A549 cells identified 2,051 NRF2-bound regions encompassing 2,395 sites, with significant enrichment for ARE motifs near genes involved in redox balance.^[36]^[37] Core antioxidant targets of NRF2 include NAD(P)H quinone dehydrogenase 1 (NQO1), which reduces quinones to prevent reactive oxygen species (ROS) formation; superoxide dismutase 2 (SOD2), a mitochondrial enzyme that converts superoxide to hydrogen peroxide; and catalase (CAT), which decomposes hydrogen peroxide into water and oxygen. These genes are inducibly upregulated by NRF2 under oxidative stress, contributing to ROS scavenging and cellular protection. NRF2 also regulates glutathione-related genes essential for maintaining redox buffering capacity, such as the catalytic subunit of glutamate-cysteine ligase (GCLC) and its modifier subunit (GCLM), which catalyze the rate-limiting step in glutathione synthesis, as well as glutathione reductase (GSR), which regenerates reduced glutathione from its oxidized form. Additionally, in heme metabolism, NRF2 induces heme oxygenase 1 (HMOX1), which catabolizes heme into biliverdin, carbon monoxide, and iron, thereby exerting antioxidant and anti-inflammatory effects.^[38]^[39]^[40] Beyond antioxidants, NRF2 governs broader pathways in detoxification and efflux. In phase II detoxification, it upregulates glutathione S-transferases (GSTs), such as GSTA1 and GSTM1, which conjugate electrophiles with glutathione for neutralization, and UDP-glucuronosyltransferase 1A (UGT1A) family members, which facilitate glucuronidation of xenobiotics for excretion. NRF2 further activates phase III transporters, including multidrug resistance-associated proteins (MRPs) like ABCC1 (MRP1) through ABCC5 (MRP5), and specifically ABCC2 (MRP2), which efflux conjugated toxins across cellular membranes to prevent intracellular accumulation. These regulatory actions enhance the cellular capacity to handle environmental toxins and chemotherapeutic agents.^[41]^[42]^[43] NRF2 integrates with other stress response pathways; for instance, during endoplasmic reticulum (ER) stress, the unfolded protein response (UPR) activates ATF4, which transcriptionally induces NRF2 to coordinate antioxidant defenses and mitigate ROS production associated with protein misfolding. Conversely, NRF2 suppresses inflammatory signaling by inhibiting NLRP3 inflammasome activation, primarily through upregulation of thioredoxin 1 (TXN) to prevent TXN-interacting protein (TXNIP) dissociation and subsequent NLRP3 oligomerization. Quantitative aspects of NRF2-ARE interactions reveal that binding affinity varies with ARE sequence variants; for example, deviations from the optimal MARE (Maf recognition element) consensus reduce affinity by up to 10-fold, influencing target gene selectivity across cellular contexts. Recent 2024 investigations have further highlighted glutathione peroxidase 4 (GPX4) as a direct NRF2 target, where NRF2-driven GPX4 expression suppresses lipid peroxidation and ferroptosis, a regulated cell death pathway implicated in stress adaptation.^[44]^[45]^[46]^[47]

Physiological Roles

Oxidative Stress Response

NRF2, encoded by the NFE2L2 gene, serves as a master regulator of the cellular oxidative stress response by translocating to the nucleus upon activation and binding to antioxidant response elements (AREs) in target gene promoters. This activation upregulates a network of genes encoding ROS scavengers, such as glutathione peroxidases (GPX, particularly GPX2) and peroxiredoxins (PRDX, including PRDX1), which directly neutralize reactive oxygen species (ROS) like hydrogen peroxide (H₂O₂) to prevent oxidative damage.^[48] Additionally, NRF2 induces expression of regenerators like thioredoxin reductase 1 (TXNRD1), which reduces oxidized thioredoxin to recycle and sustain the activity of PRDX and other antioxidants, forming a feedback loop that reactivates the NRF2 repressor KEAP1.^[48] Through these mechanisms, NRF2 maintains the glutathione (GSH)/glutathione disulfide (GSSG) ratio, a critical redox buffer, by enhancing GSH synthesis via upregulation of glutamate-cysteine ligase subunits (GCLC and GCLM), ensuring cellular redox homeostasis under oxidative challenge.^[48] At the cellular level, NRF2 activation confers cytoprotection by inhibiting apoptosis and senescence induced by ROS accumulation. By bolstering antioxidant defenses, NRF2 attenuates hydrogen peroxide-mediated apoptotic pathways, such as those involving GSK3β inactivation via p38 signaling, thereby preserving cell viability.^[49] Similarly, NRF2 reduces senescence in aging models by mitigating oxidative stress and inflammation, preventing the onset of replicative or stress-induced cellular arrest.^[50] Furthermore, NRF2 promotes mitochondrial biogenesis through crosstalk with peroxisome proliferator-activated receptor gamma coactivator 1-alpha (PGC-1α), activating a PGC-1α/p38/GSK3β/NRF2 cascade that upregulates PGC-1α expression and downstream mitochondrial genes, enhancing organelle renewal and energy production to counteract oxidative insults.^[49]^[51] In vivo studies underscore NRF2's essential role in systemic defense against oxidants. Nrf2 knockout mice exhibit heightened sensitivity to oxidative stressors, displaying exacerbated lung injury, inflammation, and mortality upon exposure to hyperoxia compared to wild-type counterparts, as evidenced by increased histological damage and impaired resolution of acute injury. This susceptibility highlights NRF2's contribution to pulmonary protection by coordinating antioxidant responses in alveolar cells. Recent investigations (2023–2025) have revealed emerging facets of NRF2's oxidative stress response, including its interplay with hypoxia-inducible factors (HIFs) under hypoxic conditions. NRF2 and HIF signaling pathways dynamically interact to modulate redox balance and adaptation to hypoxia, where NRF2 activation can stabilize HIF-1α and enhance cytoprotection in oxygen-deprived environments. In neurodegeneration, NRF2 promotes neuroprotection by inducing mitophagy, the selective autophagy of damaged mitochondria, through pathways involving PINK1/Parkin, thereby alleviating oxidative burden and reversing degeneration phenotypes in models of Parkinson's disease.^[52]

Detoxification and Metabolism

NRF2 plays a pivotal role in phase II detoxification by transcriptionally activating enzymes that conjugate xenobiotics and endogenous toxins, rendering them more water-soluble for excretion. It induces the expression of glutathione S-transferases (GSTs), such as GSTA1 and GSTM1, which catalyze the nucleophilic addition of glutathione to electrophilic compounds, including drugs and environmental toxins, thereby neutralizing their reactivity.^[53] Similarly, NRF2 upregulates UDP-glucuronosyltransferases (UGTs), exemplified by UGT1A1 and UGT1A6, which facilitate glucuronidation—a process that conjugates glucuronic acid to hydrophobic substrates like bilirubin and phenolic compounds—to enhance their renal and biliary elimination.^[54] These actions occur through antioxidant response elements (AREs) in the promoter regions of GST and UGT genes, ensuring coordinated detoxification in response to oxidative or xenobiotic stress.^[55] Beyond conjugation, NRF2 regulates membrane transporters critical for the influx and efflux of detoxification substrates. It transcriptionally activates ATP-binding cassette (ABC) transporters, particularly ABCC2 (also known as MRP2), which actively pumps glutathione- and glucuronide-conjugated toxins from hepatocytes and other cells into bile or extracellular space, preventing intracellular accumulation.^[42] This efflux mechanism is essential for clearing phase II metabolites and contributes to the overall biotransformation pathway. While direct regulation of solute carrier (SLC) transporters for substrate import is less extensively characterized, NRF2 influences nutrient uptake systems that support detoxification enzyme activity, such as those providing glutathione precursors. NRF2 integrates detoxification with broader metabolic processes, including the prevention of lipid peroxidation through induction of aldehyde dehydrogenases (ALDHs), like ALDH2, which metabolize reactive aldehydes generated from lipid oxidation into less toxic acids.^[56] This protective role extends to drug resistance, where hyperactive NRF2 in cancer cells enhances the expression of GSTs, UGTs, and ABC transporters, reducing the efficacy of chemotherapeutic agents like cisplatin and doxorubicin by accelerating their conjugation and export.^[57] Recent studies as of 2025 underscore NRF2's involvement in ferroptosis resistance in colorectal cancer by modulating lipid metabolism genes, thereby suppressing iron-dependent lipid peroxidation and promoting tumor survival under therapeutic stress.^[58]

Health and Disease

Disease Associations

Dysregulation of NFE2L2, encoding the transcription factor NRF2, has been implicated in various pathologies, with gain-of-function alterations often promoting disease progression in cancer while loss-of-function contributes to oxidative damage in neurodegenerative and metabolic disorders. In cancer, activating mutations in NFE2L2 or its negative regulator KEAP1 lead to constitutive NRF2 activation, enhancing tumor cell survival and resistance to therapies. For instance, KEAP1 loss-of-function mutations are frequent in lung squamous cell carcinoma (LUSC) and lung adenocarcinoma (LUAD), where they drive NRF2-mediated antioxidant responses that confer chemoresistance by protecting against oxidative stress induced by chemotherapeutic agents. Similarly, in bladder cancer, somatic alterations in the KEAP1-NRF2 pathway promote tumor progression and resistance to cisplatin-based treatments. An integrative analysis of over 3,600 tumors revealed that somatic alterations in KEAP1 or NFE2L2 occur in more than 10% of cases across multiple cancer types, correlating with poor prognosis and NRF2 expression signatures that serve as biomarkers.^[59]^[60]^[61]^[62] In neurodegenerative diseases, reduced NRF2 activity exacerbates oxidative stress and protein aggregation, key hallmarks of pathology. In Parkinson's disease (PD), Nrf2 deficiency cooperates with α-synuclein overexpression to aggravate dopaminergic neuron death, Lewy body formation, and neuroinflammation through impaired antioxidant defenses. Similarly, in Alzheimer's disease (AD), diminished NRF2 signaling contributes to amyloid-β and tau protein accumulation by failing to mitigate mitochondrial dysfunction and oxidative damage in neurons. Genetic polymorphisms in NFE2L2, such as variants in the antioxidant response element, have been associated with increased risk of PD and related parkinsonian disorders by reducing NRF2 binding and transcriptional activity, thereby enhancing susceptibility to oxidative insults.^[63]^[64]^[65]^[66] NFE2L2 dysregulation also plays a role in metabolic diseases, where Nrf2 deficiency worsens insulin resistance and vascular complications. In type 2 diabetes, Nrf2 knockout impairs insulin sensitivity in adipose and liver tissues by failing to counteract oxidative stress-mediated β-cell dysfunction and peripheral glucose uptake defects, leading to hyperglycemia exacerbation. For atherosclerosis, Nrf2 deficiency promotes endothelial dysfunction through increased reactive oxygen species (ROS) production, reduced nitric oxide bioavailability, and heightened inflammatory adhesion molecule expression, accelerating plaque formation in diabetic models.^[67]^[68]^[69] In immunity and inflammation, aberrant NRF2 activation influences immune cell dynamics and disease states. Hyperactivation of NRF2, often due to KEAP1 mutations, drives T-cell exhaustion in tumor microenvironments by upregulating the prostacyclin receptor PTGIR, a novel immune checkpoint that impairs CD8+ T-cell metabolism, effector function, and anti-tumor responses; this mechanism was elucidated in 2025 studies showing PTGIR silencing restores T-cell vigor. In autoimmune diseases, while Nrf2 generally suppresses inflammation, its dysregulation—such as deficiency—exacerbates conditions like systemic lupus erythematosus by promoting autoantibody production and nephritis, though context-specific hyperactivation may contribute to chronic inflammatory persistence in some models.^[70]^[71]^[72] Beyond these, NFE2L2 alterations affect other conditions, including impaired wound healing and hypoxia-related lung diseases. Nrf2 deficiency delays cutaneous wound closure by prolonging inflammation, reducing macrophage recruitment, and impairing epithelial migration and angiogenesis, as observed in diabetic and knockout models. In chronic obstructive pulmonary disease (COPD), 2024 analyses highlighted Nrf2 downregulation in response to hypoxia and cigarette smoke-induced oxidative stress, leading to alveolar epithelial damage, mucus hypersecretion, and emphysema progression; protective Nrf2 activation mitigates these defects by restoring redox balance.^[73]^[74]^[75]^[76]

Therapeutic Implications

NRF2 (encoded by NFE2L2) has emerged as a promising therapeutic target due to its role in modulating oxidative stress and inflammation, with activators primarily explored for neurodegenerative and metabolic disorders. Electrophilic compounds like sulforaphane, derived from broccoli sprouts, activate NRF2 by modifying cysteine residues on KEAP1, leading to NRF2 stabilization and translocation to the nucleus for antioxidant gene induction.^[77] Dimethyl fumarate (DMF), marketed as Tecfidera, is an FDA-approved oral therapy for relapsing-remitting multiple sclerosis (MS) that activates NRF2 through electrophilic modification of KEAP1, reducing oxidative damage and inflammation in glial cells while improving clinical outcomes in MS patients.^[78] Non-electrophilic activators, such as ADJ-310, represent a newer class that promotes NRF2 nuclear accumulation without covalent KEAP1 modification; in preclinical studies, ADJ-310 enhanced wound closure in human keratinocytes and accelerated healing in diabetic mouse models by upregulating NRF2 target genes like NQO1 and HMOX1.^[79] In contrast, NRF2 inhibitors are being developed to counteract its oncogenic role in cancer, where hyperactivation confers chemoresistance and survival advantages. Brusatol, a quassinoid from Brucea javanica seeds, destabilizes NRF2 protein by promoting its ubiquitination and proteasomal degradation, thereby sensitizing various cancer cells—including pancreatic and breast lines—to chemotherapy agents like cisplatin and paclitaxel in preclinical models.^[80] ML385, a small-molecule inhibitor, binds to the Neh1 DNA-binding domain of NRF2, disrupting its heterodimerization with MAFG and subsequent binding to antioxidant response elements (AREs), which suppresses downstream gene expression and enhances ferroptosis sensitivity in lung and head-and-neck cancers.^[81] Clinical progress includes ongoing Phase II/III trials evaluating NRF2 inducers for neurodegeneration; for instance, M102 (from Aclipse Therapeutics) activates NRF2 to protect motor neurons in amyotrophic lateral sclerosis (ALS) models, with 2025 updates showing improved survival in rodent studies and plans for human trials targeting mitochondrial dysfunction and oxidative stress.^[82] In oncology, NRF2 modulation enhances immunotherapy; 2025 research demonstrates that systemic NRF2 activation synergizes with immune checkpoint inhibitors (ICIs) like anti-PD-1 in NRF2-hyperactive lung cancers, boosting T-cell infiltration and tumor regression by alleviating immunosuppressive microenvironments.^[83] However, therapeutic challenges persist, including on-target toxicity from NRF2 overactivation, which can promote tumorigenesis or metabolic imbalances in non-target tissues, as observed in chronic dosing studies.^[84] Biomarkers such as the KEAP1/NRF2 expression ratio are being validated to predict response and monitor pathway activity, aiding patient stratification in trials for conditions like chronic kidney disease and cancer.^[85] Recent 2025 advances highlight NRF2's dual-edged potential. Additionally, hydroxytyrosol, a polyphenol from olive oil, induces ferroptosis in colorectal cancer (CRC) cells by inhibiting NRF2 signaling, downregulating GPX4 and lipid peroxidation defenses, which suppresses tumor growth in xenograft models and suggests synergy with standard chemotherapies.^[86] NRF2 (encoded by NFE2L2) engages in several key protein-protein interactions that modulate its stability, nuclear translocation, DNA binding, and transcriptional activity. The primary interaction occurs with kelch-like ECH-associated protein 1 (KEAP1), which binds the Neh2 domain of NRF2 via its ETGE and DLG motifs. This binding recruits cullin 3 (Cul3) to form an E3 ubiquitin ligase complex, targeting NRF2 for proteasomal degradation under homeostatic conditions. Oxidative or electrophilic stress modifies KEAP1's cysteine residues, disrupting this interaction and stabilizing NRF2.^[19] For transcriptional activation, NRF2 heterodimerizes with small MAF proteins (such as MAFF, MAFG, and MAFK) through its Neh1 basic leucine zipper (bZIP) domain. This complex binds to antioxidant response elements (AREs) in target gene promoters. The Neh3 domain further enhances MAF binding affinity. Additionally, NRF2 interacts with coactivators CREB-binding protein (CBP) and p300 via its Neh4 and Neh5 transactivation domains, recruiting them to chromatin for histone acetylation and gene expression.^[87] NRF2 also forms functional interactions with other transcription factors. It cross-talks with the aryl hydrocarbon receptor (AhR), where mutual regulation influences xenobiotic metabolism genes. NRF2 competes with NF-κB for CBP binding, thereby suppressing inflammatory responses. Alternative degradation pathways involve β-TrCP binding to the phosphorylated Neh6 domain of NRF2. Proteomic studies have identified over 40 additional interactors, including ATF3 and FOS family members, which conditionally modulate ARE-driven transactivation, with implications for stress responses and disease.^[88]^[87]

References

[1]
NFE2L2 NFE2 like bZIP transcription factor 2 [ (human)] - NCBI
Sep 5, 2025 · NFE2L2 encodes a transcription factor that regulates genes with antioxidant response elements, involved in response to injury and inflammation.
[2]
600492 - NUCLEAR FACTOR ERYTHROID 2-LIKE 2; NFE2L2 - OMIM
NFE2L2 encodes a transcription factor that activates genes involved in responses to cellular insults, such as oxidative stress.
[3]
An Overview of the Nrf2/ARE Pathway and Its Role in ...
Sep 4, 2021 · Nrf2 is a basic region leucine-zipper transcription factor that plays a pivotal role in the coordinated gene expression of antioxidant and detoxifying enzymes.
[4]
NRF2, a Transcription Factor for Stress Response and Beyond - PMC
NFE2L2 gene contains ARE within its promoter region that renders NRF2 the ability to directly activate its own transcription, providing a positive feedback ...
[5]
NFE2L2 and ferroptosis resistance in cancer therapy - PMC
Oct 25, 2024 · These genes regulate a variety of crucial cellular processes, including antioxidant defense, detoxification, cell proliferation and death, ...
[6]
Nrf2 at the Heart of Oxidative Stress and Cardiac Protection - PubMed
Feb 1, 2018 · The NFE2L2 gene encodes the transcription factor Nrf2 best known for regulating the expression of antioxidant and detoxification genes.
[7]
The Regulation of NFE2L2 (NRF2) Signalling and Epithelial-to ...
NFE2L2 is a transcription factor which activates signalling pathways that ensure the maintenance of the cellular redox state by activating phase II ...<|control11|><|separator|>
[8]
Genetic associations of Nrf2-encoding NFE2L2 variants with ...
Dec 12, 2014 · The transcription factor Nrf2, encoded by the NFE2L2 gene, is an important regulator of the cellular protection against oxidative stress.
[9]
600492 - NUCLEAR FACTOR ERYTHROID 2-LIKE 2; NFE2L2 - OMIM
Cytogenetic location: 2q31.2 Genomic coordinates (GRCh38) : 2:177,230,303-177,264,727 (from NCBI) ... Although the genes encoding NFE2, NFE2L1, and NFE2L2 are ...
[10]
NRF2, a Transcription Factor for Stress Response and Beyond - MDPI
NFE2L2 gene contains ARE within its promoter region that renders NRF2 the ability to directly activate its own transcription, providing a positive feedback ...Nrf2, A Transcription Factor... · 4. Nrf2 Regulates Metabolic... · 8. Nrf2 Regulates...
[11]
Gene: NFE2L2 (ENSG00000116044) - Summary - Homo_sapiens
Chromosome 2: 177,218,667-177,392,756 reverse strand. GRCh38:CM000664.2. About this gene. This gene has 67 transcripts (splice variants), 212 orthologues, 3 ...
[12]
https://www.ensembl.org/Homo_sapiens/Gene/Compara_Ortholog?g=ENSG00000116044
[13]
POLYMORPHISMS OF NRF2, AN ANTIOXIDATIVE GENE ... - NIH
These promoter polymorphisms were predicted to have functional significance, and rs6721961 affects basal Nrf2 expression and function. Thus, different ...<|control11|><|separator|>
[14]
GTEx Portal
The Genotype-Tissue Expression (GTEx) project is an ongoing effort to build a comprehensive public resource to study tissue-specific gene expression and ...
[15]
Nrf2 and Nrf2-Related Proteins in Development and Developmental ...
The expression and inducibility of anti-oxidant defenses through activation of NFE2-related factor 2 (Nrf2) and related proteins is an essential process ...
[16]
q16236 · nf2l2_human - UniProt
NF2L2 is a transcription factor that responds to oxidative stress by promoting cytoprotective genes, and also regulates innate immune response.Missing: exons introns
[17]
The circadian clock regulates rhythmic activation of the NRF2 ...
Mar 15, 2014 · Here we show that an E-box-mediated circadian rhythm of NRF2 protein is essential in regulating the rhythmic expression of antioxidant genes.Missing: glucocorticoids | Show results with:glucocorticoids
[18]
Glucocorticoid receptor signaling represses the antioxidant ...
May 5, 2017 · We found that glucocorticoids, dexamethasone and betamethasone, antagonize diethyl maleate-induced activation of NRF2 target genes in a GR-dependent manner.
[19]
NFE2L2 Gene - GeneCards | NF2L2 Protein
Protein attributes for NFE2L2 Gene. Size: 605 amino acids; Molecular mass: 67827 Da. Protein existence level: PE1 ... Exon structure for NFE2L2 gene. Exon ...Missing: introns | Show results with:introns
[20]
Structural basis of Keap1 interactions with Nrf2 - PubMed Central - NIH
Keap1 uses a two-site binding mechanism to capture Nrf2, with the Neh2 domain spanning between two Kelch domains for efficient ubiquitination.
[21]
Nrf2, the Major Regulator of the Cellular Oxidative Stress Response ...
Our results show that Nrf2 is partially disordered, with transiently structured elements in its Neh2, Neh7, and Neh1 domains.
[22]
Nrf2 and Oxidative Stress: A General Overview of Mechanisms and ...
Nrf2 contains seven conserved Neh domains. The Neh2 domain contains two motifs (29DLG31 and 79ETGE82) wherein Keap1 binds as a substrate adaptor ...
[23]
Overlooked and valuable facts to know in the NRF2/KEAP1 field
Nov 1, 2022 · They serve as a consensus sequence for transcription factors such as NRF1, NRF2, AhR, AP1, AP2, and NF-κB induced in response to various stimuli ...
[24]
Molecular basis for the disruption of Keap1–Nrf2 interaction ... - Nature
May 14, 2021 · The Neh2 domain of Nrf2 (98 amino acids) is intrinsically disordered and, upon binding to the large molecular weight protein Keap1 dimer ( ...
[25]
The Molecular Mechanisms Regulating the KEAP1-NRF2 Pathway
Apr 13, 2020 · The KEAP1-NRF2 pathway is the principal protective response to oxidative and electrophilic stresses. Under homeostatic conditions, KEAP1 ...Missing: subcellular paper
[26]
The Nrf2 cell defence pathway: Keap1-dependent and
Mar 15, 2013 · The importance of Keap1 in this process is clear, as the half life of Nrf2 increased from approximately 7.5–15 min. in unstressed COS1 cells ...
[27]
Multiple Nuclear Localization Signals Function in the Nuclear Import ...
A number of reports have indicated that phosphorylation at Ser40 of Nrf2 (in the cytoplasm) by protein kinase C, which appears to be concomitant with its ...
[28]
Keap1 Regulates the Oxidation-Sensitive Shuttling of Nrf2 into and ...
We find that Keap1 sequesters Nrf2 in the cytoplasm, not by docking it to the actin cytoskeleton but instead through an active Crm1/exportin-dependent nuclear ...
[29]
The KEAP1-NRF2 pathway: Targets for therapy and role in cancer
In this graphical review, we present an updated overview of the KEAP1-NRF2 signaling, focusing on the progress made within the past ten years. Specifically, we ...Missing: canonical | Show results with:canonical
[30]
Post-translational modifications of Keap1: the state of the art - Frontiers
The Keap1-Nrf2 signaling pathway plays a crucial role in cellular defense against oxidative stress-induced damage. Its activation entails the expression and ...Introduction · Structure and function of... · Post-translational... · Conclusion
[31]
MiR-28 regulates Nrf2 expression through a Keap1 ... - PubMed
Jun 3, 2011 · MiR-28 reduces Nrf2 mRNA and protein levels by targeting the 3'UTR of Nrf2 mRNA, independently of Keap1.
[32]
Regulation of Nrf2 Signaling - PMC - PubMed Central - NIH
There is evidence for the autoregulation of Nrf2 signaling. On the one hand, due to the presence of ARE-like sequences in the promoter region of the Nrf2 gene, ...
[33]
When NRF2 Talks, Who's Listening? | Antioxidants & Redox Signaling
In this review, we focus on recent evidence for transcriptional cross-talk between NRF2 and the arylhydrocarbon receptor (AhR), NF-κB, p53, and Notch pathways, ...
[34]
NRF2 and p53: Januses in cancer? - Oncotarget
Nov 6, 2012 · We will discuss evidence for crosstalk between these transcription factors and possible therapeutic strategies arising from these observations.
[35]
Targeting epigenetic and post-translational modifications of NRF2
Apr 21, 2025 · Nrf2 is controlled by two distinct β-TrCP recognition motifs in its Neh6 domain, one of which can be modulated by GSK-3 activity. Oncogene ...Missing: feedback loops
[36]
NRF2 activation by cysteine as a survival mechanism for triple ...
Apr 10, 2024 · Here we show that TNBC cells of the mesenchymal stem-like subtype (MSL) utilize forced cystine uptake to induce activation of the transcription factor NRF2.
[37]
Genome-wide global identification of NRF2 binding sites in A549 ...
Our ChIP-Seq data identified 2,051 coding and non-coding regions that represented 2,395 binding sites. We then identified the cell-signaling networks associated ...
[38]
Global mapping of binding sites for Nrf2 identifies novel targets in ...
Two adjacent AP-1-like binding sites form the electrophile-responsive element of the murine glutathione S-transferase Ya subunit gene. Proc. Natl Acad. Sci ...
[39]
The NRF2-Dependent Transcriptional Regulation of Antioxidant ...
The most widely studied NRF targets include genes encoding NQO1, HO-1, GSTs, and PRDxs (reviewed in [77]), but NRF2 knockdown has also been associated with the ...
[40]
[PDF] A core NRF2 gene set defined through comprehensive ... - bioRxiv
Overall, every single core NRF2 gene was a part of previous. NRF2 gene sets with 8 genes (GCLM, SRXN1, GCLC, GSR, NQO1, SLC7A11, AKR1C3,. ME1) that were a ...<|separator|>
[41]
Nrf2 signaling in heart failure: expression of Nrf2, Keap1, antioxidant ...
Mar 4, 2022 · Antioxidant genes showed decreased expression in both types of heart failure, including NQO1, SOD1, GPX3, GPX4, GSR, PRDX1, and TXNRD1.Nrf2 Signaling In Heart... · Nrf2 And Keap1 Gene... · Validating Nrf2 Gene...
[42]
The complexity of the Nrf2 pathway: Beyond the antioxidant response
Nrf2 coordinates the basal and inducible expression of antioxidant and phase II detoxification enzymes to adapt to different stress conditions.Missing: ABCC | Show results with:ABCC
[43]
Role of Nrf2 in the regulation of the Mrp2 (ABCC2) gene - PMC - NIH
Nrf2 appears to regulate Mrp2 gene expression via an ARE element located at the proximal region of its promoter in response to exposure to xenobiotics.Nrf2 Expression Plasmids · Results · Bha Induces Gene Expression...Missing: UGT1A | Show results with:UGT1A
[44]
Nrf2 enhances resistance of cancer cells to chemotherapeutic drugs ...
Aug 7, 2025 · Here, we report that inhibition of NF-E2-related factor 2 (Nrf2) may be a promising strategy to combat chemoresistance. Nrf2 is a critical ...Missing: ABCC | Show results with:ABCC
[45]
ATF4-Dependent NRF2 Transcriptional Regulation Promotes ... - NIH
Mar 1, 2020 · ATF4-dependent NRF2 transcriptional regulation promotes antioxidant protection during endoplasmic reticulum stress.Missing: inflammation NLRP3
[46]
Nrf2 inhibits NLRP3 inflammasome activation through regulating ...
Jan 15, 2018 · Our results suggested that Nrf2 acted as a protective regulator against NLRP3 inflammasome activation by regulating the Trx1/TXNIP complex, ...Missing: suppresses | Show results with:suppresses
[47]
The Effects of Sequence Variation on Genome-wide NRF2 Binding ...
Jan 29, 2016 · The impact of systematic ARE variations on NRF2 binding. In previous studies, the MARE sequence variations forming the basis for selective Maf- ...
[48]
New insights into crosstalk between Nrf2 pathway and ferroptosis in ...
Nov 18, 2024 · This review provides new insights from recent discoveries involving the modulation of Nrf2 and ferroptosis in a range of lung diseases.
[49]
https://doi.org/10.1038/s41598-017-04593-w
[50]
https://pubmed.ncbi.nlm.nih.gov/28168414/
[51]
https://pmc.ncbi.nlm.nih.gov/articles/PMC6527603/
[52]
Interaction Between the Nrf2 and PGC-1α Signaling Pathways - NIH
In this review, we summarized recent data on the Nrf2 role in mitochondrial biogenesis and its interaction with PGC-1α in the context of extending longevity.
[53]
Targeting the Interplay Between Autophagy and the Nrf2 Pathway in ...
Autophagy enhances Nrf2 activity, which mitigates oxidative stress, and preserves DAergic neurons. In turn, Nrf2 activation further stimulates autophagy and ...2. Autophagy · 3. Nrf2 Pathway · 3.2. 2. P62
[54]
An important function of Nrf2 in combating oxidative stress - PNAS
Nrf2 mediates the induction of detoxifying genes, NAD(P)H quinone oxidoreductase 1 and glutathione S-transferases (GSTs), by the phenolic antioxidant butylated ...
[55]
Nrf2-Keap1 signaling pathway regulates human UGT1A1 ... - PubMed
Mar 23, 2007 · We elucidate here that an anti-oxidative response leads to induction of UGT1A1 through the Nrf2-Keap1 pathway.
[56]
Interaction between Oxidative Stress Sensor Nrf2 and Xenobiotic ...
Aim of this study was to elucidate the regulation of UGTs expressed in the mucosa of the gastrointestinal tract by xenobiotics and the main mediator of ...
[57]
Identification of Nrf2-regulated Genes Induced by ... - AACR Journals
An Nrf2/small Maf heterodimer mediates the induction of Phase II detoxifying enzyme genes through antioxidant response elements. Biochem. Biophys. Res ...
[58]
Nrf2 Enhances Cell Proliferation and Resistance to Anticancer ...
May 18, 2009 · These results indicate that the Nrf2 system is essential for both cancer cell proliferation and resistance to anticancer drugs.Abstract · Materials and Methods · Results · Discussion
[59]
https://pmc.ncbi.nlm.nih.gov/articles/PMC12361507/
[60]
Integrative analysis of KEAP1/NFE2L2 alterations across 3600+ ...
Aug 18, 2025 · Moreover, our analysis revealed two genes involved in the NRF2 pathway selectively upregulated in LUAD but not LUSC in the presence of KEAP1 ...
[61]
Genetic alterations of Keap1 confers chemotherapeutic resistance ...
Aug 9, 2022 · Keap1 mutations regulate Nrf2 activity and lead to chemoresistance in cancers. Yet the underlying molecular mechanisms of chemoresistance ...
[62]
Integrative analysis of KEAP1/NFE2L2 alterations across 3600+ ...
Aug 18, 2025 · Our study identified novel NRF2-related functionalities associated with specific genetic alterations and revealed a KEAP1/NFE2L2 expression ...
[63]
KEAP1-NRF2 Interaction in Cancer: Competitive Interactors ... - MDPI
This review provides an overview of the KEAP1 competitive protein interactors identified thus far and explores their involvement in cancer development.
[64]
An Overview of the Nrf2/ARE Pathway and Its Role in ... - MDPI
In the NFE2L2 promoter are two ARE-like sequences, through which Nrf2 can enhance its own expression [16], as well as the xenobiotic response element (XRE)-like ...
[65]
Nrf2—a therapeutic target for the treatment of neurodegenerative ...
Nrf2 activation has been shown to mitigate a number of pathologic mechanisms associated with Alzheimer's disease, Parkinson's disease, amyotrophic lateral ...
[66]
A Polymorphic Antioxidant Response Element Links NRF2/sMAF ...
Apr 26, 2016 · Article. A Polymorphic Antioxidant Response Element Links NRF2/sMAF Binding to Enhanced MAPT Expression and Reduced Risk of Parkinsonian ...
[67]
Association of Nrf2-encoding NFE2L2 haplotypes with Parkinson's ...
Mar 2, 2010 · After correction for multiple testing none of the tag SNPs alone significantly affected risk of PD (table 3). The A allele of tag SNP 6 ( ...Tag Snp Selection · Statistical Analyses · Author Information<|separator|>
[68]
Nrf2 as a Potential Mediator of Cardiovascular Risk in Metabolic ...
The present mini-review will provide a comprehensive overview of the role of Nrf2 as a contributing factor to cardiovascular risk in metabolic diseases.
[69]
Exploring the links between polyphenols, Nrf2, and diabetes: A review
Numerous connections between Nrf2 and insulin function suggest that impairment of one can cause dysfunction of the other, creating a detrimental cycle. ...
[70]
Nrf2 as a Potential Mediator of Cardiovascular Risk in Metabolic ...
... (Nrf2) activator dh404 protects against diabetes-induced endothelial dysfunction. ... From endothelial dysfunction to atherosclerosis. Autoimmun. Rev. 9 830 ...
[71]
The prostacyclin receptor PTGIR is a NRF2-dependent regulator of ...
Jun 27, 2025 · These findings identify PTGIR as a NRF2-dependent immune checkpoint that regulates balance between effector and exhausted CD8 + T cell states.Missing: autoimmunity | Show results with:autoimmunity
[72]
The prostacyclin receptor PTGIR is a NRF2-dependent regulator of ...
These findings identify PTGIR as a NRF2-dependent immune checkpoint that regulates balance between effector and exhausted CD8 + T cell states.Missing: hyperactivation autoimmunity
[73]
The Potential of Nrf2 Activation as a Therapeutic Target in Systemic ...
Nrf2 deficiency leads to lupus-like autoimmune syndrome and nephritis in aged female mice, suggesting a direct role for Nrf2 deficiency in the pathogenesis of ...
[74]
Regulation of Wound Healing by the NRF2 Transcription Factor ...
Aug 8, 2019 · The prolonged wound inflammation in NRF2-deficient mice suggested a possible role for NRF2 in immune cells. Immune cells of the myeloid lineage ...
[75]
An Essential Role of NRF2 in Diabetic Wound Healing
Dec 30, 2015 · This study indicates an essential role for NRF2 in diabetic wound healing and the therapeutic benefits of activating NRF2 in this disease.Research Design And Methods · Pharmacological Nrf2... · Nrf2 Activation Promotes...Missing: defects | Show results with:defects
[76]
Protective factors against oxidative stress in COPD: focus on Nrf2 ...
The main mechanism driving the pathogenesis of COPD is oxidative stress. Antioxidant genes, regulated by the Nrf2/ARE signaling pathway, play a protective role ...Missing: wound hypoxia
[77]
Altered hypoxia-induced cellular responses and inflammatory profile ...
Jul 16, 2024 · In this study we aimed to investigate if there are differences in how primary distal lung fibroblasts obtained from COPD patients and healthy subjects respond ...Missing: wound | Show results with:wound
[78]
Sulforaphane and Other Nutrigenomic Nrf2 Activators: Can the ... - NIH
Broccoli-derived sulforaphane emerges as a phytochemical with this capability, with oral doses capable of favourably modifying genes associated with ...
[79]
Evidence of activation of the Nrf2 pathway in multiple sclerosis ...
Background: Delayed-release dimethyl fumarate (DMF) is an approved oral treatment for relapsing forms of multiple sclerosis (MS).
[80]
Non-electrophilic NRF2 activators promote wound healing in human ...
Oct 24, 2024 · NRF2 is highly expressed in the skin and is known to play a critical role in diabetic wound healing, a serious disease process for which ...Missing: defects hypoxia
[81]
Brusatol enhances the efficacy of chemotherapy by inhibiting the ...
Jan 4, 2011 · Because brusatol is capable of inhibiting Nrf2 in many cell types, it can be used to enhance the efficacy of a wide variety of chemotherapeutic ...
[82]
Small molecule inhibitor of NRF2 selectively intervenes therapeutic ...
Taken together, these results suggest that ML385 binds to the Neh1 DNA binding domain of NRF2, blocks the binding of the NRF2-MAFG complex to the ARE sequence ...
[83]
https://www.nature.com/articles/s41416-025-03162-7
[84]
Systemic activation of NRF2 contributes to the therapeutic efficacy of ...
Sep 1, 2025 · Tumours with genetic activation of NRF2 are resistant to all current anti-cancer therapies, including ICIs, and therefore most lung cancer ...
[85]
Advances and challenges in therapeutic targeting of NRF2
Aug 15, 2022 · NRF2 is emerging as an attractive therapeutic approach to counteract oxidative stress, inflammation, and metabolic imbalances.
[86]
Current Landscape of NRF2 Biomarkers in Clinical Trials - PMC
Nuclear factor erythroid 2 related factor 2 (NRF2) is a transcription factor that regulates the expression of over 300 target genes with roles in antioxidant ...
[87]
Targeting NRF2 uncovered an intrinsic susceptibility of acute ...
May 17, 2023 · Our previous study showed that chemotherapy increased NRF2 levels in AML cells, and NRF2 activation contributes to chemotherapy resistance [31].Nrf2 Target Gene Analysis... · Nrf2-Regulated Gpx4 Was... · Nrf2 Inhibition (ml385)...
[88]
Hydroxytyrosol induced ferroptosis through Nrf2 signaling pathway ...
Jul 1, 2025 · Ferroptosis is intimately connected to the to the imbalance of oxidative stress in the body, and Nrf2 plays a key role in regulating ferroptosis ...