{"gene":"DUOX1","run_date":"2026-06-09T23:54:42","timeline":{"discoveries":[{"year":2020,"finding":"Cryo-EM structures of mouse DUOX1-DUOXA1 complex (with and without NADPH substrate) reveal atomic details of DUOX1-DUOXA1 interaction, a lipid-mediated NADPH-binding pocket, and the electron transfer path. A dimer-of-dimers configuration was identified as an inactive state, suggesting an oligomerization-dependent regulatory mechanism.","method":"Cryo-EM structural determination, biochemical assays","journal":"Nature structural & molecular biology","confidence":"High","confidence_rationale":"Tier 1 / Strong — cryo-EM structures with and without substrate, plus biochemical validation, multiple orthogonal methods in single rigorous study","pmids":["32929281"],"is_preprint":false},{"year":2009,"finding":"DUOX1 activity depends on calcium and functional EF-hand motifs. Uniquely among DUOX isoforms, DUOX1 (but not DUOX2) activity is stimulated by cAMP/forskolin via PKA-mediated phosphorylation on serine 955. This differential regulation was confirmed in human thyroid cells co-expressing DUOX1 with its maturation factor DUOXA1.","method":"Co-expression functional assay with DuoxA1/DuoxA2, pharmacological stimulation (forskolin, phorbol esters), site-specific mutagenesis of phosphorylation sites, membrane expression analysis","journal":"The Journal of biological chemistry","confidence":"High","confidence_rationale":"Tier 1 / Strong — in vitro enzymatic assay with mutagenesis, confirmed in human thyroid cells, multiple orthogonal methods in one rigorous study","pmids":["19144650"],"is_preprint":false},{"year":2009,"finding":"The isolated human DUOX1 peroxidase domain (hDUOX1 aa 1–593) does not bind heme and has no intrinsic peroxidase activity, in contrast to the C. elegans ortholog. Neither DUOX1 domain showed significant superoxide dismutase activity, suggesting the N-terminal motif does not directly convert superoxide to H2O2 in isolation.","method":"Baculovirus expression and purification of recombinant peroxidase domains, heme-binding assays, peroxidase activity assays, superoxide dismutase activity assays","journal":"The Journal of biological chemistry","confidence":"High","confidence_rationale":"Tier 1 / Strong — purified recombinant protein, multiple in vitro enzymatic assays, rigorous biochemical characterization","pmids":["19460756"],"is_preprint":false},{"year":2010,"finding":"DUOX1 binds to inositol 1,4,5-trisphosphate receptor 1 (IP3R1) in primary human CD4+ T cells and is required for early TCR-stimulated H2O2 production. DUOX1-derived H2O2 inactivates SHP-2 phosphatase, promoting ZAP-70 phosphorylation at Tyr-319 and its association with Lck and CD3ζ, thereby creating a positive feedback loop that sustains TCR signaling, store-operated Ca2+ entry, and ERK activation.","method":"Co-immunoprecipitation (DUOX1-IP3R1), siRNA/shRNA knockdown, TCR signaling assays, phosphorylation analysis, calcium flux measurement, cytokine production assays","journal":"Science signaling","confidence":"High","confidence_rationale":"Tier 2 / Strong — reciprocal Co-IP, stable and transient knockdown with defined signaling phenotype, multiple orthogonal methods","pmids":["20682913"],"is_preprint":false},{"year":2013,"finding":"ATP activates DUOX1 via purinergic P2Y2 receptor stimulation, recruiting Src and DUOX1 into a signaling complex. DUOX1-derived H2O2 oxidizes cysteine residues within Src and ADAM17, activating both; ADAM17 then sheds EGFR ligands, leading to EGFR transactivation and downstream wound responses in airway epithelial cells.","method":"siRNA/shRNA knockdown, thiol-specific biotin labeling for cysteine oxidation detection, Co-IP (P2Y2R/Src/DUOX1 complex), EGFR phosphorylation assays, cell migration assays","journal":"PloS one","confidence":"High","confidence_rationale":"Tier 2 / Strong — reciprocal Co-IP, stable shRNA, thiol-labeling mass spectrometry, multiple orthogonal readouts replicated across labs","pmids":["23349873"],"is_preprint":false},{"year":2016,"finding":"ATP-dependent EGFR transactivation in airway epithelial cells involves DUOX1-dependent sequential sulfenylation and S-glutathionylation of cysteine residues within EGFR and Src. The intermediate sulfenylation (not S-glutathionylation) is the activating modification; C797S EGFR variant abolishes H2O2-induced EGFR kinase activation, identifying Cys-797 as the redox-sensitive regulatory site.","method":"Recombinant protein kinase activity assays with H2O2/GSSG, C797S mutagenesis, redox-labeling strategies, DUOX1 siRNA/shRNA knockdown","journal":"The Journal of biological chemistry","confidence":"High","confidence_rationale":"Tier 1 / Strong — in vitro kinase assay with mutagenesis, multiple redox-labeling strategies, confirmed with DUOX1 knockdown","pmids":["27650496"],"is_preprint":false},{"year":2014,"finding":"DUOX1 mediates ATP-stimulated transient H2O2 production and S-glutathionylation of multiple proteins in airway epithelial cells, including β-actin, peroxiredoxin 1, Src, and MAPK phosphatase 1. These DUOX1-dependent S-glutathionylation events regulate cytoskeletal dynamics and MAPK signaling involved in cell migration.","method":"Stable shRNA knockdown, primary tracheal epithelial cells from DUOX1-deficient mice, biotin-tagged GSH labeling, avidin purification, global proteomics, H2O2 measurement","journal":"Redox biology","confidence":"High","confidence_rationale":"Tier 2 / Strong — genetic KO mouse cells plus stable shRNA, proteomics with biotin-GSH, multiple orthogonal methods","pmids":["24624333"],"is_preprint":false},{"year":2009,"finding":"ATP-mediated DUOX1 activation in airway epithelial cells leads to H2O2-dependent activation of ERK1/2 and NF-κB pathways, intracellular oxidant signaling, and EGFR ligand shedding by ADAM17, culminating in IL-8/CXCL8 production in response to bacterial stimuli.","method":"DUOX1 siRNA knockdown, catalase (extracellular and intracellular scavenging), ERK/NF-κB pathway inhibitors, ADAM17 activation assays, IL-8 ELISA","journal":"The Journal of biological chemistry","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — siRNA knockdown plus pharmacological inhibitors, single lab, multiple readouts","pmids":["19386603"],"is_preprint":false},{"year":2015,"finding":"Ionizing radiation induces DUOX1-dependent H2O2 production in thyroid cells via a pathway involving p38 MAPK activation and IL-13 upregulation, which in turn sustains DUOX1 expression for days after irradiation. DUOX1-derived H2O2 causes persistent DNA double-strand breaks; catalase pretreatment or DUOX1 siRNA knockdown abrogates this damage.","method":"siRNA knockdown, p38 MAPK inhibitors, catalase treatment, DNA damage assays (γH2AX), dose-response H2O2 measurement, human thyroid tissue analysis","journal":"Proceedings of the National Academy of Sciences of the United States of America","confidence":"High","confidence_rationale":"Tier 2 / Strong — siRNA knockdown with multiple rescue conditions, human and cell line models, multiple orthogonal methods","pmids":["25848056"],"is_preprint":false},{"year":2021,"finding":"DUOX1-derived H2O2 sustains TGF-β1/Smad3 signaling in lung fibroblasts by preventing phospho-Smad3 degradation. Mechanistically, DUOX1 inhibits the interaction between phospho-Smad3 and the ubiquitin ligase NEDD4L, thereby preventing NEDD4L-mediated ubiquitination and proteasomal degradation of phospho-Smad3.","method":"DUOX1-deficient mouse models, primary human/mouse lung fibroblasts, Co-IP (pSmad3/NEDD4L interaction), ubiquitination assays, Smad3 phosphorylation kinetics","journal":"The European respiratory journal","confidence":"High","confidence_rationale":"Tier 2 / Strong — genetic KO mice, primary cells, Co-IP with mechanistic ubiquitination assays, multiple orthogonal methods","pmids":["32764116"],"is_preprint":false},{"year":2016,"finding":"DUOX1 mediates persistent EGFR activation and cysteine oxidation within EGFR and Src in airway epithelial cells during allergic asthma. DUOX1 deficiency attenuated HDM-induced mucous metaplasia, subepithelial fibrosis, neutrophilic inflammation, type 2 cytokine production (IL-33, IL-13), and central airway resistance in mice.","method":"DUOX1-deficient mice, HDM mouse model, siRNA intratracheal delivery, EGFR cysteine oxidation assays, nasal epithelial cells from asthmatic subjects, EGFR/Src phosphorylation analysis","journal":"JCI insight","confidence":"High","confidence_rationale":"Tier 2 / Strong — genetic KO mouse model plus siRNA intervention, human patient cells, multiple mechanistic and phenotypic readouts","pmids":["27812543"],"is_preprint":false},{"year":2010,"finding":"DUOX1 is the main source of H2O2 in urothelial cells, as demonstrated using Duox1 knockout mice. TRPV4 calcium channel activation elicits a calcium signal that stimulates DUOX1-dependent H2O2 production, and Duox1 knockout animals display altered pressure responses in the urinary bladder.","method":"Duox1 gene-deficient mouse model, H2O2 measurement, TRPV4 pharmacological activation, bladder pressure measurements","journal":"Free radical biology & medicine","confidence":"High","confidence_rationale":"Tier 2 / Strong — genetic KO mouse with functional phenotypic readout, multiple methods","pmids":["21146788"],"is_preprint":false},{"year":2007,"finding":"Duox1 is the main source of H2O2 in the rat thyroid cell line PCCl3, as demonstrated by siRNA knockdown reducing H2O2 production in parallel with Duox1 protein. Re-expression of rat Duox1 fully rescued H2O2 production, while human Duox1 partial rescue was also observed.","method":"siRNA knockdown, lentiviral re-expression rescue, H2O2 production assays, Western blot","journal":"Experimental cell research","confidence":"High","confidence_rationale":"Tier 2 / Strong — siRNA knockdown with rescue experiment, multiple methods confirming protein-H2O2 link","pmids":["17643428"],"is_preprint":false},{"year":2024,"finding":"DUOX1 is sorted to the apical plasma membrane in epithelial cells via its maturation factor DUOXA1. N-glycosylation of DUOXA1 is required for apical targeting of DUOX1; impairment of DUOXA1 N-glycosylation results in DUOX1 mistargeting to the basolateral membrane. DUOX2 apical sorting by DUOXA2 is regulated by a distinct mechanism involving DUOXA2's C-terminal region rather than N-glycosylation.","method":"MDCK epithelial cell co-expression system, N-glycosylation mutants of DUOXA1/DUOXA2, immunofluorescence localization, domain-swap experiments","journal":"Genes to cells","confidence":"High","confidence_rationale":"Tier 2 / Strong — mutagenesis combined with defined localization readout in polarized epithelial cells, multiple constructs tested","pmids":["39126279"],"is_preprint":false},{"year":2021,"finding":"DUOX1 and DUOX2 function as NAADP-forming enzymes that convert NAADPH to NAADP in vitro under physiological conditions. In T cells, DUOX1 and DUOX2 are required for global Ca2+ signaling (4–12 min post-stimulation), while DUOX2 specifically drives early Ca2+ microdomain formation (first 15 s). Duoxa1/Duoxa2 double KO but not single Nox1 or Nox2 KO reduced local and global Ca2+ signaling.","method":"In vitro NAADP-forming enzyme assay, Duox1/Duox2/Duoxa1/Duoxa2 knockout mouse T cells, Ca2+ imaging (local microdomains and global), genetic epistasis with single and double KOs","journal":"Science signaling","confidence":"High","confidence_rationale":"Tier 1 / Strong — in vitro enzymatic assay plus multiple genetic KO models with Ca2+ imaging, replication across multiple KO lines","pmids":["34784249"],"is_preprint":false},{"year":2014,"finding":"Testosterone activates Duox1 in epidermal keratinocytes through GPRC6A, a membrane receptor that senses testosterone and activates Gq protein, leading to IP3 generation, intracellular calcium mobilization, and subsequent Duox1-dependent H2O2 generation. GPRC6A silencing abolished testosterone-induced calcium and H2O2 responses.","method":"GPRC6A siRNA knockdown, calcium imaging, H2O2 measurement, IP3 quantification, caspase-3 activation assays, 3D skin equivalent model","journal":"The Journal of biological chemistry","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — siRNA knockdown with multiple downstream readouts, single lab","pmids":["25164816"],"is_preprint":false},{"year":2017,"finding":"Autophagy protein ATG5 is required for apical membrane localization of DUOX1 in airway epithelial cells during chronic IL-13 stimulation. ATG5 depletion significantly reduced intracellular superoxide production without diminishing total DUOX1 protein levels, indicating that non-canonical autophagy regulates DUOX1 trafficking to the apical membrane rather than its expression.","method":"ATG5 siRNA, DUOX1 siRNA, confocal immunofluorescence localization, electron paramagnetic resonance spectroscopy for superoxide, OVA mouse model, LC3BII Western blot","journal":"Redox biology","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — siRNA knockdown with localization and functional readout, supported by mouse model, single lab","pmids":["28982074"],"is_preprint":false},{"year":2018,"finding":"Duox1-derived H2O2 negatively regulates proliferative activity (but not Ig isotype production) in primary splenic B cells upon BCR stimulation. IL-4 costimulation boosts Duox1 expression and H2O2 production; Duox1-/- CD19+ B cells show enhanced proliferation, and catalase treatment of WT B cells mimics this effect. Immunized Duox1-/- mice show increased B cell expansion in germinal centers.","method":"Duox1 knockout mice, BCR stimulation assays, H2O2 measurement, proliferation assays, catalase treatment, in vivo immunization, germinal center analysis","journal":"Journal of immunology","confidence":"High","confidence_rationale":"Tier 2 / Strong — genetic KO mice with in vitro and in vivo validation, pharmacological rescue with catalase, multiple orthogonal readouts","pmids":["30559322"],"is_preprint":false},{"year":2015,"finding":"Duox1-derived H2O2 promotes late-phase neutrophil recruitment to wounds in zebrafish by inducing Cxcl8 expression via JNK/c-JUN/AP-1 signaling. This H2O2-driven cxcl8 induction is accompanied by histone modifications (increased H3K4me3, H3K9ac; decreased H3K9me3) at the cxcl8 promoter. ERK and NF-κB signaling were not involved.","method":"Zebrafish duox1 morpholino knockdown, wound assays, JNK inhibitors, ChIP for histone modifications, neutrophil recruitment quantification","journal":"Journal of immunology","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — zebrafish ortholog model with pathway inhibitors and ChIP, single lab","pmids":["25582859"],"is_preprint":false},{"year":2019,"finding":"DUOX1 missense mutation p.R1307Q impairs H2O2 generation and reduces DUOX1 mRNA and protein expression. A companion DUOXA1 mutation p.R56W similarly reduces DUOX1 expression and H2O2 generation, demonstrating that intact DUOXA1 is required for full DUOX1 activity; both mutations can cause congenital hypothyroidism.","method":"Patient sequencing, functional H2O2 generation assays in transfected cells, qRT-PCR, Western blot","journal":"Frontiers in endocrinology","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — functional assay in transfected cells with patient-derived mutations, single lab","pmids":["31428054"],"is_preprint":false},{"year":2016,"finding":"Loss of DUOX1 expression in lung epithelial cells promotes epithelial-to-mesenchymal transition (EMT), characterized by loss of E-cadherin, gain of vimentin/collagen, increased migration, anchorage-independent growth, and enhanced cancer stem cell markers (CD133, ALDH1). Conversely, overexpression of DUOX1 in A549 cells reverses EMT features, and DUOX1 loss confers resistance to EGFR tyrosine kinase inhibitors.","method":"shRNA stable knockdown, DUOX1 overexpression, E-cadherin/vimentin Western blot, migration assays, anchorage-independent growth, in vivo xenograft invasion assay, erlotinib resistance assays","journal":"Oncogenesis","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — stable shRNA and overexpression with in vitro and in vivo readouts, single lab","pmids":["27694834"],"is_preprint":false},{"year":2026,"finding":"DUOX1 in macrophages contributes to profibrotic activation through oxidative activation of Src kinase via cysteine oxidation. Myeloid-specific conditional DUOX1 deletion attenuated pulmonary fibrosis, impaired MoMac recruitment, reduced collagen production, and improved oxygen saturation. DUOX1 also contributes to in vitro macrophage migration and EGFR ligand production involved in macrophage-fibroblast cross-talk.","method":"LysM-Cre conditional DUOX1 knockout, BMDM migration assays, cysteine oxidation assays, collagen production measurement, Src kinase activation assays, saracatinib pharmacological inhibition","journal":"American journal of respiratory cell and molecular biology","confidence":"High","confidence_rationale":"Tier 2 / Strong — conditional genetic KO with multiple in vivo and in vitro mechanistic readouts and pharmacological validation","pmids":["40986746"],"is_preprint":false},{"year":2026,"finding":"The GPRC6A-Duox1 axis regulates hair cycle progression downstream of testosterone signaling. GPRC6A-deficient and Duox1-deficient keratinocytes fail to generate H2O2 in response to testosterone. Both GPRC6A KO and Duox1 KO mice show extended anagen phase, increased Ki-67 expression, longer hair, and resistance to testosterone-mediated hair loss.","method":"GPRC6A and Duox1 knockout mice and keratinocytes, H2O2 measurement, apoptosis assays, Ki-67 immunostaining, hair cycle staging, topical testosterone application","journal":"Tissue engineering and regenerative medicine","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — genetic KO mouse models with multiple readouts, single lab","pmids":["42047995"],"is_preprint":false},{"year":2018,"finding":"DUOX1-derived H2O2 regulates sodium transport (ENaC activity) in H441 bronchiolar epithelial cells. Dexamethasone-induced dome formation is inhibited by extracellular catalase or NADPH oxidase inhibitor DPI. H2O2 (0.2 mmol/L) acutely activates amiloride-sensitive ENaC currents in dome-forming cells, and there is a negative feedback loop where H2O2 inhibits ENaC gene transcription.","method":"Nystatin-perforated patch-clamp electrophysiology, catalase treatment, DPI inhibition, DUOX1 immunocytochemistry, RT-PCR, dome formation assays","journal":"Acta physiologica","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — electrophysiology with pharmacological inhibition, single lab, moderate mechanistic depth","pmids":["30052308"],"is_preprint":false},{"year":2022,"finding":"Macrophage-intrinsic DUOX1 contributes to type 2 inflammation and mucus metaplasia during chronic HDM-driven allergic airway disease. Cell-type-specific conditional deletion revealed that epithelial DUOX1 drives acute HDM responses and most features of chronic inflammation, while macrophage DUOX1 additionally contributes to macrophage recruitment, type 2 cytokine production, and mucus metaplasia.","method":"Conditional cell-type-specific DUOX1 knockout (epithelial and macrophage lineages), HDM mouse model, cytokine measurement, mucus staining, flow cytometry","journal":"Mucosal immunology","confidence":"High","confidence_rationale":"Tier 2 / Strong — cell-type-specific conditional KO with defined mechanistic cellular phenotypes, multiple readouts","pmids":["35654836"],"is_preprint":false},{"year":2007,"finding":"DUOX1 expression and its maturation factor DUOXA1 are co-upregulated during differentiation of human fetal lung cells into alveolar type II cells (induced by DCI treatment), with DUOX1 protein localizing near the apical cell pole. DUOX1 knockdown by siRNA blocked increases in H2O2 production and acid secretion in differentiated type II cells, establishing DUOX1 as responsible for alveolar H2O2 and acid production.","method":"Human fetal lung primary cell culture, DCI differentiation protocol, siRNA knockdown, H2O2 measurement, intracellular acid measurement, immunofluorescence localization","journal":"American journal of physiology. Lung cellular and molecular physiology","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — siRNA knockdown with localization and functional assays in primary cells, single lab","pmids":["17337509"],"is_preprint":false},{"year":2023,"finding":"HIF-2α transcriptionally promotes DUOX1 expression (confirmed by dual luciferase assay), and arsenic-induced upregulation of DUOX1 suppresses GPX4 expression, promoting ferroptosis in kidney cells. DUOX1 siRNA knockdown significantly upregulated GPX4 and attenuated ferroptosis.","method":"siRNA knockdown, dual luciferase reporter assay for HIF-2α binding to DUOX1 promoter, GPX4 Western blot, ferroptosis markers, in vivo arsenic exposure model","journal":"The Science of the total environment","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — dual luciferase + siRNA + in vivo model, single lab, mechanistic pathway partially defined","pmids":["37879473"],"is_preprint":false}],"current_model":"DUOX1 is a calcium-dependent NADPH oxidase that co-assembles with its maturation factor DUOXA1 (which directs apical membrane targeting via N-glycosylation) to form an active enzyme complex whose cryo-EM structure reveals a lipid-mediated NADPH-binding pocket and an oligomerization-dependent inactive dimer-of-dimers state; its H2O2 output is regulated by PKA-mediated phosphorylation on Ser-955 and by calcium/EF-hand motifs, and acts through reversible cysteine oxidation (sulfenylation, S-glutathionylation) of targets including Src, ADAM17, EGFR (Cys-797), SHP-2, β-actin, and peroxiredoxin 1, thereby mediating positive feedback in TCR and EGFR signaling, innate airway epithelial host defense, TGF-β1/Smad3 amplification in fibrosis (by blocking NEDD4L-mediated phospho-Smad3 ubiquitination), NAADP synthesis for T cell Ca2+ signaling, and testosterone/GPRC6A-driven H2O2 production in keratinocytes."},"narrative":{"mechanistic_narrative":"DUOX1 is a calcium-dependent NADPH oxidase that generates H2O2 at the apical surface of epithelial cells and serves as a signaling hub that converts diverse stimuli into redox-based control of downstream targets [PMID:32929281, PMID:19144650, PMID:17643428]. Its activity depends on calcium acting through EF-hand motifs and is uniquely potentiated among DUOX isoforms by cAMP/PKA-mediated phosphorylation at Ser-955 [PMID:19144650]; cryo-EM of the DUOX1-DUOXA1 complex resolves a lipid-mediated NADPH-binding pocket, the electron-transfer path, and an inactive dimer-of-dimers state indicating oligomerization-dependent regulation [PMID:32929281]. Functional maturation and apical plasma-membrane delivery require the partner factor DUOXA1, whose N-glycosylation directs correct apical targeting, while non-canonical autophagy (ATG5) further controls DUOX1 trafficking to the apical membrane [PMID:39126279, PMID:28982074, PMID:31428054]. DUOX1-derived H2O2 acts largely through reversible cysteine oxidation of signaling proteins: in airway epithelia, ATP/P2Y2-recruited DUOX1 sulfenylates and S-glutathionylates Src, ADAM17, and EGFR at Cys-797 to drive EGFR transactivation, cytoskeletal remodeling, and inflammatory output, and in T cells it inactivates SHP-2 to sustain ZAP-70/Lck-dependent TCR signaling and calcium entry [PMID:20682913, PMID:23349873, PMID:27650496, PMID:24624333]. Through these redox circuits DUOX1 amplifies type 2 airway inflammation and EGFR-driven asthma pathology, sustains TGF-β1/Smad3 fibrotic signaling by blocking NEDD4L-mediated phospho-Smad3 degradation, and contributes to macrophage profibrotic activation via oxidative Src activation [PMID:32764116, PMID:27812543, PMID:40986746, PMID:35654836]. DUOX1 and DUOX2 also act as NAADP-forming enzymes required for T-cell calcium signaling, and DUOX1 mediates testosterone/GPRC6A-driven H2O2 production controlling hair-cycle progression [PMID:34784249, PMID:42047995]. Loss-of-function missense mutations in DUOX1 (p.R1307Q) and DUOXA1 (p.R56W) impair H2O2 generation and can cause congenital hypothyroidism [PMID:31428054].","teleology":[{"year":2007,"claim":"Establishing that DUOX1 is genuinely responsible for cellular H2O2 output was required before any signaling role could be assigned; knockdown-with-rescue and differentiation studies pinned H2O2 production to DUOX1 protein.","evidence":"siRNA knockdown with lentiviral re-expression rescue in rat thyroid cells, and DCI-differentiated human fetal alveolar type II cells with apical localization","pmids":["17643428","17337509"],"confidence":"High","gaps":["Did not resolve how DUOX1 activity is switched on at the molecular level","Apical localization mechanism not defined"]},{"year":2009,"claim":"Defining the activation logic distinguished DUOX1 from DUOX2 and identified its regulatory inputs: calcium via EF-hands and uniquely cAMP/PKA phosphorylation at Ser-955.","evidence":"Co-expression functional assays with DUOXA1/DUOXA2, forskolin/phorbol stimulation, and site-specific phosphorylation mutagenesis in human thyroid cells","pmids":["19144650"],"confidence":"High","gaps":["Kinase that mediates PKA-independent inputs not mapped","Structural basis of EF-hand coupling to catalysis unresolved at the time"]},{"year":2009,"claim":"Whether the N-terminal peroxidase-homology domain of DUOX1 carries intrinsic enzymatic activity was tested directly; the isolated human domain neither binds heme nor shows peroxidase or SOD activity, constraining models of H2O2 generation.","evidence":"Baculovirus-expressed recombinant peroxidase domains with heme-binding, peroxidase, and SOD assays","pmids":["19460756"],"confidence":"High","gaps":["Function of the N-terminal domain in the full-length enzyme not established","Did not test domain behavior within the assembled complex"]},{"year":2009,"claim":"The first signaling role connected DUOX1-derived H2O2 to innate airway responses, linking ATP stimulation to ADAM17/EGFR shedding, ERK/NF-κB activation, and IL-8 production.","evidence":"DUOX1 siRNA knockdown with intracellular/extracellular catalase scavenging and pathway inhibitors in airway epithelial cells","pmids":["19386603"],"confidence":"Medium","gaps":["Direct molecular targets of H2O2 not identified","Single-lab pharmacology-dependent readouts"]},{"year":2010,"claim":"DUOX1 was shown to be a direct amplifier of TCR signaling, defining a redox positive-feedback loop in T cells via SHP-2 inactivation.","evidence":"DUOX1-IP3R1 Co-IP, siRNA/shRNA knockdown, phosphorylation and calcium-flux analysis in primary human CD4+ T cells","pmids":["20682913"],"confidence":"High","gaps":["Direct sulfenylation site on SHP-2 not mapped","Mechanism of DUOX1-IP3R1 coupling unresolved"]},{"year":2010,"claim":"Genetic deletion established DUOX1 as the dominant urothelial H2O2 source downstream of TRPV4 calcium signaling, with a bladder pressure-response phenotype.","evidence":"Duox1 knockout mice, H2O2 measurement, TRPV4 activation, and bladder pressure measurements","pmids":["21146788"],"confidence":"High","gaps":["Downstream redox targets in urothelium not identified","Physiological signaling output incompletely defined"]},{"year":2013,"claim":"The redox mechanism of EGFR transactivation was resolved at the protein level, showing DUOX1 oxidizes cysteines in Src and ADAM17 within a P2Y2-recruited complex.","evidence":"Reciprocal Co-IP, thiol-specific labeling, knockdown, and EGFR phosphorylation/migration assays in airway epithelial cells","pmids":["23349873"],"confidence":"High","gaps":["Specific oxidized cysteines on ADAM17 not pinpointed","Kinetics of complex assembly not defined"]},{"year":2014,"claim":"Proteome-scale S-glutathionylation mapping defined the breadth of DUOX1 redox targets controlling cytoskeleton and MAPK signaling.","evidence":"Stable shRNA and DUOX1-deficient mouse tracheal cells with biotin-GSH labeling and global proteomics","pmids":["24624333"],"confidence":"High","gaps":["Functional consequence of each glutathionylation event not individually validated","Reversal/deglutathionylation kinetics not addressed"]},{"year":2014,"claim":"A hormone-sensing input was identified, linking testosterone via GPRC6A/Gq/IP3/calcium to DUOX1 H2O2 production in keratinocytes.","evidence":"GPRC6A siRNA knockdown with calcium imaging, IP3 quantification, and H2O2 measurement in a 3D skin model","pmids":["25164816"],"confidence":"Medium","gaps":["Single-lab siRNA evidence","Downstream redox targets in keratinocytes not identified"]},{"year":2015,"claim":"DUOX1 was implicated in damage signaling, showing radiation-induced p38/IL-13-sustained DUOX1 expression drives persistent DNA double-strand breaks in thyroid cells.","evidence":"siRNA knockdown with p38 inhibitors and catalase rescue, γH2AX assays, and human thyroid tissue analysis","pmids":["25848056"],"confidence":"High","gaps":["Mechanism by which apical H2O2 reaches nuclear DNA not defined","Long-term carcinogenic consequence not established"]},{"year":2015,"claim":"An ortholog model defined an epigenetic mechanism for DUOX-driven inflammation, linking H2O2 to JNK/AP-1 and histone marks at the cxcl8 promoter.","evidence":"Zebrafish duox1 morpholino knockdown, JNK inhibitors, and ChIP for histone modifications at wounds","pmids":["25582859"],"confidence":"Medium","gaps":["Conservation of the histone-mark mechanism in mammals not shown","Morpholino-based knockdown only"]},{"year":2016,"claim":"The activating redox modification of EGFR was pinpointed: sulfenylation (not S-glutathionylation) of Cys-797 is the kinase-activating event.","evidence":"Recombinant kinase assays with H2O2/GSSG, C797S mutagenesis, redox labeling, and DUOX1 knockdown","pmids":["27650496"],"confidence":"High","gaps":["In vivo occupancy of Cys-797 sulfenylation not quantified","Reversal mechanism not detailed"]},{"year":2016,"claim":"In vivo asthma modeling established DUOX1 as a driver of type 2 airway pathology via sustained EGFR/Src cysteine oxidation.","evidence":"DUOX1-deficient mice and intratracheal siRNA in an HDM model, with EGFR oxidation assays and asthmatic patient cells","pmids":["27812543"],"confidence":"High","gaps":["Relative contribution of epithelial vs other cell types not yet separated","Causal cytokine hierarchy incompletely defined"]},{"year":2016,"claim":"A context-dependent tumor-suppressive role emerged, with DUOX1 loss promoting EMT, stemness, and EGFR-TKI resistance in lung epithelial cells.","evidence":"shRNA knockdown and overexpression with EMT markers, migration, anchorage-independent growth, and xenograft assays","pmids":["27694834"],"confidence":"Medium","gaps":["Mechanism linking DUOX1 loss to EMT transcriptional program unresolved","Single-lab in vitro/xenograft evidence"]},{"year":2017,"claim":"Trafficking control of DUOX1 was extended to non-canonical autophagy, with ATG5 required for apical membrane localization independent of protein levels.","evidence":"ATG5 and DUOX1 siRNA, confocal localization, EPR superoxide measurement, and OVA mouse model","pmids":["28982074"],"confidence":"Medium","gaps":["Molecular machinery linking ATG5 to DUOX1 trafficking unknown","Single-lab siRNA evidence"]},{"year":2018,"claim":"DUOX1 was shown to negatively regulate adaptive immunity, restraining B-cell proliferation downstream of BCR/IL-4.","evidence":"Duox1 knockout mice, BCR stimulation, catalase rescue, and in vivo germinal center analysis","pmids":["30559322"],"confidence":"High","gaps":["Redox targets restraining B-cell proliferation not identified","Distinction from antibody-isotype output mechanistically unexplained"]},{"year":2018,"claim":"An ion-transport regulatory role was defined, with DUOX1 H2O2 acutely activating ENaC while transcriptionally repressing it in a feedback loop.","evidence":"Perforated patch-clamp electrophysiology with catalase/DPI inhibition in H441 bronchiolar cells","pmids":["30052308"],"confidence":"Medium","gaps":["Direct ENaC redox target site not mapped","Single-lab pharmacology-based attribution"]},{"year":2019,"claim":"Patient-derived mutations established a human disease link and reinforced DUOXA1 dependence, with DUOX1 p.R1307Q and DUOXA1 p.R56W impairing H2O2 generation in congenital hypothyroidism.","evidence":"Patient sequencing with functional H2O2 assays, qRT-PCR, and Western blot in transfected cells","pmids":["31428054"],"confidence":"Medium","gaps":["Single-lab functional validation","Genotype-phenotype penetrance not fully established"]},{"year":2021,"claim":"A fibrosis-amplifying mechanism was defined, with DUOX1 H2O2 stabilizing phospho-Smad3 by blocking NEDD4L-mediated ubiquitination.","evidence":"DUOX1-deficient mice, primary lung fibroblasts, pSmad3/NEDD4L Co-IP, and ubiquitination assays","pmids":["32764116"],"confidence":"High","gaps":["Direct redox target controlling pSmad3-NEDD4L interaction not identified","Whether effect requires apical vs intracellular H2O2 unresolved"]},{"year":2021,"claim":"An unexpected enzymatic activity was demonstrated, with DUOX1/DUOX2 acting as NAADP-forming enzymes required for T-cell calcium signaling.","evidence":"In vitro NAADP-forming enzyme assay and Duox/Duoxa knockout mouse T cells with local and global Ca2+ imaging","pmids":["34784249"],"confidence":"High","gaps":["Relationship between NAADP synthesis and H2O2-generating activity not reconciled","Structural basis of NAADP formation not defined"]},{"year":2022,"claim":"Cell-type-specific deletion dissected epithelial versus macrophage DUOX1 contributions to chronic allergic airway disease.","evidence":"Conditional epithelial and macrophage DUOX1 knockouts in chronic HDM model with cytokine, mucus, and flow-cytometry readouts","pmids":["35654836"],"confidence":"High","gaps":["Macrophage-intrinsic redox targets not yet defined here","Crosstalk signals between compartments not fully mapped"]},{"year":2024,"claim":"The apical-targeting mechanism was resolved at the molecular level, showing DUOXA1 N-glycosylation directs DUOX1 to the apical membrane, distinct from DUOXA2's mechanism.","evidence":"MDCK co-expression with N-glycosylation mutants, immunofluorescence localization, and domain-swap experiments","pmids":["39126279"],"confidence":"High","gaps":["Glycan-reading trafficking machinery not identified","How glycosylation status is regulated physiologically unknown"]},{"year":2026,"claim":"Macrophage-intrinsic DUOX1 was mechanistically tied to profibrotic activation via oxidative Src activation and macrophage-fibroblast crosstalk.","evidence":"LysM-Cre conditional DUOX1 knockout with BMDM migration, cysteine oxidation, collagen, and saracatinib inhibition","pmids":["40986746"],"confidence":"High","gaps":["Oxidized Src cysteine residues in macrophages not pinpointed","Therapeutic targetability not established"]},{"year":2026,"claim":"The testosterone/GPRC6A-DUOX1 axis was extended to an in vivo physiological output, controlling hair-cycle progression and androgenetic hair loss.","evidence":"GPRC6A and Duox1 knockout mice and keratinocytes with H2O2, apoptosis, Ki-67, and hair-cycle staging after topical testosterone","pmids":["42047995"],"confidence":"Medium","gaps":["Downstream redox effectors in follicle keratinocytes not identified","Single-lab mouse evidence"]},{"year":null,"claim":"How DUOX1's H2O2-generating and NAADP-forming activities are coordinated, and which specific cysteine targets mediate each tissue-specific outcome, remains unresolved.","evidence":"","pmids":[],"confidence":"High","gaps":["No unified model reconciling NAADP synthesis with NADPH-oxidase output","Tissue-specific redox-target catalogs incomplete","Regulation of inactive dimer-of-dimers state in vivo not defined"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0016491","term_label":"oxidoreductase activity","supporting_discovery_ids":[0,1,12]},{"term_id":"GO:0140657","term_label":"ATP-dependent activity","supporting_discovery_ids":[0]},{"term_id":"GO:0140096","term_label":"catalytic activity, acting on a protein","supporting_discovery_ids":[3,4,5,6,21]},{"term_id":"GO:0016740","term_label":"transferase activity","supporting_discovery_ids":[14]}],"localization":[{"term_id":"GO:0005886","term_label":"plasma membrane","supporting_discovery_ids":[13,16,25]}],"pathway":[{"term_id":"R-HSA-162582","term_label":"Signal Transduction","supporting_discovery_ids":[3,4,5,9]},{"term_id":"R-HSA-168256","term_label":"Immune System","supporting_discovery_ids":[3,10,17,24]},{"term_id":"R-HSA-8953897","term_label":"Cellular responses to stimuli","supporting_discovery_ids":[6,8]},{"term_id":"R-HSA-1643685","term_label":"Disease","supporting_discovery_ids":[9,10,19,21]}],"complexes":["DUOX1-DUOXA1 complex"],"partners":["DUOXA1","IP3R1","SRC","ADAM17","EGFR","SHP-2","NEDD4L","P2Y2"],"other_free_text":[]}},"prefetch_data":{"uniprot":{"accession":"Q9NRD9","full_name":"Dual oxidase 1","aliases":["Large NOX 1","Long NOX 1","NADPH thyroid oxidase 1","Thyroid oxidase 1"],"length_aa":1551,"mass_kda":177.2,"function":"Generates hydrogen peroxide (H2O2) which is required for the activity of thyroid peroxidase/TPO and lactoperoxidase/LPO (PubMed:10806195, PubMed:15972824). Plays a role in thyroid hormone synthesis (PubMed:10806195). Also required for lactoperoxidase-mediated antimicrobial defense at the surface of mucosa (PubMed:12824283). Antimicrobial agent hypothiocyanite (OSCN-), which is produced by LPO from DUOX1-derived H2O2, promotes influenza virus inactivation by reducing viral binding to and entry into host cells (PubMed:34168077). Promotes antiviral immunity by increasing airway cytokine levels, promoting innate immune cell recruitment and reducing airway epithelial cell apoptosis (By similarity). In response to bacterial stimuli, activated by ATP and promotes EGFR/ERK signaling, ADAM17 activation, and EGFR ligand shedding, leading to enhanced CXCL8/IL8 expression and secretion in airway epithelia (PubMed:19386603). Synthesizes NAADP from its reduced NAADPH form which promotes Ca(2+) signaling during T cell activation (PubMed:34784249). In addition to its oxidase activity, has also been shown to have peroxidase activity through its N-terminal peroxidase-like domain (PubMed:11514595). However, another study showed that the isolated peroxidase-like domain does not bind heme and has no intrinsic peroxidase activity (PubMed:19460756)","subcellular_location":"Apical cell membrane","url":"https://www.uniprot.org/uniprotkb/Q9NRD9/entry"},"depmap":{"release":"DepMap","has_data":true,"is_common_essential":false,"resolved_as":"","url":"https://depmap.org/portal/gene/DUOX1","classification":"Not Classified","n_dependent_lines":3,"n_total_lines":1208,"dependency_fraction":0.0024834437086092716},"opencell":{"profiled":false,"resolved_as":"","ensg_id":"","cell_line_id":"","localizations":[],"interactors":[],"url":"https://opencell.sf.czbiohub.org/search/DUOX1","total_profiled":1310},"omim":[{"mim_id":"617792","title":"THIOREDOXIN DOMAIN-CONTAINING PROTEIN 11; TXNDC11","url":"https://www.omim.org/entry/617792"},{"mim_id":"615904","title":"PEROXIDASIN-LIKE PROTEIN; PXDNL","url":"https://www.omim.org/entry/615904"},{"mim_id":"614647","title":"COENZYME Q6, MONOOXYGENASE; COQ6","url":"https://www.omim.org/entry/614647"},{"mim_id":"612772","title":"DUAL OXIDASE MATURATION FACTOR 2; DUOXA2","url":"https://www.omim.org/entry/612772"},{"mim_id":"612771","title":"DUAL OXIDASE MATURATION FACTOR 1; DUOXA1","url":"https://www.omim.org/entry/612771"}],"hpa":{"profiled":true,"resolved_as":"","reliability":"","locations":[],"tissue_specificity":"Tissue enhanced","tissue_distribution":"Detected in many","driving_tissues":[{"tissue":"epididymis","ntpm":77.7},{"tissue":"esophagus","ntpm":69.8},{"tissue":"skin 1","ntpm":98.9}],"url":"https://www.proteinatlas.org/search/DUOX1"},"hgnc":{"alias_symbol":["NOXEF1","THOX1","LNOX1"],"prev_symbol":[]},"alphafold":{"accession":"Q9NRD9","domains":[{"cath_id":"1.10.640.10","chopping":"50-552","consensus_level":"high","plddt":96.0494,"start":50,"end":552},{"cath_id":"2.30.29.30","chopping":"642-756","consensus_level":"medium","plddt":82.9999,"start":642,"end":756},{"cath_id":"1.10.238","chopping":"763-850","consensus_level":"medium","plddt":73.9436,"start":763,"end":850},{"cath_id":"1.10.238.10","chopping":"852-926","consensus_level":"medium","plddt":70.8285,"start":852,"end":926},{"cath_id":"2.40.30.10","chopping":"1271-1374","consensus_level":"medium","plddt":86.4005,"start":1271,"end":1374},{"cath_id":"3.40.50.80","chopping":"1376-1551","consensus_level":"medium","plddt":88.2422,"start":1376,"end":1551},{"cath_id":"1.20.120","chopping":"1044-1268","consensus_level":"high","plddt":94.9059,"start":1044,"end":1268}],"viewer_url":"https://alphafold.ebi.ac.uk/entry/Q9NRD9","model_url":"https://alphafold.ebi.ac.uk/files/AF-Q9NRD9-F1-model_v6.cif","pae_url":"https://alphafold.ebi.ac.uk/files/AF-Q9NRD9-F1-predicted_aligned_error_v6.png","plddt_mean":85.62},"mouse_models":{"mgi_url":"https://www.informatics.jax.org/marker/summary?nomen=DUOX1","jax_strain_url":"https://www.jax.org/strain/search?query=DUOX1"},"sequence":{"accession":"Q9NRD9","fasta_url":"https://rest.uniprot.org/uniprotkb/Q9NRD9.fasta","uniprot_url":"https://www.uniprot.org/uniprotkb/Q9NRD9/entry","alphafold_viewer_url":"https://alphafold.ebi.ac.uk/entry/Q9NRD9"}},"corpus_meta":[{"pmid":"16111680","id":"PMC_16111680","title":"Differential 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Duox1.","date":"2010","source":"Free radical biology & medicine","url":"https://pubmed.ncbi.nlm.nih.gov/21146788","citation_count":79,"is_preprint":false},{"pmid":"19460756","id":"PMC_19460756","title":"Caenorhabditis elegans and human dual oxidase 1 (DUOX1) \"peroxidase\" domains: insights into heme binding and catalytic activity.","date":"2009","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/19460756","citation_count":67,"is_preprint":false},{"pmid":"27812543","id":"PMC_27812543","title":"DUOX1 mediates persistent epithelial EGFR activation, mucous cell metaplasia, and airway remodeling during allergic asthma.","date":"2016","source":"JCI insight","url":"https://pubmed.ncbi.nlm.nih.gov/27812543","citation_count":62,"is_preprint":false},{"pmid":"23349873","id":"PMC_23349873","title":"ATP-mediated transactivation of the epidermal growth factor receptor in airway epithelial cells involves DUOX1-dependent oxidation of Src and ADAM17.","date":"2013","source":"PloS one","url":"https://pubmed.ncbi.nlm.nih.gov/23349873","citation_count":60,"is_preprint":false},{"pmid":"17337509","id":"PMC_17337509","title":"Developmental regulation of DUOX1 expression and function in human fetal lung epithelial cells.","date":"2007","source":"American journal of physiology. 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Lung cellular and molecular physiology","url":"https://pubmed.ncbi.nlm.nih.gov/27612966","citation_count":15,"is_preprint":false},{"pmid":"33301419","id":"PMC_33301419","title":"Downregulation of epithelial DUOX1 in chronic obstructive pulmonary disease.","date":"2021","source":"JCI insight","url":"https://pubmed.ncbi.nlm.nih.gov/33301419","citation_count":13,"is_preprint":false},{"pmid":"29849884","id":"PMC_29849884","title":"DUOX1 Silencing in Mammary Cell Alters the Response to Genotoxic Stress.","date":"2018","source":"Oxidative medicine and cellular longevity","url":"https://pubmed.ncbi.nlm.nih.gov/29849884","citation_count":11,"is_preprint":false},{"pmid":"35654836","id":"PMC_35654836","title":"Macrophage-intrinsic DUOX1 contributes to type 2 inflammation and mucus metaplasia during allergic airway disease.","date":"2022","source":"Mucosal immunology","url":"https://pubmed.ncbi.nlm.nih.gov/35654836","citation_count":11,"is_preprint":false},{"pmid":"30559322","id":"PMC_30559322","title":"Duox1 Regulates Primary B Cell Function under the Influence of IL-4 through BCR-Mediated Generation of Hydrogen Peroxide.","date":"2018","source":"Journal of immunology (Baltimore, Md. : 1950)","url":"https://pubmed.ncbi.nlm.nih.gov/30559322","citation_count":8,"is_preprint":false},{"pmid":"39126279","id":"PMC_39126279","title":"The NADPH oxidases DUOX1 and DUOX2 are sorted to the apical plasma membrane in epithelial cells via their respective maturation factors DUOXA1 and DUOXA2.","date":"2024","source":"Genes to cells : devoted to molecular & cellular mechanisms","url":"https://pubmed.ncbi.nlm.nih.gov/39126279","citation_count":7,"is_preprint":false},{"pmid":"36625485","id":"PMC_36625485","title":"Diet-induced obesity worsens allergen-induced type 2/type 17 inflammation in airways by enhancing DUOX1 activation.","date":"2023","source":"American journal of physiology. Lung cellular and molecular physiology","url":"https://pubmed.ncbi.nlm.nih.gov/36625485","citation_count":5,"is_preprint":false},{"pmid":"33563887","id":"PMC_33563887","title":"Knockdown of Dual Oxidase 1 (DUOX1) Promotes Wound Healing by Regulating Reactive Oxygen Species (ROS) by Activation of Nuclear Kactor kappa B (NF-κB) Signaling.","date":"2021","source":"Medical science monitor : international medical journal of experimental and clinical research","url":"https://pubmed.ncbi.nlm.nih.gov/33563887","citation_count":5,"is_preprint":false},{"pmid":"32712621","id":"PMC_32712621","title":"Vitamin D Attenuates Hypoxia-Induced Injury in Rat Primary Neuron Cells through Downregulation of the Dual Oxidase 1 (DUOX1) Gene.","date":"2020","source":"Medical science monitor : international medical journal of experimental and clinical research","url":"https://pubmed.ncbi.nlm.nih.gov/32712621","citation_count":4,"is_preprint":false},{"pmid":"40621626","id":"PMC_40621626","title":"Geniposide Suppresses Tumor Progression Through DUOX1-Mediated Ferroptosis in Hepatocellular Carcinoma.","date":"2025","source":"The American journal of Chinese medicine","url":"https://pubmed.ncbi.nlm.nih.gov/40621626","citation_count":3,"is_preprint":false},{"pmid":"26504019","id":"PMC_26504019","title":"Site-specific Effects of DUOX1-Related Peroxidase on Intercellular Apoptosis Signaling.","date":"2015","source":"Anticancer research","url":"https://pubmed.ncbi.nlm.nih.gov/26504019","citation_count":3,"is_preprint":false},{"pmid":"36835420","id":"PMC_36835420","title":"DUOX1 Gene Missense Mutation Confers Susceptibility on Type 2 Amiodarone-Induced Thyrotoxicosis.","date":"2023","source":"International journal of molecular sciences","url":"https://pubmed.ncbi.nlm.nih.gov/36835420","citation_count":2,"is_preprint":false},{"pmid":"39447011","id":"PMC_39447011","title":"Eosinophil-airway epithelial cell crosstalk reveals the eosinophil-mediated DUOX1 upregulation in a murine allergic inflammation setting.","date":"2025","source":"Journal of leukocyte biology","url":"https://pubmed.ncbi.nlm.nih.gov/39447011","citation_count":2,"is_preprint":false},{"pmid":"36978957","id":"PMC_36978957","title":"Dual Role of DUOX1-Derived Reactive Oxygen Species in Melanoma.","date":"2023","source":"Antioxidants (Basel, Switzerland)","url":"https://pubmed.ncbi.nlm.nih.gov/36978957","citation_count":2,"is_preprint":false},{"pmid":"40088033","id":"PMC_40088033","title":"DUOX1 inhibits the progression of rheumatoid arthritis by regulating the NF-κB pathway in vitro.","date":"2025","source":"Allergologia et immunopathologia","url":"https://pubmed.ncbi.nlm.nih.gov/40088033","citation_count":2,"is_preprint":false},{"pmid":"30483939","id":"PMC_30483939","title":"Functional Annotation and Analysis of Dual Oxidase 1 (DUOX1): a Potential Anti-pyocyanin Immune Component.","date":"2018","source":"Interdisciplinary sciences, computational life sciences","url":"https://pubmed.ncbi.nlm.nih.gov/30483939","citation_count":2,"is_preprint":false},{"pmid":"30052308","id":"PMC_30052308","title":"DUOX1-mediated hydrogen peroxide release regulates sodium transport in H441 bronchiolar epithelial cells.","date":"2018","source":"Acta physiologica (Oxford, England)","url":"https://pubmed.ncbi.nlm.nih.gov/30052308","citation_count":1,"is_preprint":false},{"pmid":"40986746","id":"PMC_40986746","title":"The NADPH oxidase DUOX1 contributes to profibrotic macrophage activation and pulmonary fibrosis.","date":"2026","source":"American journal of respiratory cell and molecular biology","url":"https://pubmed.ncbi.nlm.nih.gov/40986746","citation_count":0,"is_preprint":false},{"pmid":"42047995","id":"PMC_42047995","title":"GPRC6A-Duox1 Axis Regulates the Hair Cycle Through H2O2 Generation.","date":"2026","source":"Tissue engineering and regenerative medicine","url":"https://pubmed.ncbi.nlm.nih.gov/42047995","citation_count":0,"is_preprint":false},{"pmid":"41910349","id":"PMC_41910349","title":"Corrigendum to: DUOX1 inhibits the progression of rheumatoid arthritis by regulating the NF-κB pathway in vitro.","date":"2026","source":"Allergologia et immunopathologia","url":"https://pubmed.ncbi.nlm.nih.gov/41910349","citation_count":0,"is_preprint":false}],"cost":{"stage1":{"model":"claude-sonnet-4-6","input_tokens":29817,"output_tokens":7305,"usd":0.099513,"stage1_stop_reason":"end_turn"},"stage2":{"model":"claude-opus-4-8","input_tokens":16482,"output_tokens":6772,"usd":0.125855,"stage2_stop_reason":"end_turn"},"total_usd":0.225368,"stage1_batch_id":"msgbatch_01AsR7g8dGTEiEw9U83bJeWn","stage2_batch_id":"msgbatch_01YV5ZyxtFNUZEjxRcpd9jzb","note":"batch pricing = 50% of standard"},"stage1_raw":"```json\n{\n  \"discoveries\": [\n    {\n      \"year\": 2020,\n      \"finding\": \"Cryo-EM structures of mouse DUOX1-DUOXA1 complex (with and without NADPH substrate) reveal atomic details of DUOX1-DUOXA1 interaction, a lipid-mediated NADPH-binding pocket, and the electron transfer path. A dimer-of-dimers configuration was identified as an inactive state, suggesting an oligomerization-dependent regulatory mechanism.\",\n      \"method\": \"Cryo-EM structural determination, biochemical assays\",\n      \"journal\": \"Nature structural & molecular biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Strong — cryo-EM structures with and without substrate, plus biochemical validation, multiple orthogonal methods in single rigorous study\",\n      \"pmids\": [\"32929281\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2009,\n      \"finding\": \"DUOX1 activity depends on calcium and functional EF-hand motifs. Uniquely among DUOX isoforms, DUOX1 (but not DUOX2) activity is stimulated by cAMP/forskolin via PKA-mediated phosphorylation on serine 955. This differential regulation was confirmed in human thyroid cells co-expressing DUOX1 with its maturation factor DUOXA1.\",\n      \"method\": \"Co-expression functional assay with DuoxA1/DuoxA2, pharmacological stimulation (forskolin, phorbol esters), site-specific mutagenesis of phosphorylation sites, membrane expression analysis\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Strong — in vitro enzymatic assay with mutagenesis, confirmed in human thyroid cells, multiple orthogonal methods in one rigorous study\",\n      \"pmids\": [\"19144650\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2009,\n      \"finding\": \"The isolated human DUOX1 peroxidase domain (hDUOX1 aa 1–593) does not bind heme and has no intrinsic peroxidase activity, in contrast to the C. elegans ortholog. Neither DUOX1 domain showed significant superoxide dismutase activity, suggesting the N-terminal motif does not directly convert superoxide to H2O2 in isolation.\",\n      \"method\": \"Baculovirus expression and purification of recombinant peroxidase domains, heme-binding assays, peroxidase activity assays, superoxide dismutase activity assays\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Strong — purified recombinant protein, multiple in vitro enzymatic assays, rigorous biochemical characterization\",\n      \"pmids\": [\"19460756\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2010,\n      \"finding\": \"DUOX1 binds to inositol 1,4,5-trisphosphate receptor 1 (IP3R1) in primary human CD4+ T cells and is required for early TCR-stimulated H2O2 production. DUOX1-derived H2O2 inactivates SHP-2 phosphatase, promoting ZAP-70 phosphorylation at Tyr-319 and its association with Lck and CD3ζ, thereby creating a positive feedback loop that sustains TCR signaling, store-operated Ca2+ entry, and ERK activation.\",\n      \"method\": \"Co-immunoprecipitation (DUOX1-IP3R1), siRNA/shRNA knockdown, TCR signaling assays, phosphorylation analysis, calcium flux measurement, cytokine production assays\",\n      \"journal\": \"Science signaling\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — reciprocal Co-IP, stable and transient knockdown with defined signaling phenotype, multiple orthogonal methods\",\n      \"pmids\": [\"20682913\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2013,\n      \"finding\": \"ATP activates DUOX1 via purinergic P2Y2 receptor stimulation, recruiting Src and DUOX1 into a signaling complex. DUOX1-derived H2O2 oxidizes cysteine residues within Src and ADAM17, activating both; ADAM17 then sheds EGFR ligands, leading to EGFR transactivation and downstream wound responses in airway epithelial cells.\",\n      \"method\": \"siRNA/shRNA knockdown, thiol-specific biotin labeling for cysteine oxidation detection, Co-IP (P2Y2R/Src/DUOX1 complex), EGFR phosphorylation assays, cell migration assays\",\n      \"journal\": \"PloS one\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — reciprocal Co-IP, stable shRNA, thiol-labeling mass spectrometry, multiple orthogonal readouts replicated across labs\",\n      \"pmids\": [\"23349873\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2016,\n      \"finding\": \"ATP-dependent EGFR transactivation in airway epithelial cells involves DUOX1-dependent sequential sulfenylation and S-glutathionylation of cysteine residues within EGFR and Src. The intermediate sulfenylation (not S-glutathionylation) is the activating modification; C797S EGFR variant abolishes H2O2-induced EGFR kinase activation, identifying Cys-797 as the redox-sensitive regulatory site.\",\n      \"method\": \"Recombinant protein kinase activity assays with H2O2/GSSG, C797S mutagenesis, redox-labeling strategies, DUOX1 siRNA/shRNA knockdown\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Strong — in vitro kinase assay with mutagenesis, multiple redox-labeling strategies, confirmed with DUOX1 knockdown\",\n      \"pmids\": [\"27650496\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2014,\n      \"finding\": \"DUOX1 mediates ATP-stimulated transient H2O2 production and S-glutathionylation of multiple proteins in airway epithelial cells, including β-actin, peroxiredoxin 1, Src, and MAPK phosphatase 1. These DUOX1-dependent S-glutathionylation events regulate cytoskeletal dynamics and MAPK signaling involved in cell migration.\",\n      \"method\": \"Stable shRNA knockdown, primary tracheal epithelial cells from DUOX1-deficient mice, biotin-tagged GSH labeling, avidin purification, global proteomics, H2O2 measurement\",\n      \"journal\": \"Redox biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — genetic KO mouse cells plus stable shRNA, proteomics with biotin-GSH, multiple orthogonal methods\",\n      \"pmids\": [\"24624333\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2009,\n      \"finding\": \"ATP-mediated DUOX1 activation in airway epithelial cells leads to H2O2-dependent activation of ERK1/2 and NF-κB pathways, intracellular oxidant signaling, and EGFR ligand shedding by ADAM17, culminating in IL-8/CXCL8 production in response to bacterial stimuli.\",\n      \"method\": \"DUOX1 siRNA knockdown, catalase (extracellular and intracellular scavenging), ERK/NF-κB pathway inhibitors, ADAM17 activation assays, IL-8 ELISA\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — siRNA knockdown plus pharmacological inhibitors, single lab, multiple readouts\",\n      \"pmids\": [\"19386603\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2015,\n      \"finding\": \"Ionizing radiation induces DUOX1-dependent H2O2 production in thyroid cells via a pathway involving p38 MAPK activation and IL-13 upregulation, which in turn sustains DUOX1 expression for days after irradiation. DUOX1-derived H2O2 causes persistent DNA double-strand breaks; catalase pretreatment or DUOX1 siRNA knockdown abrogates this damage.\",\n      \"method\": \"siRNA knockdown, p38 MAPK inhibitors, catalase treatment, DNA damage assays (γH2AX), dose-response H2O2 measurement, human thyroid tissue analysis\",\n      \"journal\": \"Proceedings of the National Academy of Sciences of the United States of America\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — siRNA knockdown with multiple rescue conditions, human and cell line models, multiple orthogonal methods\",\n      \"pmids\": [\"25848056\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"DUOX1-derived H2O2 sustains TGF-β1/Smad3 signaling in lung fibroblasts by preventing phospho-Smad3 degradation. Mechanistically, DUOX1 inhibits the interaction between phospho-Smad3 and the ubiquitin ligase NEDD4L, thereby preventing NEDD4L-mediated ubiquitination and proteasomal degradation of phospho-Smad3.\",\n      \"method\": \"DUOX1-deficient mouse models, primary human/mouse lung fibroblasts, Co-IP (pSmad3/NEDD4L interaction), ubiquitination assays, Smad3 phosphorylation kinetics\",\n      \"journal\": \"The European respiratory journal\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — genetic KO mice, primary cells, Co-IP with mechanistic ubiquitination assays, multiple orthogonal methods\",\n      \"pmids\": [\"32764116\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2016,\n      \"finding\": \"DUOX1 mediates persistent EGFR activation and cysteine oxidation within EGFR and Src in airway epithelial cells during allergic asthma. DUOX1 deficiency attenuated HDM-induced mucous metaplasia, subepithelial fibrosis, neutrophilic inflammation, type 2 cytokine production (IL-33, IL-13), and central airway resistance in mice.\",\n      \"method\": \"DUOX1-deficient mice, HDM mouse model, siRNA intratracheal delivery, EGFR cysteine oxidation assays, nasal epithelial cells from asthmatic subjects, EGFR/Src phosphorylation analysis\",\n      \"journal\": \"JCI insight\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — genetic KO mouse model plus siRNA intervention, human patient cells, multiple mechanistic and phenotypic readouts\",\n      \"pmids\": [\"27812543\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2010,\n      \"finding\": \"DUOX1 is the main source of H2O2 in urothelial cells, as demonstrated using Duox1 knockout mice. TRPV4 calcium channel activation elicits a calcium signal that stimulates DUOX1-dependent H2O2 production, and Duox1 knockout animals display altered pressure responses in the urinary bladder.\",\n      \"method\": \"Duox1 gene-deficient mouse model, H2O2 measurement, TRPV4 pharmacological activation, bladder pressure measurements\",\n      \"journal\": \"Free radical biology & medicine\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — genetic KO mouse with functional phenotypic readout, multiple methods\",\n      \"pmids\": [\"21146788\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2007,\n      \"finding\": \"Duox1 is the main source of H2O2 in the rat thyroid cell line PCCl3, as demonstrated by siRNA knockdown reducing H2O2 production in parallel with Duox1 protein. Re-expression of rat Duox1 fully rescued H2O2 production, while human Duox1 partial rescue was also observed.\",\n      \"method\": \"siRNA knockdown, lentiviral re-expression rescue, H2O2 production assays, Western blot\",\n      \"journal\": \"Experimental cell research\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — siRNA knockdown with rescue experiment, multiple methods confirming protein-H2O2 link\",\n      \"pmids\": [\"17643428\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"DUOX1 is sorted to the apical plasma membrane in epithelial cells via its maturation factor DUOXA1. N-glycosylation of DUOXA1 is required for apical targeting of DUOX1; impairment of DUOXA1 N-glycosylation results in DUOX1 mistargeting to the basolateral membrane. DUOX2 apical sorting by DUOXA2 is regulated by a distinct mechanism involving DUOXA2's C-terminal region rather than N-glycosylation.\",\n      \"method\": \"MDCK epithelial cell co-expression system, N-glycosylation mutants of DUOXA1/DUOXA2, immunofluorescence localization, domain-swap experiments\",\n      \"journal\": \"Genes to cells\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — mutagenesis combined with defined localization readout in polarized epithelial cells, multiple constructs tested\",\n      \"pmids\": [\"39126279\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"DUOX1 and DUOX2 function as NAADP-forming enzymes that convert NAADPH to NAADP in vitro under physiological conditions. In T cells, DUOX1 and DUOX2 are required for global Ca2+ signaling (4–12 min post-stimulation), while DUOX2 specifically drives early Ca2+ microdomain formation (first 15 s). Duoxa1/Duoxa2 double KO but not single Nox1 or Nox2 KO reduced local and global Ca2+ signaling.\",\n      \"method\": \"In vitro NAADP-forming enzyme assay, Duox1/Duox2/Duoxa1/Duoxa2 knockout mouse T cells, Ca2+ imaging (local microdomains and global), genetic epistasis with single and double KOs\",\n      \"journal\": \"Science signaling\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Strong — in vitro enzymatic assay plus multiple genetic KO models with Ca2+ imaging, replication across multiple KO lines\",\n      \"pmids\": [\"34784249\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2014,\n      \"finding\": \"Testosterone activates Duox1 in epidermal keratinocytes through GPRC6A, a membrane receptor that senses testosterone and activates Gq protein, leading to IP3 generation, intracellular calcium mobilization, and subsequent Duox1-dependent H2O2 generation. GPRC6A silencing abolished testosterone-induced calcium and H2O2 responses.\",\n      \"method\": \"GPRC6A siRNA knockdown, calcium imaging, H2O2 measurement, IP3 quantification, caspase-3 activation assays, 3D skin equivalent model\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — siRNA knockdown with multiple downstream readouts, single lab\",\n      \"pmids\": [\"25164816\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2017,\n      \"finding\": \"Autophagy protein ATG5 is required for apical membrane localization of DUOX1 in airway epithelial cells during chronic IL-13 stimulation. ATG5 depletion significantly reduced intracellular superoxide production without diminishing total DUOX1 protein levels, indicating that non-canonical autophagy regulates DUOX1 trafficking to the apical membrane rather than its expression.\",\n      \"method\": \"ATG5 siRNA, DUOX1 siRNA, confocal immunofluorescence localization, electron paramagnetic resonance spectroscopy for superoxide, OVA mouse model, LC3BII Western blot\",\n      \"journal\": \"Redox biology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — siRNA knockdown with localization and functional readout, supported by mouse model, single lab\",\n      \"pmids\": [\"28982074\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2018,\n      \"finding\": \"Duox1-derived H2O2 negatively regulates proliferative activity (but not Ig isotype production) in primary splenic B cells upon BCR stimulation. IL-4 costimulation boosts Duox1 expression and H2O2 production; Duox1-/- CD19+ B cells show enhanced proliferation, and catalase treatment of WT B cells mimics this effect. Immunized Duox1-/- mice show increased B cell expansion in germinal centers.\",\n      \"method\": \"Duox1 knockout mice, BCR stimulation assays, H2O2 measurement, proliferation assays, catalase treatment, in vivo immunization, germinal center analysis\",\n      \"journal\": \"Journal of immunology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — genetic KO mice with in vitro and in vivo validation, pharmacological rescue with catalase, multiple orthogonal readouts\",\n      \"pmids\": [\"30559322\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2015,\n      \"finding\": \"Duox1-derived H2O2 promotes late-phase neutrophil recruitment to wounds in zebrafish by inducing Cxcl8 expression via JNK/c-JUN/AP-1 signaling. This H2O2-driven cxcl8 induction is accompanied by histone modifications (increased H3K4me3, H3K9ac; decreased H3K9me3) at the cxcl8 promoter. ERK and NF-κB signaling were not involved.\",\n      \"method\": \"Zebrafish duox1 morpholino knockdown, wound assays, JNK inhibitors, ChIP for histone modifications, neutrophil recruitment quantification\",\n      \"journal\": \"Journal of immunology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — zebrafish ortholog model with pathway inhibitors and ChIP, single lab\",\n      \"pmids\": [\"25582859\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"DUOX1 missense mutation p.R1307Q impairs H2O2 generation and reduces DUOX1 mRNA and protein expression. A companion DUOXA1 mutation p.R56W similarly reduces DUOX1 expression and H2O2 generation, demonstrating that intact DUOXA1 is required for full DUOX1 activity; both mutations can cause congenital hypothyroidism.\",\n      \"method\": \"Patient sequencing, functional H2O2 generation assays in transfected cells, qRT-PCR, Western blot\",\n      \"journal\": \"Frontiers in endocrinology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — functional assay in transfected cells with patient-derived mutations, single lab\",\n      \"pmids\": [\"31428054\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2016,\n      \"finding\": \"Loss of DUOX1 expression in lung epithelial cells promotes epithelial-to-mesenchymal transition (EMT), characterized by loss of E-cadherin, gain of vimentin/collagen, increased migration, anchorage-independent growth, and enhanced cancer stem cell markers (CD133, ALDH1). Conversely, overexpression of DUOX1 in A549 cells reverses EMT features, and DUOX1 loss confers resistance to EGFR tyrosine kinase inhibitors.\",\n      \"method\": \"shRNA stable knockdown, DUOX1 overexpression, E-cadherin/vimentin Western blot, migration assays, anchorage-independent growth, in vivo xenograft invasion assay, erlotinib resistance assays\",\n      \"journal\": \"Oncogenesis\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — stable shRNA and overexpression with in vitro and in vivo readouts, single lab\",\n      \"pmids\": [\"27694834\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2026,\n      \"finding\": \"DUOX1 in macrophages contributes to profibrotic activation through oxidative activation of Src kinase via cysteine oxidation. Myeloid-specific conditional DUOX1 deletion attenuated pulmonary fibrosis, impaired MoMac recruitment, reduced collagen production, and improved oxygen saturation. DUOX1 also contributes to in vitro macrophage migration and EGFR ligand production involved in macrophage-fibroblast cross-talk.\",\n      \"method\": \"LysM-Cre conditional DUOX1 knockout, BMDM migration assays, cysteine oxidation assays, collagen production measurement, Src kinase activation assays, saracatinib pharmacological inhibition\",\n      \"journal\": \"American journal of respiratory cell and molecular biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — conditional genetic KO with multiple in vivo and in vitro mechanistic readouts and pharmacological validation\",\n      \"pmids\": [\"40986746\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2026,\n      \"finding\": \"The GPRC6A-Duox1 axis regulates hair cycle progression downstream of testosterone signaling. GPRC6A-deficient and Duox1-deficient keratinocytes fail to generate H2O2 in response to testosterone. Both GPRC6A KO and Duox1 KO mice show extended anagen phase, increased Ki-67 expression, longer hair, and resistance to testosterone-mediated hair loss.\",\n      \"method\": \"GPRC6A and Duox1 knockout mice and keratinocytes, H2O2 measurement, apoptosis assays, Ki-67 immunostaining, hair cycle staging, topical testosterone application\",\n      \"journal\": \"Tissue engineering and regenerative medicine\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — genetic KO mouse models with multiple readouts, single lab\",\n      \"pmids\": [\"42047995\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2018,\n      \"finding\": \"DUOX1-derived H2O2 regulates sodium transport (ENaC activity) in H441 bronchiolar epithelial cells. Dexamethasone-induced dome formation is inhibited by extracellular catalase or NADPH oxidase inhibitor DPI. H2O2 (0.2 mmol/L) acutely activates amiloride-sensitive ENaC currents in dome-forming cells, and there is a negative feedback loop where H2O2 inhibits ENaC gene transcription.\",\n      \"method\": \"Nystatin-perforated patch-clamp electrophysiology, catalase treatment, DPI inhibition, DUOX1 immunocytochemistry, RT-PCR, dome formation assays\",\n      \"journal\": \"Acta physiologica\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — electrophysiology with pharmacological inhibition, single lab, moderate mechanistic depth\",\n      \"pmids\": [\"30052308\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"Macrophage-intrinsic DUOX1 contributes to type 2 inflammation and mucus metaplasia during chronic HDM-driven allergic airway disease. Cell-type-specific conditional deletion revealed that epithelial DUOX1 drives acute HDM responses and most features of chronic inflammation, while macrophage DUOX1 additionally contributes to macrophage recruitment, type 2 cytokine production, and mucus metaplasia.\",\n      \"method\": \"Conditional cell-type-specific DUOX1 knockout (epithelial and macrophage lineages), HDM mouse model, cytokine measurement, mucus staining, flow cytometry\",\n      \"journal\": \"Mucosal immunology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — cell-type-specific conditional KO with defined mechanistic cellular phenotypes, multiple readouts\",\n      \"pmids\": [\"35654836\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2007,\n      \"finding\": \"DUOX1 expression and its maturation factor DUOXA1 are co-upregulated during differentiation of human fetal lung cells into alveolar type II cells (induced by DCI treatment), with DUOX1 protein localizing near the apical cell pole. DUOX1 knockdown by siRNA blocked increases in H2O2 production and acid secretion in differentiated type II cells, establishing DUOX1 as responsible for alveolar H2O2 and acid production.\",\n      \"method\": \"Human fetal lung primary cell culture, DCI differentiation protocol, siRNA knockdown, H2O2 measurement, intracellular acid measurement, immunofluorescence localization\",\n      \"journal\": \"American journal of physiology. Lung cellular and molecular physiology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — siRNA knockdown with localization and functional assays in primary cells, single lab\",\n      \"pmids\": [\"17337509\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"HIF-2α transcriptionally promotes DUOX1 expression (confirmed by dual luciferase assay), and arsenic-induced upregulation of DUOX1 suppresses GPX4 expression, promoting ferroptosis in kidney cells. DUOX1 siRNA knockdown significantly upregulated GPX4 and attenuated ferroptosis.\",\n      \"method\": \"siRNA knockdown, dual luciferase reporter assay for HIF-2α binding to DUOX1 promoter, GPX4 Western blot, ferroptosis markers, in vivo arsenic exposure model\",\n      \"journal\": \"The Science of the total environment\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — dual luciferase + siRNA + in vivo model, single lab, mechanistic pathway partially defined\",\n      \"pmids\": [\"37879473\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"DUOX1 is a calcium-dependent NADPH oxidase that co-assembles with its maturation factor DUOXA1 (which directs apical membrane targeting via N-glycosylation) to form an active enzyme complex whose cryo-EM structure reveals a lipid-mediated NADPH-binding pocket and an oligomerization-dependent inactive dimer-of-dimers state; its H2O2 output is regulated by PKA-mediated phosphorylation on Ser-955 and by calcium/EF-hand motifs, and acts through reversible cysteine oxidation (sulfenylation, S-glutathionylation) of targets including Src, ADAM17, EGFR (Cys-797), SHP-2, β-actin, and peroxiredoxin 1, thereby mediating positive feedback in TCR and EGFR signaling, innate airway epithelial host defense, TGF-β1/Smad3 amplification in fibrosis (by blocking NEDD4L-mediated phospho-Smad3 ubiquitination), NAADP synthesis for T cell Ca2+ signaling, and testosterone/GPRC6A-driven H2O2 production in keratinocytes.\"\n}\n```","stage2_raw":"{\n  \"mechanistic_narrative\": \"DUOX1 is a calcium-dependent NADPH oxidase that generates H2O2 at the apical surface of epithelial cells and serves as a signaling hub that converts diverse stimuli into redox-based control of downstream targets [#0, #1, #12]. Its activity depends on calcium acting through EF-hand motifs and is uniquely potentiated among DUOX isoforms by cAMP/PKA-mediated phosphorylation at Ser-955 [#1]; cryo-EM of the DUOX1-DUOXA1 complex resolves a lipid-mediated NADPH-binding pocket, the electron-transfer path, and an inactive dimer-of-dimers state indicating oligomerization-dependent regulation [#0]. Functional maturation and apical plasma-membrane delivery require the partner factor DUOXA1, whose N-glycosylation directs correct apical targeting, while non-canonical autophagy (ATG5) further controls DUOX1 trafficking to the apical membrane [#13, #16, #19]. DUOX1-derived H2O2 acts largely through reversible cysteine oxidation of signaling proteins: in airway epithelia, ATP/P2Y2-recruited DUOX1 sulfenylates and S-glutathionylates Src, ADAM17, and EGFR at Cys-797 to drive EGFR transactivation, cytoskeletal remodeling, and inflammatory output, and in T cells it inactivates SHP-2 to sustain ZAP-70/Lck-dependent TCR signaling and calcium entry [#3, #4, #5, #6]. Through these redox circuits DUOX1 amplifies type 2 airway inflammation and EGFR-driven asthma pathology, sustains TGF-β1/Smad3 fibrotic signaling by blocking NEDD4L-mediated phospho-Smad3 degradation, and contributes to macrophage profibrotic activation via oxidative Src activation [#9, #10, #21, #24]. DUOX1 and DUOX2 also act as NAADP-forming enzymes required for T-cell calcium signaling, and DUOX1 mediates testosterone/GPRC6A-driven H2O2 production controlling hair-cycle progression [#14, #22]. Loss-of-function missense mutations in DUOX1 (p.R1307Q) and DUOXA1 (p.R56W) impair H2O2 generation and can cause congenital hypothyroidism [#19].\",\n  \"teleology\": [\n    {\n      \"year\": 2007,\n      \"claim\": \"Establishing that DUOX1 is genuinely responsible for cellular H2O2 output was required before any signaling role could be assigned; knockdown-with-rescue and differentiation studies pinned H2O2 production to DUOX1 protein.\",\n      \"evidence\": \"siRNA knockdown with lentiviral re-expression rescue in rat thyroid cells, and DCI-differentiated human fetal alveolar type II cells with apical localization\",\n      \"pmids\": [\"17643428\", \"17337509\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Did not resolve how DUOX1 activity is switched on at the molecular level\", \"Apical localization mechanism not defined\"]\n    },\n    {\n      \"year\": 2009,\n      \"claim\": \"Defining the activation logic distinguished DUOX1 from DUOX2 and identified its regulatory inputs: calcium via EF-hands and uniquely cAMP/PKA phosphorylation at Ser-955.\",\n      \"evidence\": \"Co-expression functional assays with DUOXA1/DUOXA2, forskolin/phorbol stimulation, and site-specific phosphorylation mutagenesis in human thyroid cells\",\n      \"pmids\": [\"19144650\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Kinase that mediates PKA-independent inputs not mapped\", \"Structural basis of EF-hand coupling to catalysis unresolved at the time\"]\n    },\n    {\n      \"year\": 2009,\n      \"claim\": \"Whether the N-terminal peroxidase-homology domain of DUOX1 carries intrinsic enzymatic activity was tested directly; the isolated human domain neither binds heme nor shows peroxidase or SOD activity, constraining models of H2O2 generation.\",\n      \"evidence\": \"Baculovirus-expressed recombinant peroxidase domains with heme-binding, peroxidase, and SOD assays\",\n      \"pmids\": [\"19460756\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Function of the N-terminal domain in the full-length enzyme not established\", \"Did not test domain behavior within the assembled complex\"]\n    },\n    {\n      \"year\": 2009,\n      \"claim\": \"The first signaling role connected DUOX1-derived H2O2 to innate airway responses, linking ATP stimulation to ADAM17/EGFR shedding, ERK/NF-κB activation, and IL-8 production.\",\n      \"evidence\": \"DUOX1 siRNA knockdown with intracellular/extracellular catalase scavenging and pathway inhibitors in airway epithelial cells\",\n      \"pmids\": [\"19386603\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Direct molecular targets of H2O2 not identified\", \"Single-lab pharmacology-dependent readouts\"]\n    },\n    {\n      \"year\": 2010,\n      \"claim\": \"DUOX1 was shown to be a direct amplifier of TCR signaling, defining a redox positive-feedback loop in T cells via SHP-2 inactivation.\",\n      \"evidence\": \"DUOX1-IP3R1 Co-IP, siRNA/shRNA knockdown, phosphorylation and calcium-flux analysis in primary human CD4+ T cells\",\n      \"pmids\": [\"20682913\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Direct sulfenylation site on SHP-2 not mapped\", \"Mechanism of DUOX1-IP3R1 coupling unresolved\"]\n    },\n    {\n      \"year\": 2010,\n      \"claim\": \"Genetic deletion established DUOX1 as the dominant urothelial H2O2 source downstream of TRPV4 calcium signaling, with a bladder pressure-response phenotype.\",\n      \"evidence\": \"Duox1 knockout mice, H2O2 measurement, TRPV4 activation, and bladder pressure measurements\",\n      \"pmids\": [\"21146788\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Downstream redox targets in urothelium not identified\", \"Physiological signaling output incompletely defined\"]\n    },\n    {\n      \"year\": 2013,\n      \"claim\": \"The redox mechanism of EGFR transactivation was resolved at the protein level, showing DUOX1 oxidizes cysteines in Src and ADAM17 within a P2Y2-recruited complex.\",\n      \"evidence\": \"Reciprocal Co-IP, thiol-specific labeling, knockdown, and EGFR phosphorylation/migration assays in airway epithelial cells\",\n      \"pmids\": [\"23349873\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Specific oxidized cysteines on ADAM17 not pinpointed\", \"Kinetics of complex assembly not defined\"]\n    },\n    {\n      \"year\": 2014,\n      \"claim\": \"Proteome-scale S-glutathionylation mapping defined the breadth of DUOX1 redox targets controlling cytoskeleton and MAPK signaling.\",\n      \"evidence\": \"Stable shRNA and DUOX1-deficient mouse tracheal cells with biotin-GSH labeling and global proteomics\",\n      \"pmids\": [\"24624333\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Functional consequence of each glutathionylation event not individually validated\", \"Reversal/deglutathionylation kinetics not addressed\"]\n    },\n    {\n      \"year\": 2014,\n      \"claim\": \"A hormone-sensing input was identified, linking testosterone via GPRC6A/Gq/IP3/calcium to DUOX1 H2O2 production in keratinocytes.\",\n      \"evidence\": \"GPRC6A siRNA knockdown with calcium imaging, IP3 quantification, and H2O2 measurement in a 3D skin model\",\n      \"pmids\": [\"25164816\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Single-lab siRNA evidence\", \"Downstream redox targets in keratinocytes not identified\"]\n    },\n    {\n      \"year\": 2015,\n      \"claim\": \"DUOX1 was implicated in damage signaling, showing radiation-induced p38/IL-13-sustained DUOX1 expression drives persistent DNA double-strand breaks in thyroid cells.\",\n      \"evidence\": \"siRNA knockdown with p38 inhibitors and catalase rescue, γH2AX assays, and human thyroid tissue analysis\",\n      \"pmids\": [\"25848056\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Mechanism by which apical H2O2 reaches nuclear DNA not defined\", \"Long-term carcinogenic consequence not established\"]\n    },\n    {\n      \"year\": 2015,\n      \"claim\": \"An ortholog model defined an epigenetic mechanism for DUOX-driven inflammation, linking H2O2 to JNK/AP-1 and histone marks at the cxcl8 promoter.\",\n      \"evidence\": \"Zebrafish duox1 morpholino knockdown, JNK inhibitors, and ChIP for histone modifications at wounds\",\n      \"pmids\": [\"25582859\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Conservation of the histone-mark mechanism in mammals not shown\", \"Morpholino-based knockdown only\"]\n    },\n    {\n      \"year\": 2016,\n      \"claim\": \"The activating redox modification of EGFR was pinpointed: sulfenylation (not S-glutathionylation) of Cys-797 is the kinase-activating event.\",\n      \"evidence\": \"Recombinant kinase assays with H2O2/GSSG, C797S mutagenesis, redox labeling, and DUOX1 knockdown\",\n      \"pmids\": [\"27650496\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"In vivo occupancy of Cys-797 sulfenylation not quantified\", \"Reversal mechanism not detailed\"]\n    },\n    {\n      \"year\": 2016,\n      \"claim\": \"In vivo asthma modeling established DUOX1 as a driver of type 2 airway pathology via sustained EGFR/Src cysteine oxidation.\",\n      \"evidence\": \"DUOX1-deficient mice and intratracheal siRNA in an HDM model, with EGFR oxidation assays and asthmatic patient cells\",\n      \"pmids\": [\"27812543\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Relative contribution of epithelial vs other cell types not yet separated\", \"Causal cytokine hierarchy incompletely defined\"]\n    },\n    {\n      \"year\": 2016,\n      \"claim\": \"A context-dependent tumor-suppressive role emerged, with DUOX1 loss promoting EMT, stemness, and EGFR-TKI resistance in lung epithelial cells.\",\n      \"evidence\": \"shRNA knockdown and overexpression with EMT markers, migration, anchorage-independent growth, and xenograft assays\",\n      \"pmids\": [\"27694834\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Mechanism linking DUOX1 loss to EMT transcriptional program unresolved\", \"Single-lab in vitro/xenograft evidence\"]\n    },\n    {\n      \"year\": 2017,\n      \"claim\": \"Trafficking control of DUOX1 was extended to non-canonical autophagy, with ATG5 required for apical membrane localization independent of protein levels.\",\n      \"evidence\": \"ATG5 and DUOX1 siRNA, confocal localization, EPR superoxide measurement, and OVA mouse model\",\n      \"pmids\": [\"28982074\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Molecular machinery linking ATG5 to DUOX1 trafficking unknown\", \"Single-lab siRNA evidence\"]\n    },\n    {\n      \"year\": 2018,\n      \"claim\": \"DUOX1 was shown to negatively regulate adaptive immunity, restraining B-cell proliferation downstream of BCR/IL-4.\",\n      \"evidence\": \"Duox1 knockout mice, BCR stimulation, catalase rescue, and in vivo germinal center analysis\",\n      \"pmids\": [\"30559322\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Redox targets restraining B-cell proliferation not identified\", \"Distinction from antibody-isotype output mechanistically unexplained\"]\n    },\n    {\n      \"year\": 2018,\n      \"claim\": \"An ion-transport regulatory role was defined, with DUOX1 H2O2 acutely activating ENaC while transcriptionally repressing it in a feedback loop.\",\n      \"evidence\": \"Perforated patch-clamp electrophysiology with catalase/DPI inhibition in H441 bronchiolar cells\",\n      \"pmids\": [\"30052308\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Direct ENaC redox target site not mapped\", \"Single-lab pharmacology-based attribution\"]\n    },\n    {\n      \"year\": 2019,\n      \"claim\": \"Patient-derived mutations established a human disease link and reinforced DUOXA1 dependence, with DUOX1 p.R1307Q and DUOXA1 p.R56W impairing H2O2 generation in congenital hypothyroidism.\",\n      \"evidence\": \"Patient sequencing with functional H2O2 assays, qRT-PCR, and Western blot in transfected cells\",\n      \"pmids\": [\"31428054\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Single-lab functional validation\", \"Genotype-phenotype penetrance not fully established\"]\n    },\n    {\n      \"year\": 2021,\n      \"claim\": \"A fibrosis-amplifying mechanism was defined, with DUOX1 H2O2 stabilizing phospho-Smad3 by blocking NEDD4L-mediated ubiquitination.\",\n      \"evidence\": \"DUOX1-deficient mice, primary lung fibroblasts, pSmad3/NEDD4L Co-IP, and ubiquitination assays\",\n      \"pmids\": [\"32764116\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Direct redox target controlling pSmad3-NEDD4L interaction not identified\", \"Whether effect requires apical vs intracellular H2O2 unresolved\"]\n    },\n    {\n      \"year\": 2021,\n      \"claim\": \"An unexpected enzymatic activity was demonstrated, with DUOX1/DUOX2 acting as NAADP-forming enzymes required for T-cell calcium signaling.\",\n      \"evidence\": \"In vitro NAADP-forming enzyme assay and Duox/Duoxa knockout mouse T cells with local and global Ca2+ imaging\",\n      \"pmids\": [\"34784249\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Relationship between NAADP synthesis and H2O2-generating activity not reconciled\", \"Structural basis of NAADP formation not defined\"]\n    },\n    {\n      \"year\": 2022,\n      \"claim\": \"Cell-type-specific deletion dissected epithelial versus macrophage DUOX1 contributions to chronic allergic airway disease.\",\n      \"evidence\": \"Conditional epithelial and macrophage DUOX1 knockouts in chronic HDM model with cytokine, mucus, and flow-cytometry readouts\",\n      \"pmids\": [\"35654836\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Macrophage-intrinsic redox targets not yet defined here\", \"Crosstalk signals between compartments not fully mapped\"]\n    },\n    {\n      \"year\": 2024,\n      \"claim\": \"The apical-targeting mechanism was resolved at the molecular level, showing DUOXA1 N-glycosylation directs DUOX1 to the apical membrane, distinct from DUOXA2's mechanism.\",\n      \"evidence\": \"MDCK co-expression with N-glycosylation mutants, immunofluorescence localization, and domain-swap experiments\",\n      \"pmids\": [\"39126279\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Glycan-reading trafficking machinery not identified\", \"How glycosylation status is regulated physiologically unknown\"]\n    },\n    {\n      \"year\": 2026,\n      \"claim\": \"Macrophage-intrinsic DUOX1 was mechanistically tied to profibrotic activation via oxidative Src activation and macrophage-fibroblast crosstalk.\",\n      \"evidence\": \"LysM-Cre conditional DUOX1 knockout with BMDM migration, cysteine oxidation, collagen, and saracatinib inhibition\",\n      \"pmids\": [\"40986746\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Oxidized Src cysteine residues in macrophages not pinpointed\", \"Therapeutic targetability not established\"]\n    },\n    {\n      \"year\": 2026,\n      \"claim\": \"The testosterone/GPRC6A-DUOX1 axis was extended to an in vivo physiological output, controlling hair-cycle progression and androgenetic hair loss.\",\n      \"evidence\": \"GPRC6A and Duox1 knockout mice and keratinocytes with H2O2, apoptosis, Ki-67, and hair-cycle staging after topical testosterone\",\n      \"pmids\": [\"42047995\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Downstream redox effectors in follicle keratinocytes not identified\", \"Single-lab mouse evidence\"]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"How DUOX1's H2O2-generating and NAADP-forming activities are coordinated, and which specific cysteine targets mediate each tissue-specific outcome, remains unresolved.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"High\",\n      \"gaps\": [\"No unified model reconciling NAADP synthesis with NADPH-oxidase output\", \"Tissue-specific redox-target catalogs incomplete\", \"Regulation of inactive dimer-of-dimers state in vivo not defined\"]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0016491\", \"supporting_discovery_ids\": [0, 1, 12]},\n      {\"term_id\": \"GO:0140657\", \"supporting_discovery_ids\": [0]},\n      {\"term_id\": \"GO:0140096\", \"supporting_discovery_ids\": [3, 4, 5, 6, 21]},\n      {\"term_id\": \"GO:0016740\", \"supporting_discovery_ids\": [14]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005886\", \"supporting_discovery_ids\": [13, 16, 25]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-162582\", \"supporting_discovery_ids\": [3, 4, 5, 9]},\n      {\"term_id\": \"R-HSA-168256\", \"supporting_discovery_ids\": [3, 10, 17, 24]},\n      {\"term_id\": \"R-HSA-8953897\", \"supporting_discovery_ids\": [6, 8]},\n      {\"term_id\": \"R-HSA-1643685\", \"supporting_discovery_ids\": [9, 10, 19, 21]}\n    ],\n    \"complexes\": [\"DUOX1-DUOXA1 complex\"],\n    \"partners\": [\"DUOXA1\", \"IP3R1\", \"Src\", \"ADAM17\", \"EGFR\", \"SHP-2\", \"NEDD4L\", \"P2Y2\"],\n    \"other_free_text\": []\n  }\n}","audit_flag":null,"evaluation":{"pairwise":"win","faith_supported":7,"faith_total":7,"faith_pct":100.0}}