{"gene":"ALOX15","run_date":"2026-06-09T22:02:43","timeline":{"discoveries":[{"year":2023,"finding":"ALOX15 catalyzes conversion of arachidonic acid to 15-HpETE, which promotes cardiomyocyte ferroptosis by binding Pgc1α to the ubiquitin ligase RNF34, leading to ubiquitin-dependent Pgc1α degradation, attenuated mitochondrial biogenesis, and abnormal mitochondrial morphology. Myocardial-specific knockout of Alox15 alleviated I/R injury; ML351 (specific ALOX15 inhibitor) rescued cardiac function.","method":"Mouse cardiac-specific knockout, metabolomics, loss/gain-of-function experiments, protein interaction assays, pharmacological inhibition (ML351)","journal":"Circulation","confidence":"High","confidence_rationale":"Tier 2 / Strong — multiple orthogonal methods (KO, metabolomics, protein interaction, pharmacological inhibition) in a rigorous single study with clear mechanistic pathway placement","pmids":["36987924"],"is_preprint":false},{"year":2022,"finding":"ALOX15 is the primary mediator of ischemia-provoked phospholipid peroxidation in cardiomyocytes, specifically oxidizing PUFA-containing phosphatidylethanolamines to generate ferroptotic lipid signals; identified by multi-omics and confirmed by chemogenetic approaches.","method":"Multi-omics analysis, chemogenetic approaches, animal and in vitro models","journal":"Signal transduction and targeted therapy","confidence":"High","confidence_rationale":"Tier 2 / Strong — multi-omics plus chemogenetic confirmation, multiple experimental systems","pmids":["35970840"],"is_preprint":false},{"year":2004,"finding":"Alox15 (12/15-lipoxygenase) negatively regulates peak bone mineral density in mice; crossbreeding with Alox15 knockout mice confirmed this skeletal role, and pharmacological inhibitors of the enzyme improved bone density and strength in two rodent osteoporosis models.","method":"Genetic mapping, Alox15 knockout mouse crossbreeding, pharmacological inhibition in rodent models","journal":"Science (New York, N.Y.)","confidence":"High","confidence_rationale":"Tier 2 / Strong — knockout confirmation plus pharmacological validation in two independent animal models","pmids":["14716014"],"is_preprint":false},{"year":2000,"finding":"15-LOX-1 (ALOX15) enzymatic activity in colorectal cancer cells metabolizes linoleic acid to 13-S-HODE, which induces apoptosis; NSAIDs (sulindac, NS-398) increase 15-LOX-1 protein expression and enzymatic activity, and pharmacological inhibition of 15-LOX-1 blocked NSAID-induced apoptosis, which was rescued by exogenous 13-S-HODE but not linoleic acid.","method":"15-LOX-1 protein expression assay, enzymatic activity measurement, 13-S-HODE quantification, pharmacological inhibition (caffeic acid), exogenous metabolite rescue in colorectal cancer cell lines (RKO, HT-29)","journal":"Journal of the National Cancer Institute","confidence":"High","confidence_rationale":"Tier 2 / Strong — enzyme activity confirmed, metabolite quantified, inhibitor plus rescue experiment in two cell lines establish the mechanistic pathway","pmids":["10904086"],"is_preprint":false},{"year":2016,"finding":"Evolutionary alteration of ALOX15 reaction specificity—from 12-lipoxygenating in lower primates to 15-lipoxygenating in higher primates (humans, chimpanzees, orangutans)—optimizes lipoxin synthase activity; 15-lipoxygenating ALOX15 variants have >5-fold higher lipoxin synthase activity than 12-lipoxygenating variants. Triad residues (e.g., Ile418) are key specificity determinants confirmed by mutagenesis and QM/MM calculations.","method":"Cloning and expression of multiple ALOX15 orthologs, lipoxin synthase activity assays, site-directed mutagenesis (humIle418Ala), QM/MM energy calculations, molecular dynamics simulations","journal":"Proceedings of the National Academy of Sciences of the United States of America","confidence":"High","confidence_rationale":"Tier 1 / Strong — in vitro reconstitution with mutagenesis, computational validation, and functional quantification across multiple orthologs","pmids":["27412860"],"is_preprint":false},{"year":2017,"finding":"Mammalian ALOX15 orthologs exhibit pronounced dual positional specificity with DHA, forming similar amounts of 14- and 17-HpDHA; ALOX15 orthologs prefer DHA and EPA over arachidonic acid when equimolar amounts are supplied. The reaction specificity strongly depends on fatty acid substrate chemistry.","method":"Recombinant expression of eight mammalian ALOX15 orthologs as his-tag fusion proteins, substrate oxygenation assays with multiple PUFAs (AA, EPA, DHA), product characterization","journal":"Biochimica et biophysica acta. Molecular and cell biology of lipids","confidence":"High","confidence_rationale":"Tier 1 / Strong — systematic in vitro enzymatic characterization across eight species orthologs with multiple substrates","pmids":["28400162"],"is_preprint":false},{"year":2015,"finding":"Mutagenesis of triad determinants (Leu353, Ile593, Ala404) in rat Alox15 alters reaction specificity with free fatty acids and complex ester lipids; Leu353Phe mutation converts the enzyme from 12-lipoxygenating to 15-lipoxygenating activity. The Ala404Gly exchange induced subtle alterations in enantioselectivity, partially supporting the Coffa/Brash concept.","method":"Recombinant rat Alox15 expression (prokaryotic and eukaryotic systems), multiple site-directed mutagenesis, enzymatic oxygenation assays with free fatty acids and phospholipid substrates (biomembranes, HDL)","journal":"Archives of biochemistry and biophysics","confidence":"High","confidence_rationale":"Tier 1 / Strong — systematic mutagenesis with in vitro enzymatic characterization across multiple substrates and multiple mutants","pmids":["25731857"],"is_preprint":false},{"year":2020,"finding":"15-LOX-1 (ALOX15) peroxidates linoleic acid in phosphatidylinositol-3-phosphates (PI3P_LA → PI3P_13-HODE), which decreases PI3P binding to SNX17 and LRP5, inhibits LRP5 recycling from endosomes to the plasma membrane, and promotes LRP5 lysosomal degradation, thereby suppressing Wnt/β-catenin signaling and colorectal carcinogenesis.","method":"Transgenic mouse intestinal expression of ALOX15, Apc-mutant mouse model, azoxymethane CRC model, in vitro reconstitution of lipid peroxidation, PI3P-SNX17-LRP5 interaction assays, endosomal trafficking assays","journal":"Cell reports","confidence":"High","confidence_rationale":"Tier 2 / Strong — multiple in vivo models plus defined biochemical mechanism of PI3P modification and LRP5 trafficking","pmids":["32814052"],"is_preprint":false},{"year":2006,"finding":"A C-to-T SNP at position c.-292 in the ALOX15 promoter creates a novel binding site for transcription factor SPI1 (PU.1); alleles carrying c.-292T transcribe twice as efficiently as c.-292C alleles in macrophages that constitutively express SPI1, but not in lung epithelial cells lacking SPI1. EMSA confirmed selective SPI1 binding to the mutant promoter; primary macrophages from heterozygous carriers expressed 3× more ALOX15 mRNA.","method":"Luciferase transcription assays, EMSA (electrophoretic mobility shift assay), SPI1 core-binding site mutagenesis, qRT-PCR in primary human macrophages from genotyped donors","journal":"Human mutation","confidence":"High","confidence_rationale":"Tier 1 / Moderate — EMSA + promoter mutagenesis + functional transcription assays + primary human cell validation in a single study","pmids":["16320347"],"is_preprint":false},{"year":2009,"finding":"ALOX15 transcription in colon cancer cells is suppressed by recruitment of the NuRD repressor complex (containing MTA1 and HDAC1) to the region -120 to -391 of the 15-LOX-1 promoter; HDAC inhibitors (depsipeptide) reduce NuRD recruitment and reactivate 15-LOX-1 transcription. MTA1 expression in colorectal cancer is negatively correlated with 15-LOX-1 expression.","method":"siRNA knockdown of HDAC1, HDAC2, MTA1; luciferase reporter assays with 15-LOX-1 promoter constructs; chromatin immunoprecipitation; HDAC inhibitor treatment (depsipeptide)","journal":"Oncogene","confidence":"High","confidence_rationale":"Tier 2 / Moderate — ChIP, siRNA knockdown, luciferase reporter assays, and in vivo correlation in a single study with multiple orthogonal methods","pmids":["19198625"],"is_preprint":false},{"year":2006,"finding":"GATA-6 transcription factor overexpression contributes to 15-LOX-1 transcriptional silencing in colon cancer cells; GATA-6 siRNA knockdown alone is insufficient but significantly contributes to restoring 15-LOX-1 expression and inducing apoptosis when combined with NSAID (NS-398) or HDAC inhibitor (sodium butyrate) treatment.","method":"GATA-6 siRNA transfection in Caco-2 and HCT-116 cells, 15-LOX-1 expression measurement, apoptosis assays, NS-398 and sodium butyrate treatments, paired human colorectal cancer tissue analysis","journal":"FASEB journal","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — siRNA knockdown with functional readouts in two cell lines plus human tissue validation, but single lab study","pmids":["17167069"],"is_preprint":false},{"year":2007,"finding":"A coding SNP T560M in ALOX15 results in a near-null variant with 20-fold reduction in catalytic activity compared to wild-type enzyme, as demonstrated by in vitro enzymatic assays.","method":"ALOX15 resequencing, in vitro enzymatic activity assay of T560M vs T560 variants, genotyping in population studies","journal":"Atherosclerosis","confidence":"Medium","confidence_rationale":"Tier 1 / Weak — in vitro enzymatic characterization of single SNP variant, single study","pmids":["17959182"],"is_preprint":false},{"year":2010,"finding":"ALOX15 (15-lipoxygenase) is present in sperm cytoplasmic droplets (CDs) and required for normal epididymal sperm maturation; Alox15 knockout male mice have significantly more spermatozoa with retained CDs containing intact mitochondria in both caput and cauda epididymidis, compared to wild-type where CDs contain only hollow membrane vesicles with no intact mitochondria.","method":"Alox15 knockout mouse analysis, differential interference contrast microscopy (DIC), transmission electron microscopy (TEM) of epididymal sperm from Alox15 vs wild-type males","journal":"Cell and tissue research","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — knockout model with direct ultrastructural phenotypic readout showing organelle localization and functional consequence","pmids":["20449608"],"is_preprint":false},{"year":2016,"finding":"Systemic inactivation of the Alox15 gene normalizes reduced fertility and sperm motility in male mice heterozygous for catalytically inactive Gpx4 (Sec46Ala-Gpx4+/-); Alox15 deficiency rescues sperm midpiece structural alterations and motility defects, demonstrating that ALOX15-mediated lipid oxidation is functionally antagonized by GPX4 in male reproductive function.","method":"Genetic cross of Gpx4 knock-in mice with Alox15 knockout mice, fertility assessment, sperm motility analysis, ultrastructural analysis","journal":"The Journal of biological chemistry","confidence":"High","confidence_rationale":"Tier 2 / Strong — epistasis by genetic double-mutant rescue experiment with defined fertility and motility phenotypic readouts","pmids":["27634046"],"is_preprint":false},{"year":2014,"finding":"Alox15 deficiency does NOT rescue homozygous Gpx4 catalytically inactive knock-in mice (Sec46Ala-Gpx4+/+) from embryonic lethality (E6-7), indicating that Alox15-mediated lipid peroxidation is not the primary cause of embryonic lethality in GPX4-null mice.","method":"Genetic cross of Sec46Ala-Gpx4 knock-in mice with Alox15 knockout mice, embryonic viability assessment, embryo staging","journal":"Antioxidants & redox signaling","confidence":"High","confidence_rationale":"Tier 2 / Strong — epistasis via genetic double-mutant showing clear negative result (no rescue), rigorous embryonic staging","pmids":["25313597"],"is_preprint":false},{"year":2010,"finding":"Alox15 gene disruption in ApoE-/- mice protects against nonalcoholic fatty liver disease; Alox15 deficiency reduces hepatic steatosis, inflammation, macrophage infiltration, attenuates glucose intolerance and insulin resistance, upregulates insulin receptor substrate-2, and exerts opposite effects on hepatic JNK and AMPK phosphorylation. Hepatocytes from Alox15-deficient mice are less vulnerable to TNFα-induced programmed cell death.","method":"ApoE-/-/Alox15-/- double knockout mice, hepatocyte isolation and TNFα challenge, JNK and AMPK phosphorylation assays, glucose/insulin tolerance tests, gene expression analysis","journal":"Hepatology (Baltimore, Md.)","confidence":"High","confidence_rationale":"Tier 2 / Strong — genetic double-KO model with multiple orthogonal functional readouts and defined signaling pathway placement","pmids":["20967760"],"is_preprint":false},{"year":2013,"finding":"12/15-lipoxygenase (Alox15) is expressed in macrophages but not B and T cells of NOD mice; deletion of Alox15 decreases proinflammatory cytokine expression in macrophages, splenocytes from NOD-Alox15(null) mice cannot transfer diabetes in adoptive transfer, and absence of Alox15 maintains islet health and beta cell mass, demonstrating that Alox15 regulates macrophage inflammation driving autoimmunity in type 1 diabetes.","method":"qRT-PCR cell-type expression analysis, adoptive transfer model, Western blotting, beta cell mass quantification in NOD-Alox15(null) mice","journal":"PloS one","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — KO model with adoptive transfer epistasis and multiple cellular readouts, single lab","pmids":["23437231"],"is_preprint":false},{"year":2022,"finding":"Vagus nerve stimulation (VNS) promotes resolution of inflammation and increases specialized proresolving mediators (SPMs) from DHA and n-3 DPA metabolomes; this VNS-mediated shift toward proresolving lipid mediator profile is inverted in Alox15-deficient mice, establishing Alox15 as a required effector in the neural-immune inflammatory reflex pathway.","method":"Electrical VNS in zymosan-induced peritonitis mouse model, lipid mediator metabololipidomics, Alox15-deficient mouse comparison, α7nAChR-deficient mouse comparison","journal":"Proceedings of the National Academy of Sciences of the United States of America","confidence":"High","confidence_rationale":"Tier 2 / Strong — genetic epistasis using Alox15 KO mice plus metabololipidomics in a defined in vivo model with clear pathway placement","pmids":["35622894"],"is_preprint":false},{"year":2021,"finding":"Alox15-derived 15-HEPE (15-hydroxy eicosapentaenoic acid) is the anti-colitis mediator responsible for the protective effect of increased n-3 PUFA in fat1 transgenic mice; Alox15 gene inactivation in fat1 mice abolishes their protection from DSS- and TNBS-induced colitis, and intraperitoneal 15S-HEPE administration protects wild-type mice from colitis.","method":"fat1×Alox15-/- genetic cross, DSS and TNBS colitis models, Alox15 lipid metabolite analysis, 15S-HEPE intraperitoneal treatment in wild-type mice","journal":"FASEB journal","confidence":"High","confidence_rationale":"Tier 2 / Strong — genetic epistasis with double KO plus pharmacological rescue with defined metabolite in two colitis models","pmids":["33710695"],"is_preprint":false},{"year":2018,"finding":"Alox15/ALOX15B are expressed in cardiac fibroblasts; hypoxia significantly increases ALOX15 and ALOX15B gene and protein expression in fibroblasts from failing human hearts. Conditioned medium from hypoxic fibroblasts decreases beating frequency of human iPSC-derived cardiomyocytes in an ALOX15-dependent manner (effect blocked by ALOX15 inhibition).","method":"Gene expression in isolated cardiac fibroblasts under hypoxia vs normoxia, 15-HETE measurement, paracrine co-culture with iPSC-derived cardiomyocytes, calcium imaging, ALOX15 inhibition","journal":"PloS one","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — functional paracrine assay with ALOX15 inhibition confirms dependency, single lab study","pmids":["30138423"],"is_preprint":false},{"year":2017,"finding":"ALOX15 (Alox15) is expressed in rat brain, with highest mRNA and protein expression in the prefrontal cortex; it is localized to dendrites by immuno-electron microscopy. Alox15 inhibition or antisense knockdown in prefrontal cortex significantly decreased resolvin D1 levels, blocked hippocampo-prefrontal cortex long-term potentiation, and increased errors in T-maze spatial working memory, establishing Alox15 as mediating DHA-to-resolvin D1 conversion required for synaptic plasticity and memory.","method":"RT-PCR, Western blot, immunohistochemistry, immuno-electron microscopy, LC-MS for resolvin D1, pharmacological inhibition and antisense knockdown of Alox15 in prefrontal cortex, LTP recording, T-maze behavioral test","journal":"Molecular neurobiology","confidence":"High","confidence_rationale":"Tier 2 / Strong — multiple orthogonal methods including subcellular localization (immuno-EM), metabolite measurement, and functional behavioral/electrophysiological readouts","pmids":["28181190"],"is_preprint":false},{"year":2017,"finding":"Alox15 expression in neuroblastoma cells is regulated by histone acetylation; HDAC inhibitors (TSA, sodium butyrate, Class I inhibitors MS-275 and depsipeptide) upregulate Alox15 mRNA. The p300 histone acetyltransferase (HAT) inhibitor C646 modulates TSA- and retinoic acid-induced Alox15 upregulation, indicating p300 HAT activity is required for Alox15 transcriptional induction during neuronal differentiation.","method":"HDAC inhibitor treatment of SH-SY5Y cells, Class I-specific HDAC inhibitors, p300 HAT inhibitor (C646) co-treatment, retinoic acid differentiation, qRT-PCR, primary murine cortical neuron developmental expression","journal":"Neurochemical research","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — pharmacological dissection of specific HDAC classes and HAT requirement, single lab, no ChIP","pmids":["29235036"],"is_preprint":false},{"year":2017,"finding":"ALOX15 upregulation leads to hepatic overproduction of 13-HODE, which induces oxidative stress, ER stress, lipid metabolic disorder, and apoptosis; ALOX15 knockout prevents alcohol-induced liver damage through attenuation of these pathways. Linoleic acid (not ethanol or acetaldehyde) induces ALOX15 expression in hepatocytes.","method":"Chronic alcohol feeding mouse model, ALOX15 knockout mice, ALOX15 inhibitor (PD146176) treatment, 13-HODE exposure to Hepa-1c1c7 cells, oxidative stress, ER stress, and apoptosis markers","journal":"Scientific reports","confidence":"High","confidence_rationale":"Tier 2 / Strong — genetic KO plus pharmacological inhibition plus defined metabolite mechanistic link, multiple orthogonal readouts","pmids":["28827690"],"is_preprint":false},{"year":2022,"finding":"SSAT1 overexpression upregulates ALOX15 expression in neurons and promotes ferroptosis by decreasing GPX4 and SLC7A11 levels; SSAT1 knockdown downregulates ALOX15 and reduces lipid hydroperoxide production. ALOX15 inhibitor PD146176 partially reverses SSAT1 upregulation-induced ferroptosis, placing ALOX15 downstream of SSAT1 in neuronal ferroptosis signaling.","method":"SSAT1 knockdown/overexpression in primary cortical neurons and tMCAO/R mouse model, ALOX15 inhibitor (PD146176) treatment, iron/ROS/lipid peroxide measurements, GPX4/SLC7A11 protein levels","journal":"Neuroscience","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — epistasis placement via KD/OE plus pharmacological rescue, single lab","pmids":["35090880"],"is_preprint":false},{"year":2023,"finding":"DHODH inhibits neuronal ferroptosis after spinal cord injury via the P53/ALOX15 signaling pathway: DHODH inhibits P53 expression, which in turn suppresses ALOX15 expression, reducing lipid peroxidation and ferroptosis.","method":"Rat SCI model, erastin-induced PC12 cells, DHODH overexpression/inhibition, P53 and ALOX15 expression analysis, lipid peroxide and mitochondrial damage measurement","journal":"CNS neuroscience & therapeutics","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — in vivo and in vitro models with pathway placement, single lab study","pmids":["36942513"],"is_preprint":false},{"year":2020,"finding":"Efferocytosis by alternatively activated macrophages potentiates ALOX15 expression through LXR activation; AC uptake upregulates LXR-dependent gene expression and suppresses SREBP-2-dependent gene expression via NPC1-mediated sterol transport. LXRα/β silencing attenuates this ALOX15 potentiation. Enhanced ALOX15 expression increases capacity to synthesize SPM precursors 15-HETE and 17-HDHA and resolvin D5.","method":"Apoptotic cell efferocytosis in primary human macrophages, global transcriptome analysis, LXR agonist (T0901317), NPC1 inhibition, LXRα/β siRNA silencing, lipidomics (15-HETE, 17-HDHA, resolvin D5 measurement)","journal":"Cell death and differentiation","confidence":"High","confidence_rationale":"Tier 2 / Strong — primary human cells, transcriptomics, siRNA knockdown, sterol transport inhibition, and metabolite production all supporting a defined regulatory mechanism","pmids":["33177619"],"is_preprint":false},{"year":2008,"finding":"Alox15 is induced in the small and large intestine and liver of iron-deficient rats; increased Alox15 protein leads to production of 12-HETE, 13-HODE, and 13-HOTE (confirmed by HPLC), representing a perturbation of intestinal lipid metabolism driven by Alox15 activity during iron deficiency.","method":"Gene chip studies in iron-deficient and Belgrade rats, TaqMan qRT-PCR, polyclonal antibody immunodetection, HPLC lipid analysis","journal":"American journal of physiology. Gastrointestinal and liver physiology","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — direct lipid product identification by HPLC linked to Alox15 induction, single lab study","pmids":["18258795"],"is_preprint":false},{"year":2010,"finding":"Selenoprotein P (SelP) has in vitro lipid hydroperoxidase activity against 15-HpETE (the ALOX15 product), attenuates 15-HpETE oxidation in cellular assays, and reduces oxidative damage in a transcellular assay when 15-LOX-1 is metabolically active, identifying SelP as an antioxidant counterpart to 15-LOX-1.","method":"In vitro lipid hydroperoxidase assay with recombinant SelP and 15-HpETE, cellular 15-HpETE oxidation assay, transcellular assay with metabolically active 15-LOX-1","journal":"Prostaglandins, leukotrienes, and essential fatty acids","confidence":"Medium","confidence_rationale":"Tier 1 / Moderate — in vitro enzymatic assay with defined substrate (15-HpETE) and cellular validation, single lab","pmids":["20826080"],"is_preprint":false},{"year":2022,"finding":"The ALOX15 specificity switch during mammalian evolution follows a Triad Concept: >90% of mammalian ALOX15 orthologs catalyze 12-lipoxygenation; mammals ranked above gibbons express 15-lipoxygenating orthologs; triad determinant mutagenesis studies on 44 novel orthologs confirmed the specificity predictions. Gibbon ALOX15 represents a transition enzyme with dual specificity.","method":"Sequence analysis of 152 mammalian ALOX15 orthologs, expression of 44 novel recombinant ALOX15 orthologs, enzymatic specificity assays, extensive mutagenesis of triad determinants","journal":"Frontiers in cell and developmental biology","confidence":"High","confidence_rationale":"Tier 1 / Strong — large-scale systematic in vitro enzymatic characterization plus mutagenesis across 44 expressed proteins from diverse species","pmids":["35531094"],"is_preprint":false},{"year":2017,"finding":"Mutagenesis of porcine ALOX15 Val418 and Val419 sequence determinants converts the enzyme from 12-lipoxygenating to 15-lipoxygenating activity; QM/MM calculations showed that the Val418Ile+Val419Met double mutant has a 6.0 kcal/mol higher barrier for 12-lipoxygenation than 15-lipoxygenation, explaining the specificity switch by altering enzyme-substrate complex geometry.","method":"Site-directed mutagenesis of porcine ALOX15 crystal structure, molecular docking, MD simulations, QM/MM calculations","journal":"Chemistry (Weinheim an der Bergstrasse, Germany)","confidence":"High","confidence_rationale":"Tier 1 / Strong — mutagenesis on a crystallized enzyme combined with QM/MM calculations providing mechanistic basis for catalytic specificity","pmids":["29154477"],"is_preprint":false},{"year":2020,"finding":"Gln596 of mammalian ALOX15 does not directly interact with the carboxylate of arachidonic acid; instead, mutations at Gln596 destabilize secondary/tertiary structure and impair allosteric enzyme properties by disturbing the electrostatic interaction network. MD simulations suggest dimer interface geometry depends on substrate bound in the active site, and Gln596Ala impairs allosteric communication between monomers.","method":"Site-directed mutagenesis of rabbit and human ALOX15 at Gln596, structural stability assays, steady-state kinetics, MD simulations, substrate docking","journal":"Biochimica et biophysica acta. Molecular and cell biology of lipids","confidence":"Medium","confidence_rationale":"Tier 1 / Moderate — mutagenesis with kinetics and MD simulations in a single lab study","pmids":["32151768"],"is_preprint":false},{"year":2022,"finding":"N-substituted 5-(1H-indol-2-yl)-2-methoxyanilines are allosteric inhibitors of the linoleate oxygenase activity of rabbit and human ALOX15; they selectively inhibit oxygenation of linoleic acid at submicromolar concentrations without affecting arachidonic acid oxygenation. Steady-state kinetics, mutagenesis, and MD simulations of the ALOX15 dimer model indicate binding at the active site of one monomer induces conformational changes in the other monomer, compromising productive linoleic acid-enzyme complex formation.","method":"In vitro enzymatic assays (steady-state kinetics), site-directed mutagenesis, MD simulations of ALOX15 dimer model, synthesis of inhibitor series","journal":"Journal of medicinal chemistry","confidence":"High","confidence_rationale":"Tier 1 / Strong — systematic kinetics + mutagenesis + MD simulations establishing allosteric mechanism in a single rigorous study","pmids":["35073698"],"is_preprint":false},{"year":2021,"finding":"Rabbit ALOX15 and human ALOX15B exist as two conformers with distinct electrophoretic mobilities in native PAGE; MD simulations and mutagenesis suggest motional flexibility in aqueous solution impacting enzymatic properties, consistent with a dynamic monomer-dimer equilibrium with substrate-dependent conformational heterogeneity.","method":"Native polyacrylamide electrophoresis, in silico docking, MD simulations, site-directed mutagenesis, kinetic measurements","journal":"International journal of molecular sciences","confidence":"Medium","confidence_rationale":"Tier 1 / Moderate — direct conformational observation by native PAGE plus computational and mutagenesis support, single lab","pmids":["33807076"],"is_preprint":false},{"year":2004,"finding":"15-LOX-1 overexpression in HCT-116 colon cancer cells induces ERK1/2 phosphorylation, decreases p21 (Cip/WAF1) expression, and increases cell growth; its product 13-S-HODE reverses NaBT-induced p21 expression. LOX inhibitor (NDGA) decreased ERK1/2 phosphorylation and increased p21 expression in 15-LOX-1 overexpressing cells.","method":"15-LOX-1 transfection in HCT-116 cells, ERK1/2 phosphorylation assay, p21 expression analysis, NDGA inhibitor treatment, sodium butyrate treatment","journal":"Prostaglandins & other lipid mediators","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — gain-of-function plus pharmacological inhibition with defined signaling readouts, single lab","pmids":["15165036"],"is_preprint":false},{"year":2017,"finding":"Intestinally-targeted transgenic expression of ALOX15 in mice inhibits DSS-induced colitis from promoting azoxymethane-induced colorectal tumorigenesis; ALOX15 suppresses TNF-α, IL-1β/NF-κB, and IL-6/STAT3 inflammatory signaling pathways that drive colorectal cancer promotion.","method":"ALOX15 intestinal transgenic mouse model, DSS-colitis model, azoxymethane carcinogen model, cytokine/signaling pathway analysis","journal":"Prostaglandins & other lipid mediators","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — in vivo transgenic model with defined pathway readouts, single lab","pmids":["28089732"],"is_preprint":false},{"year":2018,"finding":"Systemic Alox15 deficiency in female mice protects against DSS-induced colitis by sustaining expression of the tight junction protein ZO-1 and maintaining intestinal epithelial barrier function; exogenous 12S-HETE (the major mouse Alox15 product) enantioselectively compromises transepithelial electric resistance in colon epithelial cells. Transgenic overexpression of human ALOX15 intensified inflammatory symptoms.","method":"Alox15-/- mouse DSS colitis model, 12S-HETE epithelial barrier assay (transepithelial electric resistance), ZO-1 immunostaining, ALOX15 transgenic overexpression, 12-HETE in vivo quantification","journal":"Biochimica et biophysica acta. Molecular and cell biology of lipids","confidence":"High","confidence_rationale":"Tier 2 / Strong — KO and transgenic overexpression models plus direct mechanistic link to tight junction protein and barrier function via metabolite assay","pmids":["29702245"],"is_preprint":false},{"year":2013,"finding":"IFN-γ suppresses ALOX15 expression during resolution of PMA-induced skin inflammation; neutralization of endogenous IFN-γ boosts ALOX15 expression and LXA4 production, promoting proresolution effects. Exogenous LXA4 (an ALOX15 product) recapitulates the proresolution effect of IFN-γ neutralization.","method":"In vivo PMA-induced skin inflammation model in mice, IFN-γ neutralizing antibody, ALOX15 expression analysis, exogenous LXA4 treatment","journal":"Mediators of inflammation","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — in vivo neutralization experiment with ALOX15 expression readout and metabolite rescue, single lab","pmids":["23818745"],"is_preprint":false},{"year":2018,"finding":"Spinal TLR4 activation by intrathecal KLA increases 15-LOX-1 expression specifically in microglia (not astrocytes) and increases 12-lipoxygenase-mediated hepoxilin production in lumbar spinal cord; intrathecal pretreatment with 15-LOX-1-specific inhibitors (ML127, ML351, CDC) or a selective antibody against rat 15-LOX-1 completely abrogated TLR4-induced tactile allodynia, establishing spinal microglial ALOX15 as mediating TLR4-dependent neuropathic pain.","method":"Intrathecal KLA administration, lipidomics of spinal cord, primary spinal microglia/astrocyte cultures, selective ALOX15 antibody neutralization, ML127/ML351/CDC pharmacological inhibition, HEK-293T heterologous expression of rat 15-LOX-1 for inhibitor testing","journal":"Pain","confidence":"High","confidence_rationale":"Tier 2 / Strong — cell-type-specific expression plus multiple selective pharmacological inhibitors with behavioral readouts and in vitro enzymatic confirmation","pmids":["30130298"],"is_preprint":false},{"year":2014,"finding":"15-LOX-1 re-expression in colon cancer cells (HCT116, HT29LMM, LoVo) under hypoxia inhibits HIF-1α protein expression and stability, reduces VEGF expression, angiogenesis, cancer cell migration, invasion, and survival under hypoxia, demonstrating that 15-LOX-1 suppresses hypoxia-driven metastatic mechanisms.","method":"15-LOX-1 re-expression in three colon cancer cell lines under hypoxia, HIF-1α protein stability assay, VEGF ELISA, angiogenesis assay, migration and invasion assays","journal":"Cancer medicine","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — gain-of-function in three cell lines with multiple functional readouts, single lab","pmids":["24634093"],"is_preprint":false},{"year":2024,"finding":"FGF21 deficiency results in abnormal activation of the ALOX15/15-HETE pathway, triggering innate immunity-dominated pro-inflammatory responses in liver grafts; pharmacological recombinant FGF21 administration protects against hepatic I/R injury and suppresses the ALOX15/15-HETE axis, particularly in steatotic livers.","method":"Hepatic I/R injury and orthotopic LT mouse models, FGF21 KO mice, recombinant FGF21 treatment, ALOX15/15-HETE pathway analysis, arachidonic acid metabolomics","journal":"Nature communications","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — genetic KO and pharmacological rescue with defined metabolite pathway readout, single lab","pmids":["39362839"],"is_preprint":false},{"year":2024,"finding":"PEBP1 forms a complex with ALOX15 in macrophages under psychological stress, mediating membrane phospholipid peroxidation that compromises macrophage phagocytic capacity; inhibition of the ALOX15/PEBP1 interaction (e.g., by isochlorogenic acid C, ICAC) attenuates phospholipid peroxidation and restores phagocytic function in the tumor microenvironment.","method":"Co-immunoprecipitation, microscale thermophoresis (MST) binding assay, molecular docking, LC-MS/MS phospholipidomics, flow cytometry phagocytosis assay, in vivo breast cancer model","journal":"Phytomedicine","confidence":"High","confidence_rationale":"Tier 2 / Strong — direct protein-protein interaction confirmed by Co-IP and MST, metabolite-function link established by phospholipidomics and phagocytosis assay","pmids":["38492368"],"is_preprint":false},{"year":2021,"finding":"Alox15 deficiency in chronic kidney disease (5/6 nephrectomy) mice improves renal function and inhibits interstitial fibrosis; mediator lipidomics revealed significantly higher PGD2 in Alox15-/- CKD kidneys, and exogenous PGD2 dose-dependently suppresses TGF-β1-induced type I collagen and αSMA in renal epithelial cells, suggesting ALOX15 suppresses PGD2, which is anti-fibrotic.","method":"Alox15-/- mouse 5/6 nephrectomy CKD model, mediator lipidomics, PGD2 treatment of NRK-52E and HK-2 cells, fibrosis marker quantification","journal":"Clinical and experimental nephrology","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — KO mouse model plus in vitro metabolite rescue experiment, single lab","pmids":["33595729"],"is_preprint":false},{"year":2023,"finding":"Bmal1 inhibits RSL3-induced ferroptosis in AML cells by recruiting EZH2 to the EBF3 promoter to enhance its methylation and suppress EBF3 expression; EBF3 binds to the ALOX15 promoter to enhance ALOX15 expression, promoting ferroptosis. Thus ALOX15 is downstream of the Bmal1/EZH2/EBF3 axis in ferroptosis regulation in AML.","method":"Bmal1/EBF3 knockdown/overexpression in HL-60 and NB4 cells, EZH2 ChIP at EBF3 promoter, EBF3 ChIP at ALOX15 promoter, subcutaneous AML xenograft model, ferroptosis assays (MDA, GSH, lipid peroxidation, iron)","journal":"Cancer science","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — ChIP at multiple promoters establishing pathway, supported by in vivo xenograft validation, single lab","pmids":["37271497"],"is_preprint":false},{"year":2025,"finding":"ALOX15 promotes lipid accumulation in hepatocytes through the PPARγ/CD36 pathway; ALOX15 overexpression increases intracellular fatty acid uptake and lipotoxicity, which are reversed by PPARγ antagonist (GW9662) or CD36 inhibitor. GLP-1 receptor agonist liraglutide improves hepatic lipid accumulation in MASLD mice by suppressing the ALOX15/PPARγ/CD36 pathway.","method":"ALOX15 lentiviral overexpression and siRNA KD in HepG2 cells, palmitic acid lipotoxicity model, PPARγ antagonist (GW9662) and CD36 inhibitor treatment, ALOX15 metabolomics (12-HpETE, 15-HpETE), transcriptomics, HFD/STZ mouse model","journal":"Antioxidants & redox signaling","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — gain- and loss-of-function with defined downstream pathway inhibitors and in vivo model, single lab","pmids":["39815992"],"is_preprint":false},{"year":2024,"finding":"Alpha-tocopherol (Vitamin E) downregulates Alox15 expression and inhibits ferroptosis after spinal cord injury; mutation of the 87th residue of Alox15 weakens the inhibitory effect of alpha-tocopherol on Alox15, identifying this residue as important for the alpha-tocopherol/Alox15 interaction.","method":"SCI rat model, alpha-tocopherol treatment, Alox15 expression analysis, Alox15 87th residue mutation, immunofluorescence localization, in vitro ROS and lipid peroxidation assays","journal":"Biomedicine & pharmacotherapy","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — residue mutagenesis supports direct interaction, combined in vivo/in vitro, single lab","pmids":["38754264"],"is_preprint":false},{"year":2024,"finding":"Lactate-induced HIF1A transcriptional activation (via histone lactylation of the HIF1A promoter) exacerbates severe acute pancreatitis by inducing ferroptosis through the ACSL4/LPCAT3/ALOX15 pathway; HIF1A absence or overexpression changes SAP severity by modulating this ferroptosis pathway. Elevated lactate from glycolytic abnormalities is the upstream driver.","method":"l-Arg SAP mouse model, sh-HIF1A mice, CHIP-qPCR for HIF1A promoter lactylation, luciferase reporter, ACSL4/LPCAT3/ALOX15 expression analysis, WB for lipid peroxidation markers","journal":"Journal of cellular biochemistry","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — ChIP-qPCR and luciferase reporter establish HIF1A regulation, pathway placement via KO mouse, single lab","pmids":["39676583"],"is_preprint":false},{"year":2023,"finding":"In AML cells, EBF3 binds to the ALOX15 promoter and enhances its expression, promoting ferroptosis; this regulatory axis is confirmed by EBF3 ChIP at the ALOX15 promoter and rescue experiments with EBF3 knockdown.","method":"ChIP-qPCR of EBF3 binding at ALOX15 promoter, EBF3 overexpression/knockdown, ALOX15 ferroptosis assays in AML cells","journal":"Cancer science","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — ChIP at ALOX15 promoter with functional rescue, part of a larger study (same PMID as Bmal1/EZH2/EBF3/ALOX15)","pmids":["37271497"],"is_preprint":false},{"year":2024,"finding":"PER1 directly binds to the SREBF2 promoter to repress SREBF2 transcription; SREBF2 in turn binds to the ALOX15 promoter to repress ALOX15 transcription; PER1 overexpression thus increases ALOX15 expression and promotes ferroptosis in granulosa cells in PCOS.","method":"PER1/SREBF2 promoter binding assays (ChIP implied from 'binds to promoter'), PER1 overexpression/SREBF2 overexpression/ALOX15 silencing in granulosa cells, ferroptosis marker measurement, PCOS mouse model","journal":"Biochimica et biophysica acta. Molecular basis of disease","confidence":"Low","confidence_rationale":"Tier 3 / Moderate — promoter binding described but specific ChIP methodology not clearly confirmed in abstract; pathway placement supported by rescue experiments","pmids":["38653359"],"is_preprint":false},{"year":2025,"finding":"ALOX15 modulates ferroptosis in doxorubicin-induced cardiotoxicity via the ROS-mediated MAPK signaling pathway; ALOX15 loss of function ameliorates Dox-induced ferroptosis by inhibiting ROS-mediated MAPK activation. Linoleic acid increases susceptibility to Dox-induced damage, abolished by ALOX15 inhibition.","method":"Dox mouse and H9C2 cell models, ALOX15 inhibition/silencing/overexpression, ROS measurement, MAPK phosphorylation assays, ferroptosis markers","journal":"Antioxidants & redox signaling","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — gain- and loss-of-function with defined downstream signaling readout (MAPK phosphorylation), single lab","pmids":["40455659"],"is_preprint":false},{"year":2025,"finding":"Wogonin directly binds to and inhibits ALOX15, suppressing cardiomyocyte ferroptosis in septic cardiomyopathy; pharmacological ALOX15 inhibition (ML351) recapitulates wogonin's cardioprotection. In LPS-challenged cardiomyocytes, ALOX15 overexpression or its downstream metabolite 15-HpETE abolishes the anti-ferroptotic effects of wogonin.","method":"CLP septic cardiomyopathy mouse model, molecular docking, direct binding assay, ML351 pharmacological inhibition, ALOX15 overexpression in HL-1 cardiomyocytes, 15-HpETE rescue experiment","journal":"Acta pharmacologica Sinica","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — direct binding assay plus pharmacological inhibitor recapitulation and metabolite rescue, single lab","pmids":["40205009"],"is_preprint":false},{"year":2023,"finding":"Macrophage-derived exosomal miRNA-660-5p is transported into cervical cancer cells where it attenuates ALOX15 expression to suppress ferroptosis; macrophage miRNA-660-5p upregulation depends on autocrine IL4/IL13-activated STAT6 pathway.","method":"Exosome isolation and transfer, miRNA-660-5p transfection/inhibition, ALOX15 expression analysis, ferroptosis assays in cervical cancer cells, STAT6 pathway analysis in macrophages","journal":"Acta pharmaceutica Sinica. B","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — defined miRNA-mediated ALOX15 suppression mechanism via exosome transfer with upstream STAT6 pathway, single lab","pmids":["37425043"],"is_preprint":false},{"year":2023,"finding":"Knock-in mice expressing the arachidonic acid 15-lipoxygenating Leu353Phe Alox15 mutant (functionally humanized) develop normally and are fertile but show modified plasma oxylipidomes; aged male Alox15-KI mice develop dysfunctional erythropoiesis (reduced hematocrit, lower erythrocyte counts, attenuated hemoglobin) with improved ex vivo osmotic resistance of red blood cells, in a sex-specific manner.","method":"Alox15 Leu353Phe knock-in mouse generation, plasma oxylipidomics, hematological parameter measurement, osmotic resistance assay","journal":"Cellular & molecular biology letters","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — knock-in mouse with defined functional humanization and erythropoietic phenotype, single lab","pmids":["38030974"],"is_preprint":false}],"current_model":"ALOX15 (15-lipoxygenase-1) is a lipid-peroxidizing enzyme that oxygenates polyunsaturated fatty acids—principally arachidonic acid and linoleic acid—to generate bioactive hydroperoxy lipid metabolites (15-HpETE, 13-HODE, 15-HETE, 15-HEPE, and SPM precursors); its reaction specificity is determined by active-site triad residues and evolved from 12-lipoxygenating in lower mammals to 15-lipoxygenating in Hominidae, optimizing lipoxin biosynthesis. ALOX15-derived 15-HpETE drives cardiomyocyte ferroptosis by targeting Pgc1α for ubiquitin-dependent degradation via RNF34, and ALOX15-mediated PI3P peroxidation (PI3P_13-HODE) suppresses LRP5 endosomal recycling to inhibit Wnt/β-catenin signaling in colonic epithelium. Its transcription is silenced in cancer cells by the NuRD repressor complex (HDAC1/MTA1) and by GATA-6 overexpression, and is activated in macrophages by a SPI1-binding promoter SNP and by LXR following efferocytosis. ALOX15 functions as a pro-ferroptotic enzyme downstream of SSAT1/SAT1, p53, and Bmal1/EBF3 transcriptional axes, is antagonized by GPX4, and mediates neural-immune resolution via Alox15-derived SPMs in the vagal inflammatory reflex."},"narrative":{"mechanistic_narrative":"ALOX15 (15-lipoxygenase-1) is a non-heme iron lipid-peroxidizing enzyme that oxygenates polyunsaturated fatty acids and their esterified derivatives to generate bioactive hydroperoxy and hydroxy lipid mediators governing inflammation resolution, ferroptosis, and tissue homeostasis [PMID:36987924, PMID:10904086, PMID:33177619]. The enzyme acts on free arachidonic acid, linoleic acid, EPA, and DHA, with marked substrate preference for n-3 PUFAs, producing 15-HpETE/15-HETE, 13-HODE, 15-HEPE, and DHA-derived 14-/17-HpDHA that serve as precursors for specialized pro-resolving mediators including lipoxins and resolvins [PMID:10904086, PMID:28400162, PMID:33710695, PMID:33177619]. Its positional specificity is set by an active-site triad (e.g. Leu353/Ile418, Val418/Val419, Ala404), and during mammalian/primate evolution mutation of these determinants converted the ancestral 12-lipoxygenating enzyme into the 15-lipoxygenating form found in higher primates, optimizing lipoxin synthase activity [PMID:27412860, PMID:25731857, PMID:35531094, PMID:29154477]. Beyond free fatty acids, ALOX15 peroxidizes membrane and organelle phospholipids—oxidizing PUFA-phosphatidylethanolamines to drive ferroptotic signals and linoleate within PI3P to suppress LRP5 endosomal recycling and Wnt/β-catenin signaling—and forms a complex with PEBP1 to direct phospholipid peroxidation [PMID:35970840, PMID:32814052, PMID:38492368]. Through 15-HpETE generation it executes ferroptotic cell death programs, in cardiomyocytes by promoting RNF34-dependent Pgc1α degradation and impaired mitochondrial biogenesis, an activity antagonized by GPX4 [PMID:36987924, PMID:27634046]. ALOX15 sits downstream of multiple transcriptional and metabolic ferroptosis axes including SSAT1, p53/DHODH, and Bmal1/EZH2/EBF3, while its own transcription is repressed by the NuRD (MTA1/HDAC1) complex and GATA-6 in cancer cells, activated by an SPI1-binding promoter SNP and LXR in macrophages, and tuned by histone acetylation [PMID:16320347, PMID:19198625, PMID:35090880, PMID:36942513, PMID:37271497, PMID:33177619]. Physiologically, ALOX15-derived mediators drive resolution of inflammation in vagal neural-immune circuits, synaptic plasticity and memory, intestinal barrier integrity, and skin and colitis resolution, while its peroxidation activity also contributes to NAFLD, alcoholic liver injury, neuropathic pain, autoimmune diabetes, and reduced bone mineral density [PMID:14716014, PMID:35622894, PMID:28181190, PMID:29702245, PMID:23818745, PMID:30130298].","teleology":[{"year":2000,"claim":"Established that ALOX15 is not merely a metabolic enzyme but a pro-apoptotic effector in colon cancer cells via its linoleic acid product 13-S-HODE, answering whether its catalytic output has direct cell-fate consequences.","evidence":"Enzyme activity assays, 13-S-HODE quantification, inhibitor plus metabolite rescue in RKO and HT-29 colorectal cancer cells","pmids":["10904086"],"confidence":"High","gaps":["Downstream apoptotic effectors of 13-S-HODE not defined","Does not address phospholipid substrates or in vivo relevance"]},{"year":2004,"claim":"Extended ALOX15 function beyond cancer to skeletal physiology, showing the enzyme negatively regulates peak bone mineral density and is a pharmacological target for bone strength.","evidence":"Genetic mapping, Alox15 knockout crossbreeding, and inhibitor treatment in two rodent osteoporosis models","pmids":["14716014"],"confidence":"High","gaps":["Lipid mediator linking ALOX15 to osteoblast/osteoclast biology not identified","Cell type responsible not pinpointed"]},{"year":2006,"claim":"Defined cis- and trans-acting transcriptional control of ALOX15, showing a promoter SNP creates an SPI1 site driving macrophage-selective expression, and GATA-6 contributes to silencing in cancer.","evidence":"Luciferase assays, EMSA, promoter mutagenesis, primary human macrophage qRT-PCR; GATA-6 siRNA in Caco-2/HCT-116 cells","pmids":["16320347","17167069"],"confidence":"High","gaps":["How SPI1 and repressors are coordinated across cell types unresolved","GATA-6 effect insufficient alone"]},{"year":2009,"claim":"Identified the chromatin machinery silencing ALOX15 in cancer, placing the NuRD complex (MTA1/HDAC1) on the promoter and explaining HDAC-inhibitor reactivation.","evidence":"siRNA knockdown of HDAC1/MTA1, ChIP, luciferase reporters, depsipeptide in colon cancer cells","pmids":["19198625"],"confidence":"High","gaps":["Signals recruiting NuRD to the promoter not defined","Generality beyond colon cancer untested"]},{"year":2016,"claim":"Resolved the structural basis and evolutionary logic of ALOX15 reaction specificity, showing triad residues convert 12- to 15-lipoxygenation and optimize lipoxin synthesis in higher primates.","evidence":"Ortholog cloning/expression, lipoxin synthase assays, site-directed mutagenesis, QM/MM and MD simulations across many species","pmids":["27412860","25731857","35531094","29154477"],"confidence":"High","gaps":["Physiological consequence of the evolutionary switch in vivo not directly tested at this stage","Specificity with esterified substrates less fully characterized"]},{"year":2017,"claim":"Broadened the substrate scope, showing ALOX15 prefers EPA and DHA over arachidonic acid and produces dual-specificity SPM precursors, reframing it as a key generator of n-3-derived resolution mediators.","evidence":"Recombinant expression of eight mammalian orthologs with multiple PUFA substrates and product characterization","pmids":["28400162"],"confidence":"High","gaps":["In vivo substrate selection under physiological PUFA ratios not established","Tissue-specific product spectra unknown"]},{"year":2017,"claim":"Demonstrated CNS roles for ALOX15, localizing it to prefrontal cortical dendrites and showing its DHA-to-resolvin D1 conversion is required for synaptic plasticity and working memory.","evidence":"Immuno-EM, LC-MS for resolvin D1, inhibition/antisense knockdown, LTP recording, and T-maze testing in rat","pmids":["28181190"],"confidence":"High","gaps":["Receptor for resolvin D1 mediating LTP not identified","Human relevance untested"]},{"year":2020,"claim":"Established that ALOX15 oxidizes esterified lipids in trafficking compartments, peroxidizing PI3P-linoleate to block LRP5 recycling and suppress Wnt/β-catenin signaling in colon.","evidence":"Transgenic and Apc/azoxymethane mouse models, in vitro lipid peroxidation, PI3P-SNX17-LRP5 interaction and endosomal trafficking assays","pmids":["32814052"],"confidence":"High","gaps":["Whether PI3P peroxidation occurs in non-intestinal tissues unknown","Selectivity for PI3P vs other phosphoinositides not fully mapped"]},{"year":2020,"claim":"Connected ALOX15 expression to macrophage resolution programs, showing efferocytosis potentiates ALOX15 via LXR to enhance SPM precursor synthesis.","evidence":"Primary human macrophage efferocytosis, transcriptomics, LXR agonist, NPC1 inhibition, LXR siRNA, lipidomics","pmids":["33177619"],"confidence":"High","gaps":["Direct LXR binding to the ALOX15 promoter not shown","In vivo contribution to resolution not measured here"]},{"year":2022,"claim":"Positioned ALOX15 as the executioner of ischemic phospholipid peroxidation and ferroptotic signaling in cardiomyocytes, identifying its preferred PE-PUFA substrates.","evidence":"Multi-omics and chemogenetic approaches in animal and in vitro cardiac models","pmids":["35970840"],"confidence":"High","gaps":["Specific oxidized PE species driving ferroptosis not exhaustively defined","Relationship to GPX4 antagonism in heart not quantified"]},{"year":2022,"claim":"Defined ALOX15 as a required effector in the neural-immune inflammatory reflex, linking vagus nerve stimulation to SPM-driven resolution.","evidence":"Electrical VNS in zymosan peritonitis with metabololipidomics and Alox15-deficient mouse comparison","pmids":["35622894"],"confidence":"High","gaps":["Cellular source of ALOX15 in the reflex not pinpointed","Receptors transducing the SPM signal not defined"]},{"year":2023,"claim":"Defined the cardiac ferroptosis mechanism downstream of ALOX15, showing 15-HpETE drives RNF34-dependent Pgc1α degradation and mitochondrial collapse during I/R injury.","evidence":"Cardiac-specific knockout, metabolomics, protein interaction assays, and ML351 inhibition in mice","pmids":["36987924"],"confidence":"High","gaps":["How 15-HpETE engages the Pgc1α-RNF34 interaction biochemically unresolved","Whether the same axis operates in non-cardiac ferroptosis untested"]},{"year":2024,"claim":"Identified a direct protein partner, showing PEBP1 complexes with ALOX15 to direct membrane phospholipid peroxidation and impair macrophage phagocytosis under stress.","evidence":"Co-IP, microscale thermophoresis, docking, phospholipidomics, and phagocytosis assays in a breast cancer model","pmids":["38492368"],"confidence":"High","gaps":["Structural basis of the ALOX15-PEBP1 interface not solved","Whether PEBP1 redirects positional specificity unknown"]},{"year":2024,"claim":"Mapped the transcriptional ferroptosis circuitry feeding ALOX15, placing it downstream of Bmal1/EZH2/EBF3, p53/DHODH, SSAT1, and SREBF2/PER1 regulatory axes.","evidence":"ChIP, knockdown/overexpression, and ferroptosis assays across AML, neuronal, granulosa, and SCI models","pmids":["37271497","35090880","36942513","38653359"],"confidence":"Medium","gaps":["Direct vs indirect promoter regulation by several factors not uniformly confirmed","PER1/SREBF2 axis rests on Low-confidence binding evidence"]},{"year":null,"claim":"It remains unresolved how ALOX15 selects between resolution-promoting (SPM precursor) and cytotoxic (ferroptotic phospholipid peroxidation) outputs in a given cell, and what governs the switch between these opposing functions.","evidence":"No single discovery in the corpus reconciles the pro-resolving and pro-ferroptotic activities mechanistically","pmids":[],"confidence":"Low","gaps":["No unifying model linking substrate channeling, PEBP1 complex, and product fate","Determinants of free-fatty-acid vs esterified-lipid oxidation in vivo unknown"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0016491","term_label":"oxidoreductase activity","supporting_discovery_ids":[0,3,4,5,6,28,29]},{"term_id":"GO:0140096","term_label":"catalytic activity, acting on a protein","supporting_discovery_ids":[1,7,40]},{"term_id":"GO:0008289","term_label":"lipid binding","supporting_discovery_ids":[5,7]},{"term_id":"GO:0016740","term_label":"transferase activity","supporting_discovery_ids":[4]}],"localization":[{"term_id":"GO:0005829","term_label":"cytosol","supporting_discovery_ids":[12]},{"term_id":"GO:0005768","term_label":"endosome","supporting_discovery_ids":[7]},{"term_id":"GO:0005886","term_label":"plasma membrane","supporting_discovery_ids":[1,35]}],"pathway":[{"term_id":"R-HSA-5357801","term_label":"Programmed Cell Death","supporting_discovery_ids":[0,1,13,23,42]},{"term_id":"R-HSA-168256","term_label":"Immune System","supporting_discovery_ids":[17,25,36,37]},{"term_id":"R-HSA-1430728","term_label":"Metabolism","supporting_discovery_ids":[3,5,26,43]},{"term_id":"R-HSA-162582","term_label":"Signal Transduction","supporting_discovery_ids":[7,33]},{"term_id":"R-HSA-74160","term_label":"Gene expression (Transcription)","supporting_discovery_ids":[8,9,42,25]}],"complexes":[],"partners":["PEBP1","RNF34","GPX4","SNX17","LRP5"],"other_free_text":[]}},"prefetch_data":{"uniprot":{"accession":"P16050","full_name":"Polyunsaturated fatty acid lipoxygenase ALOX15","aliases":["12/15-lipoxygenase","Arachidonate 12-lipoxygenase, leukocyte-type","12-LOX","Arachidonate 15-lipoxygenase","15-LOX","15-LOX-1","Arachidonate omega-6 lipoxygenase","Hepoxilin A3 synthase Alox15","Linoleate 13S-lipoxygenase"],"length_aa":662,"mass_kda":74.8,"function":"Non-heme iron-containing dioxygenase that catalyzes the stereo-specific peroxidation of free and esterified polyunsaturated fatty acids generating a spectrum of bioactive lipid mediators (PubMed:17052953, PubMed:1944593, PubMed:24282679, PubMed:25293588, PubMed:32404334, PubMed:8334154). It inserts peroxyl groups at C12 or C15 of arachidonate ((5Z,8Z,11Z,14Z)-eicosatetraenoate) producing both 12-hydroperoxyeicosatetraenoate/12-HPETE and 15-hydroperoxyeicosatetraenoate/15-HPETE (PubMed:17052953, PubMed:1944593, PubMed:24282679, PubMed:8334154). It may then act on 12-HPETE to produce hepoxilins, which may show pro-inflammatory properties (By similarity). Can also peroxidize linoleate ((9Z,12Z)-octadecadienoate) to 13-hydroperoxyoctadecadienoate/13-HPODE (PubMed:8334154). May participate in the sequential oxidations of DHA ((4Z,7Z,10Z,13Z,16Z,19Z)-docosahexaenoate) to generate specialized pro-resolving mediators (SPMs)like resolvin D5 ((7S,17S)-diHPDHA) and (7S,14S)-diHPDHA, that actively down-regulate the immune response and have anti-aggregation properties with platelets (PubMed:32404334). Can convert epoxy fatty acids to hydroperoxy-epoxides derivatives followed by an intramolecular nucleophilic substitution leading to the formation of monocyclic endoperoxides (PubMed:25293588). Plays an important role during the maintenance of self-tolerance by peroxidizing membrane-bound phosphatidylethanolamine which can then signal the sorting process for clearance of apoptotic cells during inflammation and prevent an autoimmune response. In addition to its role in the immune and inflammatory responses, this enzyme may play a role in epithelial wound healing in the cornea through production of lipoxin A4 (LXA(4)) and docosahexaenoic acid-derived neuroprotectin D1 (NPD1; 10R,17S-HDHA), both lipid autacoids exhibit anti-inflammatory and neuroprotective properties. Furthermore, it may regulate actin polymerization which is crucial for several biological processes such as the phagocytosis of apoptotic cells. It is also implicated in the generation of endogenous ligands for peroxisome proliferator activated receptor (PPAR-gamma), hence modulating macrophage development and function. It may also exert a negative effect on skeletal development by regulating bone mass through this pathway. As well as participates in ER stress and downstream inflammation in adipocytes, pancreatic islets, and liver (By similarity). 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Inhibition.","date":"2018","source":"Archives of Iranian medicine","url":"https://pubmed.ncbi.nlm.nih.gov/29940744","citation_count":12,"is_preprint":false},{"pmid":"39457099","id":"PMC_39457099","title":"SAT1/ALOX15 Signaling Pathway Is Involved in Ferroptosis After Skeletal Muscle Contusion.","date":"2024","source":"International journal of molecular sciences","url":"https://pubmed.ncbi.nlm.nih.gov/39457099","citation_count":11,"is_preprint":false},{"pmid":"39815992","id":"PMC_39815992","title":"ALOX15 Aggravates Metabolic Dysfunction-Associated Steatotic Liver Disease in Mice with Type 2 Diabetes via Activating the PPARγ/CD36 Axis.","date":"2025","source":"Antioxidants & redox signaling","url":"https://pubmed.ncbi.nlm.nih.gov/39815992","citation_count":11,"is_preprint":false},{"pmid":"31663148","id":"PMC_31663148","title":"15-LOX-1 has diverse roles in the resensitization of resistant cancer cell lines to doxorubicin.","date":"2019","source":"Journal of cellular physiology","url":"https://pubmed.ncbi.nlm.nih.gov/31663148","citation_count":11,"is_preprint":false},{"pmid":"31717983","id":"PMC_31717983","title":"Deficiency of 15-LOX-1 Induces Radioresistance through Downregulation of MacroH2A2 in Colorectal Cancer.","date":"2019","source":"Cancers","url":"https://pubmed.ncbi.nlm.nih.gov/31717983","citation_count":11,"is_preprint":false},{"pmid":"29235036","id":"PMC_29235036","title":"Expression of DHA-Metabolizing Enzyme Alox15 is Regulated by Selective Histone Acetylation in Neuroblastoma Cells.","date":"2017","source":"Neurochemical research","url":"https://pubmed.ncbi.nlm.nih.gov/29235036","citation_count":10,"is_preprint":false},{"pmid":"32151768","id":"PMC_32151768","title":"A role of Gln596 in fine-tuning mammalian ALOX15 specificity, protein stability and allosteric properties.","date":"2020","source":"Biochimica et biophysica acta. Molecular and cell biology of lipids","url":"https://pubmed.ncbi.nlm.nih.gov/32151768","citation_count":10,"is_preprint":false},{"pmid":"39675198","id":"PMC_39675198","title":"ALOX15-Driven Ferroptosis: The key target in Dihydrotanshinone I's epigenetic Battle in hepatic stellate cells against liver fibrosis.","date":"2024","source":"International immunopharmacology","url":"https://pubmed.ncbi.nlm.nih.gov/39675198","citation_count":10,"is_preprint":false},{"pmid":"33595729","id":"PMC_33595729","title":"Deletion of Alox15 improves kidney dysfunction and inhibits fibrosis by increased PGD2 in the kidney.","date":"2021","source":"Clinical and experimental nephrology","url":"https://pubmed.ncbi.nlm.nih.gov/33595729","citation_count":10,"is_preprint":false},{"pmid":"28812459","id":"PMC_28812459","title":"ALOX15 Immunohistochemistry Aids in the Diagnosis of Eosinophilic Esophagitis on Pauci-eosinophilic Biopsies in Children.","date":"2017","source":"Pediatric and developmental pathology : the official journal of the Society for Pediatric Pathology and the Paediatric Pathology Society","url":"https://pubmed.ncbi.nlm.nih.gov/28812459","citation_count":10,"is_preprint":false},{"pmid":"31565963","id":"PMC_31565963","title":"Association between ALOX15 gene polymorphism and brick-tea type skeletal fluorosis in Tibetans, Kazaks and Han, China.","date":"2019","source":"International journal of environmental health research","url":"https://pubmed.ncbi.nlm.nih.gov/31565963","citation_count":9,"is_preprint":false},{"pmid":"38213304","id":"PMC_38213304","title":"Development of novel ALOX15 inhibitors combining dual machine learning filtering and fragment substitution optimisation approaches, molecular docking and dynamic simulation methods.","date":"2024","source":"Journal of enzyme inhibition and medicinal chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/38213304","citation_count":9,"is_preprint":false},{"pmid":"35073698","id":"PMC_35073698","title":"N-Substituted 5-(1H-Indol-2-yl)-2-methoxyanilines Are Allosteric Inhibitors of the Linoleate Oxygenase Activity of Selected Mammalian ALOX15 Orthologs: Mechanism of Action.","date":"2022","source":"Journal of medicinal chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/35073698","citation_count":9,"is_preprint":false},{"pmid":"27696115","id":"PMC_27696115","title":"The Analgesic and Anxiolytic Effect of Souvenaid, a Novel Nutraceutical, Is Mediated by Alox15 Activity in the Prefrontal Cortex.","date":"2016","source":"Molecular neurobiology","url":"https://pubmed.ncbi.nlm.nih.gov/27696115","citation_count":9,"is_preprint":false},{"pmid":"29154477","id":"PMC_29154477","title":"Mutagenesis of Sequence Determinants of Truncated Porcine ALOX15 Induces Changes in the Reaction Specificity by Altering the Catalytic Mechanism of Initial Hydrogen Abstraction.","date":"2017","source":"Chemistry (Weinheim an der Bergstrasse, Germany)","url":"https://pubmed.ncbi.nlm.nih.gov/29154477","citation_count":9,"is_preprint":false},{"pmid":"40205009","id":"PMC_40205009","title":"Wogonin attenuates septic cardiomyopathy by suppressing ALOX15-mediated ferroptosis.","date":"2025","source":"Acta pharmacologica Sinica","url":"https://pubmed.ncbi.nlm.nih.gov/40205009","citation_count":8,"is_preprint":false},{"pmid":"40387519","id":"PMC_40387519","title":"ALOX15-Mediated Neuron Ferroptosis Was Involved in Diabetic Peripheral Neuropathic Pain.","date":"2025","source":"CNS neuroscience & therapeutics","url":"https://pubmed.ncbi.nlm.nih.gov/40387519","citation_count":8,"is_preprint":false},{"pmid":"33807076","id":"PMC_33807076","title":"Conformational Heterogeneity and Cooperative Effects of Mammalian ALOX15.","date":"2021","source":"International journal of molecular sciences","url":"https://pubmed.ncbi.nlm.nih.gov/33807076","citation_count":8,"is_preprint":false},{"pmid":"37154013","id":"PMC_37154013","title":"Silencing of ALOX15 reduces ferroptosis and inflammation induced by cerebral ischemia-reperfusion by regulating PHD2/HIF2α signaling pathway.","date":"2023","source":"Biotechnology & genetic engineering reviews","url":"https://pubmed.ncbi.nlm.nih.gov/37154013","citation_count":8,"is_preprint":false},{"pmid":"35571613","id":"PMC_35571613","title":"Targeting Circulating lncRNA ENST00000538705.1 Relieves Acute Coronary Syndrome via Modulating ALOX15.","date":"2022","source":"Disease markers","url":"https://pubmed.ncbi.nlm.nih.gov/35571613","citation_count":8,"is_preprint":false},{"pmid":"38030974","id":"PMC_38030974","title":"Knock-in mice expressing a humanized arachidonic acid 15-lipoxygenase (Alox15) carry a partly dysfunctional erythropoietic system.","date":"2023","source":"Cellular & molecular biology letters","url":"https://pubmed.ncbi.nlm.nih.gov/38030974","citation_count":8,"is_preprint":false},{"pmid":"28492741","id":"PMC_28492741","title":"Effect of Alox-15 Polymorphism on GCF Levels of Lipoxin-A4 in Chronic Periodontitis: A Preliminary Study.","date":"2017","source":"Brazilian dental journal","url":"https://pubmed.ncbi.nlm.nih.gov/28492741","citation_count":8,"is_preprint":false},{"pmid":"38653359","id":"PMC_38653359","title":"Rhythm gene PER1 mediates ferroptosis and lipid metabolism through SREBF2/ALOX15 axis in polycystic ovary syndrome.","date":"2024","source":"Biochimica et biophysica acta. Molecular basis of disease","url":"https://pubmed.ncbi.nlm.nih.gov/38653359","citation_count":7,"is_preprint":false},{"pmid":"22668814","id":"PMC_22668814","title":"Association of ALOX12 and ALOX15 gene polymorphisms with age at menarche and natural menopause in Chinese women.","date":"2012","source":"Menopause (New York, N.Y.)","url":"https://pubmed.ncbi.nlm.nih.gov/22668814","citation_count":7,"is_preprint":false},{"pmid":"32606775","id":"PMC_32606775","title":"Eicosapentaenoic acid's metabolism of 15-LOX-1 promotes the expression of miR-101 thus inhibits Cox2 pathway in colon cancer.","date":"2020","source":"OncoTargets and therapy","url":"https://pubmed.ncbi.nlm.nih.gov/32606775","citation_count":7,"is_preprint":false},{"pmid":"22301860","id":"PMC_22301860","title":"Association of ALOX15 gene polymorphisms with obesity-related phenotypes in Chinese nuclear families with male offspring.","date":"2012","source":"Acta pharmacologica Sinica","url":"https://pubmed.ncbi.nlm.nih.gov/22301860","citation_count":7,"is_preprint":false},{"pmid":"40455659","id":"PMC_40455659","title":"ALOX15 Modulates Ferroptosis via the Reactive Oxygen Species-Mediated MAPK Pathway in Doxorubicin-Induced Cardiotoxicity.","date":"2025","source":"Antioxidants & redox signaling","url":"https://pubmed.ncbi.nlm.nih.gov/40455659","citation_count":6,"is_preprint":false}],"cost":{"stage1":{"model":"claude-sonnet-4-6","input_tokens":56786,"output_tokens":14310,"usd":0.192504,"stage1_stop_reason":"end_turn"},"stage2":{"model":"claude-opus-4-8","input_tokens":26633,"output_tokens":5077,"usd":0.130045,"stage2_stop_reason":"end_turn"},"total_usd":0.322549,"stage1_batch_id":"msgbatch_0179Z2pxRYBsXi2xADneafRw","stage2_batch_id":"msgbatch_01LaY9ZaVdgjWArcZo9eqQE5","note":"batch pricing = 50% of standard"},"stage1_raw":"```json\n{\n  \"discoveries\": [\n    {\n      \"year\": 2023,\n      \"finding\": \"ALOX15 catalyzes conversion of arachidonic acid to 15-HpETE, which promotes cardiomyocyte ferroptosis by binding Pgc1α to the ubiquitin ligase RNF34, leading to ubiquitin-dependent Pgc1α degradation, attenuated mitochondrial biogenesis, and abnormal mitochondrial morphology. Myocardial-specific knockout of Alox15 alleviated I/R injury; ML351 (specific ALOX15 inhibitor) rescued cardiac function.\",\n      \"method\": \"Mouse cardiac-specific knockout, metabolomics, loss/gain-of-function experiments, protein interaction assays, pharmacological inhibition (ML351)\",\n      \"journal\": \"Circulation\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — multiple orthogonal methods (KO, metabolomics, protein interaction, pharmacological inhibition) in a rigorous single study with clear mechanistic pathway placement\",\n      \"pmids\": [\"36987924\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"ALOX15 is the primary mediator of ischemia-provoked phospholipid peroxidation in cardiomyocytes, specifically oxidizing PUFA-containing phosphatidylethanolamines to generate ferroptotic lipid signals; identified by multi-omics and confirmed by chemogenetic approaches.\",\n      \"method\": \"Multi-omics analysis, chemogenetic approaches, animal and in vitro models\",\n      \"journal\": \"Signal transduction and targeted therapy\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — multi-omics plus chemogenetic confirmation, multiple experimental systems\",\n      \"pmids\": [\"35970840\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2004,\n      \"finding\": \"Alox15 (12/15-lipoxygenase) negatively regulates peak bone mineral density in mice; crossbreeding with Alox15 knockout mice confirmed this skeletal role, and pharmacological inhibitors of the enzyme improved bone density and strength in two rodent osteoporosis models.\",\n      \"method\": \"Genetic mapping, Alox15 knockout mouse crossbreeding, pharmacological inhibition in rodent models\",\n      \"journal\": \"Science (New York, N.Y.)\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — knockout confirmation plus pharmacological validation in two independent animal models\",\n      \"pmids\": [\"14716014\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2000,\n      \"finding\": \"15-LOX-1 (ALOX15) enzymatic activity in colorectal cancer cells metabolizes linoleic acid to 13-S-HODE, which induces apoptosis; NSAIDs (sulindac, NS-398) increase 15-LOX-1 protein expression and enzymatic activity, and pharmacological inhibition of 15-LOX-1 blocked NSAID-induced apoptosis, which was rescued by exogenous 13-S-HODE but not linoleic acid.\",\n      \"method\": \"15-LOX-1 protein expression assay, enzymatic activity measurement, 13-S-HODE quantification, pharmacological inhibition (caffeic acid), exogenous metabolite rescue in colorectal cancer cell lines (RKO, HT-29)\",\n      \"journal\": \"Journal of the National Cancer Institute\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — enzyme activity confirmed, metabolite quantified, inhibitor plus rescue experiment in two cell lines establish the mechanistic pathway\",\n      \"pmids\": [\"10904086\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2016,\n      \"finding\": \"Evolutionary alteration of ALOX15 reaction specificity—from 12-lipoxygenating in lower primates to 15-lipoxygenating in higher primates (humans, chimpanzees, orangutans)—optimizes lipoxin synthase activity; 15-lipoxygenating ALOX15 variants have >5-fold higher lipoxin synthase activity than 12-lipoxygenating variants. Triad residues (e.g., Ile418) are key specificity determinants confirmed by mutagenesis and QM/MM calculations.\",\n      \"method\": \"Cloning and expression of multiple ALOX15 orthologs, lipoxin synthase activity assays, site-directed mutagenesis (humIle418Ala), QM/MM energy calculations, molecular dynamics simulations\",\n      \"journal\": \"Proceedings of the National Academy of Sciences of the United States of America\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Strong — in vitro reconstitution with mutagenesis, computational validation, and functional quantification across multiple orthologs\",\n      \"pmids\": [\"27412860\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2017,\n      \"finding\": \"Mammalian ALOX15 orthologs exhibit pronounced dual positional specificity with DHA, forming similar amounts of 14- and 17-HpDHA; ALOX15 orthologs prefer DHA and EPA over arachidonic acid when equimolar amounts are supplied. The reaction specificity strongly depends on fatty acid substrate chemistry.\",\n      \"method\": \"Recombinant expression of eight mammalian ALOX15 orthologs as his-tag fusion proteins, substrate oxygenation assays with multiple PUFAs (AA, EPA, DHA), product characterization\",\n      \"journal\": \"Biochimica et biophysica acta. Molecular and cell biology of lipids\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Strong — systematic in vitro enzymatic characterization across eight species orthologs with multiple substrates\",\n      \"pmids\": [\"28400162\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2015,\n      \"finding\": \"Mutagenesis of triad determinants (Leu353, Ile593, Ala404) in rat Alox15 alters reaction specificity with free fatty acids and complex ester lipids; Leu353Phe mutation converts the enzyme from 12-lipoxygenating to 15-lipoxygenating activity. The Ala404Gly exchange induced subtle alterations in enantioselectivity, partially supporting the Coffa/Brash concept.\",\n      \"method\": \"Recombinant rat Alox15 expression (prokaryotic and eukaryotic systems), multiple site-directed mutagenesis, enzymatic oxygenation assays with free fatty acids and phospholipid substrates (biomembranes, HDL)\",\n      \"journal\": \"Archives of biochemistry and biophysics\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Strong — systematic mutagenesis with in vitro enzymatic characterization across multiple substrates and multiple mutants\",\n      \"pmids\": [\"25731857\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"15-LOX-1 (ALOX15) peroxidates linoleic acid in phosphatidylinositol-3-phosphates (PI3P_LA → PI3P_13-HODE), which decreases PI3P binding to SNX17 and LRP5, inhibits LRP5 recycling from endosomes to the plasma membrane, and promotes LRP5 lysosomal degradation, thereby suppressing Wnt/β-catenin signaling and colorectal carcinogenesis.\",\n      \"method\": \"Transgenic mouse intestinal expression of ALOX15, Apc-mutant mouse model, azoxymethane CRC model, in vitro reconstitution of lipid peroxidation, PI3P-SNX17-LRP5 interaction assays, endosomal trafficking assays\",\n      \"journal\": \"Cell reports\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — multiple in vivo models plus defined biochemical mechanism of PI3P modification and LRP5 trafficking\",\n      \"pmids\": [\"32814052\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2006,\n      \"finding\": \"A C-to-T SNP at position c.-292 in the ALOX15 promoter creates a novel binding site for transcription factor SPI1 (PU.1); alleles carrying c.-292T transcribe twice as efficiently as c.-292C alleles in macrophages that constitutively express SPI1, but not in lung epithelial cells lacking SPI1. EMSA confirmed selective SPI1 binding to the mutant promoter; primary macrophages from heterozygous carriers expressed 3× more ALOX15 mRNA.\",\n      \"method\": \"Luciferase transcription assays, EMSA (electrophoretic mobility shift assay), SPI1 core-binding site mutagenesis, qRT-PCR in primary human macrophages from genotyped donors\",\n      \"journal\": \"Human mutation\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Moderate — EMSA + promoter mutagenesis + functional transcription assays + primary human cell validation in a single study\",\n      \"pmids\": [\"16320347\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2009,\n      \"finding\": \"ALOX15 transcription in colon cancer cells is suppressed by recruitment of the NuRD repressor complex (containing MTA1 and HDAC1) to the region -120 to -391 of the 15-LOX-1 promoter; HDAC inhibitors (depsipeptide) reduce NuRD recruitment and reactivate 15-LOX-1 transcription. MTA1 expression in colorectal cancer is negatively correlated with 15-LOX-1 expression.\",\n      \"method\": \"siRNA knockdown of HDAC1, HDAC2, MTA1; luciferase reporter assays with 15-LOX-1 promoter constructs; chromatin immunoprecipitation; HDAC inhibitor treatment (depsipeptide)\",\n      \"journal\": \"Oncogene\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — ChIP, siRNA knockdown, luciferase reporter assays, and in vivo correlation in a single study with multiple orthogonal methods\",\n      \"pmids\": [\"19198625\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2006,\n      \"finding\": \"GATA-6 transcription factor overexpression contributes to 15-LOX-1 transcriptional silencing in colon cancer cells; GATA-6 siRNA knockdown alone is insufficient but significantly contributes to restoring 15-LOX-1 expression and inducing apoptosis when combined with NSAID (NS-398) or HDAC inhibitor (sodium butyrate) treatment.\",\n      \"method\": \"GATA-6 siRNA transfection in Caco-2 and HCT-116 cells, 15-LOX-1 expression measurement, apoptosis assays, NS-398 and sodium butyrate treatments, paired human colorectal cancer tissue analysis\",\n      \"journal\": \"FASEB journal\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — siRNA knockdown with functional readouts in two cell lines plus human tissue validation, but single lab study\",\n      \"pmids\": [\"17167069\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2007,\n      \"finding\": \"A coding SNP T560M in ALOX15 results in a near-null variant with 20-fold reduction in catalytic activity compared to wild-type enzyme, as demonstrated by in vitro enzymatic assays.\",\n      \"method\": \"ALOX15 resequencing, in vitro enzymatic activity assay of T560M vs T560 variants, genotyping in population studies\",\n      \"journal\": \"Atherosclerosis\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 1 / Weak — in vitro enzymatic characterization of single SNP variant, single study\",\n      \"pmids\": [\"17959182\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2010,\n      \"finding\": \"ALOX15 (15-lipoxygenase) is present in sperm cytoplasmic droplets (CDs) and required for normal epididymal sperm maturation; Alox15 knockout male mice have significantly more spermatozoa with retained CDs containing intact mitochondria in both caput and cauda epididymidis, compared to wild-type where CDs contain only hollow membrane vesicles with no intact mitochondria.\",\n      \"method\": \"Alox15 knockout mouse analysis, differential interference contrast microscopy (DIC), transmission electron microscopy (TEM) of epididymal sperm from Alox15 vs wild-type males\",\n      \"journal\": \"Cell and tissue research\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — knockout model with direct ultrastructural phenotypic readout showing organelle localization and functional consequence\",\n      \"pmids\": [\"20449608\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2016,\n      \"finding\": \"Systemic inactivation of the Alox15 gene normalizes reduced fertility and sperm motility in male mice heterozygous for catalytically inactive Gpx4 (Sec46Ala-Gpx4+/-); Alox15 deficiency rescues sperm midpiece structural alterations and motility defects, demonstrating that ALOX15-mediated lipid oxidation is functionally antagonized by GPX4 in male reproductive function.\",\n      \"method\": \"Genetic cross of Gpx4 knock-in mice with Alox15 knockout mice, fertility assessment, sperm motility analysis, ultrastructural analysis\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — epistasis by genetic double-mutant rescue experiment with defined fertility and motility phenotypic readouts\",\n      \"pmids\": [\"27634046\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2014,\n      \"finding\": \"Alox15 deficiency does NOT rescue homozygous Gpx4 catalytically inactive knock-in mice (Sec46Ala-Gpx4+/+) from embryonic lethality (E6-7), indicating that Alox15-mediated lipid peroxidation is not the primary cause of embryonic lethality in GPX4-null mice.\",\n      \"method\": \"Genetic cross of Sec46Ala-Gpx4 knock-in mice with Alox15 knockout mice, embryonic viability assessment, embryo staging\",\n      \"journal\": \"Antioxidants & redox signaling\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — epistasis via genetic double-mutant showing clear negative result (no rescue), rigorous embryonic staging\",\n      \"pmids\": [\"25313597\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2010,\n      \"finding\": \"Alox15 gene disruption in ApoE-/- mice protects against nonalcoholic fatty liver disease; Alox15 deficiency reduces hepatic steatosis, inflammation, macrophage infiltration, attenuates glucose intolerance and insulin resistance, upregulates insulin receptor substrate-2, and exerts opposite effects on hepatic JNK and AMPK phosphorylation. Hepatocytes from Alox15-deficient mice are less vulnerable to TNFα-induced programmed cell death.\",\n      \"method\": \"ApoE-/-/Alox15-/- double knockout mice, hepatocyte isolation and TNFα challenge, JNK and AMPK phosphorylation assays, glucose/insulin tolerance tests, gene expression analysis\",\n      \"journal\": \"Hepatology (Baltimore, Md.)\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — genetic double-KO model with multiple orthogonal functional readouts and defined signaling pathway placement\",\n      \"pmids\": [\"20967760\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2013,\n      \"finding\": \"12/15-lipoxygenase (Alox15) is expressed in macrophages but not B and T cells of NOD mice; deletion of Alox15 decreases proinflammatory cytokine expression in macrophages, splenocytes from NOD-Alox15(null) mice cannot transfer diabetes in adoptive transfer, and absence of Alox15 maintains islet health and beta cell mass, demonstrating that Alox15 regulates macrophage inflammation driving autoimmunity in type 1 diabetes.\",\n      \"method\": \"qRT-PCR cell-type expression analysis, adoptive transfer model, Western blotting, beta cell mass quantification in NOD-Alox15(null) mice\",\n      \"journal\": \"PloS one\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — KO model with adoptive transfer epistasis and multiple cellular readouts, single lab\",\n      \"pmids\": [\"23437231\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"Vagus nerve stimulation (VNS) promotes resolution of inflammation and increases specialized proresolving mediators (SPMs) from DHA and n-3 DPA metabolomes; this VNS-mediated shift toward proresolving lipid mediator profile is inverted in Alox15-deficient mice, establishing Alox15 as a required effector in the neural-immune inflammatory reflex pathway.\",\n      \"method\": \"Electrical VNS in zymosan-induced peritonitis mouse model, lipid mediator metabololipidomics, Alox15-deficient mouse comparison, α7nAChR-deficient mouse comparison\",\n      \"journal\": \"Proceedings of the National Academy of Sciences of the United States of America\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — genetic epistasis using Alox15 KO mice plus metabololipidomics in a defined in vivo model with clear pathway placement\",\n      \"pmids\": [\"35622894\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"Alox15-derived 15-HEPE (15-hydroxy eicosapentaenoic acid) is the anti-colitis mediator responsible for the protective effect of increased n-3 PUFA in fat1 transgenic mice; Alox15 gene inactivation in fat1 mice abolishes their protection from DSS- and TNBS-induced colitis, and intraperitoneal 15S-HEPE administration protects wild-type mice from colitis.\",\n      \"method\": \"fat1×Alox15-/- genetic cross, DSS and TNBS colitis models, Alox15 lipid metabolite analysis, 15S-HEPE intraperitoneal treatment in wild-type mice\",\n      \"journal\": \"FASEB journal\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — genetic epistasis with double KO plus pharmacological rescue with defined metabolite in two colitis models\",\n      \"pmids\": [\"33710695\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2018,\n      \"finding\": \"Alox15/ALOX15B are expressed in cardiac fibroblasts; hypoxia significantly increases ALOX15 and ALOX15B gene and protein expression in fibroblasts from failing human hearts. Conditioned medium from hypoxic fibroblasts decreases beating frequency of human iPSC-derived cardiomyocytes in an ALOX15-dependent manner (effect blocked by ALOX15 inhibition).\",\n      \"method\": \"Gene expression in isolated cardiac fibroblasts under hypoxia vs normoxia, 15-HETE measurement, paracrine co-culture with iPSC-derived cardiomyocytes, calcium imaging, ALOX15 inhibition\",\n      \"journal\": \"PloS one\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — functional paracrine assay with ALOX15 inhibition confirms dependency, single lab study\",\n      \"pmids\": [\"30138423\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2017,\n      \"finding\": \"ALOX15 (Alox15) is expressed in rat brain, with highest mRNA and protein expression in the prefrontal cortex; it is localized to dendrites by immuno-electron microscopy. Alox15 inhibition or antisense knockdown in prefrontal cortex significantly decreased resolvin D1 levels, blocked hippocampo-prefrontal cortex long-term potentiation, and increased errors in T-maze spatial working memory, establishing Alox15 as mediating DHA-to-resolvin D1 conversion required for synaptic plasticity and memory.\",\n      \"method\": \"RT-PCR, Western blot, immunohistochemistry, immuno-electron microscopy, LC-MS for resolvin D1, pharmacological inhibition and antisense knockdown of Alox15 in prefrontal cortex, LTP recording, T-maze behavioral test\",\n      \"journal\": \"Molecular neurobiology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — multiple orthogonal methods including subcellular localization (immuno-EM), metabolite measurement, and functional behavioral/electrophysiological readouts\",\n      \"pmids\": [\"28181190\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2017,\n      \"finding\": \"Alox15 expression in neuroblastoma cells is regulated by histone acetylation; HDAC inhibitors (TSA, sodium butyrate, Class I inhibitors MS-275 and depsipeptide) upregulate Alox15 mRNA. The p300 histone acetyltransferase (HAT) inhibitor C646 modulates TSA- and retinoic acid-induced Alox15 upregulation, indicating p300 HAT activity is required for Alox15 transcriptional induction during neuronal differentiation.\",\n      \"method\": \"HDAC inhibitor treatment of SH-SY5Y cells, Class I-specific HDAC inhibitors, p300 HAT inhibitor (C646) co-treatment, retinoic acid differentiation, qRT-PCR, primary murine cortical neuron developmental expression\",\n      \"journal\": \"Neurochemical research\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — pharmacological dissection of specific HDAC classes and HAT requirement, single lab, no ChIP\",\n      \"pmids\": [\"29235036\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2017,\n      \"finding\": \"ALOX15 upregulation leads to hepatic overproduction of 13-HODE, which induces oxidative stress, ER stress, lipid metabolic disorder, and apoptosis; ALOX15 knockout prevents alcohol-induced liver damage through attenuation of these pathways. Linoleic acid (not ethanol or acetaldehyde) induces ALOX15 expression in hepatocytes.\",\n      \"method\": \"Chronic alcohol feeding mouse model, ALOX15 knockout mice, ALOX15 inhibitor (PD146176) treatment, 13-HODE exposure to Hepa-1c1c7 cells, oxidative stress, ER stress, and apoptosis markers\",\n      \"journal\": \"Scientific reports\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — genetic KO plus pharmacological inhibition plus defined metabolite mechanistic link, multiple orthogonal readouts\",\n      \"pmids\": [\"28827690\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"SSAT1 overexpression upregulates ALOX15 expression in neurons and promotes ferroptosis by decreasing GPX4 and SLC7A11 levels; SSAT1 knockdown downregulates ALOX15 and reduces lipid hydroperoxide production. ALOX15 inhibitor PD146176 partially reverses SSAT1 upregulation-induced ferroptosis, placing ALOX15 downstream of SSAT1 in neuronal ferroptosis signaling.\",\n      \"method\": \"SSAT1 knockdown/overexpression in primary cortical neurons and tMCAO/R mouse model, ALOX15 inhibitor (PD146176) treatment, iron/ROS/lipid peroxide measurements, GPX4/SLC7A11 protein levels\",\n      \"journal\": \"Neuroscience\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — epistasis placement via KD/OE plus pharmacological rescue, single lab\",\n      \"pmids\": [\"35090880\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"DHODH inhibits neuronal ferroptosis after spinal cord injury via the P53/ALOX15 signaling pathway: DHODH inhibits P53 expression, which in turn suppresses ALOX15 expression, reducing lipid peroxidation and ferroptosis.\",\n      \"method\": \"Rat SCI model, erastin-induced PC12 cells, DHODH overexpression/inhibition, P53 and ALOX15 expression analysis, lipid peroxide and mitochondrial damage measurement\",\n      \"journal\": \"CNS neuroscience & therapeutics\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — in vivo and in vitro models with pathway placement, single lab study\",\n      \"pmids\": [\"36942513\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"Efferocytosis by alternatively activated macrophages potentiates ALOX15 expression through LXR activation; AC uptake upregulates LXR-dependent gene expression and suppresses SREBP-2-dependent gene expression via NPC1-mediated sterol transport. LXRα/β silencing attenuates this ALOX15 potentiation. Enhanced ALOX15 expression increases capacity to synthesize SPM precursors 15-HETE and 17-HDHA and resolvin D5.\",\n      \"method\": \"Apoptotic cell efferocytosis in primary human macrophages, global transcriptome analysis, LXR agonist (T0901317), NPC1 inhibition, LXRα/β siRNA silencing, lipidomics (15-HETE, 17-HDHA, resolvin D5 measurement)\",\n      \"journal\": \"Cell death and differentiation\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — primary human cells, transcriptomics, siRNA knockdown, sterol transport inhibition, and metabolite production all supporting a defined regulatory mechanism\",\n      \"pmids\": [\"33177619\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2008,\n      \"finding\": \"Alox15 is induced in the small and large intestine and liver of iron-deficient rats; increased Alox15 protein leads to production of 12-HETE, 13-HODE, and 13-HOTE (confirmed by HPLC), representing a perturbation of intestinal lipid metabolism driven by Alox15 activity during iron deficiency.\",\n      \"method\": \"Gene chip studies in iron-deficient and Belgrade rats, TaqMan qRT-PCR, polyclonal antibody immunodetection, HPLC lipid analysis\",\n      \"journal\": \"American journal of physiology. Gastrointestinal and liver physiology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — direct lipid product identification by HPLC linked to Alox15 induction, single lab study\",\n      \"pmids\": [\"18258795\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2010,\n      \"finding\": \"Selenoprotein P (SelP) has in vitro lipid hydroperoxidase activity against 15-HpETE (the ALOX15 product), attenuates 15-HpETE oxidation in cellular assays, and reduces oxidative damage in a transcellular assay when 15-LOX-1 is metabolically active, identifying SelP as an antioxidant counterpart to 15-LOX-1.\",\n      \"method\": \"In vitro lipid hydroperoxidase assay with recombinant SelP and 15-HpETE, cellular 15-HpETE oxidation assay, transcellular assay with metabolically active 15-LOX-1\",\n      \"journal\": \"Prostaglandins, leukotrienes, and essential fatty acids\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 1 / Moderate — in vitro enzymatic assay with defined substrate (15-HpETE) and cellular validation, single lab\",\n      \"pmids\": [\"20826080\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"The ALOX15 specificity switch during mammalian evolution follows a Triad Concept: >90% of mammalian ALOX15 orthologs catalyze 12-lipoxygenation; mammals ranked above gibbons express 15-lipoxygenating orthologs; triad determinant mutagenesis studies on 44 novel orthologs confirmed the specificity predictions. Gibbon ALOX15 represents a transition enzyme with dual specificity.\",\n      \"method\": \"Sequence analysis of 152 mammalian ALOX15 orthologs, expression of 44 novel recombinant ALOX15 orthologs, enzymatic specificity assays, extensive mutagenesis of triad determinants\",\n      \"journal\": \"Frontiers in cell and developmental biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Strong — large-scale systematic in vitro enzymatic characterization plus mutagenesis across 44 expressed proteins from diverse species\",\n      \"pmids\": [\"35531094\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2017,\n      \"finding\": \"Mutagenesis of porcine ALOX15 Val418 and Val419 sequence determinants converts the enzyme from 12-lipoxygenating to 15-lipoxygenating activity; QM/MM calculations showed that the Val418Ile+Val419Met double mutant has a 6.0 kcal/mol higher barrier for 12-lipoxygenation than 15-lipoxygenation, explaining the specificity switch by altering enzyme-substrate complex geometry.\",\n      \"method\": \"Site-directed mutagenesis of porcine ALOX15 crystal structure, molecular docking, MD simulations, QM/MM calculations\",\n      \"journal\": \"Chemistry (Weinheim an der Bergstrasse, Germany)\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Strong — mutagenesis on a crystallized enzyme combined with QM/MM calculations providing mechanistic basis for catalytic specificity\",\n      \"pmids\": [\"29154477\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"Gln596 of mammalian ALOX15 does not directly interact with the carboxylate of arachidonic acid; instead, mutations at Gln596 destabilize secondary/tertiary structure and impair allosteric enzyme properties by disturbing the electrostatic interaction network. MD simulations suggest dimer interface geometry depends on substrate bound in the active site, and Gln596Ala impairs allosteric communication between monomers.\",\n      \"method\": \"Site-directed mutagenesis of rabbit and human ALOX15 at Gln596, structural stability assays, steady-state kinetics, MD simulations, substrate docking\",\n      \"journal\": \"Biochimica et biophysica acta. Molecular and cell biology of lipids\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 1 / Moderate — mutagenesis with kinetics and MD simulations in a single lab study\",\n      \"pmids\": [\"32151768\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"N-substituted 5-(1H-indol-2-yl)-2-methoxyanilines are allosteric inhibitors of the linoleate oxygenase activity of rabbit and human ALOX15; they selectively inhibit oxygenation of linoleic acid at submicromolar concentrations without affecting arachidonic acid oxygenation. Steady-state kinetics, mutagenesis, and MD simulations of the ALOX15 dimer model indicate binding at the active site of one monomer induces conformational changes in the other monomer, compromising productive linoleic acid-enzyme complex formation.\",\n      \"method\": \"In vitro enzymatic assays (steady-state kinetics), site-directed mutagenesis, MD simulations of ALOX15 dimer model, synthesis of inhibitor series\",\n      \"journal\": \"Journal of medicinal chemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Strong — systematic kinetics + mutagenesis + MD simulations establishing allosteric mechanism in a single rigorous study\",\n      \"pmids\": [\"35073698\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"Rabbit ALOX15 and human ALOX15B exist as two conformers with distinct electrophoretic mobilities in native PAGE; MD simulations and mutagenesis suggest motional flexibility in aqueous solution impacting enzymatic properties, consistent with a dynamic monomer-dimer equilibrium with substrate-dependent conformational heterogeneity.\",\n      \"method\": \"Native polyacrylamide electrophoresis, in silico docking, MD simulations, site-directed mutagenesis, kinetic measurements\",\n      \"journal\": \"International journal of molecular sciences\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 1 / Moderate — direct conformational observation by native PAGE plus computational and mutagenesis support, single lab\",\n      \"pmids\": [\"33807076\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2004,\n      \"finding\": \"15-LOX-1 overexpression in HCT-116 colon cancer cells induces ERK1/2 phosphorylation, decreases p21 (Cip/WAF1) expression, and increases cell growth; its product 13-S-HODE reverses NaBT-induced p21 expression. LOX inhibitor (NDGA) decreased ERK1/2 phosphorylation and increased p21 expression in 15-LOX-1 overexpressing cells.\",\n      \"method\": \"15-LOX-1 transfection in HCT-116 cells, ERK1/2 phosphorylation assay, p21 expression analysis, NDGA inhibitor treatment, sodium butyrate treatment\",\n      \"journal\": \"Prostaglandins & other lipid mediators\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — gain-of-function plus pharmacological inhibition with defined signaling readouts, single lab\",\n      \"pmids\": [\"15165036\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2017,\n      \"finding\": \"Intestinally-targeted transgenic expression of ALOX15 in mice inhibits DSS-induced colitis from promoting azoxymethane-induced colorectal tumorigenesis; ALOX15 suppresses TNF-α, IL-1β/NF-κB, and IL-6/STAT3 inflammatory signaling pathways that drive colorectal cancer promotion.\",\n      \"method\": \"ALOX15 intestinal transgenic mouse model, DSS-colitis model, azoxymethane carcinogen model, cytokine/signaling pathway analysis\",\n      \"journal\": \"Prostaglandins & other lipid mediators\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — in vivo transgenic model with defined pathway readouts, single lab\",\n      \"pmids\": [\"28089732\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2018,\n      \"finding\": \"Systemic Alox15 deficiency in female mice protects against DSS-induced colitis by sustaining expression of the tight junction protein ZO-1 and maintaining intestinal epithelial barrier function; exogenous 12S-HETE (the major mouse Alox15 product) enantioselectively compromises transepithelial electric resistance in colon epithelial cells. Transgenic overexpression of human ALOX15 intensified inflammatory symptoms.\",\n      \"method\": \"Alox15-/- mouse DSS colitis model, 12S-HETE epithelial barrier assay (transepithelial electric resistance), ZO-1 immunostaining, ALOX15 transgenic overexpression, 12-HETE in vivo quantification\",\n      \"journal\": \"Biochimica et biophysica acta. Molecular and cell biology of lipids\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — KO and transgenic overexpression models plus direct mechanistic link to tight junction protein and barrier function via metabolite assay\",\n      \"pmids\": [\"29702245\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2013,\n      \"finding\": \"IFN-γ suppresses ALOX15 expression during resolution of PMA-induced skin inflammation; neutralization of endogenous IFN-γ boosts ALOX15 expression and LXA4 production, promoting proresolution effects. Exogenous LXA4 (an ALOX15 product) recapitulates the proresolution effect of IFN-γ neutralization.\",\n      \"method\": \"In vivo PMA-induced skin inflammation model in mice, IFN-γ neutralizing antibody, ALOX15 expression analysis, exogenous LXA4 treatment\",\n      \"journal\": \"Mediators of inflammation\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — in vivo neutralization experiment with ALOX15 expression readout and metabolite rescue, single lab\",\n      \"pmids\": [\"23818745\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2018,\n      \"finding\": \"Spinal TLR4 activation by intrathecal KLA increases 15-LOX-1 expression specifically in microglia (not astrocytes) and increases 12-lipoxygenase-mediated hepoxilin production in lumbar spinal cord; intrathecal pretreatment with 15-LOX-1-specific inhibitors (ML127, ML351, CDC) or a selective antibody against rat 15-LOX-1 completely abrogated TLR4-induced tactile allodynia, establishing spinal microglial ALOX15 as mediating TLR4-dependent neuropathic pain.\",\n      \"method\": \"Intrathecal KLA administration, lipidomics of spinal cord, primary spinal microglia/astrocyte cultures, selective ALOX15 antibody neutralization, ML127/ML351/CDC pharmacological inhibition, HEK-293T heterologous expression of rat 15-LOX-1 for inhibitor testing\",\n      \"journal\": \"Pain\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — cell-type-specific expression plus multiple selective pharmacological inhibitors with behavioral readouts and in vitro enzymatic confirmation\",\n      \"pmids\": [\"30130298\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2014,\n      \"finding\": \"15-LOX-1 re-expression in colon cancer cells (HCT116, HT29LMM, LoVo) under hypoxia inhibits HIF-1α protein expression and stability, reduces VEGF expression, angiogenesis, cancer cell migration, invasion, and survival under hypoxia, demonstrating that 15-LOX-1 suppresses hypoxia-driven metastatic mechanisms.\",\n      \"method\": \"15-LOX-1 re-expression in three colon cancer cell lines under hypoxia, HIF-1α protein stability assay, VEGF ELISA, angiogenesis assay, migration and invasion assays\",\n      \"journal\": \"Cancer medicine\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — gain-of-function in three cell lines with multiple functional readouts, single lab\",\n      \"pmids\": [\"24634093\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"FGF21 deficiency results in abnormal activation of the ALOX15/15-HETE pathway, triggering innate immunity-dominated pro-inflammatory responses in liver grafts; pharmacological recombinant FGF21 administration protects against hepatic I/R injury and suppresses the ALOX15/15-HETE axis, particularly in steatotic livers.\",\n      \"method\": \"Hepatic I/R injury and orthotopic LT mouse models, FGF21 KO mice, recombinant FGF21 treatment, ALOX15/15-HETE pathway analysis, arachidonic acid metabolomics\",\n      \"journal\": \"Nature communications\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — genetic KO and pharmacological rescue with defined metabolite pathway readout, single lab\",\n      \"pmids\": [\"39362839\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"PEBP1 forms a complex with ALOX15 in macrophages under psychological stress, mediating membrane phospholipid peroxidation that compromises macrophage phagocytic capacity; inhibition of the ALOX15/PEBP1 interaction (e.g., by isochlorogenic acid C, ICAC) attenuates phospholipid peroxidation and restores phagocytic function in the tumor microenvironment.\",\n      \"method\": \"Co-immunoprecipitation, microscale thermophoresis (MST) binding assay, molecular docking, LC-MS/MS phospholipidomics, flow cytometry phagocytosis assay, in vivo breast cancer model\",\n      \"journal\": \"Phytomedicine\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — direct protein-protein interaction confirmed by Co-IP and MST, metabolite-function link established by phospholipidomics and phagocytosis assay\",\n      \"pmids\": [\"38492368\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"Alox15 deficiency in chronic kidney disease (5/6 nephrectomy) mice improves renal function and inhibits interstitial fibrosis; mediator lipidomics revealed significantly higher PGD2 in Alox15-/- CKD kidneys, and exogenous PGD2 dose-dependently suppresses TGF-β1-induced type I collagen and αSMA in renal epithelial cells, suggesting ALOX15 suppresses PGD2, which is anti-fibrotic.\",\n      \"method\": \"Alox15-/- mouse 5/6 nephrectomy CKD model, mediator lipidomics, PGD2 treatment of NRK-52E and HK-2 cells, fibrosis marker quantification\",\n      \"journal\": \"Clinical and experimental nephrology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — KO mouse model plus in vitro metabolite rescue experiment, single lab\",\n      \"pmids\": [\"33595729\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"Bmal1 inhibits RSL3-induced ferroptosis in AML cells by recruiting EZH2 to the EBF3 promoter to enhance its methylation and suppress EBF3 expression; EBF3 binds to the ALOX15 promoter to enhance ALOX15 expression, promoting ferroptosis. Thus ALOX15 is downstream of the Bmal1/EZH2/EBF3 axis in ferroptosis regulation in AML.\",\n      \"method\": \"Bmal1/EBF3 knockdown/overexpression in HL-60 and NB4 cells, EZH2 ChIP at EBF3 promoter, EBF3 ChIP at ALOX15 promoter, subcutaneous AML xenograft model, ferroptosis assays (MDA, GSH, lipid peroxidation, iron)\",\n      \"journal\": \"Cancer science\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — ChIP at multiple promoters establishing pathway, supported by in vivo xenograft validation, single lab\",\n      \"pmids\": [\"37271497\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"ALOX15 promotes lipid accumulation in hepatocytes through the PPARγ/CD36 pathway; ALOX15 overexpression increases intracellular fatty acid uptake and lipotoxicity, which are reversed by PPARγ antagonist (GW9662) or CD36 inhibitor. GLP-1 receptor agonist liraglutide improves hepatic lipid accumulation in MASLD mice by suppressing the ALOX15/PPARγ/CD36 pathway.\",\n      \"method\": \"ALOX15 lentiviral overexpression and siRNA KD in HepG2 cells, palmitic acid lipotoxicity model, PPARγ antagonist (GW9662) and CD36 inhibitor treatment, ALOX15 metabolomics (12-HpETE, 15-HpETE), transcriptomics, HFD/STZ mouse model\",\n      \"journal\": \"Antioxidants & redox signaling\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — gain- and loss-of-function with defined downstream pathway inhibitors and in vivo model, single lab\",\n      \"pmids\": [\"39815992\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"Alpha-tocopherol (Vitamin E) downregulates Alox15 expression and inhibits ferroptosis after spinal cord injury; mutation of the 87th residue of Alox15 weakens the inhibitory effect of alpha-tocopherol on Alox15, identifying this residue as important for the alpha-tocopherol/Alox15 interaction.\",\n      \"method\": \"SCI rat model, alpha-tocopherol treatment, Alox15 expression analysis, Alox15 87th residue mutation, immunofluorescence localization, in vitro ROS and lipid peroxidation assays\",\n      \"journal\": \"Biomedicine & pharmacotherapy\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — residue mutagenesis supports direct interaction, combined in vivo/in vitro, single lab\",\n      \"pmids\": [\"38754264\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"Lactate-induced HIF1A transcriptional activation (via histone lactylation of the HIF1A promoter) exacerbates severe acute pancreatitis by inducing ferroptosis through the ACSL4/LPCAT3/ALOX15 pathway; HIF1A absence or overexpression changes SAP severity by modulating this ferroptosis pathway. Elevated lactate from glycolytic abnormalities is the upstream driver.\",\n      \"method\": \"l-Arg SAP mouse model, sh-HIF1A mice, CHIP-qPCR for HIF1A promoter lactylation, luciferase reporter, ACSL4/LPCAT3/ALOX15 expression analysis, WB for lipid peroxidation markers\",\n      \"journal\": \"Journal of cellular biochemistry\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — ChIP-qPCR and luciferase reporter establish HIF1A regulation, pathway placement via KO mouse, single lab\",\n      \"pmids\": [\"39676583\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"In AML cells, EBF3 binds to the ALOX15 promoter and enhances its expression, promoting ferroptosis; this regulatory axis is confirmed by EBF3 ChIP at the ALOX15 promoter and rescue experiments with EBF3 knockdown.\",\n      \"method\": \"ChIP-qPCR of EBF3 binding at ALOX15 promoter, EBF3 overexpression/knockdown, ALOX15 ferroptosis assays in AML cells\",\n      \"journal\": \"Cancer science\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — ChIP at ALOX15 promoter with functional rescue, part of a larger study (same PMID as Bmal1/EZH2/EBF3/ALOX15)\",\n      \"pmids\": [\"37271497\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"PER1 directly binds to the SREBF2 promoter to repress SREBF2 transcription; SREBF2 in turn binds to the ALOX15 promoter to repress ALOX15 transcription; PER1 overexpression thus increases ALOX15 expression and promotes ferroptosis in granulosa cells in PCOS.\",\n      \"method\": \"PER1/SREBF2 promoter binding assays (ChIP implied from 'binds to promoter'), PER1 overexpression/SREBF2 overexpression/ALOX15 silencing in granulosa cells, ferroptosis marker measurement, PCOS mouse model\",\n      \"journal\": \"Biochimica et biophysica acta. Molecular basis of disease\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 / Moderate — promoter binding described but specific ChIP methodology not clearly confirmed in abstract; pathway placement supported by rescue experiments\",\n      \"pmids\": [\"38653359\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"ALOX15 modulates ferroptosis in doxorubicin-induced cardiotoxicity via the ROS-mediated MAPK signaling pathway; ALOX15 loss of function ameliorates Dox-induced ferroptosis by inhibiting ROS-mediated MAPK activation. Linoleic acid increases susceptibility to Dox-induced damage, abolished by ALOX15 inhibition.\",\n      \"method\": \"Dox mouse and H9C2 cell models, ALOX15 inhibition/silencing/overexpression, ROS measurement, MAPK phosphorylation assays, ferroptosis markers\",\n      \"journal\": \"Antioxidants & redox signaling\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — gain- and loss-of-function with defined downstream signaling readout (MAPK phosphorylation), single lab\",\n      \"pmids\": [\"40455659\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"Wogonin directly binds to and inhibits ALOX15, suppressing cardiomyocyte ferroptosis in septic cardiomyopathy; pharmacological ALOX15 inhibition (ML351) recapitulates wogonin's cardioprotection. In LPS-challenged cardiomyocytes, ALOX15 overexpression or its downstream metabolite 15-HpETE abolishes the anti-ferroptotic effects of wogonin.\",\n      \"method\": \"CLP septic cardiomyopathy mouse model, molecular docking, direct binding assay, ML351 pharmacological inhibition, ALOX15 overexpression in HL-1 cardiomyocytes, 15-HpETE rescue experiment\",\n      \"journal\": \"Acta pharmacologica Sinica\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — direct binding assay plus pharmacological inhibitor recapitulation and metabolite rescue, single lab\",\n      \"pmids\": [\"40205009\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"Macrophage-derived exosomal miRNA-660-5p is transported into cervical cancer cells where it attenuates ALOX15 expression to suppress ferroptosis; macrophage miRNA-660-5p upregulation depends on autocrine IL4/IL13-activated STAT6 pathway.\",\n      \"method\": \"Exosome isolation and transfer, miRNA-660-5p transfection/inhibition, ALOX15 expression analysis, ferroptosis assays in cervical cancer cells, STAT6 pathway analysis in macrophages\",\n      \"journal\": \"Acta pharmaceutica Sinica. B\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — defined miRNA-mediated ALOX15 suppression mechanism via exosome transfer with upstream STAT6 pathway, single lab\",\n      \"pmids\": [\"37425043\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"Knock-in mice expressing the arachidonic acid 15-lipoxygenating Leu353Phe Alox15 mutant (functionally humanized) develop normally and are fertile but show modified plasma oxylipidomes; aged male Alox15-KI mice develop dysfunctional erythropoiesis (reduced hematocrit, lower erythrocyte counts, attenuated hemoglobin) with improved ex vivo osmotic resistance of red blood cells, in a sex-specific manner.\",\n      \"method\": \"Alox15 Leu353Phe knock-in mouse generation, plasma oxylipidomics, hematological parameter measurement, osmotic resistance assay\",\n      \"journal\": \"Cellular & molecular biology letters\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — knock-in mouse with defined functional humanization and erythropoietic phenotype, single lab\",\n      \"pmids\": [\"38030974\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"ALOX15 (15-lipoxygenase-1) is a lipid-peroxidizing enzyme that oxygenates polyunsaturated fatty acids—principally arachidonic acid and linoleic acid—to generate bioactive hydroperoxy lipid metabolites (15-HpETE, 13-HODE, 15-HETE, 15-HEPE, and SPM precursors); its reaction specificity is determined by active-site triad residues and evolved from 12-lipoxygenating in lower mammals to 15-lipoxygenating in Hominidae, optimizing lipoxin biosynthesis. ALOX15-derived 15-HpETE drives cardiomyocyte ferroptosis by targeting Pgc1α for ubiquitin-dependent degradation via RNF34, and ALOX15-mediated PI3P peroxidation (PI3P_13-HODE) suppresses LRP5 endosomal recycling to inhibit Wnt/β-catenin signaling in colonic epithelium. Its transcription is silenced in cancer cells by the NuRD repressor complex (HDAC1/MTA1) and by GATA-6 overexpression, and is activated in macrophages by a SPI1-binding promoter SNP and by LXR following efferocytosis. ALOX15 functions as a pro-ferroptotic enzyme downstream of SSAT1/SAT1, p53, and Bmal1/EBF3 transcriptional axes, is antagonized by GPX4, and mediates neural-immune resolution via Alox15-derived SPMs in the vagal inflammatory reflex.\"\n}\n```","stage2_raw":"{\n  \"mechanistic_narrative\": \"ALOX15 (15-lipoxygenase-1) is a non-heme iron lipid-peroxidizing enzyme that oxygenates polyunsaturated fatty acids and their esterified derivatives to generate bioactive hydroperoxy and hydroxy lipid mediators governing inflammation resolution, ferroptosis, and tissue homeostasis [#0, #3, #25]. The enzyme acts on free arachidonic acid, linoleic acid, EPA, and DHA, with marked substrate preference for n-3 PUFAs, producing 15-HpETE/15-HETE, 13-HODE, 15-HEPE, and DHA-derived 14-/17-HpDHA that serve as precursors for specialized pro-resolving mediators including lipoxins and resolvins [#3, #5, #18, #25]. Its positional specificity is set by an active-site triad (e.g. Leu353/Ile418, Val418/Val419, Ala404), and during mammalian/primate evolution mutation of these determinants converted the ancestral 12-lipoxygenating enzyme into the 15-lipoxygenating form found in higher primates, optimizing lipoxin synthase activity [#4, #6, #28, #29]. Beyond free fatty acids, ALOX15 peroxidizes membrane and organelle phospholipids—oxidizing PUFA-phosphatidylethanolamines to drive ferroptotic signals and linoleate within PI3P to suppress LRP5 endosomal recycling and Wnt/\\u03b2-catenin signaling—and forms a complex with PEBP1 to direct phospholipid peroxidation [#1, #7, #40]. Through 15-HpETE generation it executes ferroptotic cell death programs, in cardiomyocytes by promoting RNF34-dependent Pgc1\\u03b1 degradation and impaired mitochondrial biogenesis, an activity antagonized by GPX4 [#0, #13]. ALOX15 sits downstream of multiple transcriptional and metabolic ferroptosis axes including SSAT1, p53/DHODH, and Bmal1/EZH2/EBF3, while its own transcription is repressed by the NuRD (MTA1/HDAC1) complex and GATA-6 in cancer cells, activated by an SPI1-binding promoter SNP and LXR in macrophages, and tuned by histone acetylation [#8, #9, #23, #24, #42, #25]. Physiologically, ALOX15-derived mediators drive resolution of inflammation in vagal neural-immune circuits, synaptic plasticity and memory, intestinal barrier integrity, and skin and colitis resolution, while its peroxidation activity also contributes to NAFLD, alcoholic liver injury, neuropathic pain, autoimmune diabetes, and reduced bone mineral density [#2, #17, #20, #35, #36, #37].\"\n,\n  \"teleology\": [\n    {\n      \"year\": 2000,\n      \"claim\": \"Established that ALOX15 is not merely a metabolic enzyme but a pro-apoptotic effector in colon cancer cells via its linoleic acid product 13-S-HODE, answering whether its catalytic output has direct cell-fate consequences.\",\n      \"evidence\": \"Enzyme activity assays, 13-S-HODE quantification, inhibitor plus metabolite rescue in RKO and HT-29 colorectal cancer cells\",\n      \"pmids\": [\"10904086\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Downstream apoptotic effectors of 13-S-HODE not defined\", \"Does not address phospholipid substrates or in vivo relevance\"]\n    },\n    {\n      \"year\": 2004,\n      \"claim\": \"Extended ALOX15 function beyond cancer to skeletal physiology, showing the enzyme negatively regulates peak bone mineral density and is a pharmacological target for bone strength.\",\n      \"evidence\": \"Genetic mapping, Alox15 knockout crossbreeding, and inhibitor treatment in two rodent osteoporosis models\",\n      \"pmids\": [\"14716014\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Lipid mediator linking ALOX15 to osteoblast/osteoclast biology not identified\", \"Cell type responsible not pinpointed\"]\n    },\n    {\n      \"year\": 2006,\n      \"claim\": \"Defined cis- and trans-acting transcriptional control of ALOX15, showing a promoter SNP creates an SPI1 site driving macrophage-selective expression, and GATA-6 contributes to silencing in cancer.\",\n      \"evidence\": \"Luciferase assays, EMSA, promoter mutagenesis, primary human macrophage qRT-PCR; GATA-6 siRNA in Caco-2/HCT-116 cells\",\n      \"pmids\": [\"16320347\", \"17167069\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"How SPI1 and repressors are coordinated across cell types unresolved\", \"GATA-6 effect insufficient alone\"]\n    },\n    {\n      \"year\": 2009,\n      \"claim\": \"Identified the chromatin machinery silencing ALOX15 in cancer, placing the NuRD complex (MTA1/HDAC1) on the promoter and explaining HDAC-inhibitor reactivation.\",\n      \"evidence\": \"siRNA knockdown of HDAC1/MTA1, ChIP, luciferase reporters, depsipeptide in colon cancer cells\",\n      \"pmids\": [\"19198625\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Signals recruiting NuRD to the promoter not defined\", \"Generality beyond colon cancer untested\"]\n    },\n    {\n      \"year\": 2016,\n      \"claim\": \"Resolved the structural basis and evolutionary logic of ALOX15 reaction specificity, showing triad residues convert 12- to 15-lipoxygenation and optimize lipoxin synthesis in higher primates.\",\n      \"evidence\": \"Ortholog cloning/expression, lipoxin synthase assays, site-directed mutagenesis, QM/MM and MD simulations across many species\",\n      \"pmids\": [\"27412860\", \"25731857\", \"35531094\", \"29154477\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Physiological consequence of the evolutionary switch in vivo not directly tested at this stage\", \"Specificity with esterified substrates less fully characterized\"]\n    },\n    {\n      \"year\": 2017,\n      \"claim\": \"Broadened the substrate scope, showing ALOX15 prefers EPA and DHA over arachidonic acid and produces dual-specificity SPM precursors, reframing it as a key generator of n-3-derived resolution mediators.\",\n      \"evidence\": \"Recombinant expression of eight mammalian orthologs with multiple PUFA substrates and product characterization\",\n      \"pmids\": [\"28400162\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"In vivo substrate selection under physiological PUFA ratios not established\", \"Tissue-specific product spectra unknown\"]\n    },\n    {\n      \"year\": 2017,\n      \"claim\": \"Demonstrated CNS roles for ALOX15, localizing it to prefrontal cortical dendrites and showing its DHA-to-resolvin D1 conversion is required for synaptic plasticity and working memory.\",\n      \"evidence\": \"Immuno-EM, LC-MS for resolvin D1, inhibition/antisense knockdown, LTP recording, and T-maze testing in rat\",\n      \"pmids\": [\"28181190\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Receptor for resolvin D1 mediating LTP not identified\", \"Human relevance untested\"]\n    },\n    {\n      \"year\": 2020,\n      \"claim\": \"Established that ALOX15 oxidizes esterified lipids in trafficking compartments, peroxidizing PI3P-linoleate to block LRP5 recycling and suppress Wnt/\\u03b2-catenin signaling in colon.\",\n      \"evidence\": \"Transgenic and Apc/azoxymethane mouse models, in vitro lipid peroxidation, PI3P-SNX17-LRP5 interaction and endosomal trafficking assays\",\n      \"pmids\": [\"32814052\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Whether PI3P peroxidation occurs in non-intestinal tissues unknown\", \"Selectivity for PI3P vs other phosphoinositides not fully mapped\"]\n    },\n    {\n      \"year\": 2020,\n      \"claim\": \"Connected ALOX15 expression to macrophage resolution programs, showing efferocytosis potentiates ALOX15 via LXR to enhance SPM precursor synthesis.\",\n      \"evidence\": \"Primary human macrophage efferocytosis, transcriptomics, LXR agonist, NPC1 inhibition, LXR siRNA, lipidomics\",\n      \"pmids\": [\"33177619\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Direct LXR binding to the ALOX15 promoter not shown\", \"In vivo contribution to resolution not measured here\"]\n    },\n    {\n      \"year\": 2022,\n      \"claim\": \"Positioned ALOX15 as the executioner of ischemic phospholipid peroxidation and ferroptotic signaling in cardiomyocytes, identifying its preferred PE-PUFA substrates.\",\n      \"evidence\": \"Multi-omics and chemogenetic approaches in animal and in vitro cardiac models\",\n      \"pmids\": [\"35970840\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Specific oxidized PE species driving ferroptosis not exhaustively defined\", \"Relationship to GPX4 antagonism in heart not quantified\"]\n    },\n    {\n      \"year\": 2022,\n      \"claim\": \"Defined ALOX15 as a required effector in the neural-immune inflammatory reflex, linking vagus nerve stimulation to SPM-driven resolution.\",\n      \"evidence\": \"Electrical VNS in zymosan peritonitis with metabololipidomics and Alox15-deficient mouse comparison\",\n      \"pmids\": [\"35622894\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Cellular source of ALOX15 in the reflex not pinpointed\", \"Receptors transducing the SPM signal not defined\"]\n    },\n    {\n      \"year\": 2023,\n      \"claim\": \"Defined the cardiac ferroptosis mechanism downstream of ALOX15, showing 15-HpETE drives RNF34-dependent Pgc1\\u03b1 degradation and mitochondrial collapse during I/R injury.\",\n      \"evidence\": \"Cardiac-specific knockout, metabolomics, protein interaction assays, and ML351 inhibition in mice\",\n      \"pmids\": [\"36987924\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"How 15-HpETE engages the Pgc1\\u03b1-RNF34 interaction biochemically unresolved\", \"Whether the same axis operates in non-cardiac ferroptosis untested\"]\n    },\n    {\n      \"year\": 2024,\n      \"claim\": \"Identified a direct protein partner, showing PEBP1 complexes with ALOX15 to direct membrane phospholipid peroxidation and impair macrophage phagocytosis under stress.\",\n      \"evidence\": \"Co-IP, microscale thermophoresis, docking, phospholipidomics, and phagocytosis assays in a breast cancer model\",\n      \"pmids\": [\"38492368\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Structural basis of the ALOX15-PEBP1 interface not solved\", \"Whether PEBP1 redirects positional specificity unknown\"]\n    },\n    {\n      \"year\": 2024,\n      \"claim\": \"Mapped the transcriptional ferroptosis circuitry feeding ALOX15, placing it downstream of Bmal1/EZH2/EBF3, p53/DHODH, SSAT1, and SREBF2/PER1 regulatory axes.\",\n      \"evidence\": \"ChIP, knockdown/overexpression, and ferroptosis assays across AML, neuronal, granulosa, and SCI models\",\n      \"pmids\": [\"37271497\", \"35090880\", \"36942513\", \"38653359\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Direct vs indirect promoter regulation by several factors not uniformly confirmed\", \"PER1/SREBF2 axis rests on Low-confidence binding evidence\"]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"It remains unresolved how ALOX15 selects between resolution-promoting (SPM precursor) and cytotoxic (ferroptotic phospholipid peroxidation) outputs in a given cell, and what governs the switch between these opposing functions.\",\n      \"evidence\": \"No single discovery in the corpus reconciles the pro-resolving and pro-ferroptotic activities mechanistically\",\n      \"pmids\": [],\n      \"confidence\": \"Low\",\n      \"gaps\": [\"No unifying model linking substrate channeling, PEBP1 complex, and product fate\", \"Determinants of free-fatty-acid vs esterified-lipid oxidation in vivo unknown\"]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0016491\", \"supporting_discovery_ids\": [0, 3, 4, 5, 6, 28, 29]},\n      {\"term_id\": \"GO:0140096\", \"supporting_discovery_ids\": [1, 7, 40]},\n      {\"term_id\": \"GO:0008289\", \"supporting_discovery_ids\": [5, 7]},\n      {\"term_id\": \"GO:0016740\", \"supporting_discovery_ids\": [4]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005829\", \"supporting_discovery_ids\": [12]},\n      {\"term_id\": \"GO:0005768\", \"supporting_discovery_ids\": [7]},\n      {\"term_id\": \"GO:0005886\", \"supporting_discovery_ids\": [1, 35]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-5357801\", \"supporting_discovery_ids\": [0, 1, 13, 23, 42]},\n      {\"term_id\": \"R-HSA-168256\", \"supporting_discovery_ids\": [17, 25, 36, 37]},\n      {\"term_id\": \"R-HSA-1430728\", \"supporting_discovery_ids\": [3, 5, 26, 43]},\n      {\"term_id\": \"R-HSA-162582\", \"supporting_discovery_ids\": [7, 33]},\n      {\"term_id\": \"R-HSA-74160\", \"supporting_discovery_ids\": [8, 9, 42, 25]}\n    ],\n    \"complexes\": [],\n    \"partners\": [\"PEBP1\", \"RNF34\", \"GPX4\", \"SNX17\", \"LRP5\"],\n    \"other_free_text\": []\n  }\n}","audit_flag":null,"evaluation":{"pairwise":"win","faith_supported":7,"faith_total":7,"faith_pct":100.0}}