{"gene":"NLE1","run_date":"2026-06-10T05:19:52","timeline":{"discoveries":[{"year":2022,"finding":"NLE1 is upregulated in colorectal cancer upon SMAD4 loss via c-MYC occupying an E-box-containing region within the NLE1 promoter, thereby preventing TGFβ-mediated downregulation of NLE1.","method":"ChIP (c-MYC at NLE1 promoter E-box), patient-derived tumor organoids with ectopic c-MYC expression, TGFβ1 treatment","journal":"Cancer research","confidence":"High","confidence_rationale":"Tier 2 / Strong — ChIP demonstrating direct c-MYC binding to NLE1 promoter, combined with organoid disease modeling and multiple orthogonal functional assays in a single rigorous study","pmids":["36219392"],"is_preprint":false},{"year":2022,"finding":"NLE1 is limiting for de novo protein biosynthesis in colorectal cancer cells; its ablation activates p38/MAPK signaling, impairs autophagy (accumulation of p62- and LC3-positive structures), increases reactive oxygen species, and inhibits proliferation, migration, invasion, clonogenicity, and anchorage-independent growth.","method":"NLE1 knockout (CRISPR/KO) in colorectal cancer cell lines and organoids; protein synthesis assays; immunofluorescence for p62/LC3; ROS measurement; proliferation, migration, invasion, and colony formation assays","journal":"Cancer research","confidence":"High","confidence_rationale":"Tier 2 / Strong — multiple orthogonal loss-of-function assays with defined molecular readouts in a single comprehensive study","pmids":["36219392"],"is_preprint":false},{"year":2022,"finding":"NLE1 ablation impairs colorectal tumor growth in the colon and reduces primary tumor-derived liver metastasis in an endoscopy-guided orthotopic mouse transplantation model; deletion of TP53 further sensitizes NLE1-deficient cells to apoptosis.","method":"Orthotopic mouse transplantation model with NLE1 KO colorectal cancer cells; in vivo tumor growth and liver metastasis quantification; apoptosis assays with TP53 co-deletion","journal":"Cancer research","confidence":"High","confidence_rationale":"Tier 2 / Strong — in vivo orthotopic model with genetic epistasis (NLE1 KO + TP53 KO), multiple orthogonal endpoints","pmids":["36219392"],"is_preprint":false},{"year":2024,"finding":"NLE1 directly interacts with Notch1 to form a NLE1-Notch1 complex in brain tumor stem cells (BTSCs); the drug edaravone directly targets NLE1, impairs Notch signaling, alters stem cell marker expression, and sensitizes BTSCs to ionizing radiation-induced cell death.","method":"Global gene expression analysis, mechanistic drug-target identification of edaravone binding to NLE1, Notch pathway reporter assays, self-renewal/growth assays in BTSCs, preclinical glioblastoma mouse models with survival analysis","journal":"Stem cell reports","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — mechanistic drug-target identification and Notch pathway assays, single lab, abstract does not detail precise biochemical binding method","pmids":["39423824"],"is_preprint":false},{"year":2017,"finding":"NLE1 (notchless homolog 1) expression is reduced in the somatosensory cortex of WAG/Rij rats (genetic absence epilepsy model) compared to controls, coinciding with reduced Notch1 expression and altered neuron-to-glia ratio; acute Notch1 agonist (Jagged1) suppressed spike-and-wave discharges while Notch antagonist (DAPT) facilitated them, placing NLE1 as a modulator of the Notch1 signaling pathway in cortical development.","method":"Immunohistochemistry, qRT-PCR for NLE1 and Notch1 mRNA/protein in brain regions; in vivo EEG with pharmacological Notch1 agonist/antagonist administration","journal":"Brain structure & function","confidence":"Medium","confidence_rationale":"Tier 3 / Moderate — immunohistochemistry and in vivo pharmacological epistasis, single lab, indirect mechanistic link between NLE1 and Notch1","pmids":["28210849"],"is_preprint":false},{"year":2023,"finding":"NLE1 promotes NSCLC development by activating E2F1-mediated transcription of CDK1; PI3K/Akt signaling acts downstream of NLE1. CDK1 overexpression or Akt pathway activator treatment reversed NLE1 knockdown-induced NSCLC inhibition.","method":"NLE1 knockdown in NSCLC cell lines (in vitro loss-of-function); in vivo xenograft model; CDK1 rescue overexpression; Akt activator rescue experiment; cell growth, migration, and apoptosis assays","journal":"Frontiers in oncology","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — genetic epistasis via rescue experiments (CDK1 OE and Akt activator), loss-of-function in vitro and in vivo, single lab","pmids":["36818671"],"is_preprint":false},{"year":2021,"finding":"NLE1 knockdown in melanoma cells inhibits proliferation, increases apoptotic sensitivity, and impairs migration in vitro, and inhibits tumor formation in vivo; the apoptosis induction requires apoptosis-related proteins and NLE1 activates the PI3K/AKT signaling pathway.","method":"shRNA knockdown of NLE1 in melanoma cell lines; proliferation, apoptosis, migration assays; mouse xenograft model; western blotting for PI3K/AKT pathway components and apoptosis-related proteins","journal":"Experimental cell research","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — loss-of-function with in vitro and in vivo readouts and PI3K/AKT pathway characterization, single lab","pmids":["34019907"],"is_preprint":false},{"year":2025,"finding":"NLE1 (WD40 protein) promotes hepatocellular carcinoma cell proliferation as demonstrated by proliferation assays (CCK-8 and colony formation); enrichment analysis linked elevated NLE1 expression to proliferative gene programs.","method":"CCK-8 proliferation assay, colony formation assay, genome-wide CRISPR screen data, multiomics enrichment analysis","journal":"Clinical Medicine Insights. Oncology","confidence":"Low","confidence_rationale":"Tier 3 / Weak — single lab, proliferation assays without detailed pathway mechanistic follow-up in abstract","pmids":["40625398"],"is_preprint":false}],"current_model":"NLE1 (Notchless Homolog 1) is a WD40-repeat ribosome biogenesis factor that is transcriptionally activated by c-MYC binding to an E-box in its promoter (opposing TGFβ/SMAD4-mediated repression), limits de novo protein biosynthesis, forms a complex with Notch1 to modulate Notch signaling in stem cells, and promotes tumor cell proliferation, survival, and metastasis in colorectal cancer, NSCLC, melanoma, and glioblastoma partly through PI3K/AKT signaling and E2F1/CDK1 transcriptional activation."},"narrative":{"mechanistic_narrative":"NLE1 (Notchless Homolog 1) is a WD40-repeat protein that limits de novo protein biosynthesis and supports tumor cell proliferation, survival, and metastasis across multiple cancers [PMID:36219392]. In colorectal cancer, NLE1 is transcriptionally controlled at an E-box in its promoter: c-MYC binds this element and prevents TGFβ-mediated downregulation, so SMAD4 loss leads to NLE1 upregulation [PMID:36219392]. NLE1 is rate-limiting for protein synthesis, and its ablation activates p38/MAPK signaling, impairs autophagy with accumulation of p62- and LC3-positive structures, raises reactive oxygen species, and blocks proliferation, migration, invasion, and anchorage-independent growth [PMID:36219392]. In vivo, NLE1 loss impairs orthotopic colorectal tumor growth and liver metastasis, and co-deletion of TP53 sensitizes NLE1-deficient cells to apoptosis [PMID:36219392]. Beyond its biosynthetic role, NLE1 modulates Notch signaling: it forms a direct complex with Notch1 in brain tumor stem cells, where the NLE1-targeting drug edaravone impairs Notch signaling and sensitizes cells to radiation [PMID:39423824]. In NSCLC and melanoma, NLE1 drives tumor phenotypes through PI3K/AKT signaling, and in NSCLC it activates E2F1-mediated transcription of CDK1, with CDK1 overexpression or Akt activation rescuing the effects of NLE1 knockdown [PMID:36818671, PMID:34019907].","teleology":[{"year":2017,"claim":"Initial in vivo evidence placed NLE1 within the Notch1 signaling axis, linking its expression to cortical development and a neurological phenotype.","evidence":"Immunohistochemistry/qRT-PCR for NLE1 and Notch1 plus in vivo EEG with Notch agonist/antagonist in an absence epilepsy rat model","pmids":["28210849"],"confidence":"Medium","gaps":["Correlative association between NLE1 and Notch1 without direct physical or biochemical interaction shown","No molecular mechanism connecting NLE1 to spike-and-wave discharges","Causality of NLE1 reduction not tested by gene manipulation"]},{"year":2021,"claim":"Loss-of-function in melanoma established NLE1 as a pro-tumorigenic factor acting through PI3K/AKT signaling, extending its role beyond Notch.","evidence":"shRNA knockdown in melanoma cell lines with proliferation/apoptosis/migration assays, xenografts, and PI3K/AKT western blotting","pmids":["34019907"],"confidence":"Medium","gaps":["Mechanism by which NLE1 activates PI3K/AKT not defined","No direct binding partner identified","Single lab, single cancer type"]},{"year":2022,"claim":"A comprehensive colorectal cancer study defined the transcriptional control of NLE1 and its function as a limiting factor for protein biosynthesis driving tumor growth and metastasis.","evidence":"ChIP for c-MYC at the NLE1 promoter E-box, organoid and CRISPR-KO models, protein synthesis/autophagy/ROS assays, and an orthotopic metastasis model with TP53 epistasis","pmids":["36219392"],"confidence":"High","gaps":["Direct molecular substrate of NLE1 in the protein biosynthesis machinery not biochemically defined","Mechanism linking NLE1 loss to p38/MAPK activation and autophagy impairment unresolved","Whether the biosynthetic role and Notch role are mechanistically connected is unknown"]},{"year":2023,"claim":"NSCLC studies placed E2F1/CDK1 transcriptional activation and PI3K/AKT signaling downstream of NLE1, defining an effector axis via rescue epistasis.","evidence":"NLE1 knockdown in NSCLC cells and xenografts with CDK1 overexpression and Akt activator rescue experiments","pmids":["36818671"],"confidence":"Medium","gaps":["How NLE1 activates E2F1 transcription mechanistically not shown","Ordering of PI3K/AKT relative to E2F1/CDK1 not fully resolved","No direct NLE1-E2F1 interaction demonstrated"]},{"year":2024,"claim":"Direct interaction with Notch1 was demonstrated and exploited pharmacologically, establishing NLE1 as a druggable Notch-pathway component in brain tumor stem cells.","evidence":"Drug-target identification of edaravone binding NLE1, Notch reporter and self-renewal assays in BTSCs, and preclinical glioblastoma survival models","pmids":["39423824"],"confidence":"Medium","gaps":["Precise biochemical NLE1-Notch1 binding interface not detailed","Whether edaravone-NLE1 binding is direct and specific not fully characterized","Relationship of the Notch1 complex to NLE1's protein-biosynthesis role unaddressed"]},{"year":2025,"claim":"NLE1's pro-proliferative role was extended to hepatocellular carcinoma, reinforcing its broad pan-cancer growth function.","evidence":"CCK-8 and colony formation proliferation assays with CRISPR screen and multiomics enrichment analysis","pmids":["40625398"],"confidence":"Low","gaps":["Proliferation assays without pathway mechanistic follow-up","No direct partner or substrate identified in HCC","Correlative enrichment analysis only"]},{"year":null,"claim":"How NLE1's molecular role in protein biosynthesis mechanistically connects to its Notch1, PI3K/AKT, and E2F1/CDK1 functions remains unresolved.","evidence":"","pmids":[],"confidence":"Medium","gaps":["No structural or biochemical definition of NLE1's molecular activity","No unifying model linking the biosynthetic and signaling functions","Direct substrates and stable complex composition unknown"]}],"mechanism_profile":{"molecular_activity":[],"localization":[],"pathway":[{"term_id":"R-HSA-392499","term_label":"Metabolism of proteins","supporting_discovery_ids":[1]},{"term_id":"R-HSA-162582","term_label":"Signal Transduction","supporting_discovery_ids":[3,5,6]},{"term_id":"R-HSA-1643685","term_label":"Disease","supporting_discovery_ids":[0,2]}],"complexes":[],"partners":["NOTCH1"],"other_free_text":[]}},"prefetch_data":{"uniprot":{"accession":"Q9NVX2","full_name":"Notchless protein homolog 1","aliases":[],"length_aa":485,"mass_kda":53.3,"function":"Plays a role in regulating Notch activity. Plays a role in regulating the expression of CDKN1A and several members of the Wnt pathway, probably via its effects on Notch activity. Required during embryogenesis for inner mass cell survival (By similarity)","subcellular_location":"Nucleus, nucleolus","url":"https://www.uniprot.org/uniprotkb/Q9NVX2/entry"},"depmap":{"release":"DepMap","has_data":true,"is_common_essential":true,"resolved_as":"","url":"https://depmap.org/portal/gene/NLE1","classification":"Common Essential","n_dependent_lines":1187,"n_total_lines":1208,"dependency_fraction":0.9826158940397351},"opencell":{"profiled":false,"resolved_as":"","ensg_id":"","cell_line_id":"","localizations":[],"interactors":[],"url":"https://opencell.sf.czbiohub.org/search/NLE1","total_profiled":1310},"omim":[{"mim_id":"620924","title":"NOTCHLESS HOMOLOG 1; NLE1","url":"https://www.omim.org/entry/620924"}],"hpa":{"profiled":true,"resolved_as":"","reliability":"Supported","locations":[{"location":"Nucleoli","reliability":"Supported"},{"location":"Nucleoplasm","reliability":"Additional"}],"tissue_specificity":"Low tissue specificity","tissue_distribution":"Detected in all","driving_tissues":[],"url":"https://www.proteinatlas.org/search/NLE1"},"hgnc":{"alias_symbol":["FLJ10458","Rsa4"],"prev_symbol":["NLE"]},"alphafold":{"accession":"Q9NVX2","domains":[{"cath_id":"3.10.20.90","chopping":"17-95","consensus_level":"high","plddt":88.9599,"start":17,"end":95},{"cath_id":"2.130.10.10","chopping":"106-485","consensus_level":"high","plddt":96.3393,"start":106,"end":485}],"viewer_url":"https://alphafold.ebi.ac.uk/entry/Q9NVX2","model_url":"https://alphafold.ebi.ac.uk/files/AF-Q9NVX2-F1-model_v6.cif","pae_url":"https://alphafold.ebi.ac.uk/files/AF-Q9NVX2-F1-predicted_aligned_error_v6.png","plddt_mean":93.69},"mouse_models":{"mgi_url":"https://www.informatics.jax.org/marker/summary?nomen=NLE1","jax_strain_url":"https://www.jax.org/strain/search?query=NLE1"},"sequence":{"accession":"Q9NVX2","fasta_url":"https://rest.uniprot.org/uniprotkb/Q9NVX2.fasta","uniprot_url":"https://www.uniprot.org/uniprotkb/Q9NVX2/entry","alphafold_viewer_url":"https://alphafold.ebi.ac.uk/entry/Q9NVX2"}},"corpus_meta":[{"pmid":"11948125","id":"PMC_11948125","title":"Bombesin receptor subtypes in human cancers: detection with the universal radioligand (125)I-[D-TYR(6), beta-ALA(11), PHE(13), NLE(14)] bombesin(6-14).","date":"2002","source":"Clinical cancer research : an official journal of the American Association for Cancer Research","url":"https://pubmed.ncbi.nlm.nih.gov/11948125","citation_count":281,"is_preprint":false},{"pmid":"14980384","id":"PMC_14980384","title":"Effect of I.C.V. injection of AT4 receptor ligands, NLE1-angiotensin IV and LVV-hemorphin 7, on spatial learning in rats.","date":"2004","source":"Neuroscience","url":"https://pubmed.ncbi.nlm.nih.gov/14980384","citation_count":111,"is_preprint":false},{"pmid":"20501122","id":"PMC_20501122","title":"Autoradiography of CCK receptors in the rat brain using [(3)H]Boc[Nle(28)(31)]CCK(27)-(33) and [(125)I]bolton-hunter CCK(8). 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its ablation activates p38/MAPK signaling, impairs autophagy (accumulation of p62- and LC3-positive structures), increases reactive oxygen species, and inhibits proliferation, migration, invasion, clonogenicity, and anchorage-independent growth.\",\n      \"method\": \"NLE1 knockout (CRISPR/KO) in colorectal cancer cell lines and organoids; protein synthesis assays; immunofluorescence for p62/LC3; ROS measurement; proliferation, migration, invasion, and colony formation assays\",\n      \"journal\": \"Cancer research\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — multiple orthogonal loss-of-function assays with defined molecular readouts in a single comprehensive study\",\n      \"pmids\": [\"36219392\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"NLE1 ablation impairs colorectal tumor growth in the colon and reduces primary tumor-derived liver metastasis in an endoscopy-guided orthotopic mouse transplantation model; deletion of TP53 further sensitizes NLE1-deficient cells to apoptosis.\",\n      \"method\": \"Orthotopic mouse transplantation model with NLE1 KO colorectal cancer cells; in vivo tumor growth and liver metastasis quantification; apoptosis assays with TP53 co-deletion\",\n      \"journal\": \"Cancer research\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — in vivo orthotopic model with genetic epistasis (NLE1 KO + TP53 KO), multiple orthogonal endpoints\",\n      \"pmids\": [\"36219392\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"NLE1 directly interacts with Notch1 to form a NLE1-Notch1 complex in brain tumor stem cells (BTSCs); the drug edaravone directly targets NLE1, impairs Notch signaling, alters stem cell marker expression, and sensitizes BTSCs to ionizing radiation-induced cell death.\",\n      \"method\": \"Global gene expression analysis, mechanistic drug-target identification of edaravone binding to NLE1, Notch pathway reporter assays, self-renewal/growth assays in BTSCs, preclinical glioblastoma mouse models with survival analysis\",\n      \"journal\": \"Stem cell reports\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — mechanistic drug-target identification and Notch pathway assays, single lab, abstract does not detail precise biochemical binding method\",\n      \"pmids\": [\"39423824\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2017,\n      \"finding\": \"NLE1 (notchless homolog 1) expression is reduced in the somatosensory cortex of WAG/Rij rats (genetic absence epilepsy model) compared to controls, coinciding with reduced Notch1 expression and altered neuron-to-glia ratio; acute Notch1 agonist (Jagged1) suppressed spike-and-wave discharges while Notch antagonist (DAPT) facilitated them, placing NLE1 as a modulator of the Notch1 signaling pathway in cortical development.\",\n      \"method\": \"Immunohistochemistry, qRT-PCR for NLE1 and Notch1 mRNA/protein in brain regions; in vivo EEG with pharmacological Notch1 agonist/antagonist administration\",\n      \"journal\": \"Brain structure & function\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 3 / Moderate — immunohistochemistry and in vivo pharmacological epistasis, single lab, indirect mechanistic link between NLE1 and Notch1\",\n      \"pmids\": [\"28210849\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"NLE1 promotes NSCLC development by activating E2F1-mediated transcription of CDK1; PI3K/Akt signaling acts downstream of NLE1. CDK1 overexpression or Akt pathway activator treatment reversed NLE1 knockdown-induced NSCLC inhibition.\",\n      \"method\": \"NLE1 knockdown in NSCLC cell lines (in vitro loss-of-function); in vivo xenograft model; CDK1 rescue overexpression; Akt activator rescue experiment; cell growth, migration, and apoptosis assays\",\n      \"journal\": \"Frontiers in oncology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — genetic epistasis via rescue experiments (CDK1 OE and Akt activator), loss-of-function in vitro and in vivo, single lab\",\n      \"pmids\": [\"36818671\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"NLE1 knockdown in melanoma cells inhibits proliferation, increases apoptotic sensitivity, and impairs migration in vitro, and inhibits tumor formation in vivo; the apoptosis induction requires apoptosis-related proteins and NLE1 activates the PI3K/AKT signaling pathway.\",\n      \"method\": \"shRNA knockdown of NLE1 in melanoma cell lines; proliferation, apoptosis, migration assays; mouse xenograft model; western blotting for PI3K/AKT pathway components and apoptosis-related proteins\",\n      \"journal\": \"Experimental cell research\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — loss-of-function with in vitro and in vivo readouts and PI3K/AKT pathway characterization, single lab\",\n      \"pmids\": [\"34019907\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"NLE1 (WD40 protein) promotes hepatocellular carcinoma cell proliferation as demonstrated by proliferation assays (CCK-8 and colony formation); enrichment analysis linked elevated NLE1 expression to proliferative gene programs.\",\n      \"method\": \"CCK-8 proliferation assay, colony formation assay, genome-wide CRISPR screen data, multiomics enrichment analysis\",\n      \"journal\": \"Clinical Medicine Insights. Oncology\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 / Weak — single lab, proliferation assays without detailed pathway mechanistic follow-up in abstract\",\n      \"pmids\": [\"40625398\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"NLE1 (Notchless Homolog 1) is a WD40-repeat ribosome biogenesis factor that is transcriptionally activated by c-MYC binding to an E-box in its promoter (opposing TGFβ/SMAD4-mediated repression), limits de novo protein biosynthesis, forms a complex with Notch1 to modulate Notch signaling in stem cells, and promotes tumor cell proliferation, survival, and metastasis in colorectal cancer, NSCLC, melanoma, and glioblastoma partly through PI3K/AKT signaling and E2F1/CDK1 transcriptional activation.\"\n}\n```","stage2_raw":"{\n  \"mechanistic_narrative\": \"NLE1 (Notchless Homolog 1) is a WD40-repeat protein that limits de novo protein biosynthesis and supports tumor cell proliferation, survival, and metastasis across multiple cancers [#1, #2]. In colorectal cancer, NLE1 is transcriptionally controlled at an E-box in its promoter: c-MYC binds this element and prevents TGFβ-mediated downregulation, so SMAD4 loss leads to NLE1 upregulation [#0]. NLE1 is rate-limiting for protein synthesis, and its ablation activates p38/MAPK signaling, impairs autophagy with accumulation of p62- and LC3-positive structures, raises reactive oxygen species, and blocks proliferation, migration, invasion, and anchorage-independent growth [#1]. In vivo, NLE1 loss impairs orthotopic colorectal tumor growth and liver metastasis, and co-deletion of TP53 sensitizes NLE1-deficient cells to apoptosis [#2]. Beyond its biosynthetic role, NLE1 modulates Notch signaling: it forms a direct complex with Notch1 in brain tumor stem cells, where the NLE1-targeting drug edaravone impairs Notch signaling and sensitizes cells to radiation [#3]. In NSCLC and melanoma, NLE1 drives tumor phenotypes through PI3K/AKT signaling, and in NSCLC it activates E2F1-mediated transcription of CDK1, with CDK1 overexpression or Akt activation rescuing the effects of NLE1 knockdown [#5, #6].\",\n  \"teleology\": [\n    {\n      \"year\": 2017,\n      \"claim\": \"Initial in vivo evidence placed NLE1 within the Notch1 signaling axis, linking its expression to cortical development and a neurological phenotype.\",\n      \"evidence\": \"Immunohistochemistry/qRT-PCR for NLE1 and Notch1 plus in vivo EEG with Notch agonist/antagonist in an absence epilepsy rat model\",\n      \"pmids\": [\"28210849\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\n        \"Correlative association between NLE1 and Notch1 without direct physical or biochemical interaction shown\",\n        \"No molecular mechanism connecting NLE1 to spike-and-wave discharges\",\n        \"Causality of NLE1 reduction not tested by gene manipulation\"\n      ]\n    },\n    {\n      \"year\": 2021,\n      \"claim\": \"Loss-of-function in melanoma established NLE1 as a pro-tumorigenic factor acting through PI3K/AKT signaling, extending its role beyond Notch.\",\n      \"evidence\": \"shRNA knockdown in melanoma cell lines with proliferation/apoptosis/migration assays, xenografts, and PI3K/AKT western blotting\",\n      \"pmids\": [\"34019907\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\n        \"Mechanism by which NLE1 activates PI3K/AKT not defined\",\n        \"No direct binding partner identified\",\n        \"Single lab, single cancer type\"\n      ]\n    },\n    {\n      \"year\": 2022,\n      \"claim\": \"A comprehensive colorectal cancer study defined the transcriptional control of NLE1 and its function as a limiting factor for protein biosynthesis driving tumor growth and metastasis.\",\n      \"evidence\": \"ChIP for c-MYC at the NLE1 promoter E-box, organoid and CRISPR-KO models, protein synthesis/autophagy/ROS assays, and an orthotopic metastasis model with TP53 epistasis\",\n      \"pmids\": [\"36219392\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\n        \"Direct molecular substrate of NLE1 in the protein biosynthesis machinery not biochemically defined\",\n        \"Mechanism linking NLE1 loss to p38/MAPK activation and autophagy impairment unresolved\",\n        \"Whether the biosynthetic role and Notch role are mechanistically connected is unknown\"\n      ]\n    },\n    {\n      \"year\": 2023,\n      \"claim\": \"NSCLC studies placed E2F1/CDK1 transcriptional activation and PI3K/AKT signaling downstream of NLE1, defining an effector axis via rescue epistasis.\",\n      \"evidence\": \"NLE1 knockdown in NSCLC cells and xenografts with CDK1 overexpression and Akt activator rescue experiments\",\n      \"pmids\": [\"36818671\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\n        \"How NLE1 activates E2F1 transcription mechanistically not shown\",\n        \"Ordering of PI3K/AKT relative to E2F1/CDK1 not fully resolved\",\n        \"No direct NLE1-E2F1 interaction demonstrated\"\n      ]\n    },\n    {\n      \"year\": 2024,\n      \"claim\": \"Direct interaction with Notch1 was demonstrated and exploited pharmacologically, establishing NLE1 as a druggable Notch-pathway component in brain tumor stem cells.\",\n      \"evidence\": \"Drug-target identification of edaravone binding NLE1, Notch reporter and self-renewal assays in BTSCs, and preclinical glioblastoma survival models\",\n      \"pmids\": [\"39423824\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\n        \"Precise biochemical NLE1-Notch1 binding interface not detailed\",\n        \"Whether edaravone-NLE1 binding is direct and specific not fully characterized\",\n        \"Relationship of the Notch1 complex to NLE1's protein-biosynthesis role unaddressed\"\n      ]\n    },\n    {\n      \"year\": 2025,\n      \"claim\": \"NLE1's pro-proliferative role was extended to hepatocellular carcinoma, reinforcing its broad pan-cancer growth function.\",\n      \"evidence\": \"CCK-8 and colony formation proliferation assays with CRISPR screen and multiomics enrichment analysis\",\n      \"pmids\": [\"40625398\"],\n      \"confidence\": \"Low\",\n      \"gaps\": [\n        \"Proliferation assays without pathway mechanistic follow-up\",\n        \"No direct partner or substrate identified in HCC\",\n        \"Correlative enrichment analysis only\"\n      ]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"How NLE1's molecular role in protein biosynthesis mechanistically connects to its Notch1, PI3K/AKT, and E2F1/CDK1 functions remains unresolved.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\n        \"No structural or biochemical definition of NLE1's molecular activity\",\n        \"No unifying model linking the biosynthetic and signaling functions\",\n        \"Direct substrates and stable complex composition unknown\"\n      ]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [],\n    \"localization\": [],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-392499\", \"supporting_discovery_ids\": [1]},\n      {\"term_id\": \"R-HSA-162582\", \"supporting_discovery_ids\": [3, 5, 6]},\n      {\"term_id\": \"R-HSA-1643685\", \"supporting_discovery_ids\": [0, 2]}\n    ],\n    \"complexes\": [],\n    \"partners\": [\"NOTCH1\"],\n    \"other_free_text\": []\n  }\n}","audit_flag":null,"evaluation":{"pairwise":"win","faith_supported":6,"faith_total":6,"faith_pct":100.0}}