{"gene":"NTN1","run_date":"2026-06-10T05:19:52","timeline":{"discoveries":[{"year":2019,"finding":"The microRNA let-7 directly targets the 3'-UTR of Ntn1 mRNA, represses Ntn1 expression, and reduces secretion of Ntn1 protein in Schwann cells, thereby reducing axon outgrowth in dorsal root ganglion neurons.","method":"Quantitative RT-PCR, biochemical assays, and luciferase reporter gene assay targeting the 3'-UTR of Ntn1","journal":"The Journal of biological chemistry","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — direct 3'-UTR reporter assay plus functional readout (axon outgrowth), single lab, two orthogonal methods","pmids":["30626732"],"is_preprint":false},{"year":2018,"finding":"Netrin-1 (NTN1) promotes migration of immortalized GnRH neurons (GN11 cells), and rare heterozygous NTN1 mutations found in congenital hypogonadotropic hypogonadism patients alter intracellular signaling and cell morphology, consistent with a role in GnRH neuron ontogeny.","method":"In vitro migration assay of GN11 cells; in vitro characterization of patient-derived rare sequence variants showing altered intracellular signaling and cell morphology","journal":"Human molecular genetics","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — cell migration functional assay plus in vitro variant characterization, single lab, two orthogonal methods","pmids":["29202173"],"is_preprint":false},{"year":2023,"finding":"m5C methylation of NTN1 mRNA by NSUN2 stabilizes NTN1 mRNA; the lncRNA DIAPH2-AS1 protects NSUN2 from ubiquitin-proteasomal degradation (by masking K577/K579 of NSUN2), thereby increasing NTN1 expression and promoting neural invasion of gastric cancer cells.","method":"RNA pulldown, RNA immunoprecipitation, ubiquitination assays, in vitro and in vivo invasion/migration models","journal":"Cell death & disease","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — reciprocal pulldown/RIP plus functional rescue experiments, single lab, multiple orthogonal methods","pmids":["37037818"],"is_preprint":false},{"year":2013,"finding":"IGF-1 upregulates Ntn1 transcription in the neonatal murine cochlea via both the PI3K/Akt and MEK/ERK pathways, as demonstrated by pathway inhibitor experiments, with the transcriptional response limited to within 12 hours of IGF-1 treatment.","method":"cDNA microarray, qRT-PCR, and pharmacological inhibition of PI3K/Akt and MEK/ERK pathways","journal":"Neuroscience letters","confidence":"Low","confidence_rationale":"Tier 3 / Weak — pathway inhibitors used but indirect readout (mRNA levels), single lab, single study","pmids":["24333914"],"is_preprint":false},{"year":2022,"finding":"The lncRNA SIMALR partners with HIF1α to regulate NTN1 promoter activity; NTN1 knockdown in LPS/IFNγ-stimulated macrophages induces apoptosis, and the apoptotic phenotype caused by SIMALR knockdown is rescued by recombinant NTN1, establishing NTN1 as a downstream survival factor in inflammatory macrophages.","method":"RNA immunoprecipitation, luciferase NTN1 promoter reporter assay, siRNA knockdown, Annexin V apoptosis assay, recombinant NTN1 rescue experiment","journal":"Arteriosclerosis, thrombosis, and vascular biology","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — promoter reporter assay, RIP, functional rescue, multiple orthogonal methods, single lab","pmids":["36546321"],"is_preprint":false},{"year":2019,"finding":"The rs4791331-T allele in the NTN1 locus disrupts a transcription factor binding motif, reduces NTN1 enhancer/promoter activity (measured by luciferase reporter), lowers NTN1 expression, and exerts anti-apoptotic effects and altered cell cycle progression; ntn1a knockout zebrafish display wider intermaxillary fissures, consistent with a role for NTN1 in craniofacial development.","method":"Luciferase enhancer reporter assay, CRISPR/Cas9 ntn1a knockout zebrafish, cell cycle and apoptosis assays, qRT-PCR","journal":"European journal of human genetics : EJHG","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — luciferase reporter plus in vivo KO model plus cellular assays, single lab, multiple methods","pmids":["31780810"],"is_preprint":false},{"year":2025,"finding":"In cerebellar granule neuron germinal zone exit, Pard3 polarity protein and JamC adhesion molecule promote, while the Siah2 ubiquitin ligase inhibits, surface recruitment of the Netrin-1 receptor DCC; this gating of DCC surface availability controls Netrin-1-mediated repulsion of maturing neurons from the germinal zone, demonstrating that cell polarity integrates adhesive and guidance cues for germinal zone exit.","method":"Genetic epistasis analysis in mouse cerebellum, live imaging, loss-of-function and rescue experiments for Pard3, JamC, Siah2, and DCC; direct measurement of DCC receptor surface recruitment","journal":"Nature communications","confidence":"High","confidence_rationale":"Tier 2 / Strong — genetic epistasis with multiple components (Pard3, JamC, Siah2, DCC), direct receptor surface recruitment assay, in vivo mouse model, peer-reviewed replication of preprint","pmids":["39774925","39399669"],"is_preprint":false},{"year":2019,"finding":"NTN1 silencing in C2C12 myoblasts promotes cell proliferation and inhibits differentiation, whereas NTN1 overexpression inhibits proliferation and promotes differentiation; these effects are linked to the AMPK pathway, suggesting NTN1 regulates muscle cell fate via AMPK signaling.","method":"siRNA knockdown, gene overexpression, proliferation and differentiation assays in C2C12 cells, Western blotting","journal":"Animals : an open access journal from MDPI","confidence":"Low","confidence_rationale":"Tier 3 / Weak — functional cell assays with pathway inference, single lab, single study, no direct mechanistic confirmation of AMPK link","pmids":["31461826"],"is_preprint":false},{"year":2025,"finding":"NTN1 overexpression suppresses apoptosis, inflammation, and ROS in LPS-treated kidney tubular cells (HK-2); mechanistically, NTN1 interacts with MAP1B (identified by co-immunoprecipitation), promotes MAP1B-dependent autophagy (increased Atg5, LC3II/I ratio), and MAP1B in turn interacts with DAPK1 to regulate membrane blebbing and cell survival.","method":"Co-immunoprecipitation, Western blotting, MAP1B phosphorylation inhibitors, DAPK1 knockdown/overexpression, ROS measurement","journal":"Open medicine (Warsaw, Poland)","confidence":"Low","confidence_rationale":"Tier 3 / Weak — single Co-IP for NTN1-MAP1B interaction, functional assays but single lab, no in vitro reconstitution","pmids":["42158103"],"is_preprint":false},{"year":2025,"finding":"NTN1 promotes pancreatic tumorigenesis and sympathetic innervation through its receptor NEO1; NTN1/NEO1 signaling activates focal adhesion kinase (FAK) to enhance EMT (ZEB1, SOX9 upregulation) and cancer stemness. NTN1 is upregulated by KrasG12D mutation and β-adrenergic signaling via the MAPK pathway. Ntn1 knockout in KrasG12D mice decreased sympathetic innervation and pancreatic intraepithelial neoplasia. A NTN1-neutralizing antibody or NEO1 knockdown reduced tumor progression in a liver metastasis model.","method":"KrasG12D pancreatic organoid qRT-PCR screen; in vivo Ntn1 KO mouse model (Pdx1-Cre;LSL-KrasG12D/+); ex vivo celiac ganglia axonogenesis assay; recombinant NTN1 treatment; NTN1-neutralizing antibody; Neo1 shRNA knockdown; Western blotting for FAK phosphorylation","journal":"bioRxiv","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — multiple orthogonal in vivo and in vitro methods in single preprint, genetic KO plus pharmacological neutralization plus receptor KD, functional phenotypes measured","pmids":["bio_10.1101_2025.07.22.666009"],"is_preprint":true},{"year":2025,"finding":"Genetic NTN1 overexpression stabilizes the blood-retina barrier (BRB) in an Unc5b-dependent manner: NTN1 overexpression rescued vascular leak in oxygen-induced retinopathy and laser-induced choroidal neovascularization models, while Unc5b-independent effects on retinal angiogenesis were minimal and transient.","method":"Genetic NTN1 overexpression in mice; vascular leak assays in OIR and CNV models; electroretinogram (ERG); genetic epistasis with Unc5b","journal":"bioRxiv","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — genetic overexpression with receptor epistasis (Unc5b), multiple functional assays, single lab preprint","pmids":["bio_10.1101_2025.06.14.658755"],"is_preprint":true},{"year":2025,"finding":"NTN1 is expressed in optic fissure fusion pioneer cells in the chicken embryo, as revealed by a CRISPR/Cas9-generated NTN1-T2A-eGFP reporter line in which GFP faithfully replicated endogenous NTN1 expression; transcriptomic profiling of these pioneer cells identified new optic fissure closure markers and novel pathways for developmental tissue fusion.","method":"CRISPR/Cas9 knock-in of T2A-eGFP reporter into NTN1 locus; spectral fluorescence cell-sorting; RNA-seq transcriptomic profiling of pioneer cells","journal":"Scientific reports","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — direct knock-in reporter line faithfully replicating endogenous expression, FACS isolation plus transcriptomics, peer-reviewed publication of preprint","pmids":["40128351"],"is_preprint":false},{"year":2025,"finding":"NTN1 attenuates Aβ1-42-induced neuroinflammation and memory impairment in rats by inhibiting microglial activation, suppressing NLRP3 inflammasome activation, preventing IL-1β and IL-18 release, and blocking NF-κB upstream signaling in the hippocampus and cortex.","method":"Microinjection of recombinant NTN1 in Aβ1-42 rat model; Morris water maze; Western blotting for NLRP3, NF-κB pathway components, caspase-1, IL-1β, IL-18","journal":"Frontiers in aging neuroscience","confidence":"Low","confidence_rationale":"Tier 3 / Weak — in vivo pharmacological protein administration with behavioral and protein readouts, single lab, no direct molecular mechanism established for NTN1's anti-inflammatory action","pmids":["40357234"],"is_preprint":false}],"current_model":"NTN1 (Netrin-1) is a secreted axon guidance cue that signals through receptors DCC, NEO1, and UNC5B to direct neuronal migration and axon pathfinding, regulate germinal zone exit via a cell-polarity-gated DCC surface recruitment circuit (controlled by Pard3/JamC vs. Siah2), stabilize the blood-retina barrier through Unc5b, promote pancreatic tumor innervation and EMT through NEO1/FAK signaling, and act as a macrophage survival factor downstream of HIF1α; its expression is post-transcriptionally regulated by let-7 (via 3'-UTR binding) and transcriptionally by IGF-1 through PI3K/Akt and MEK/ERK pathways, while its mRNA stability is enhanced by NSUN2-mediated m5C modification."},"narrative":{"mechanistic_narrative":"NTN1 (Netrin-1) is a secreted guidance cue that signals through cell-surface receptors to direct neuronal migration, axon outgrowth, and tissue morphogenesis, and that is co-opted in cancer and vascular and inflammatory settings [PMID:39774925, PMID:39399669, PMID:bio_10.1101_2025.07.22.666009]. In cerebellar granule neuron germinal zone exit, Netrin-1-mediated repulsion of maturing neurons is gated by surface availability of the receptor DCC, which is promoted by the polarity protein Pard3 and the adhesion molecule JamC and antagonized by the ubiquitin ligase Siah2 — integrating adhesive and guidance cues with cell polarity [PMID:39774925, PMID:39399669]. In peripheral neurons, Netrin-1 secreted by Schwann cells supports axon outgrowth in dorsal root ganglion neurons, and in cell-based assays it promotes migration of GnRH neurons, with rare heterozygous NTN1 variants from congenital hypogonadotropic hypogonadism patients altering intracellular signaling and morphology [PMID:30626732, PMID:29202173]. Beyond the nervous system, Netrin-1 acts through distinct receptors in disease contexts: NEO1 signaling activates FAK to drive EMT and stemness during pancreatic tumorigenesis and sympathetic innervation [PMID:bio_10.1101_2025.07.22.666009], and Unc5b-dependent signaling stabilizes the blood-retina barrier [PMID:bio_10.1101_2025.06.14.658755]. NTN1 also functions as a survival factor for inflammatory macrophages downstream of a HIF1α/SIMALR transcriptional circuit [PMID:36546321]. NTN1 expression is controlled at multiple levels: transcriptionally through enhancer/promoter elements [PMID:31780810], post-transcriptionally by let-7 binding the 3'-UTR [PMID:30626732], and via NSUN2-mediated m5C methylation that stabilizes NTN1 mRNA to promote gastric cancer neural invasion [PMID:37037818].","teleology":[{"year":2013,"claim":"Established that Ntn1 is an inducible target of growth-factor signaling, linking IGF-1 to upregulation of a guidance cue in developing sensory tissue.","evidence":"cDNA microarray, qRT-PCR, and pharmacological inhibition of PI3K/Akt and MEK/ERK pathways in neonatal murine cochlea","pmids":["24333914"],"confidence":"Low","gaps":["Indirect mRNA-level readout without identification of the responsible transcription factor","Does not establish a functional consequence for cochlear development"]},{"year":2018,"claim":"Connected NTN1 to GnRH neuron biology and human disease by showing it promotes GnRH neuron migration and that patient-derived variants perturb signaling.","evidence":"GN11 cell migration assays and in vitro characterization of rare variants from congenital hypogonadotropic hypogonadism patients","pmids":["29202173"],"confidence":"Medium","gaps":["Migration assayed in an immortalized cell line, not in vivo GnRH neuron tracts","Receptor mediating the migratory effect not defined"]},{"year":2019,"claim":"Defined a post-transcriptional brake on NTN1, showing let-7 directly represses Ntn1 to limit axon outgrowth.","evidence":"3'-UTR luciferase reporter, qRT-PCR, and DRG axon outgrowth assays in Schwann cells","pmids":["30626732"],"confidence":"Medium","gaps":["Does not address upstream regulation of let-7 itself","Single-lab functional readout"]},{"year":2019,"claim":"Identified cis-regulatory control of NTN1 and an in vivo developmental role, linking a risk allele to reduced enhancer activity and craniofacial phenotypes.","evidence":"Luciferase enhancer reporter, CRISPR/Cas9 ntn1a knockout zebrafish, cell cycle and apoptosis assays","pmids":["31780810"],"confidence":"Medium","gaps":["Transcription factor binding the disrupted motif not conclusively identified","Zebrafish phenotype not mechanistically tied to receptor signaling"]},{"year":2022,"claim":"Placed NTN1 downstream of a HIF1α/lncRNA transcriptional circuit and established it as a macrophage survival factor.","evidence":"RIP, NTN1 promoter luciferase reporter, siRNA knockdown, Annexin V apoptosis, and recombinant NTN1 rescue in stimulated macrophages","pmids":["36546321"],"confidence":"Medium","gaps":["Receptor transducing the survival signal in macrophages not defined","In vitro stimulation model only"]},{"year":2023,"claim":"Revealed epitranscriptomic control of NTN1, with NSUN2-deposited m5C stabilizing NTN1 mRNA to drive cancer neural invasion.","evidence":"RNA pulldown, RIP, ubiquitination assays, and in vitro/in vivo invasion models in gastric cancer","pmids":["37037818"],"confidence":"Medium","gaps":["m5C site mapping on NTN1 mRNA not detailed","Downstream receptor signaling driving invasion not addressed"]},{"year":2025,"claim":"Resolved how Netrin-1 guidance is spatially gated during neuronal migration, showing cell polarity controls DCC surface availability for germinal zone exit.","evidence":"Genetic epistasis (Pard3, JamC, Siah2, DCC), live imaging, and direct DCC surface recruitment assays in mouse cerebellum","pmids":["39774925","39399669"],"confidence":"High","gaps":["Molecular mechanism by which Siah2 vs Pard3/JamC differentially route DCC not fully resolved","Whether the same gating operates in other migrating neuron populations untested"]},{"year":2025,"claim":"Defined a NEO1/FAK signaling axis through which NTN1 drives pancreatic tumorigenesis and tumor innervation.","evidence":"KrasG12D organoid screen, Ntn1 KO mouse model, ex vivo axonogenesis assay, neutralizing antibody and Neo1 shRNA (preprint)","pmids":["bio_10.1101_2025.07.22.666009"],"confidence":"Medium","gaps":["Preprint, not peer-reviewed","How NEO1 engages FAK mechanistically not established"]},{"year":2025,"claim":"Showed NTN1 stabilizes the blood-retina barrier through Unc5b, distinguishing barrier maintenance from angiogenic effects.","evidence":"Genetic NTN1 overexpression in mice, vascular leak assays in OIR and CNV models, Unc5b epistasis (preprint)","pmids":["bio_10.1101_2025.06.14.658755"],"confidence":"Medium","gaps":["Preprint, not peer-reviewed","Endothelial signaling downstream of Unc5b not characterized"]},{"year":2025,"claim":"Demonstrated faithful NTN1 expression in optic fissure fusion pioneer cells and used a reporter to profile their transcriptome.","evidence":"CRISPR/Cas9 NTN1-T2A-eGFP knock-in chicken reporter, FACS isolation, and RNA-seq","pmids":["40128351"],"confidence":"Medium","gaps":["Functional requirement of NTN1 for optic fissure fusion not demonstrated","Receptor partners in pioneer cells unknown"]},{"year":2025,"claim":"Reported anti-inflammatory and neuroprotective actions of NTN1 in an Alzheimer's model via suppression of NLRP3/NF-κB signaling.","evidence":"Microinjection of recombinant NTN1 in Aβ1-42 rat model, Morris water maze, and Western blotting for inflammasome components","pmids":["40357234"],"confidence":"Low","gaps":["No direct molecular mechanism linking NTN1 to NLRP3/NF-κB suppression","Receptor mediating the anti-inflammatory effect not identified"]},{"year":2025,"claim":"Suggested NTN1 regulates myoblast proliferation and differentiation through AMPK, but the link is inferential.","evidence":"siRNA knockdown, overexpression, and proliferation/differentiation assays in C2C12 cells","pmids":["31461826"],"confidence":"Low","gaps":["AMPK link inferred, not directly confirmed","Receptor and signaling intermediates undefined"]},{"year":null,"claim":"How NTN1's distinct receptors (DCC, NEO1, UNC5B) and intracellular partners are selected across these diverse contexts to produce migration, survival, barrier, or oncogenic outputs remains unresolved.","evidence":"","pmids":[],"confidence":"Medium","gaps":["No unified model linking receptor choice to downstream effector pathways","Tissue-specific co-receptor and adaptor requirements largely uncharacterized"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0048018","term_label":"receptor ligand activity","supporting_discovery_ids":[6,9,10]},{"term_id":"GO:0098772","term_label":"molecular function regulator activity","supporting_discovery_ids":[4,9,10]}],"localization":[{"term_id":"GO:0005576","term_label":"extracellular region","supporting_discovery_ids":[0,4,12]}],"pathway":[{"term_id":"R-HSA-1266738","term_label":"Developmental Biology","supporting_discovery_ids":[6,11]},{"term_id":"R-HSA-1643685","term_label":"Disease","supporting_discovery_ids":[2,9]},{"term_id":"R-HSA-162582","term_label":"Signal Transduction","supporting_discovery_ids":[9,10]}],"complexes":[],"partners":["DCC","NEO1","UNC5B","MAP1B"],"other_free_text":[]}},"prefetch_data":{"uniprot":{"accession":"O95631","full_name":"Netrin-1","aliases":["Epididymis tissue protein Li 131P"],"length_aa":604,"mass_kda":67.7,"function":"Netrins control guidance of CNS commissural axons and peripheral motor axons. Its association with either DCC or some UNC5 receptors will lead to axon attraction or repulsion, respectively. Binding to UNC5C might cause dissociation of UNC5C from polymerized TUBB3 in microtubules and thereby lead to increased microtubule dynamics and axon repulsion (PubMed:28483977). Involved in dorsal root ganglion axon projection towards the spinal cord (PubMed:28483977). It also serves as a survival factor via its association with its receptors which prevent the initiation of apoptosis. Involved in tumorigenesis by regulating apoptosis (PubMed:15343335)","subcellular_location":"Secreted; Cytoplasm","url":"https://www.uniprot.org/uniprotkb/O95631/entry"},"depmap":{"release":"DepMap","has_data":true,"is_common_essential":false,"resolved_as":"","url":"https://depmap.org/portal/gene/NTN1","classification":"Not Classified","n_dependent_lines":2,"n_total_lines":1208,"dependency_fraction":0.0016556291390728477},"opencell":{"profiled":true,"resolved_as":"NET1","ensg_id":"ENSG00000173848","cell_line_id":"CID000580","localizations":[{"compartment":"nucleoplasm","grade":3}],"interactors":[{"gene":"KPNB1","stoichiometry":0.2}],"url":"https://opencell.sf.czbiohub.org/target/CID000580","total_profiled":1310},"omim":[{"mim_id":"618689","title":"NETRIN G2; NTNG2","url":"https://www.omim.org/entry/618689"},{"mim_id":"618264","title":"MIRROR MOVEMENTS 4; MRMV4","url":"https://www.omim.org/entry/618264"},{"mim_id":"616466","title":"UNC5 NETRIN RECEPTOR D; UNC5D","url":"https://www.omim.org/entry/616466"},{"mim_id":"615029","title":"PRECEREBELLIN 4; CBLN4","url":"https://www.omim.org/entry/615029"},{"mim_id":"614688","title":"PONTINE TEGMENTAL CAP DYSPLASIA; PTCD","url":"https://www.omim.org/entry/614688"}],"hpa":{"profiled":true,"resolved_as":"","reliability":"Approved","locations":[{"location":"Cytosol","reliability":"Approved"},{"location":"Nucleoplasm","reliability":"Additional"},{"location":"Actin filaments","reliability":"Additional"}],"tissue_specificity":"Tissue enhanced","tissue_distribution":"Detected in many","driving_tissues":[{"tissue":"blood vessel","ntpm":35.5},{"tissue":"heart muscle","ntpm":56.1}],"url":"https://www.proteinatlas.org/search/NTN1"},"hgnc":{"alias_symbol":["NTN1L","NET1"],"prev_symbol":[]},"alphafold":{"accession":"O95631","domains":[{"cath_id":"2.60.120.260","chopping":"55-284","consensus_level":"high","plddt":93.2089,"start":55,"end":284},{"cath_id":"2.10.25.10","chopping":"404-451","consensus_level":"medium","plddt":94.5227,"start":404,"end":451},{"cath_id":"2.40.50.120","chopping":"483-597","consensus_level":"high","plddt":87.5411,"start":483,"end":597}],"viewer_url":"https://alphafold.ebi.ac.uk/entry/O95631","model_url":"https://alphafold.ebi.ac.uk/files/AF-O95631-F1-model_v6.cif","pae_url":"https://alphafold.ebi.ac.uk/files/AF-O95631-F1-predicted_aligned_error_v6.png","plddt_mean":88.5},"mouse_models":{"mgi_url":"https://www.informatics.jax.org/marker/summary?nomen=NTN1","jax_strain_url":"https://www.jax.org/strain/search?query=NTN1"},"sequence":{"accession":"O95631","fasta_url":"https://rest.uniprot.org/uniprotkb/O95631.fasta","uniprot_url":"https://www.uniprot.org/uniprotkb/O95631/entry","alphafold_viewer_url":"https://alphafold.ebi.ac.uk/entry/O95631"}},"corpus_meta":[{"pmid":"30626732","id":"PMC_30626732","title":"The microRNAs let-7 and miR-9 down-regulate the axon-guidance genes Ntn1 and Dcc during peripheral nerve regeneration.","date":"2019","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/30626732","citation_count":47,"is_preprint":false},{"pmid":"37037818","id":"PMC_37037818","title":"Long noncoding RNA DIAPH2-AS1 promotes neural invasion of gastric cancer via stabilizing NSUN2 to enhance the m5C modification of NTN1.","date":"2023","source":"Cell death & disease","url":"https://pubmed.ncbi.nlm.nih.gov/37037818","citation_count":46,"is_preprint":false},{"pmid":"29202173","id":"PMC_29202173","title":"DCC/NTN1 complex mutations in patients with congenital hypogonadotropic hypogonadism impair GnRH neuron development.","date":"2018","source":"Human molecular genetics","url":"https://pubmed.ncbi.nlm.nih.gov/29202173","citation_count":36,"is_preprint":false},{"pmid":"24333914","id":"PMC_24333914","title":"Insulin-like growth factor 1 induces the transcription of Gap43 and Ntn1 during hair cell protection in the neonatal murine cochlea.","date":"2013","source":"Neuroscience letters","url":"https://pubmed.ncbi.nlm.nih.gov/24333914","citation_count":31,"is_preprint":false},{"pmid":"36546321","id":"PMC_36546321","title":"Human Macrophage Long Intergenic Noncoding RNA, SIMALR, Suppresses Inflammatory Macrophage Apoptosis via NTN1 (Netrin-1).","date":"2022","source":"Arteriosclerosis, thrombosis, and vascular biology","url":"https://pubmed.ncbi.nlm.nih.gov/36546321","citation_count":17,"is_preprint":false},{"pmid":"31780810","id":"PMC_31780810","title":"The functional variant of NTN1 contributes to the risk of nonsyndromic cleft lip with or without cleft palate.","date":"2019","source":"European journal of human genetics : EJHG","url":"https://pubmed.ncbi.nlm.nih.gov/31780810","citation_count":17,"is_preprint":false},{"pmid":"31461826","id":"PMC_31461826","title":"NTN1 Affects Porcine Intramuscular Fat Content by Affecting the Expression of Myogenic Regulatory Factors.","date":"2019","source":"Animals : an open access journal from MDPI","url":"https://pubmed.ncbi.nlm.nih.gov/31461826","citation_count":10,"is_preprint":false},{"pmid":"28489749","id":"PMC_28489749","title":"Association between PAX7 and NTN1 gene polymorphisms and nonsyndromic orofacial clefts in a northern Chinese population.","date":"2017","source":"Medicine","url":"https://pubmed.ncbi.nlm.nih.gov/28489749","citation_count":10,"is_preprint":false},{"pmid":"36879001","id":"PMC_36879001","title":"Identification of putative regulatory single-nucleotide variants in NTN1 gene associated with NSCL/P.","date":"2023","source":"Journal of human genetics","url":"https://pubmed.ncbi.nlm.nih.gov/36879001","citation_count":6,"is_preprint":false},{"pmid":"30397880","id":"PMC_30397880","title":"NLRP1 and NTN1, Deregulated Blood Differentially Methylated Regions in Mild Cognitive Impairment Patients.","date":"2018","source":"Journal of molecular neuroscience : 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Dcc.","date":"2024","source":"Research square","url":"https://pubmed.ncbi.nlm.nih.gov/39399669","citation_count":1,"is_preprint":false},{"pmid":"40604203","id":"PMC_40604203","title":"Therapeutic effects of miR-937-3p by targeting NTN1 expression and regulating apoptosis in an Aβ-induced neuronal cell death.","date":"2025","source":"Scientific reports","url":"https://pubmed.ncbi.nlm.nih.gov/40604203","citation_count":0,"is_preprint":false},{"pmid":"42158103","id":"PMC_42158103","title":"NTN1 regulates autophagy through the MAP1B/DAPK1 axis to ameliorate acute kidney injury in vitro.","date":"2026","source":"Open medicine (Warsaw, Poland)","url":"https://pubmed.ncbi.nlm.nih.gov/42158103","citation_count":0,"is_preprint":false},{"pmid":"39648562","id":"PMC_39648562","title":"A Novel De Novo Missense Variant in Netrin-1 (NTN1) Associated With Chorioretinal Coloboma, Sensorineural Hearing Loss and Polydactyly.","date":"2024","source":"Clinical genetics","url":"https://pubmed.ncbi.nlm.nih.gov/39648562","citation_count":0,"is_preprint":false},{"pmid":null,"id":"bio_10.1101_2025.07.22.666009","title":"Netrin-1 promotes pancreatic tumorigenesis and innervation through NEO1","date":"2025-07-26","source":"bioRxiv","url":"https://doi.org/10.1101/2025.07.22.666009","citation_count":0,"is_preprint":true},{"pmid":null,"id":"bio_10.1101_2025.06.14.658755","title":"Restoring compromised Blood-Retina-Barrier integrity with Netrin-1 overexpression","date":"2025-06-16","source":"bioRxiv","url":"https://doi.org/10.1101/2025.06.14.658755","citation_count":0,"is_preprint":true},{"pmid":null,"id":"bio_10.1101_2024.10.01.616014","title":"A stable  <i>Netrin-1</i>  fluorescent reporter chicken reveals cell-specific molecular signatures during optic fissure closure","date":"2024-10-02","source":"bioRxiv","url":"https://doi.org/10.1101/2024.10.01.616014","citation_count":0,"is_preprint":true},{"pmid":null,"id":"bio_10.1101_2025.01.21.25320889","title":"Variants in<i>CALD1</i>,<i>ESRP1</i>, and<i>RBFOX1</i>are associated with orofacial cleft risk","date":"2025-01-22","source":"bioRxiv","url":"https://doi.org/10.1101/2025.01.21.25320889","citation_count":0,"is_preprint":true}],"cost":{"stage1":{"model":"claude-sonnet-4-6","input_tokens":14974,"output_tokens":3613,"usd":0.049558,"stage1_stop_reason":"end_turn"},"stage2":{"model":"claude-opus-4-8","input_tokens":11252,"output_tokens":3768,"usd":0.07523,"stage2_stop_reason":"end_turn"},"total_usd":0.124788,"stage1_batch_id":"msgbatch_013a8chtv3AWyTqv4z4sEF4C","stage2_batch_id":"msgbatch_01AHsuW63vinsvuTgzmQ3n62","note":"batch pricing = 50% of standard"},"stage1_raw":"```json\n{\n  \"discoveries\": [\n    {\n      \"year\": 2019,\n      \"finding\": \"The microRNA let-7 directly targets the 3'-UTR of Ntn1 mRNA, represses Ntn1 expression, and reduces secretion of Ntn1 protein in Schwann cells, thereby reducing axon outgrowth in dorsal root ganglion neurons.\",\n      \"method\": \"Quantitative RT-PCR, biochemical assays, and luciferase reporter gene assay targeting the 3'-UTR of Ntn1\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — direct 3'-UTR reporter assay plus functional readout (axon outgrowth), single lab, two orthogonal methods\",\n      \"pmids\": [\"30626732\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2018,\n      \"finding\": \"Netrin-1 (NTN1) promotes migration of immortalized GnRH neurons (GN11 cells), and rare heterozygous NTN1 mutations found in congenital hypogonadotropic hypogonadism patients alter intracellular signaling and cell morphology, consistent with a role in GnRH neuron ontogeny.\",\n      \"method\": \"In vitro migration assay of GN11 cells; in vitro characterization of patient-derived rare sequence variants showing altered intracellular signaling and cell morphology\",\n      \"journal\": \"Human molecular genetics\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — cell migration functional assay plus in vitro variant characterization, single lab, two orthogonal methods\",\n      \"pmids\": [\"29202173\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"m5C methylation of NTN1 mRNA by NSUN2 stabilizes NTN1 mRNA; the lncRNA DIAPH2-AS1 protects NSUN2 from ubiquitin-proteasomal degradation (by masking K577/K579 of NSUN2), thereby increasing NTN1 expression and promoting neural invasion of gastric cancer cells.\",\n      \"method\": \"RNA pulldown, RNA immunoprecipitation, ubiquitination assays, in vitro and in vivo invasion/migration models\",\n      \"journal\": \"Cell death & disease\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — reciprocal pulldown/RIP plus functional rescue experiments, single lab, multiple orthogonal methods\",\n      \"pmids\": [\"37037818\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2013,\n      \"finding\": \"IGF-1 upregulates Ntn1 transcription in the neonatal murine cochlea via both the PI3K/Akt and MEK/ERK pathways, as demonstrated by pathway inhibitor experiments, with the transcriptional response limited to within 12 hours of IGF-1 treatment.\",\n      \"method\": \"cDNA microarray, qRT-PCR, and pharmacological inhibition of PI3K/Akt and MEK/ERK pathways\",\n      \"journal\": \"Neuroscience letters\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 / Weak — pathway inhibitors used but indirect readout (mRNA levels), single lab, single study\",\n      \"pmids\": [\"24333914\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"The lncRNA SIMALR partners with HIF1α to regulate NTN1 promoter activity; NTN1 knockdown in LPS/IFNγ-stimulated macrophages induces apoptosis, and the apoptotic phenotype caused by SIMALR knockdown is rescued by recombinant NTN1, establishing NTN1 as a downstream survival factor in inflammatory macrophages.\",\n      \"method\": \"RNA immunoprecipitation, luciferase NTN1 promoter reporter assay, siRNA knockdown, Annexin V apoptosis assay, recombinant NTN1 rescue experiment\",\n      \"journal\": \"Arteriosclerosis, thrombosis, and vascular biology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — promoter reporter assay, RIP, functional rescue, multiple orthogonal methods, single lab\",\n      \"pmids\": [\"36546321\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"The rs4791331-T allele in the NTN1 locus disrupts a transcription factor binding motif, reduces NTN1 enhancer/promoter activity (measured by luciferase reporter), lowers NTN1 expression, and exerts anti-apoptotic effects and altered cell cycle progression; ntn1a knockout zebrafish display wider intermaxillary fissures, consistent with a role for NTN1 in craniofacial development.\",\n      \"method\": \"Luciferase enhancer reporter assay, CRISPR/Cas9 ntn1a knockout zebrafish, cell cycle and apoptosis assays, qRT-PCR\",\n      \"journal\": \"European journal of human genetics : EJHG\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — luciferase reporter plus in vivo KO model plus cellular assays, single lab, multiple methods\",\n      \"pmids\": [\"31780810\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"In cerebellar granule neuron germinal zone exit, Pard3 polarity protein and JamC adhesion molecule promote, while the Siah2 ubiquitin ligase inhibits, surface recruitment of the Netrin-1 receptor DCC; this gating of DCC surface availability controls Netrin-1-mediated repulsion of maturing neurons from the germinal zone, demonstrating that cell polarity integrates adhesive and guidance cues for germinal zone exit.\",\n      \"method\": \"Genetic epistasis analysis in mouse cerebellum, live imaging, loss-of-function and rescue experiments for Pard3, JamC, Siah2, and DCC; direct measurement of DCC receptor surface recruitment\",\n      \"journal\": \"Nature communications\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — genetic epistasis with multiple components (Pard3, JamC, Siah2, DCC), direct receptor surface recruitment assay, in vivo mouse model, peer-reviewed replication of preprint\",\n      \"pmids\": [\"39774925\", \"39399669\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"NTN1 silencing in C2C12 myoblasts promotes cell proliferation and inhibits differentiation, whereas NTN1 overexpression inhibits proliferation and promotes differentiation; these effects are linked to the AMPK pathway, suggesting NTN1 regulates muscle cell fate via AMPK signaling.\",\n      \"method\": \"siRNA knockdown, gene overexpression, proliferation and differentiation assays in C2C12 cells, Western blotting\",\n      \"journal\": \"Animals : an open access journal from MDPI\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 / Weak — functional cell assays with pathway inference, single lab, single study, no direct mechanistic confirmation of AMPK link\",\n      \"pmids\": [\"31461826\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"NTN1 overexpression suppresses apoptosis, inflammation, and ROS in LPS-treated kidney tubular cells (HK-2); mechanistically, NTN1 interacts with MAP1B (identified by co-immunoprecipitation), promotes MAP1B-dependent autophagy (increased Atg5, LC3II/I ratio), and MAP1B in turn interacts with DAPK1 to regulate membrane blebbing and cell survival.\",\n      \"method\": \"Co-immunoprecipitation, Western blotting, MAP1B phosphorylation inhibitors, DAPK1 knockdown/overexpression, ROS measurement\",\n      \"journal\": \"Open medicine (Warsaw, Poland)\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 / Weak — single Co-IP for NTN1-MAP1B interaction, functional assays but single lab, no in vitro reconstitution\",\n      \"pmids\": [\"42158103\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"NTN1 promotes pancreatic tumorigenesis and sympathetic innervation through its receptor NEO1; NTN1/NEO1 signaling activates focal adhesion kinase (FAK) to enhance EMT (ZEB1, SOX9 upregulation) and cancer stemness. NTN1 is upregulated by KrasG12D mutation and β-adrenergic signaling via the MAPK pathway. Ntn1 knockout in KrasG12D mice decreased sympathetic innervation and pancreatic intraepithelial neoplasia. A NTN1-neutralizing antibody or NEO1 knockdown reduced tumor progression in a liver metastasis model.\",\n      \"method\": \"KrasG12D pancreatic organoid qRT-PCR screen; in vivo Ntn1 KO mouse model (Pdx1-Cre;LSL-KrasG12D/+); ex vivo celiac ganglia axonogenesis assay; recombinant NTN1 treatment; NTN1-neutralizing antibody; Neo1 shRNA knockdown; Western blotting for FAK phosphorylation\",\n      \"journal\": \"bioRxiv\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — multiple orthogonal in vivo and in vitro methods in single preprint, genetic KO plus pharmacological neutralization plus receptor KD, functional phenotypes measured\",\n      \"pmids\": [\"bio_10.1101_2025.07.22.666009\"],\n      \"is_preprint\": true\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"Genetic NTN1 overexpression stabilizes the blood-retina barrier (BRB) in an Unc5b-dependent manner: NTN1 overexpression rescued vascular leak in oxygen-induced retinopathy and laser-induced choroidal neovascularization models, while Unc5b-independent effects on retinal angiogenesis were minimal and transient.\",\n      \"method\": \"Genetic NTN1 overexpression in mice; vascular leak assays in OIR and CNV models; electroretinogram (ERG); genetic epistasis with Unc5b\",\n      \"journal\": \"bioRxiv\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — genetic overexpression with receptor epistasis (Unc5b), multiple functional assays, single lab preprint\",\n      \"pmids\": [\"bio_10.1101_2025.06.14.658755\"],\n      \"is_preprint\": true\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"NTN1 is expressed in optic fissure fusion pioneer cells in the chicken embryo, as revealed by a CRISPR/Cas9-generated NTN1-T2A-eGFP reporter line in which GFP faithfully replicated endogenous NTN1 expression; transcriptomic profiling of these pioneer cells identified new optic fissure closure markers and novel pathways for developmental tissue fusion.\",\n      \"method\": \"CRISPR/Cas9 knock-in of T2A-eGFP reporter into NTN1 locus; spectral fluorescence cell-sorting; RNA-seq transcriptomic profiling of pioneer cells\",\n      \"journal\": \"Scientific reports\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — direct knock-in reporter line faithfully replicating endogenous expression, FACS isolation plus transcriptomics, peer-reviewed publication of preprint\",\n      \"pmids\": [\"40128351\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"NTN1 attenuates Aβ1-42-induced neuroinflammation and memory impairment in rats by inhibiting microglial activation, suppressing NLRP3 inflammasome activation, preventing IL-1β and IL-18 release, and blocking NF-κB upstream signaling in the hippocampus and cortex.\",\n      \"method\": \"Microinjection of recombinant NTN1 in Aβ1-42 rat model; Morris water maze; Western blotting for NLRP3, NF-κB pathway components, caspase-1, IL-1β, IL-18\",\n      \"journal\": \"Frontiers in aging neuroscience\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 / Weak — in vivo pharmacological protein administration with behavioral and protein readouts, single lab, no direct molecular mechanism established for NTN1's anti-inflammatory action\",\n      \"pmids\": [\"40357234\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"NTN1 (Netrin-1) is a secreted axon guidance cue that signals through receptors DCC, NEO1, and UNC5B to direct neuronal migration and axon pathfinding, regulate germinal zone exit via a cell-polarity-gated DCC surface recruitment circuit (controlled by Pard3/JamC vs. Siah2), stabilize the blood-retina barrier through Unc5b, promote pancreatic tumor innervation and EMT through NEO1/FAK signaling, and act as a macrophage survival factor downstream of HIF1α; its expression is post-transcriptionally regulated by let-7 (via 3'-UTR binding) and transcriptionally by IGF-1 through PI3K/Akt and MEK/ERK pathways, while its mRNA stability is enhanced by NSUN2-mediated m5C modification.\"\n}\n```","stage2_raw":"{\n  \"mechanistic_narrative\": \"NTN1 (Netrin-1) is a secreted guidance cue that signals through cell-surface receptors to direct neuronal migration, axon outgrowth, and tissue morphogenesis, and that is co-opted in cancer and vascular and inflammatory settings [#6, #9]. In cerebellar granule neuron germinal zone exit, Netrin-1-mediated repulsion of maturing neurons is gated by surface availability of the receptor DCC, which is promoted by the polarity protein Pard3 and the adhesion molecule JamC and antagonized by the ubiquitin ligase Siah2 — integrating adhesive and guidance cues with cell polarity [#6]. In peripheral neurons, Netrin-1 secreted by Schwann cells supports axon outgrowth in dorsal root ganglion neurons, and in cell-based assays it promotes migration of GnRH neurons, with rare heterozygous NTN1 variants from congenital hypogonadotropic hypogonadism patients altering intracellular signaling and morphology [#0, #1]. Beyond the nervous system, Netrin-1 acts through distinct receptors in disease contexts: NEO1 signaling activates FAK to drive EMT and stemness during pancreatic tumorigenesis and sympathetic innervation [#9], and Unc5b-dependent signaling stabilizes the blood-retina barrier [#10]. NTN1 also functions as a survival factor for inflammatory macrophages downstream of a HIF1\\u03b1/SIMALR transcriptional circuit [#4]. NTN1 expression is controlled at multiple levels: transcriptionally through enhancer/promoter elements [#5], post-transcriptionally by let-7 binding the 3'-UTR [#0], and via NSUN2-mediated m5C methylation that stabilizes NTN1 mRNA to promote gastric cancer neural invasion [#2].\",\n  \"teleology\": [\n    {\n      \"year\": 2013,\n      \"claim\": \"Established that Ntn1 is an inducible target of growth-factor signaling, linking IGF-1 to upregulation of a guidance cue in developing sensory tissue.\",\n      \"evidence\": \"cDNA microarray, qRT-PCR, and pharmacological inhibition of PI3K/Akt and MEK/ERK pathways in neonatal murine cochlea\",\n      \"pmids\": [\"24333914\"],\n      \"confidence\": \"Low\",\n      \"gaps\": [\"Indirect mRNA-level readout without identification of the responsible transcription factor\", \"Does not establish a functional consequence for cochlear development\"]\n    },\n    {\n      \"year\": 2018,\n      \"claim\": \"Connected NTN1 to GnRH neuron biology and human disease by showing it promotes GnRH neuron migration and that patient-derived variants perturb signaling.\",\n      \"evidence\": \"GN11 cell migration assays and in vitro characterization of rare variants from congenital hypogonadotropic hypogonadism patients\",\n      \"pmids\": [\"29202173\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Migration assayed in an immortalized cell line, not in vivo GnRH neuron tracts\", \"Receptor mediating the migratory effect not defined\"]\n    },\n    {\n      \"year\": 2019,\n      \"claim\": \"Defined a post-transcriptional brake on NTN1, showing let-7 directly represses Ntn1 to limit axon outgrowth.\",\n      \"evidence\": \"3'-UTR luciferase reporter, qRT-PCR, and DRG axon outgrowth assays in Schwann cells\",\n      \"pmids\": [\"30626732\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Does not address upstream regulation of let-7 itself\", \"Single-lab functional readout\"]\n    },\n    {\n      \"year\": 2019,\n      \"claim\": \"Identified cis-regulatory control of NTN1 and an in vivo developmental role, linking a risk allele to reduced enhancer activity and craniofacial phenotypes.\",\n      \"evidence\": \"Luciferase enhancer reporter, CRISPR/Cas9 ntn1a knockout zebrafish, cell cycle and apoptosis assays\",\n      \"pmids\": [\"31780810\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Transcription factor binding the disrupted motif not conclusively identified\", \"Zebrafish phenotype not mechanistically tied to receptor signaling\"]\n    },\n    {\n      \"year\": 2022,\n      \"claim\": \"Placed NTN1 downstream of a HIF1\\u03b1/lncRNA transcriptional circuit and established it as a macrophage survival factor.\",\n      \"evidence\": \"RIP, NTN1 promoter luciferase reporter, siRNA knockdown, Annexin V apoptosis, and recombinant NTN1 rescue in stimulated macrophages\",\n      \"pmids\": [\"36546321\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Receptor transducing the survival signal in macrophages not defined\", \"In vitro stimulation model only\"]\n    },\n    {\n      \"year\": 2023,\n      \"claim\": \"Revealed epitranscriptomic control of NTN1, with NSUN2-deposited m5C stabilizing NTN1 mRNA to drive cancer neural invasion.\",\n      \"evidence\": \"RNA pulldown, RIP, ubiquitination assays, and in vitro/in vivo invasion models in gastric cancer\",\n      \"pmids\": [\"37037818\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"m5C site mapping on NTN1 mRNA not detailed\", \"Downstream receptor signaling driving invasion not addressed\"]\n    },\n    {\n      \"year\": 2025,\n      \"claim\": \"Resolved how Netrin-1 guidance is spatially gated during neuronal migration, showing cell polarity controls DCC surface availability for germinal zone exit.\",\n      \"evidence\": \"Genetic epistasis (Pard3, JamC, Siah2, DCC), live imaging, and direct DCC surface recruitment assays in mouse cerebellum\",\n      \"pmids\": [\"39774925\", \"39399669\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Molecular mechanism by which Siah2 vs Pard3/JamC differentially route DCC not fully resolved\", \"Whether the same gating operates in other migrating neuron populations untested\"]\n    },\n    {\n      \"year\": 2025,\n      \"claim\": \"Defined a NEO1/FAK signaling axis through which NTN1 drives pancreatic tumorigenesis and tumor innervation.\",\n      \"evidence\": \"KrasG12D organoid screen, Ntn1 KO mouse model, ex vivo axonogenesis assay, neutralizing antibody and Neo1 shRNA (preprint)\",\n      \"pmids\": [\"bio_10.1101_2025.07.22.666009\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Preprint, not peer-reviewed\", \"How NEO1 engages FAK mechanistically not established\"]\n    },\n    {\n      \"year\": 2025,\n      \"claim\": \"Showed NTN1 stabilizes the blood-retina barrier through Unc5b, distinguishing barrier maintenance from angiogenic effects.\",\n      \"evidence\": \"Genetic NTN1 overexpression in mice, vascular leak assays in OIR and CNV models, Unc5b epistasis (preprint)\",\n      \"pmids\": [\"bio_10.1101_2025.06.14.658755\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Preprint, not peer-reviewed\", \"Endothelial signaling downstream of Unc5b not characterized\"]\n    },\n    {\n      \"year\": 2025,\n      \"claim\": \"Demonstrated faithful NTN1 expression in optic fissure fusion pioneer cells and used a reporter to profile their transcriptome.\",\n      \"evidence\": \"CRISPR/Cas9 NTN1-T2A-eGFP knock-in chicken reporter, FACS isolation, and RNA-seq\",\n      \"pmids\": [\"40128351\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Functional requirement of NTN1 for optic fissure fusion not demonstrated\", \"Receptor partners in pioneer cells unknown\"]\n    },\n    {\n      \"year\": 2025,\n      \"claim\": \"Reported anti-inflammatory and neuroprotective actions of NTN1 in an Alzheimer's model via suppression of NLRP3/NF-\\u03baB signaling.\",\n      \"evidence\": \"Microinjection of recombinant NTN1 in A\\u03b21-42 rat model, Morris water maze, and Western blotting for inflammasome components\",\n      \"pmids\": [\"40357234\"],\n      \"confidence\": \"Low\",\n      \"gaps\": [\"No direct molecular mechanism linking NTN1 to NLRP3/NF-\\u03baB suppression\", \"Receptor mediating the anti-inflammatory effect not identified\"]\n    },\n    {\n      \"year\": 2025,\n      \"claim\": \"Suggested NTN1 regulates myoblast proliferation and differentiation through AMPK, but the link is inferential.\",\n      \"evidence\": \"siRNA knockdown, overexpression, and proliferation/differentiation assays in C2C12 cells\",\n      \"pmids\": [\"31461826\"],\n      \"confidence\": \"Low\",\n      \"gaps\": [\"AMPK link inferred, not directly confirmed\", \"Receptor and signaling intermediates undefined\"]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"How NTN1's distinct receptors (DCC, NEO1, UNC5B) and intracellular partners are selected across these diverse contexts to produce migration, survival, barrier, or oncogenic outputs remains unresolved.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"No unified model linking receptor choice to downstream effector pathways\", \"Tissue-specific co-receptor and adaptor requirements largely uncharacterized\"]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0048018\", \"supporting_discovery_ids\": [6, 9, 10]},\n      {\"term_id\": \"GO:0098772\", \"supporting_discovery_ids\": [4, 9, 10]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005576\", \"supporting_discovery_ids\": [0, 4, 12]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-1266738\", \"supporting_discovery_ids\": [6, 11]},\n      {\"term_id\": \"R-HSA-1643685\", \"supporting_discovery_ids\": [2, 9]},\n      {\"term_id\": \"R-HSA-162582\", \"supporting_discovery_ids\": [9, 10]}\n    ],\n    \"complexes\": [],\n    \"partners\": [\"DCC\", \"NEO1\", \"UNC5B\", \"MAP1B\"],\n    \"other_free_text\": []\n  }\n}","audit_flag":null,"evaluation":{"pairwise":"tie","faith_supported":5,"faith_total":5,"faith_pct":100.0}}