{"gene":"NLGN3","run_date":"2026-06-10T05:19:52","timeline":{"discoveries":[{"year":2003,"finding":"NLGN3 (and NLGN4) are cell-adhesion molecules localized at the synapse; missense and frameshift mutations in NLGN3 were identified in siblings with autism-spectrum disorders, implicating defective synaptogenesis as a predisposing mechanism.","method":"Genetic sequencing of affected sibling pairs; cellular localization described as postsynaptic","journal":"Nature genetics","confidence":"Medium","confidence_rationale":"Tier 3 / Strong — foundational human genetics with localization claim, replicated broadly across labs, but original paper is a genetic/sequencing study without direct biochemical reconstitution of mechanism","pmids":["12669065"],"is_preprint":false},{"year":2019,"finding":"Two autism-associated NLGN3 missense variants (p.Arg597Trp and p.Pro514Ser) reduce mature NLGN3 protein levels, impair plasma membrane localization, reduce extracellular cleavage, and induce an unfolded protein response (UPR) due to ER retention of immature protein, more severely than the p.Arg451Cys variant; control population variants (p.Ala632Thr, p.Val341Ala) have no such effect.","method":"Overexpression in HEK293 and HeLa cells; flow cytometry / immunofluorescence for plasma membrane localization; western blot for protein levels and UPR markers","journal":"Human mutation","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — multiple orthogonal cellular assays in one lab, with appropriate controls (benign variants tested in parallel)","pmids":["31184401"],"is_preprint":false},{"year":2021,"finding":"In glioma cells, NLGN3 promotes proliferation, migration, and invasion by activating PI3K-AKT, ERK1/2, and LYN signaling pathways; LYN in turn upregulates ADAM10 sheddase expression, which cleaves NLGN3, forming a positive autocrine feedback loop that promotes NLGN3 cleavage and further signaling.","method":"Knockdown and overexpression in U251 and U87 glioma cells; western blot for pathway activation; ADAM10 inhibition; proliferation/migration/invasion assays","journal":"Frontiers in cell and developmental biology","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — multiple functional assays and pathway inhibition in one lab, two cell lines","pmids":["34485271"],"is_preprint":false},{"year":2018,"finding":"ADAM10-mediated cleavage and secretion of NLGN3 from neurons is required for glioblastoma cell growth; inhibition of ADAM10 (ADAM10i) blocks neuron-conditioned-medium-induced GBM proliferation.","method":"ADAM10 inhibitor treatment of neuron-conditioned medium cultures; U251, U87-MG, and patient-derived GBM cell growth assays","journal":"Cancer medicine","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — pharmacological inhibition with functional readout, single lab","pmids":["29777576"],"is_preprint":false},{"year":2019,"finding":"NLGN3 promotes neuroblastoma cell proliferation and growth by activating the PI3K/AKT signaling pathway and FOXO family transcription activity; PI3K/AKT inhibition in NLGN3-overexpressing cells reverses the proliferative effect.","method":"MTT assay, colony formation, cell cycle analysis, BrdU incorporation, xenograft animal model; western blot for AKT phosphorylation; PI3K/AKT pathway inhibition","journal":"European journal of pharmacology","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — multiple functional assays plus pathway inhibition rescue, single lab","pmids":["31150649"],"is_preprint":false},{"year":2023,"finding":"CDK5 associates with NLGN3 in vivo and phosphorylates NLGN3 on serine 725 (S725); this phosphorylation regulates NLGN3 association with the Rho-GEF Kalirin-7, NLGN3 surface expression, and NLGN3-mediated synaptic currents in cultured neurons.","method":"HA-tagged knock-in mouse; Co-immunoprecipitation (in vivo Cdk5–NLGN3 association); in vitro phosphorylation assay; mutagenesis at S725; surface expression assay; patch-clamp electrophysiology in cultured rat neurons","journal":"The Journal of neuroscience","confidence":"High","confidence_rationale":"Tier 1 / Moderate — in vitro kinase assay plus mutagenesis plus knock-in mouse plus electrophysiological functional readout, all in one study","pmids":["37699715"],"is_preprint":false},{"year":2023,"finding":"Secreted/shed NLGN3 activates Gαi1/3-Akt-mTOR and Erk signaling in neuronal cells to protect against ischemia-reperfusion injury; Gαi1/3 silencing or knockout abolishes NLGN3-induced Akt activation and neuroprotection. In vivo, ADAM10-mediated cleavage is required for NLGN3 secretion following MCAO; ADAM10 inhibition blocks NLGN3 secretion and worsens ischemic injury.","method":"SH-SY5Y and primary cortical neurons with OGD/R; Gαi1/3 siRNA knockdown and genetic KO; ADAM10 inhibition; neuronal NLGN3 overexpression/silencing by viral vector; MCAO mouse model; western blot for signaling","journal":"Cell death & disease","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — genetic KO and knockdown with rescue in vivo plus in vitro mechanistic assays, single lab","pmids":["37880221"],"is_preprint":false},{"year":2024,"finding":"KPNB1 promotes NLGN3 expression in glioblastoma by mediating nuclear import of the transcription factor YBX1, which directly binds the NLGN3 promoter; USP7 deubiquitinase stabilizes KPNB1 by removing ubiquitin modifications; the USP7–KPNB1–YBX1–NLGN3 axis promotes GBM proliferation and migration.","method":"Co-immunoprecipitation and mass spectrometry (KPNB1–YBX1 interaction); chromatin immunoprecipitation (YBX1 binding to NLGN3 promoter); nuclear-cytoplasmic fractionation and immunofluorescence (YBX1 nuclear translocation); ubiquitination assays (USP7 effect on KPNB1); KPNB1/USP7/NLGN3 knockdown functional assays; intracranial orthotopic tumor model","journal":"Journal of experimental & clinical cancer research","confidence":"High","confidence_rationale":"Tier 1-2 / Moderate — ChIP, Co-IP/MS, ubiquitination assay, nuclear fractionation, and in vivo tumor model with multiple orthogonal methods in one study","pmids":["38254206"],"is_preprint":false},{"year":2023,"finding":"Wnt/β-catenin signaling transcriptionally upregulates NLGN3 expression in glioblastoma; secreted NLGN3 in conditioned medium and recombinant NLGN3 induce cancer stem cell (CSC) properties in neighboring GBM cells; DAB2IP suppresses CSC acquisition by targeting NLGN3.","method":"Conditioned medium and recombinant NLGN3 treatment of GBM cells; CSC sphere formation and marker assays; Wnt/β-catenin inhibitor treatment; DAB2IP knockdown/overexpression","journal":"Cell death & disease","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — multiple functional assays and pharmacological intervention, single lab","pmids":["37443071"],"is_preprint":false},{"year":2024,"finding":"NLGN3 interacts with Gαi1/3 (demonstrated by Co-IP) in cardiac endothelial cells; this NLGN3–Gαi1/3 interaction promotes endothelial cell proliferation and tube formation via PI3K-Akt-mTOR signaling; endothelial-specific NLGN3 knockdown or ADAM10 inhibition reduces angiogenesis and worsens cardiac function after myocardial infarction.","method":"Co-immunoprecipitation (NLGN3–Gαi1/3); Gαi1/3 knockout mouse MI model; endothelial-specific NLGN3 knockdown; ADAM10 inhibition; tube formation and proliferation assays","journal":"Basic research in cardiology","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — Co-IP plus genetic KO plus functional in vivo assays, single lab","pmids":["41398092"],"is_preprint":false},{"year":2024,"finding":"NLGN3 engages two distinct transsynaptic pathways: canonical interaction with neurexins (NRXNs) and a noncanonical interaction with protein tyrosine phosphatase δ (PTPδ). Selectively disrupting the NLGN3–PTPδ pathway impairs social motivation/reward (juvenile social conditioned place preference) and remote spatial memory, while disrupting the NLGN3–NRXN pathway attenuates contextual fear conditioning. The two pathways thus differentially regulate distinct higher-order brain functions.","method":"Nlgn3 knock-in mice selectively lacking interaction with NRXNs or PTPδ; social conditioned place preference, Barnes maze, and contextual fear conditioning behavioral tests","journal":"Molecular brain","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — genetic dissection using pathway-specific knock-in mice with multiple behavioral readouts, single lab","pmids":["38475840"],"is_preprint":false},{"year":2025,"finding":"Shed NLGN3 binds directly to CSPG4 on glioma cells and oligodendrocyte precursor cells (OPCs); NLGN3–CSPG4 interaction facilitates CSPG4 shedding by ADAM10, alters membrane tension, and activates PIEZO1 mechanosensitive channels, causing membrane depolarization; this NLGN3–CSPG4–PIEZO1 axis maintains OPCs in an undifferentiated stem-like state and promotes glioma proliferation.","method":"Binding/interaction assays (NLGN3–CSPG4); ADAM10 inhibition; PIEZO1 activity measurements; OPC differentiation assays; glioma proliferation assays","journal":"bioRxiv","confidence":"Low","confidence_rationale":"Tier 3 / Weak — preprint with mechanistic claims but abstract-level detail insufficient to assess full rigor; single lab, not yet peer-reviewed","pmids":["bio_10.1101_2025.07.12.664340"],"is_preprint":true},{"year":2026,"finding":"In Drosophila, the fly Nlgn3 homolog regulates sleep patterns, synaptic architecture, and vesicle dynamics. Human NLGN3 rescues the Nlg3-null phenotype. The de novo p.R175W variant and maternally inherited p.R451C variant alter synapse morphology and sleep; p.R597W alters sleep and vesicle dynamics with minimal synapse morphology effects. Overexpression data suggest p.R175W is gain-of-function while maternally inherited variants show mixed loss/gain-of-function effects.","method":"Transgenic Drosophila rescue and overexpression models; confocal imaging of synaptic morphology; sleep behavior assays; vesicle dynamics imaging","journal":"bioRxiv","confidence":"Low","confidence_rationale":"Tier 3 / Weak — preprint, Drosophila model system, abstract-level detail; cross-species ortholog study with functional validation but not peer-reviewed","pmids":["41929011"],"is_preprint":true},{"year":2023,"finding":"NLGN3 promotes neuritogenesis in developing GnRH neurons; overexpression of wild-type NLGN3 (but not loss-of-function mutant NLGN3) promotes neurite outgrowth, linking NLGN3 function to GnRH neuron maturation.","method":"Overexpression of wild-type vs. LoF mutant NLGN3 in developing GnRH cells; neuritogenesis quantification","journal":"Disease models & mechanisms","confidence":"Low","confidence_rationale":"Tier 3 / Weak — single overexpression experiment in one cell type, single lab","pmids":["36810932"],"is_preprint":false}],"current_model":"NLGN3 is a postsynaptic X-linked cell-adhesion molecule that organizes synapse assembly through canonical interactions with neurexins and a noncanonical interaction with PTPδ, is phosphorylated by CDK5 on S725 to regulate its association with the Rho-GEF Kalirin-7 and its surface expression and synaptic currents, and is shed from the cell surface by ADAM10; the shed ectodomain acts as a paracrine/autocrine factor that activates Gαi1/3–PI3K–Akt–mTOR signaling in neurons (neuroprotection), glioma cells (proliferation via LYN–ADAM10 feedback and CSC induction via Wnt/β-catenin–YBX1 transcriptional upregulation), and cardiac endothelial cells (angiogenesis), while disease-associated missense variants impair its ER processing, plasma membrane localization, and cleavage."},"narrative":{"mechanistic_narrative":"NLGN3 is an X-linked postsynaptic cell-adhesion molecule that organizes synapse assembly and, through proteolytic shedding of its ectodomain, acts as a paracrine signaling factor in both physiological and pathological contexts [PMID:12669065, PMID:37699715]. At the synapse it engages two distinct transsynaptic pathways—a canonical interaction with neurexins and a noncanonical interaction with PTPδ—that differentially regulate higher-order brain functions including social motivation, spatial memory, and contextual fear [PMID:38475840]. Its surface expression and synaptic currents are controlled by CDK5, which associates with NLGN3 in vivo and phosphorylates serine 725 to regulate NLGN3 association with the Rho-GEF Kalirin-7 [PMID:37699715]. NLGN3 is cleaved from the cell surface by ADAM10, and the shed ectodomain functions as a soluble ligand that activates Gαi1/3–PI3K–Akt–mTOR signaling: in neurons this confers neuroprotection against ischemia-reperfusion injury [PMID:37880221], and in cardiac endothelial cells it drives proliferation, tube formation, and angiogenesis after myocardial infarction [PMID:41398092]. In glioma the same shed NLGN3 axis is co-opted as a growth driver, activating PI3K-AKT, ERK, and LYN signaling, with LYN upregulating ADAM10 to form an autocrine cleavage feedback loop [PMID:34485271, PMID:29777576]; NLGN3 expression is transcriptionally amplified by Wnt/β-catenin signaling and by a USP7–KPNB1–YBX1 axis in which YBX1 binds the NLGN3 promoter, promoting cancer stem cell properties and tumor proliferation [PMID:38254206, PMID:37443071]. Autism-associated missense variants impair ER processing and maturation of NLGN3, reduce plasma membrane localization, and decrease extracellular cleavage, triggering an unfolded protein response [PMID:12669065, PMID:31184401].","teleology":[{"year":2003,"claim":"Established NLGN3 as a synaptic cell-adhesion molecule and linked it to disease, defining defective synaptogenesis as a candidate mechanism for autism-spectrum disorders.","evidence":"Genetic sequencing of affected sibling pairs with postsynaptic localization assignment","pmids":["12669065"],"confidence":"Medium","gaps":["No biochemical reconstitution of the synaptogenic mechanism","Causal mutation effect not functionally tested in this study"]},{"year":2018,"claim":"Resolved how NLGN3 promotes tumor growth non-cell-autonomously, showing that ADAM10-dependent shedding of neuronal NLGN3 is required to stimulate glioblastoma proliferation.","evidence":"ADAM10 inhibition of neuron-conditioned medium with patient-derived and cell-line GBM growth assays","pmids":["29777576"],"confidence":"Medium","gaps":["Downstream receptor on GBM cells not identified here","Single lab, pharmacological inhibition only"]},{"year":2019,"claim":"Showed that autism-associated missense variants act by disrupting NLGN3 maturation rather than synaptic binding per se, retaining immature protein in the ER and triggering a UPR.","evidence":"Overexpression in HEK293/HeLa with flow cytometry, immunofluorescence, and western blot, including benign control variants","pmids":["31184401"],"confidence":"Medium","gaps":["Heterologous overexpression, not neuronal context","Functional synaptic consequences not measured"]},{"year":2019,"claim":"Extended NLGN3's pro-proliferative role to neuroblastoma and identified PI3K/AKT-FOXO as the effector pathway through rescue by pathway inhibition.","evidence":"Proliferation, colony, cell-cycle, BrdU and xenograft assays with PI3K/AKT inhibition","pmids":["31150649"],"confidence":"Medium","gaps":["Receptor mediating NLGN3 signaling not defined","Single lab"]},{"year":2021,"claim":"Defined an autocrine feedback circuit in glioma whereby NLGN3 signaling through LYN upregulates ADAM10 to amplify its own cleavage and downstream PI3K-AKT/ERK signaling.","evidence":"Knockdown/overexpression in two glioma lines with ADAM10 inhibition and proliferation/migration/invasion assays","pmids":["34485271"],"confidence":"Medium","gaps":["Direct NLGN3 receptor not identified","Feedback loop quantification limited to two cell lines"]},{"year":2023,"claim":"Identified CDK5 as a kinase that phosphorylates NLGN3 at S725 to control its surface expression, Kalirin-7 association, and synaptic currents, providing a post-translational regulatory mechanism for synaptic function.","evidence":"Knock-in mouse Co-IP, in vitro kinase assay, S725 mutagenesis, surface expression assay, and patch-clamp electrophysiology","pmids":["37699715"],"confidence":"High","gaps":["Upstream signals activating CDK5-NLGN3 phosphorylation unknown","Link to disease variants not tested"]},{"year":2023,"claim":"Demonstrated a physiological neuroprotective role for shed NLGN3, showing it activates Gαi1/3-Akt-mTOR signaling in neurons and requires ADAM10 cleavage for protection against ischemic injury.","evidence":"OGD/R in SH-SY5Y and primary neurons with Gαi1/3 knockdown/KO, ADAM10 inhibition, viral NLGN3 manipulation, and MCAO mouse model","pmids":["37880221"],"confidence":"Medium","gaps":["NLGN3 receptor coupling to Gαi1/3 not structurally defined","Single lab"]},{"year":2023,"claim":"Connected NLGN3 to glioma stemness, showing Wnt/β-catenin transcriptionally upregulates NLGN3 and that secreted NLGN3 induces cancer stem cell properties, with DAB2IP as a suppressor.","evidence":"Conditioned medium and recombinant NLGN3 treatment, CSC sphere/marker assays, Wnt inhibition, DAB2IP manipulation","pmids":["37443071"],"confidence":"Medium","gaps":["Mechanism of DAB2IP targeting of NLGN3 unresolved","Single lab"]},{"year":2024,"claim":"Mapped a transcriptional control axis for NLGN3 in glioblastoma in which USP7 stabilizes KPNB1, which imports YBX1 to bind and activate the NLGN3 promoter.","evidence":"Co-IP/MS, ChIP, nuclear fractionation, ubiquitination assays, knockdown functional assays, and intracranial tumor model","pmids":["38254206"],"confidence":"High","gaps":["Whether this axis operates in non-tumor neurons unknown","Upstream regulation of USP7 not addressed"]},{"year":2024,"claim":"Genetically dissected NLGN3's two transsynaptic pathways, showing the noncanonical PTPδ interaction governs social reward and remote spatial memory while the neurexin interaction governs contextual fear, separating distinct behavioral outputs.","evidence":"Pathway-selective Nlgn3 knock-in mice with social conditioned place preference, Barnes maze, and fear conditioning","pmids":["38475840"],"confidence":"Medium","gaps":["Synaptic/circuit-level basis of behavioral divergence not resolved","Single lab"]},{"year":2024,"claim":"Extended the shed-NLGN3/Gαi1/3 paracrine mechanism to the cardiovascular system, showing NLGN3 binds Gαi1/3 in endothelial cells to drive angiogenesis and cardiac repair after infarction.","evidence":"Co-IP, Gαi1/3 KO MI model, endothelial-specific NLGN3 knockdown, ADAM10 inhibition, tube formation/proliferation assays","pmids":["41398092"],"confidence":"Medium","gaps":["Receptor coupling NLGN3 to Gαi1/3 not identified","Single lab"]},{"year":2025,"claim":"Proposed a mechanotransduction mechanism in which shed NLGN3 binds CSPG4 to promote its ADAM10 shedding and activate PIEZO1, maintaining OPC stemness and glioma growth.","evidence":"Binding assays, ADAM10 inhibition, PIEZO1 activity and OPC differentiation/glioma assays (preprint)","pmids":["bio_10.1101_2025.07.12.664340"],"confidence":"Low","gaps":["Preprint, not peer-reviewed; abstract-level detail insufficient to assess rigor","Direct CSPG4 binding interface undefined"]},{"year":2026,"claim":"Used a cross-species ortholog model to test variant functional consequences, showing human NLGN3 rescues fly Nlg3-null phenotypes and that disease variants exert variant-specific gain- or loss-of-function effects on synapse morphology, sleep, and vesicle dynamics.","evidence":"Transgenic Drosophila rescue/overexpression with confocal synapse imaging, sleep assays, and vesicle dynamics imaging (preprint)","pmids":["41929011"],"confidence":"Low","gaps":["Preprint, Drosophila system; relevance to mammalian synapses unconfirmed","Mechanistic basis of gain- vs loss-of-function not resolved"]},{"year":null,"claim":"The cell-surface receptor(s) that couple shed NLGN3 to Gαi1/3-PI3K-Akt-mTOR signaling across neurons, glioma, and endothelium remain unidentified, leaving the entry point of the paracrine signaling axis undefined.","evidence":"","pmids":[],"confidence":"Medium","gaps":["No receptor mediating Gαi1/3 activation identified","Whether one receptor serves all cell types is unknown"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0098631","term_label":"cell adhesion mediator activity","supporting_discovery_ids":[0,10]},{"term_id":"GO:0048018","term_label":"receptor ligand activity","supporting_discovery_ids":[6,9]},{"term_id":"GO:0060089","term_label":"molecular transducer activity","supporting_discovery_ids":[6,9]}],"localization":[{"term_id":"GO:0005886","term_label":"plasma membrane","supporting_discovery_ids":[0,1,5]},{"term_id":"GO:0005783","term_label":"endoplasmic reticulum","supporting_discovery_ids":[1]},{"term_id":"GO:0005576","term_label":"extracellular region","supporting_discovery_ids":[3,6,8]}],"pathway":[{"term_id":"R-HSA-162582","term_label":"Signal Transduction","supporting_discovery_ids":[2,6,9]},{"term_id":"R-HSA-112316","term_label":"Neuronal System","supporting_discovery_ids":[5,10]},{"term_id":"R-HSA-1643685","term_label":"Disease","supporting_discovery_ids":[0,1,2]}],"complexes":[],"partners":["NRXN1","PTPRD","CDK5","KALRN","ADAM10","GNAI1","GNAI3","CSPG4"],"other_free_text":[]}},"prefetch_data":{"uniprot":{"accession":"Q9NZ94","full_name":"Neuroligin-3","aliases":["Gliotactin homolog"],"length_aa":848,"mass_kda":93.9,"function":"Cell surface protein involved in cell-cell-interactions via its interactions with neurexin family members. Plays a role in synapse function and synaptic signal transmission, and may mediate its effects by clustering other synaptic proteins. May promote the initial formation of synapses, but is not essential for this. May also play a role in glia-glia or glia-neuron interactions in the developing peripheral nervous system (By similarity)","subcellular_location":"Cell membrane; Synapse; Cell projection, dendrite","url":"https://www.uniprot.org/uniprotkb/Q9NZ94/entry"},"depmap":{"release":"DepMap","has_data":true,"is_common_essential":false,"resolved_as":"","url":"https://depmap.org/portal/gene/NLGN3","classification":"Not Classified","n_dependent_lines":2,"n_total_lines":1208,"dependency_fraction":0.0016556291390728477},"opencell":{"profiled":false,"resolved_as":"","ensg_id":"","cell_line_id":"","localizations":[],"interactors":[],"url":"https://opencell.sf.czbiohub.org/search/NLGN3","total_profiled":1310},"omim":[{"mim_id":"606479","title":"NEUROLIGIN 2; NLGN2","url":"https://www.omim.org/entry/606479"},{"mim_id":"600568","title":"NEUROLIGIN 1; NLGN1","url":"https://www.omim.org/entry/600568"},{"mim_id":"600565","title":"NEUREXIN I; NRXN1","url":"https://www.omim.org/entry/600565"},{"mim_id":"400028","title":"NEUROLIGIN 4, Y-LINKED; NLGN4Y","url":"https://www.omim.org/entry/400028"},{"mim_id":"300495","title":"AUTISM, SUSCEPTIBILITY TO, X-LINKED 2; AUTSX2","url":"https://www.omim.org/entry/300495"}],"hpa":{"profiled":true,"resolved_as":"","reliability":"Supported","locations":[{"location":"Golgi apparatus","reliability":"Supported"},{"location":"Cell Junctions","reliability":"Supported"}],"tissue_specificity":"Tissue enhanced","tissue_distribution":"Detected in many","driving_tissues":[{"tissue":"brain","ntpm":31.9},{"tissue":"seminal vesicle","ntpm":11.4}],"url":"https://www.proteinatlas.org/search/NLGN3"},"hgnc":{"alias_symbol":["HNL3","KIAA1480","ASPGX1","AUTSX1"],"prev_symbol":[]},"alphafold":{"accession":"Q9NZ94","domains":[{"cath_id":"3.40.50.1820","chopping":"42-167_198-392","consensus_level":"medium","plddt":92.4019,"start":42,"end":392},{"cath_id":"3.40.50.1820","chopping":"399-620","consensus_level":"medium","plddt":93.5872,"start":399,"end":620}],"viewer_url":"https://alphafold.ebi.ac.uk/entry/Q9NZ94","model_url":"https://alphafold.ebi.ac.uk/files/AF-Q9NZ94-F1-model_v6.cif","pae_url":"https://alphafold.ebi.ac.uk/files/AF-Q9NZ94-F1-predicted_aligned_error_v6.png","plddt_mean":76.44},"mouse_models":{"mgi_url":"https://www.informatics.jax.org/marker/summary?nomen=NLGN3","jax_strain_url":"https://www.jax.org/strain/search?query=NLGN3"},"sequence":{"accession":"Q9NZ94","fasta_url":"https://rest.uniprot.org/uniprotkb/Q9NZ94.fasta","uniprot_url":"https://www.uniprot.org/uniprotkb/Q9NZ94/entry","alphafold_viewer_url":"https://alphafold.ebi.ac.uk/entry/Q9NZ94"}},"corpus_meta":[{"pmid":"12669065","id":"PMC_12669065","title":"Mutations of the X-linked genes encoding neuroligins NLGN3 and NLGN4 are associated with autism.","date":"2003","source":"Nature genetics","url":"https://pubmed.ncbi.nlm.nih.gov/12669065","citation_count":1343,"is_preprint":false},{"pmid":"16648374","id":"PMC_16648374","title":"Novel splice isoforms for NLGN3 and NLGN4 with possible implications in autism.","date":"2006","source":"Journal of medical genetics","url":"https://pubmed.ncbi.nlm.nih.gov/16648374","citation_count":131,"is_preprint":false},{"pmid":"24773431","id":"PMC_24773431","title":"Fmr1 and Nlgn3 knockout rats: novel tools for investigating autism spectrum disorders.","date":"2014","source":"Behavioral neuroscience","url":"https://pubmed.ncbi.nlm.nih.gov/24773431","citation_count":117,"is_preprint":false},{"pmid":"15389766","id":"PMC_15389766","title":"NLGN3/NLGN4 gene mutations are not responsible for autism in the Quebec population.","date":"2005","source":"American journal of medical genetics. Part B, Neuropsychiatric genetics : the official publication of the International Society of Psychiatric Genetics","url":"https://pubmed.ncbi.nlm.nih.gov/15389766","citation_count":89,"is_preprint":false},{"pmid":"31184401","id":"PMC_31184401","title":"Novel mutations in NLGN3 causing autism spectrum disorder and cognitive impairment.","date":"2019","source":"Human mutation","url":"https://pubmed.ncbi.nlm.nih.gov/31184401","citation_count":44,"is_preprint":false},{"pmid":"29775613","id":"PMC_29775613","title":"Suppression of NLRP3 inflammasome attenuates stress-induced depression-like behavior in NLGN3-deficient mice.","date":"2018","source":"Biochemical and biophysical research communications","url":"https://pubmed.ncbi.nlm.nih.gov/29775613","citation_count":43,"is_preprint":false},{"pmid":"18189281","id":"PMC_18189281","title":"No evidence for involvement of genetic variants in the X-linked neuroligin genes NLGN3 and NLGN4X in probands with autism spectrum disorder on high functioning level.","date":"2008","source":"American journal of medical genetics. 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Switzerland)","url":"https://pubmed.ncbi.nlm.nih.gov/23431752","citation_count":13,"is_preprint":false},{"pmid":"36810932","id":"PMC_36810932","title":"Autism-linked NLGN3 is a key regulator of gonadotropin-releasing hormone deficiency.","date":"2023","source":"Disease models & mechanisms","url":"https://pubmed.ncbi.nlm.nih.gov/36810932","citation_count":11,"is_preprint":false},{"pmid":"31705895","id":"PMC_31705895","title":"Evidence for a Contribution of the Nlgn3/Cyfip1/Fmr1 Pathway in the Pathophysiology of Autism Spectrum Disorders.","date":"2019","source":"Neuroscience","url":"https://pubmed.ncbi.nlm.nih.gov/31705895","citation_count":11,"is_preprint":false},{"pmid":"37699715","id":"PMC_37699715","title":"Regulation of NLGN3 and the Synaptic Rho-GEF Signaling Pathway by CDK5.","date":"2023","source":"The Journal of neuroscience : the official journal of the Society for 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autism","url":"https://pubmed.ncbi.nlm.nih.gov/35850732","citation_count":8,"is_preprint":false},{"pmid":"37542090","id":"PMC_37542090","title":"Impaired cecal motility and secretion alongside expansion of gut-associated lymphoid tissue in the Nlgn3R451C mouse model of autism.","date":"2023","source":"Scientific reports","url":"https://pubmed.ncbi.nlm.nih.gov/37542090","citation_count":5,"is_preprint":false},{"pmid":"35955861","id":"PMC_35955861","title":"Reference Genes across Nine Brain Areas of Wild Type and Prader-Willi Syndrome Mice: Assessing Differences in Igfbp7, Pcsk1, Nhlh2 and Nlgn3 Expression.","date":"2022","source":"International journal of molecular sciences","url":"https://pubmed.ncbi.nlm.nih.gov/35955861","citation_count":4,"is_preprint":false},{"pmid":"38475840","id":"PMC_38475840","title":"Differential contribution of canonical and noncanonical NLGN3 pathways to early social development and memory performance.","date":"2024","source":"Molecular brain","url":"https://pubmed.ncbi.nlm.nih.gov/38475840","citation_count":3,"is_preprint":false},{"pmid":"38255906","id":"PMC_38255906","title":"Faster Gastrointestinal Transit, Reduced Small Intestinal Smooth Muscle Tone and Dysmotility in the Nlgn3 Mouse Model of Autism.","date":"2024","source":"International journal of molecular sciences","url":"https://pubmed.ncbi.nlm.nih.gov/38255906","citation_count":3,"is_preprint":false},{"pmid":"41398092","id":"PMC_41398092","title":"NLGN3 contributes to angiogenesis in myocardial infarction via activation of the Gαi1/3-Akt pathway.","date":"2025","source":"Basic research in cardiology","url":"https://pubmed.ncbi.nlm.nih.gov/41398092","citation_count":0,"is_preprint":false},{"pmid":"41929011","id":"PMC_41929011","title":"NLGN3 autism variants have distinct functional impact on synapses and sleep behavior in Drosophila.","date":"2026","source":"bioRxiv : the preprint server for biology","url":"https://pubmed.ncbi.nlm.nih.gov/41929011","citation_count":0,"is_preprint":false},{"pmid":null,"id":"bio_10.1101_2025.07.12.664340","title":"Neuroligin-3 interaction with CSPG4 regulates normal and malignant glial precursors through PIEZO1","date":"2025-07-17","source":"bioRxiv","url":"https://doi.org/10.1101/2025.07.12.664340","citation_count":0,"is_preprint":true}],"cost":{"stage1":{"model":"claude-sonnet-4-6","input_tokens":18859,"output_tokens":3933,"usd":0.057786,"stage1_stop_reason":"end_turn"},"stage2":{"model":"claude-opus-4-8","input_tokens":11661,"output_tokens":4329,"usd":0.083265,"stage2_stop_reason":"end_turn"},"total_usd":0.141051,"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\": 2003,\n      \"finding\": \"NLGN3 (and NLGN4) are cell-adhesion molecules localized at the synapse; missense and frameshift mutations in NLGN3 were identified in siblings with autism-spectrum disorders, implicating defective synaptogenesis as a predisposing mechanism.\",\n      \"method\": \"Genetic sequencing of affected sibling pairs; cellular localization described as postsynaptic\",\n      \"journal\": \"Nature genetics\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 3 / Strong — foundational human genetics with localization claim, replicated broadly across labs, but original paper is a genetic/sequencing study without direct biochemical reconstitution of mechanism\",\n      \"pmids\": [\"12669065\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"Two autism-associated NLGN3 missense variants (p.Arg597Trp and p.Pro514Ser) reduce mature NLGN3 protein levels, impair plasma membrane localization, reduce extracellular cleavage, and induce an unfolded protein response (UPR) due to ER retention of immature protein, more severely than the p.Arg451Cys variant; control population variants (p.Ala632Thr, p.Val341Ala) have no such effect.\",\n      \"method\": \"Overexpression in HEK293 and HeLa cells; flow cytometry / immunofluorescence for plasma membrane localization; western blot for protein levels and UPR markers\",\n      \"journal\": \"Human mutation\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — multiple orthogonal cellular assays in one lab, with appropriate controls (benign variants tested in parallel)\",\n      \"pmids\": [\"31184401\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"In glioma cells, NLGN3 promotes proliferation, migration, and invasion by activating PI3K-AKT, ERK1/2, and LYN signaling pathways; LYN in turn upregulates ADAM10 sheddase expression, which cleaves NLGN3, forming a positive autocrine feedback loop that promotes NLGN3 cleavage and further signaling.\",\n      \"method\": \"Knockdown and overexpression in U251 and U87 glioma cells; western blot for pathway activation; ADAM10 inhibition; proliferation/migration/invasion assays\",\n      \"journal\": \"Frontiers in cell and developmental biology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — multiple functional assays and pathway inhibition in one lab, two cell lines\",\n      \"pmids\": [\"34485271\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2018,\n      \"finding\": \"ADAM10-mediated cleavage and secretion of NLGN3 from neurons is required for glioblastoma cell growth; inhibition of ADAM10 (ADAM10i) blocks neuron-conditioned-medium-induced GBM proliferation.\",\n      \"method\": \"ADAM10 inhibitor treatment of neuron-conditioned medium cultures; U251, U87-MG, and patient-derived GBM cell growth assays\",\n      \"journal\": \"Cancer medicine\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — pharmacological inhibition with functional readout, single lab\",\n      \"pmids\": [\"29777576\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"NLGN3 promotes neuroblastoma cell proliferation and growth by activating the PI3K/AKT signaling pathway and FOXO family transcription activity; PI3K/AKT inhibition in NLGN3-overexpressing cells reverses the proliferative effect.\",\n      \"method\": \"MTT assay, colony formation, cell cycle analysis, BrdU incorporation, xenograft animal model; western blot for AKT phosphorylation; PI3K/AKT pathway inhibition\",\n      \"journal\": \"European journal of pharmacology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — multiple functional assays plus pathway inhibition rescue, single lab\",\n      \"pmids\": [\"31150649\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"CDK5 associates with NLGN3 in vivo and phosphorylates NLGN3 on serine 725 (S725); this phosphorylation regulates NLGN3 association with the Rho-GEF Kalirin-7, NLGN3 surface expression, and NLGN3-mediated synaptic currents in cultured neurons.\",\n      \"method\": \"HA-tagged knock-in mouse; Co-immunoprecipitation (in vivo Cdk5–NLGN3 association); in vitro phosphorylation assay; mutagenesis at S725; surface expression assay; patch-clamp electrophysiology in cultured rat neurons\",\n      \"journal\": \"The Journal of neuroscience\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Moderate — in vitro kinase assay plus mutagenesis plus knock-in mouse plus electrophysiological functional readout, all in one study\",\n      \"pmids\": [\"37699715\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"Secreted/shed NLGN3 activates Gαi1/3-Akt-mTOR and Erk signaling in neuronal cells to protect against ischemia-reperfusion injury; Gαi1/3 silencing or knockout abolishes NLGN3-induced Akt activation and neuroprotection. In vivo, ADAM10-mediated cleavage is required for NLGN3 secretion following MCAO; ADAM10 inhibition blocks NLGN3 secretion and worsens ischemic injury.\",\n      \"method\": \"SH-SY5Y and primary cortical neurons with OGD/R; Gαi1/3 siRNA knockdown and genetic KO; ADAM10 inhibition; neuronal NLGN3 overexpression/silencing by viral vector; MCAO mouse model; western blot for signaling\",\n      \"journal\": \"Cell death & disease\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — genetic KO and knockdown with rescue in vivo plus in vitro mechanistic assays, single lab\",\n      \"pmids\": [\"37880221\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"KPNB1 promotes NLGN3 expression in glioblastoma by mediating nuclear import of the transcription factor YBX1, which directly binds the NLGN3 promoter; USP7 deubiquitinase stabilizes KPNB1 by removing ubiquitin modifications; the USP7–KPNB1–YBX1–NLGN3 axis promotes GBM proliferation and migration.\",\n      \"method\": \"Co-immunoprecipitation and mass spectrometry (KPNB1–YBX1 interaction); chromatin immunoprecipitation (YBX1 binding to NLGN3 promoter); nuclear-cytoplasmic fractionation and immunofluorescence (YBX1 nuclear translocation); ubiquitination assays (USP7 effect on KPNB1); KPNB1/USP7/NLGN3 knockdown functional assays; intracranial orthotopic tumor model\",\n      \"journal\": \"Journal of experimental & clinical cancer research\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1-2 / Moderate — ChIP, Co-IP/MS, ubiquitination assay, nuclear fractionation, and in vivo tumor model with multiple orthogonal methods in one study\",\n      \"pmids\": [\"38254206\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"Wnt/β-catenin signaling transcriptionally upregulates NLGN3 expression in glioblastoma; secreted NLGN3 in conditioned medium and recombinant NLGN3 induce cancer stem cell (CSC) properties in neighboring GBM cells; DAB2IP suppresses CSC acquisition by targeting NLGN3.\",\n      \"method\": \"Conditioned medium and recombinant NLGN3 treatment of GBM cells; CSC sphere formation and marker assays; Wnt/β-catenin inhibitor treatment; DAB2IP knockdown/overexpression\",\n      \"journal\": \"Cell death & disease\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — multiple functional assays and pharmacological intervention, single lab\",\n      \"pmids\": [\"37443071\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"NLGN3 interacts with Gαi1/3 (demonstrated by Co-IP) in cardiac endothelial cells; this NLGN3–Gαi1/3 interaction promotes endothelial cell proliferation and tube formation via PI3K-Akt-mTOR signaling; endothelial-specific NLGN3 knockdown or ADAM10 inhibition reduces angiogenesis and worsens cardiac function after myocardial infarction.\",\n      \"method\": \"Co-immunoprecipitation (NLGN3–Gαi1/3); Gαi1/3 knockout mouse MI model; endothelial-specific NLGN3 knockdown; ADAM10 inhibition; tube formation and proliferation assays\",\n      \"journal\": \"Basic research in cardiology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — Co-IP plus genetic KO plus functional in vivo assays, single lab\",\n      \"pmids\": [\"41398092\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"NLGN3 engages two distinct transsynaptic pathways: canonical interaction with neurexins (NRXNs) and a noncanonical interaction with protein tyrosine phosphatase δ (PTPδ). Selectively disrupting the NLGN3–PTPδ pathway impairs social motivation/reward (juvenile social conditioned place preference) and remote spatial memory, while disrupting the NLGN3–NRXN pathway attenuates contextual fear conditioning. The two pathways thus differentially regulate distinct higher-order brain functions.\",\n      \"method\": \"Nlgn3 knock-in mice selectively lacking interaction with NRXNs or PTPδ; social conditioned place preference, Barnes maze, and contextual fear conditioning behavioral tests\",\n      \"journal\": \"Molecular brain\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — genetic dissection using pathway-specific knock-in mice with multiple behavioral readouts, single lab\",\n      \"pmids\": [\"38475840\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"Shed NLGN3 binds directly to CSPG4 on glioma cells and oligodendrocyte precursor cells (OPCs); NLGN3–CSPG4 interaction facilitates CSPG4 shedding by ADAM10, alters membrane tension, and activates PIEZO1 mechanosensitive channels, causing membrane depolarization; this NLGN3–CSPG4–PIEZO1 axis maintains OPCs in an undifferentiated stem-like state and promotes glioma proliferation.\",\n      \"method\": \"Binding/interaction assays (NLGN3–CSPG4); ADAM10 inhibition; PIEZO1 activity measurements; OPC differentiation assays; glioma proliferation assays\",\n      \"journal\": \"bioRxiv\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 / Weak — preprint with mechanistic claims but abstract-level detail insufficient to assess full rigor; single lab, not yet peer-reviewed\",\n      \"pmids\": [\"bio_10.1101_2025.07.12.664340\"],\n      \"is_preprint\": true\n    },\n    {\n      \"year\": 2026,\n      \"finding\": \"In Drosophila, the fly Nlgn3 homolog regulates sleep patterns, synaptic architecture, and vesicle dynamics. Human NLGN3 rescues the Nlg3-null phenotype. The de novo p.R175W variant and maternally inherited p.R451C variant alter synapse morphology and sleep; p.R597W alters sleep and vesicle dynamics with minimal synapse morphology effects. Overexpression data suggest p.R175W is gain-of-function while maternally inherited variants show mixed loss/gain-of-function effects.\",\n      \"method\": \"Transgenic Drosophila rescue and overexpression models; confocal imaging of synaptic morphology; sleep behavior assays; vesicle dynamics imaging\",\n      \"journal\": \"bioRxiv\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 / Weak — preprint, Drosophila model system, abstract-level detail; cross-species ortholog study with functional validation but not peer-reviewed\",\n      \"pmids\": [\"41929011\"],\n      \"is_preprint\": true\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"NLGN3 promotes neuritogenesis in developing GnRH neurons; overexpression of wild-type NLGN3 (but not loss-of-function mutant NLGN3) promotes neurite outgrowth, linking NLGN3 function to GnRH neuron maturation.\",\n      \"method\": \"Overexpression of wild-type vs. LoF mutant NLGN3 in developing GnRH cells; neuritogenesis quantification\",\n      \"journal\": \"Disease models & mechanisms\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 / Weak — single overexpression experiment in one cell type, single lab\",\n      \"pmids\": [\"36810932\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"NLGN3 is a postsynaptic X-linked cell-adhesion molecule that organizes synapse assembly through canonical interactions with neurexins and a noncanonical interaction with PTPδ, is phosphorylated by CDK5 on S725 to regulate its association with the Rho-GEF Kalirin-7 and its surface expression and synaptic currents, and is shed from the cell surface by ADAM10; the shed ectodomain acts as a paracrine/autocrine factor that activates Gαi1/3–PI3K–Akt–mTOR signaling in neurons (neuroprotection), glioma cells (proliferation via LYN–ADAM10 feedback and CSC induction via Wnt/β-catenin–YBX1 transcriptional upregulation), and cardiac endothelial cells (angiogenesis), while disease-associated missense variants impair its ER processing, plasma membrane localization, and cleavage.\"\n}\n```","stage2_raw":"{\n  \"mechanistic_narrative\": \"NLGN3 is an X-linked postsynaptic cell-adhesion molecule that organizes synapse assembly and, through proteolytic shedding of its ectodomain, acts as a paracrine signaling factor in both physiological and pathological contexts [#0, #5]. At the synapse it engages two distinct transsynaptic pathways—a canonical interaction with neurexins and a noncanonical interaction with PTPδ—that differentially regulate higher-order brain functions including social motivation, spatial memory, and contextual fear [#10]. Its surface expression and synaptic currents are controlled by CDK5, which associates with NLGN3 in vivo and phosphorylates serine 725 to regulate NLGN3 association with the Rho-GEF Kalirin-7 [#5]. NLGN3 is cleaved from the cell surface by ADAM10, and the shed ectodomain functions as a soluble ligand that activates Gαi1/3–PI3K–Akt–mTOR signaling: in neurons this confers neuroprotection against ischemia-reperfusion injury [#6], and in cardiac endothelial cells it drives proliferation, tube formation, and angiogenesis after myocardial infarction [#9]. In glioma the same shed NLGN3 axis is co-opted as a growth driver, activating PI3K-AKT, ERK, and LYN signaling, with LYN upregulating ADAM10 to form an autocrine cleavage feedback loop [#2, #3]; NLGN3 expression is transcriptionally amplified by Wnt/β-catenin signaling and by a USP7–KPNB1–YBX1 axis in which YBX1 binds the NLGN3 promoter, promoting cancer stem cell properties and tumor proliferation [#7, #8]. Autism-associated missense variants impair ER processing and maturation of NLGN3, reduce plasma membrane localization, and decrease extracellular cleavage, triggering an unfolded protein response [#0, #1].\",\n  \"teleology\": [\n    {\n      \"year\": 2003,\n      \"claim\": \"Established NLGN3 as a synaptic cell-adhesion molecule and linked it to disease, defining defective synaptogenesis as a candidate mechanism for autism-spectrum disorders.\",\n      \"evidence\": \"Genetic sequencing of affected sibling pairs with postsynaptic localization assignment\",\n      \"pmids\": [\"12669065\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"No biochemical reconstitution of the synaptogenic mechanism\", \"Causal mutation effect not functionally tested in this study\"]\n    },\n    {\n      \"year\": 2018,\n      \"claim\": \"Resolved how NLGN3 promotes tumor growth non-cell-autonomously, showing that ADAM10-dependent shedding of neuronal NLGN3 is required to stimulate glioblastoma proliferation.\",\n      \"evidence\": \"ADAM10 inhibition of neuron-conditioned medium with patient-derived and cell-line GBM growth assays\",\n      \"pmids\": [\"29777576\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Downstream receptor on GBM cells not identified here\", \"Single lab, pharmacological inhibition only\"]\n    },\n    {\n      \"year\": 2019,\n      \"claim\": \"Showed that autism-associated missense variants act by disrupting NLGN3 maturation rather than synaptic binding per se, retaining immature protein in the ER and triggering a UPR.\",\n      \"evidence\": \"Overexpression in HEK293/HeLa with flow cytometry, immunofluorescence, and western blot, including benign control variants\",\n      \"pmids\": [\"31184401\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Heterologous overexpression, not neuronal context\", \"Functional synaptic consequences not measured\"]\n    },\n    {\n      \"year\": 2019,\n      \"claim\": \"Extended NLGN3's pro-proliferative role to neuroblastoma and identified PI3K/AKT-FOXO as the effector pathway through rescue by pathway inhibition.\",\n      \"evidence\": \"Proliferation, colony, cell-cycle, BrdU and xenograft assays with PI3K/AKT inhibition\",\n      \"pmids\": [\"31150649\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Receptor mediating NLGN3 signaling not defined\", \"Single lab\"]\n    },\n    {\n      \"year\": 2021,\n      \"claim\": \"Defined an autocrine feedback circuit in glioma whereby NLGN3 signaling through LYN upregulates ADAM10 to amplify its own cleavage and downstream PI3K-AKT/ERK signaling.\",\n      \"evidence\": \"Knockdown/overexpression in two glioma lines with ADAM10 inhibition and proliferation/migration/invasion assays\",\n      \"pmids\": [\"34485271\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Direct NLGN3 receptor not identified\", \"Feedback loop quantification limited to two cell lines\"]\n    },\n    {\n      \"year\": 2023,\n      \"claim\": \"Identified CDK5 as a kinase that phosphorylates NLGN3 at S725 to control its surface expression, Kalirin-7 association, and synaptic currents, providing a post-translational regulatory mechanism for synaptic function.\",\n      \"evidence\": \"Knock-in mouse Co-IP, in vitro kinase assay, S725 mutagenesis, surface expression assay, and patch-clamp electrophysiology\",\n      \"pmids\": [\"37699715\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Upstream signals activating CDK5-NLGN3 phosphorylation unknown\", \"Link to disease variants not tested\"]\n    },\n    {\n      \"year\": 2023,\n      \"claim\": \"Demonstrated a physiological neuroprotective role for shed NLGN3, showing it activates Gαi1/3-Akt-mTOR signaling in neurons and requires ADAM10 cleavage for protection against ischemic injury.\",\n      \"evidence\": \"OGD/R in SH-SY5Y and primary neurons with Gαi1/3 knockdown/KO, ADAM10 inhibition, viral NLGN3 manipulation, and MCAO mouse model\",\n      \"pmids\": [\"37880221\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"NLGN3 receptor coupling to Gαi1/3 not structurally defined\", \"Single lab\"]\n    },\n    {\n      \"year\": 2023,\n      \"claim\": \"Connected NLGN3 to glioma stemness, showing Wnt/β-catenin transcriptionally upregulates NLGN3 and that secreted NLGN3 induces cancer stem cell properties, with DAB2IP as a suppressor.\",\n      \"evidence\": \"Conditioned medium and recombinant NLGN3 treatment, CSC sphere/marker assays, Wnt inhibition, DAB2IP manipulation\",\n      \"pmids\": [\"37443071\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Mechanism of DAB2IP targeting of NLGN3 unresolved\", \"Single lab\"]\n    },\n    {\n      \"year\": 2024,\n      \"claim\": \"Mapped a transcriptional control axis for NLGN3 in glioblastoma in which USP7 stabilizes KPNB1, which imports YBX1 to bind and activate the NLGN3 promoter.\",\n      \"evidence\": \"Co-IP/MS, ChIP, nuclear fractionation, ubiquitination assays, knockdown functional assays, and intracranial tumor model\",\n      \"pmids\": [\"38254206\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Whether this axis operates in non-tumor neurons unknown\", \"Upstream regulation of USP7 not addressed\"]\n    },\n    {\n      \"year\": 2024,\n      \"claim\": \"Genetically dissected NLGN3's two transsynaptic pathways, showing the noncanonical PTPδ interaction governs social reward and remote spatial memory while the neurexin interaction governs contextual fear, separating distinct behavioral outputs.\",\n      \"evidence\": \"Pathway-selective Nlgn3 knock-in mice with social conditioned place preference, Barnes maze, and fear conditioning\",\n      \"pmids\": [\"38475840\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Synaptic/circuit-level basis of behavioral divergence not resolved\", \"Single lab\"]\n    },\n    {\n      \"year\": 2024,\n      \"claim\": \"Extended the shed-NLGN3/Gαi1/3 paracrine mechanism to the cardiovascular system, showing NLGN3 binds Gαi1/3 in endothelial cells to drive angiogenesis and cardiac repair after infarction.\",\n      \"evidence\": \"Co-IP, Gαi1/3 KO MI model, endothelial-specific NLGN3 knockdown, ADAM10 inhibition, tube formation/proliferation assays\",\n      \"pmids\": [\"41398092\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Receptor coupling NLGN3 to Gαi1/3 not identified\", \"Single lab\"]\n    },\n    {\n      \"year\": 2025,\n      \"claim\": \"Proposed a mechanotransduction mechanism in which shed NLGN3 binds CSPG4 to promote its ADAM10 shedding and activate PIEZO1, maintaining OPC stemness and glioma growth.\",\n      \"evidence\": \"Binding assays, ADAM10 inhibition, PIEZO1 activity and OPC differentiation/glioma assays (preprint)\",\n      \"pmids\": [\"bio_10.1101_2025.07.12.664340\"],\n      \"confidence\": \"Low\",\n      \"gaps\": [\"Preprint, not peer-reviewed; abstract-level detail insufficient to assess rigor\", \"Direct CSPG4 binding interface undefined\"]\n    },\n    {\n      \"year\": 2026,\n      \"claim\": \"Used a cross-species ortholog model to test variant functional consequences, showing human NLGN3 rescues fly Nlg3-null phenotypes and that disease variants exert variant-specific gain- or loss-of-function effects on synapse morphology, sleep, and vesicle dynamics.\",\n      \"evidence\": \"Transgenic Drosophila rescue/overexpression with confocal synapse imaging, sleep assays, and vesicle dynamics imaging (preprint)\",\n      \"pmids\": [\"41929011\"],\n      \"confidence\": \"Low\",\n      \"gaps\": [\"Preprint, Drosophila system; relevance to mammalian synapses unconfirmed\", \"Mechanistic basis of gain- vs loss-of-function not resolved\"]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"The cell-surface receptor(s) that couple shed NLGN3 to Gαi1/3-PI3K-Akt-mTOR signaling across neurons, glioma, and endothelium remain unidentified, leaving the entry point of the paracrine signaling axis undefined.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"No receptor mediating Gαi1/3 activation identified\", \"Whether one receptor serves all cell types is unknown\"]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0098631\", \"supporting_discovery_ids\": [0, 10]},\n      {\"term_id\": \"GO:0048018\", \"supporting_discovery_ids\": [6, 9]},\n      {\"term_id\": \"GO:0060089\", \"supporting_discovery_ids\": [6, 9]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005886\", \"supporting_discovery_ids\": [0, 1, 5]},\n      {\"term_id\": \"GO:0005783\", \"supporting_discovery_ids\": [1]},\n      {\"term_id\": \"GO:0005576\", \"supporting_discovery_ids\": [3, 6, 8]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-162582\", \"supporting_discovery_ids\": [2, 6, 9]},\n      {\"term_id\": \"R-HSA-112316\", \"supporting_discovery_ids\": [5, 10]},\n      {\"term_id\": \"R-HSA-1643685\", \"supporting_discovery_ids\": [0, 1, 2]}\n    ],\n    \"complexes\": [],\n    \"partners\": [\"NRXN1\", \"PTPRD\", \"CDK5\", \"KALRN\", \"ADAM10\", \"GNAI1\", \"GNAI3\", \"CSPG4\"],\n    \"other_free_text\": []\n  }\n}","audit_flag":null,"evaluation":{"pairwise":"win","faith_supported":6,"faith_total":6,"faith_pct":100.0}}