{"gene":"NRSN2","run_date":"2026-06-10T05:19:52","timeline":{"discoveries":[{"year":2015,"finding":"NRSN2 promotes non-small cell lung cancer cell growth through the PI3K/Akt/mTOR signaling pathway, as demonstrated by loss- and gain-of-function experiments in NSCLC cell lines showing effects on cell growth but not invasion.","method":"Loss- and gain-of-function assays in NSCLC cell lines, western blot for pathway components","journal":"International journal of clinical and experimental pathology","confidence":"Medium","confidence_rationale":"Tier 2 / Weak — clean KD/OE with defined cellular phenotype and pathway readout, single lab, single study","pmids":["26045763"],"is_preprint":false},{"year":2015,"finding":"NRSN2 inhibits hepatocellular carcinoma cell proliferation and promotes senescence and apoptosis by regulating PI3K/AKT signaling and the p53/p21 pathway, as shown by loss- and gain-of-function assays.","method":"CCK-8 assays, SA-β-gal staining, Annexin V/PI staining, western blot for PI3K/AKT and p53/p21 pathway components; loss- and gain-of-function in HCC cell lines","journal":"Digestive diseases and sciences","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — multiple orthogonal functional assays (proliferation, senescence, apoptosis) plus pathway western blots, single lab","pmids":["26055238"],"is_preprint":false},{"year":2016,"finding":"NRSN2 promotes malignant phenotype in ovarian cancer cells including anchorage-independent growth, invasion, and chemoresistance, and positively correlates with CD133 stem cell marker expression; Wnt canonical signaling and Twist/Akt/Erk axis are regulated by NRSN2.","method":"Loss- and gain-of-function assays, colony formation, subcutaneous tumor formation, invasion assays, western blot for Wnt/Akt/Erk pathway components","journal":"Biomedicine & pharmacotherapy","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — multiple orthogonal functional assays plus pathway readouts, single lab","pmids":["27908706"],"is_preprint":false},{"year":2017,"finding":"NRSN2 promotes osteosarcoma cell proliferation and growth through dysregulation of both PI3K/AKT/mTOR and Wnt/β-catenin signaling pathways, demonstrated by loss- and gain-of-function assays in vitro and in vivo.","method":"Loss- and gain-of-function assays in osteosarcoma cell lines, in vivo tumor growth assays, western blot for PI3K/AKT/mTOR and Wnt/β-catenin pathway components","journal":"American journal of cancer research","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — in vitro and in vivo KD/OE with defined phenotype and two pathway readouts, single lab","pmids":["28401012"],"is_preprint":false},{"year":2019,"finding":"NRSN2 promotes breast cancer cell proliferation, migration, and invasion by activating PI3K/AKT/mTOR and NF-κB signaling pathways, as shown by KD/OE experiments in vitro and in vivo.","method":"MTT assay, western blot for PI3K/AKT/mTOR and NF-κB pathway components, KD/OE in breast cancer cells, in vivo xenograft","journal":"Oncology letters","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — in vitro and in vivo functional assays with two pathway readouts, single lab","pmids":["31885716"],"is_preprint":false},{"year":2021,"finding":"miR-143 directly targets NRSN2 (binding relationship verified by dual luciferase assay); BMSC-derived extracellular vesicles deliver lncRNA MALAT1 into osteosarcoma cells where MALAT1 sponges miR-143, thereby de-repressing NRSN2 expression and activating Wnt/β-catenin signaling to promote osteosarcoma proliferation, invasion, and migration.","method":"Dual luciferase reporter assay, EV isolation and characterization, transfection of si-MALAT1 and miR-143 mimic, western blot for Wnt/β-catenin pathway proteins, in vivo xenograft","journal":"OncoTargets and therapy","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — dual luciferase validation of miR-143/NRSN2 axis, in vitro and in vivo rescue experiments, single lab","pmids":["33564242"],"is_preprint":false},{"year":2021,"finding":"miR-1913 directly targets NRSN2 (validated by dual luciferase reporter assay); hBMSC-derived exosomes deliver miR-1913 into osteosarcoma cells, suppressing NRSN2 expression and thereby inhibiting osteosarcoma cell viability, proliferation, migration, and invasion.","method":"Dual luciferase reporter assay, CCK8, colony formation, wound healing, transwell assays, RT-qPCR and western blot for NRSN2 and EMT markers, TEM and NTA for exosome characterization","journal":"American journal of translational research","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — dual luciferase validation of miR-1913/NRSN2 targeting, multiple orthogonal functional assays, single lab","pmids":["34650689"],"is_preprint":false},{"year":2022,"finding":"NRSN2 activates the ERK1/2-Bcl-xL signaling pathway; miR-206 (delivered via BMSC-derived extracellular vesicles) directly targets NRSN2 (validated by dual luciferase assay), suppressing NRSN2-mediated ERK1/2-Bcl-xL activation and inhibiting osteosarcoma progression.","method":"Dual luciferase reporter assay, western blot for p-ERK1/2, ERK1/2, Bcl-xL, EdU and colony formation assays, flow cytometry for apoptosis, immunofluorescence, in vivo xenograft","journal":"European journal of histochemistry","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — dual luciferase validation, pathway rescue by NRSN2 OE reversing EV effect, in vitro and in vivo, single lab","pmids":["35730574"],"is_preprint":false},{"year":2022,"finding":"E2F1 directly transcribes NRSN2 (confirmed by luciferase reporter and chromatin immunoprecipitation assays); the E2F1/NRSN2 axis promotes ESCC cell proliferation, migration, and invasion by activating the AKT/mTOR pathway.","method":"Luciferase reporter assay, chromatin immunoprecipitation (ChIP), loss- and gain-of-function assays, western blot for AKT/mTOR pathway, in vivo xenograft","journal":"Pathology, research and practice","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — ChIP and luciferase reporter to establish upstream E2F1 regulation, in vitro and in vivo functional assays, single lab","pmids":["35662041"],"is_preprint":false},{"year":2024,"finding":"NRSN2 promotes malignant behavior of HPV-transfected laryngeal carcinoma cells through activation of the AMPK/ULK1 pathway and consequent autophagy induction; NRSN2 knockdown suppresses AMPK/ULK1-mediated autophagy and restrains malignant behavior.","method":"RT-qPCR, immunoblotting, immunofluorescence, CCK-8, EdU staining, transwell, wound healing, flow cytometry, TEM for autophagosomes, AMPK/ULK1 pathway inhibition experiments","journal":"Cancer biology & therapy","confidence":"Medium","confidence_rationale":"Tier 2 / Weak — pathway inhibition plus KD functional assays, multiple readouts, single lab single study","pmids":["38569536"],"is_preprint":false},{"year":2024,"finding":"NRSN2 knockout mice show significant alterations in functional brain network connectivity—particularly in sensory-related (gustatory and auditory) and default mode network regions—without marked changes in brain microstructure or regional activity, linking NRSN2 to sensory processing and stress resilience circuitry.","method":"11.7 T multimodal MRI (structural and functional) in Nrsn2 homozygous knockout mice compared with wild-type at juvenile and adult stages","journal":"Phenomics","confidence":"Medium","confidence_rationale":"Tier 2 / Weak — clean KO with defined neuroimaging phenotype, multimodal MRI, single lab single study","pmids":["39723223"],"is_preprint":false},{"year":2024,"finding":"lncRNA LUESCC functions as a ceRNA by sponging miR-6785-5p to enhance NRSN2 expression, which is critical for ESCC proliferation, migration, and invasion; this axis was mechanistically established by luciferase reporter assays and rescue experiments.","method":"Luciferase reporter assay (LUESCC/miR-6785-5p and miR-6785-5p/NRSN2 interactions), KD of LUESCC, rescue by NRSN2 modulation, in vitro and in vivo functional assays","journal":"Cellular and molecular life sciences","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — dual luciferase validation of ceRNA axis, rescue experiments, in vitro and in vivo, single lab","pmids":["38457049"],"is_preprint":false}],"current_model":"NRSN2 (Neurensin-2) is a small neuronal membrane protein that, in multiple cancer contexts, acts as a pro-proliferative/oncogenic factor by activating PI3K/AKT/mTOR, Wnt/β-catenin, NF-κB, ERK1/2-Bcl-xL, and AMPK/ULK1 signaling pathways; its transcription is directly driven by E2F1 (in ESCC), it is post-transcriptionally suppressed by miRNAs (miR-143, miR-206, miR-1913, miR-6785-5p), and in the nervous system its knockout alters functional brain network connectivity in sensory and default mode network regions without gross structural changes."},"narrative":{"mechanistic_narrative":"NRSN2 (Neurensin-2) is a small protein that functions predominantly as a pro-proliferative factor across diverse epithelial and mesenchymal cancers, where it couples upstream transcriptional and microRNA regulation to multiple growth-promoting signaling cascades [PMID:26045763, PMID:28401012, PMID:35662041]. In non-small cell lung, osteosarcoma, breast, esophageal squamous cell carcinoma (ESCC), and other tumor cells, NRSN2 gain- and loss-of-function drives proliferation, migration, and invasion through activation of PI3K/AKT/mTOR signaling, frequently in concert with Wnt/β-catenin, NF-κB, ERK1/2-Bcl-xL, or AMPK/ULK1-mediated autophagy [PMID:26045763, PMID:28401012, PMID:31885716, PMID:35730574, PMID:38569536]. Its expression is controlled at two levels: direct transcriptional activation by E2F1 in ESCC [PMID:35662041], and post-transcriptional repression by multiple microRNAs (miR-143, miR-206, miR-1913, miR-6785-5p) that are themselves modulated by lncRNA ceRNA networks delivered via mesenchymal stem cell-derived extracellular vesicles [PMID:33564242, PMID:34650689, PMID:35730574, PMID:38457049]. Notably, in hepatocellular carcinoma NRSN2 acts in the opposite direction, suppressing proliferation and promoting senescence and apoptosis via the p53/p21 axis, indicating context-dependent function [PMID:26055238]. Beyond cancer, knockout mice reveal a neuronal role: loss of NRSN2 alters functional brain network connectivity in sensory and default mode network regions without gross structural change [PMID:39723223]. The biochemical activity of the NRSN2 protein itself—its molecular partners and the mechanism by which it engages these signaling pathways—has not been characterized in the available corpus.","teleology":[{"year":2015,"claim":"Established NRSN2 as a functional driver of cancer cell growth and linked it to a specific signaling cascade, defining its first mechanistic role.","evidence":"Loss- and gain-of-function assays with pathway western blots in NSCLC cell lines","pmids":["26045763"],"confidence":"Medium","gaps":["Does not show direct molecular activity of NRSN2 on PI3K/Akt components","Single cell-line context, no in vivo data","Effect on invasion not observed"]},{"year":2015,"claim":"Revealed that NRSN2 function is context-dependent, acting as a growth suppressor in hepatocellular carcinoma rather than a uniform oncogene.","evidence":"Proliferation, senescence (SA-β-gal), and apoptosis assays with p53/p21 and PI3K/AKT western blots in HCC cell lines","pmids":["26055238"],"confidence":"Medium","gaps":["Mechanism underlying opposite directionality versus other cancers unresolved","No direct interaction with p53/p21 demonstrated","Single lab"]},{"year":2016,"claim":"Extended NRSN2's oncogenic repertoire to invasion, chemoresistance, and stemness, broadening its phenotypic scope beyond proliferation.","evidence":"Anchorage-independent growth, invasion, tumor formation, CD133 correlation, and Wnt/Akt/Erk western blots in ovarian cancer cells","pmids":["27908706"],"confidence":"Medium","gaps":["Multiple pathways implicated without identifying primary effector","No upstream regulator defined","Single lab"]},{"year":2017,"claim":"Showed NRSN2 simultaneously dysregulates two canonical pathways in vivo, consolidating its pro-growth role in osteosarcoma.","evidence":"In vitro and in vivo KD/OE with PI3K/AKT/mTOR and Wnt/β-catenin western blots in osteosarcoma","pmids":["28401012"],"confidence":"Medium","gaps":["Hierarchy between the two pathways not resolved","No direct molecular mechanism of activation","Single lab"]},{"year":2019,"claim":"Added NF-κB to the set of NRSN2-activated cascades and confirmed pro-metastatic activity in vivo in breast cancer.","evidence":"MTT, in vivo xenograft, PI3K/AKT/mTOR and NF-κB western blots in breast cancer cells","pmids":["31885716"],"confidence":"Medium","gaps":["Direct link between NRSN2 and NF-κB activation not shown","No partner identified","Single lab"]},{"year":2021,"claim":"Identified the first upstream regulatory layer, showing NRSN2 is a direct microRNA target embedded in extracellular-vesicle-delivered lncRNA ceRNA networks.","evidence":"Dual luciferase validation of miR-143 and miR-1913 targeting NRSN2, with EV/exosome delivery and rescue in osteosarcoma","pmids":["33564242","34650689"],"confidence":"Medium","gaps":["Whether NRSN2 protein output is the sole effector of these axes unclear","Generalizability beyond osteosarcoma untested","Single lab per axis"]},{"year":2022,"claim":"Defined the ERK1/2-Bcl-xL pathway as an NRSN2 effector and established a complete transcriptional control point via E2F1.","evidence":"Dual luciferase and ChIP showing E2F1 transcribes NRSN2 in ESCC; miR-206/EV experiments linking NRSN2 to ERK1/2-Bcl-xL in osteosarcoma","pmids":["35730574","35662041"],"confidence":"Medium","gaps":["How NRSN2 physically engages ERK1/2 or Bcl-xL unknown","E2F1 regulation shown only in ESCC","Single lab per study"]},{"year":2024,"claim":"Connected NRSN2 to autophagy regulation through AMPK/ULK1 and reinforced the ESCC ceRNA axis with an additional lncRNA-miRNA pair.","evidence":"AMPK/ULK1 pathway inhibition and autophagosome TEM in HPV-laryngeal carcinoma; luciferase and rescue for LUESCC/miR-6785-5p/NRSN2 in ESCC","pmids":["38569536","38457049"],"confidence":"Medium","gaps":["Direct molecular mechanism linking NRSN2 to AMPK activation absent","Whether autophagy is cause or consequence of malignancy unclear","Single lab per study"]},{"year":2024,"claim":"Provided the first in vivo evidence of an endogenous neuronal function, implicating NRSN2 in functional brain network connectivity.","evidence":"11.7 T multimodal structural and functional MRI in Nrsn2 knockout versus wild-type mice","pmids":["39723223"],"confidence":"Medium","gaps":["Cellular/molecular basis of connectivity changes not defined","No behavioral correlation established","Relationship to its cancer signaling roles unknown"]},{"year":null,"claim":"The intrinsic biochemical activity of the NRSN2 protein and its direct molecular partners remain undefined, leaving open how it mechanistically engages the many signaling pathways attributed to it.","evidence":"No discovery in the corpus reports a direct binding partner, enzymatic activity, or structural mechanism for NRSN2","pmids":[],"confidence":"Low","gaps":["No direct physical interactor identified","No structural or biochemical characterization","Context-dependent oncogene-versus-suppressor switch unexplained"]}],"mechanism_profile":{"molecular_activity":[],"localization":[],"pathway":[{"term_id":"R-HSA-162582","term_label":"Signal Transduction","supporting_discovery_ids":[0,3,4,8,9]},{"term_id":"R-HSA-9612973","term_label":"Autophagy","supporting_discovery_ids":[9]},{"term_id":"R-HSA-112316","term_label":"Neuronal System","supporting_discovery_ids":[10]}],"complexes":[],"partners":[],"other_free_text":[]}},"prefetch_data":{"uniprot":{"accession":"Q9GZP1","full_name":"Neurensin-2","aliases":[],"length_aa":204,"mass_kda":22.0,"function":"May play a role in maintenance and/or transport of vesicles","subcellular_location":"Membrane","url":"https://www.uniprot.org/uniprotkb/Q9GZP1/entry"},"depmap":{"release":"DepMap","has_data":true,"is_common_essential":false,"resolved_as":"","url":"https://depmap.org/portal/gene/NRSN2","classification":"Not Classified","n_dependent_lines":0,"n_total_lines":1208,"dependency_fraction":0.0},"opencell":{"profiled":false,"resolved_as":"","ensg_id":"","cell_line_id":"","localizations":[],"interactors":[],"url":"https://opencell.sf.czbiohub.org/search/NRSN2","total_profiled":1310},"omim":[{"mim_id":"610666","title":"NEURENSIN 2; NRSN2","url":"https://www.omim.org/entry/610666"}],"hpa":{"profiled":true,"resolved_as":"","reliability":"Approved","locations":[{"location":"Vesicles","reliability":"Approved"},{"location":"Nucleoplasm","reliability":"Additional"},{"location":"Cytosol","reliability":"Additional"}],"tissue_specificity":"Tissue enhanced","tissue_distribution":"Detected in all","driving_tissues":[{"tissue":"brain","ntpm":152.5}],"url":"https://www.proteinatlas.org/search/NRSN2"},"hgnc":{"alias_symbol":["dJ1103G7.6"],"prev_symbol":["C20orf98"]},"alphafold":{"accession":"Q9GZP1","domains":[{"cath_id":"1.10.287","chopping":"64-93_101-146","consensus_level":"high","plddt":87.1584,"start":64,"end":146}],"viewer_url":"https://alphafold.ebi.ac.uk/entry/Q9GZP1","model_url":"https://alphafold.ebi.ac.uk/files/AF-Q9GZP1-F1-model_v6.cif","pae_url":"https://alphafold.ebi.ac.uk/files/AF-Q9GZP1-F1-predicted_aligned_error_v6.png","plddt_mean":64.56},"mouse_models":{"mgi_url":"https://www.informatics.jax.org/marker/summary?nomen=NRSN2","jax_strain_url":"https://www.jax.org/strain/search?query=NRSN2"},"sequence":{"accession":"Q9GZP1","fasta_url":"https://rest.uniprot.org/uniprotkb/Q9GZP1.fasta","uniprot_url":"https://www.uniprot.org/uniprotkb/Q9GZP1/entry","alphafold_viewer_url":"https://alphafold.ebi.ac.uk/entry/Q9GZP1"}},"corpus_meta":[{"pmid":"28401012","id":"PMC_28401012","title":"NRSN2 promotes osteosarcoma cell proliferation and growth through PI3K/Akt/MTOR and Wnt/β-catenin signaling.","date":"2017","source":"American journal of cancer research","url":"https://pubmed.ncbi.nlm.nih.gov/28401012","citation_count":38,"is_preprint":false},{"pmid":"33564242","id":"PMC_33564242","title":"Bone Marrow Mesenchymal Stem Cells-Derived Extracellular Vesicles Promote Proliferation, Invasion and Migration of Osteosarcoma Cells via the lncRNA MALAT1/miR-143/NRSN2/Wnt/β-Catenin Axis.","date":"2021","source":"OncoTargets and therapy","url":"https://pubmed.ncbi.nlm.nih.gov/33564242","citation_count":30,"is_preprint":false},{"pmid":"26055238","id":"PMC_26055238","title":"Down-Regulated NRSN2 Promotes Cell Proliferation and Survival Through PI3K/Akt/mTOR Pathway in Hepatocellular Carcinoma.","date":"2015","source":"Digestive diseases and sciences","url":"https://pubmed.ncbi.nlm.nih.gov/26055238","citation_count":19,"is_preprint":false},{"pmid":"26045763","id":"PMC_26045763","title":"NRSN2 promotes non-small cell lung cancer cell growth through PI3K/Akt/mTOR pathway.","date":"2015","source":"International journal of clinical and experimental pathology","url":"https://pubmed.ncbi.nlm.nih.gov/26045763","citation_count":18,"is_preprint":false},{"pmid":"31885716","id":"PMC_31885716","title":"NRSN2 promotes breast cancer metastasis by activating PI3K/AKT/mTOR and NF-κB signaling pathways.","date":"2019","source":"Oncology letters","url":"https://pubmed.ncbi.nlm.nih.gov/31885716","citation_count":17,"is_preprint":false},{"pmid":"37875515","id":"PMC_37875515","title":"Long non-coding RNA NRSN2-AS1 promotes ovarian cancer progression through targeting PTK2/β-catenin pathway.","date":"2023","source":"Cell death & disease","url":"https://pubmed.ncbi.nlm.nih.gov/37875515","citation_count":14,"is_preprint":false},{"pmid":"34650689","id":"PMC_34650689","title":"The exosomal transfer of human bone marrow mesenchymal stem cell-derived miR-1913 inhibits osteosarcoma progression by targeting NRSN2.","date":"2021","source":"American journal of translational research","url":"https://pubmed.ncbi.nlm.nih.gov/34650689","citation_count":14,"is_preprint":false},{"pmid":"34791059","id":"PMC_34791059","title":"Long non-coding RNA NRSN2-AS1 facilitates tumorigenesis and progression of ovarian cancer via miR-744-5p/PRKX axis.","date":"2022","source":"Biology of reproduction","url":"https://pubmed.ncbi.nlm.nih.gov/34791059","citation_count":13,"is_preprint":false},{"pmid":"38457049","id":"PMC_38457049","title":"LncRNA LUESCC promotes esophageal squamous cell carcinoma by targeting the miR-6785-5p/NRSN2 axis.","date":"2024","source":"Cellular and molecular life sciences : CMLS","url":"https://pubmed.ncbi.nlm.nih.gov/38457049","citation_count":13,"is_preprint":false},{"pmid":"35730574","id":"PMC_35730574","title":"Extracellular vesicles from bone mesenchymal stem cells transport microRNA-206 into osteosarcoma cells and target NRSN2 to block the ERK1/2-Bcl-xL signaling pathway.","date":"2022","source":"European journal of histochemistry : EJH","url":"https://pubmed.ncbi.nlm.nih.gov/35730574","citation_count":13,"is_preprint":false},{"pmid":"35715622","id":"PMC_35715622","title":"Long non-coding RNA NRSN2-AS1, transcribed by SOX2, promotes progression of esophageal squamous cell carcinoma by regulating the ubiquitin-degradation of PGK1.","date":"2022","source":"Clinical & experimental metastasis","url":"https://pubmed.ncbi.nlm.nih.gov/35715622","citation_count":12,"is_preprint":false},{"pmid":"27908706","id":"PMC_27908706","title":"Highly expressed NRSN2 is related to malignant phenotype in ovarian cancer.","date":"2016","source":"Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie","url":"https://pubmed.ncbi.nlm.nih.gov/27908706","citation_count":10,"is_preprint":false},{"pmid":"35662041","id":"PMC_35662041","title":"E2F1-activated NRSN2 promotes esophageal squamous cell carcinoma progression through AKT/mTOR pathway.","date":"2022","source":"Pathology, research and practice","url":"https://pubmed.ncbi.nlm.nih.gov/35662041","citation_count":5,"is_preprint":false},{"pmid":"38569536","id":"PMC_38569536","title":"NRSN2 promotes the malignant behavior of HPV-transfected laryngeal carcinoma cells through AMPK/ULK1 pathway mediated autophagy activation.","date":"2024","source":"Cancer biology & therapy","url":"https://pubmed.ncbi.nlm.nih.gov/38569536","citation_count":3,"is_preprint":false},{"pmid":"39723223","id":"PMC_39723223","title":"Alterations of Sensory-related Functional Brain Network Connectivity in Nrsn2 Homozygous Knockout Mice.","date":"2024","source":"Phenomics (Cham, Switzerland)","url":"https://pubmed.ncbi.nlm.nih.gov/39723223","citation_count":3,"is_preprint":false},{"pmid":"38996888","id":"PMC_38996888","title":"Recombinant human protein TCFL5-activated NRSN2-AS1 promotes esophageal cancer progression via the microRNA-874-5p/RELT regulatory axis.","date":"2024","source":"International journal of biological macromolecules","url":"https://pubmed.ncbi.nlm.nih.gov/38996888","citation_count":1,"is_preprint":false}],"cost":{"stage1":{"model":"claude-sonnet-4-6","input_tokens":9969,"output_tokens":3077,"usd":0.038031,"stage1_stop_reason":"end_turn"},"stage2":{"model":"claude-opus-4-8","input_tokens":10348,"output_tokens":3224,"usd":0.06617,"stage2_stop_reason":"end_turn"},"total_usd":0.104201,"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\": 2015,\n      \"finding\": \"NRSN2 promotes non-small cell lung cancer cell growth through the PI3K/Akt/mTOR signaling pathway, as demonstrated by loss- and gain-of-function experiments in NSCLC cell lines showing effects on cell growth but not invasion.\",\n      \"method\": \"Loss- and gain-of-function assays in NSCLC cell lines, western blot for pathway components\",\n      \"journal\": \"International journal of clinical and experimental pathology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Weak — clean KD/OE with defined cellular phenotype and pathway readout, single lab, single study\",\n      \"pmids\": [\"26045763\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2015,\n      \"finding\": \"NRSN2 inhibits hepatocellular carcinoma cell proliferation and promotes senescence and apoptosis by regulating PI3K/AKT signaling and the p53/p21 pathway, as shown by loss- and gain-of-function assays.\",\n      \"method\": \"CCK-8 assays, SA-β-gal staining, Annexin V/PI staining, western blot for PI3K/AKT and p53/p21 pathway components; loss- and gain-of-function in HCC cell lines\",\n      \"journal\": \"Digestive diseases and sciences\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — multiple orthogonal functional assays (proliferation, senescence, apoptosis) plus pathway western blots, single lab\",\n      \"pmids\": [\"26055238\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2016,\n      \"finding\": \"NRSN2 promotes malignant phenotype in ovarian cancer cells including anchorage-independent growth, invasion, and chemoresistance, and positively correlates with CD133 stem cell marker expression; Wnt canonical signaling and Twist/Akt/Erk axis are regulated by NRSN2.\",\n      \"method\": \"Loss- and gain-of-function assays, colony formation, subcutaneous tumor formation, invasion assays, western blot for Wnt/Akt/Erk pathway components\",\n      \"journal\": \"Biomedicine & pharmacotherapy\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — multiple orthogonal functional assays plus pathway readouts, single lab\",\n      \"pmids\": [\"27908706\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2017,\n      \"finding\": \"NRSN2 promotes osteosarcoma cell proliferation and growth through dysregulation of both PI3K/AKT/mTOR and Wnt/β-catenin signaling pathways, demonstrated by loss- and gain-of-function assays in vitro and in vivo.\",\n      \"method\": \"Loss- and gain-of-function assays in osteosarcoma cell lines, in vivo tumor growth assays, western blot for PI3K/AKT/mTOR and Wnt/β-catenin pathway components\",\n      \"journal\": \"American journal of cancer research\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — in vitro and in vivo KD/OE with defined phenotype and two pathway readouts, single lab\",\n      \"pmids\": [\"28401012\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"NRSN2 promotes breast cancer cell proliferation, migration, and invasion by activating PI3K/AKT/mTOR and NF-κB signaling pathways, as shown by KD/OE experiments in vitro and in vivo.\",\n      \"method\": \"MTT assay, western blot for PI3K/AKT/mTOR and NF-κB pathway components, KD/OE in breast cancer cells, in vivo xenograft\",\n      \"journal\": \"Oncology letters\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — in vitro and in vivo functional assays with two pathway readouts, single lab\",\n      \"pmids\": [\"31885716\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"miR-143 directly targets NRSN2 (binding relationship verified by dual luciferase assay); BMSC-derived extracellular vesicles deliver lncRNA MALAT1 into osteosarcoma cells where MALAT1 sponges miR-143, thereby de-repressing NRSN2 expression and activating Wnt/β-catenin signaling to promote osteosarcoma proliferation, invasion, and migration.\",\n      \"method\": \"Dual luciferase reporter assay, EV isolation and characterization, transfection of si-MALAT1 and miR-143 mimic, western blot for Wnt/β-catenin pathway proteins, in vivo xenograft\",\n      \"journal\": \"OncoTargets and therapy\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — dual luciferase validation of miR-143/NRSN2 axis, in vitro and in vivo rescue experiments, single lab\",\n      \"pmids\": [\"33564242\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"miR-1913 directly targets NRSN2 (validated by dual luciferase reporter assay); hBMSC-derived exosomes deliver miR-1913 into osteosarcoma cells, suppressing NRSN2 expression and thereby inhibiting osteosarcoma cell viability, proliferation, migration, and invasion.\",\n      \"method\": \"Dual luciferase reporter assay, CCK8, colony formation, wound healing, transwell assays, RT-qPCR and western blot for NRSN2 and EMT markers, TEM and NTA for exosome characterization\",\n      \"journal\": \"American journal of translational research\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — dual luciferase validation of miR-1913/NRSN2 targeting, multiple orthogonal functional assays, single lab\",\n      \"pmids\": [\"34650689\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"NRSN2 activates the ERK1/2-Bcl-xL signaling pathway; miR-206 (delivered via BMSC-derived extracellular vesicles) directly targets NRSN2 (validated by dual luciferase assay), suppressing NRSN2-mediated ERK1/2-Bcl-xL activation and inhibiting osteosarcoma progression.\",\n      \"method\": \"Dual luciferase reporter assay, western blot for p-ERK1/2, ERK1/2, Bcl-xL, EdU and colony formation assays, flow cytometry for apoptosis, immunofluorescence, in vivo xenograft\",\n      \"journal\": \"European journal of histochemistry\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — dual luciferase validation, pathway rescue by NRSN2 OE reversing EV effect, in vitro and in vivo, single lab\",\n      \"pmids\": [\"35730574\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"E2F1 directly transcribes NRSN2 (confirmed by luciferase reporter and chromatin immunoprecipitation assays); the E2F1/NRSN2 axis promotes ESCC cell proliferation, migration, and invasion by activating the AKT/mTOR pathway.\",\n      \"method\": \"Luciferase reporter assay, chromatin immunoprecipitation (ChIP), loss- and gain-of-function assays, western blot for AKT/mTOR pathway, in vivo xenograft\",\n      \"journal\": \"Pathology, research and practice\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — ChIP and luciferase reporter to establish upstream E2F1 regulation, in vitro and in vivo functional assays, single lab\",\n      \"pmids\": [\"35662041\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"NRSN2 promotes malignant behavior of HPV-transfected laryngeal carcinoma cells through activation of the AMPK/ULK1 pathway and consequent autophagy induction; NRSN2 knockdown suppresses AMPK/ULK1-mediated autophagy and restrains malignant behavior.\",\n      \"method\": \"RT-qPCR, immunoblotting, immunofluorescence, CCK-8, EdU staining, transwell, wound healing, flow cytometry, TEM for autophagosomes, AMPK/ULK1 pathway inhibition experiments\",\n      \"journal\": \"Cancer biology & therapy\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Weak — pathway inhibition plus KD functional assays, multiple readouts, single lab single study\",\n      \"pmids\": [\"38569536\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"NRSN2 knockout mice show significant alterations in functional brain network connectivity—particularly in sensory-related (gustatory and auditory) and default mode network regions—without marked changes in brain microstructure or regional activity, linking NRSN2 to sensory processing and stress resilience circuitry.\",\n      \"method\": \"11.7 T multimodal MRI (structural and functional) in Nrsn2 homozygous knockout mice compared with wild-type at juvenile and adult stages\",\n      \"journal\": \"Phenomics\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Weak — clean KO with defined neuroimaging phenotype, multimodal MRI, single lab single study\",\n      \"pmids\": [\"39723223\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"lncRNA LUESCC functions as a ceRNA by sponging miR-6785-5p to enhance NRSN2 expression, which is critical for ESCC proliferation, migration, and invasion; this axis was mechanistically established by luciferase reporter assays and rescue experiments.\",\n      \"method\": \"Luciferase reporter assay (LUESCC/miR-6785-5p and miR-6785-5p/NRSN2 interactions), KD of LUESCC, rescue by NRSN2 modulation, in vitro and in vivo functional assays\",\n      \"journal\": \"Cellular and molecular life sciences\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — dual luciferase validation of ceRNA axis, rescue experiments, in vitro and in vivo, single lab\",\n      \"pmids\": [\"38457049\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"NRSN2 (Neurensin-2) is a small neuronal membrane protein that, in multiple cancer contexts, acts as a pro-proliferative/oncogenic factor by activating PI3K/AKT/mTOR, Wnt/β-catenin, NF-κB, ERK1/2-Bcl-xL, and AMPK/ULK1 signaling pathways; its transcription is directly driven by E2F1 (in ESCC), it is post-transcriptionally suppressed by miRNAs (miR-143, miR-206, miR-1913, miR-6785-5p), and in the nervous system its knockout alters functional brain network connectivity in sensory and default mode network regions without gross structural changes.\"\n}\n```","stage2_raw":"{\n  \"mechanistic_narrative\": \"NRSN2 (Neurensin-2) is a small protein that functions predominantly as a pro-proliferative factor across diverse epithelial and mesenchymal cancers, where it couples upstream transcriptional and microRNA regulation to multiple growth-promoting signaling cascades [#0, #3, #8]. In non-small cell lung, osteosarcoma, breast, esophageal squamous cell carcinoma (ESCC), and other tumor cells, NRSN2 gain- and loss-of-function drives proliferation, migration, and invasion through activation of PI3K/AKT/mTOR signaling, frequently in concert with Wnt/\\u03b2-catenin, NF-\\u03baB, ERK1/2-Bcl-xL, or AMPK/ULK1-mediated autophagy [#0, #3, #4, #7, #9]. Its expression is controlled at two levels: direct transcriptional activation by E2F1 in ESCC [#8], and post-transcriptional repression by multiple microRNAs (miR-143, miR-206, miR-1913, miR-6785-5p) that are themselves modulated by lncRNA ceRNA networks delivered via mesenchymal stem cell-derived extracellular vesicles [#5, #6, #7, #11]. Notably, in hepatocellular carcinoma NRSN2 acts in the opposite direction, suppressing proliferation and promoting senescence and apoptosis via the p53/p21 axis, indicating context-dependent function [#1]. Beyond cancer, knockout mice reveal a neuronal role: loss of NRSN2 alters functional brain network connectivity in sensory and default mode network regions without gross structural change [#10]. The biochemical activity of the NRSN2 protein itself\\u2014its molecular partners and the mechanism by which it engages these signaling pathways\\u2014has not been characterized in the available corpus.\",\n  \"teleology\": [\n    {\n      \"year\": 2015,\n      \"claim\": \"Established NRSN2 as a functional driver of cancer cell growth and linked it to a specific signaling cascade, defining its first mechanistic role.\",\n      \"evidence\": \"Loss- and gain-of-function assays with pathway western blots in NSCLC cell lines\",\n      \"pmids\": [\"26045763\"],\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"\",\n      \"gaps\": [\"Does not show direct molecular activity of NRSN2 on PI3K/Akt components\", \"Single cell-line context, no in vivo data\", \"Effect on invasion not observed\"]\n    },\n    {\n      \"year\": 2015,\n      \"claim\": \"Revealed that NRSN2 function is context-dependent, acting as a growth suppressor in hepatocellular carcinoma rather than a uniform oncogene.\",\n      \"evidence\": \"Proliferation, senescence (SA-\\u03b2-gal), and apoptosis assays with p53/p21 and PI3K/AKT western blots in HCC cell lines\",\n      \"pmids\": [\"26055238\"],\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"\",\n      \"gaps\": [\"Mechanism underlying opposite directionality versus other cancers unresolved\", \"No direct interaction with p53/p21 demonstrated\", \"Single lab\"]\n    },\n    {\n      \"year\": 2016,\n      \"claim\": \"Extended NRSN2's oncogenic repertoire to invasion, chemoresistance, and stemness, broadening its phenotypic scope beyond proliferation.\",\n      \"evidence\": \"Anchorage-independent growth, invasion, tumor formation, CD133 correlation, and Wnt/Akt/Erk western blots in ovarian cancer cells\",\n      \"pmids\": [\"27908706\"],\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"\",\n      \"gaps\": [\"Multiple pathways implicated without identifying primary effector\", \"No upstream regulator defined\", \"Single lab\"]\n    },\n    {\n      \"year\": 2017,\n      \"claim\": \"Showed NRSN2 simultaneously dysregulates two canonical pathways in vivo, consolidating its pro-growth role in osteosarcoma.\",\n      \"evidence\": \"In vitro and in vivo KD/OE with PI3K/AKT/mTOR and Wnt/\\u03b2-catenin western blots in osteosarcoma\",\n      \"pmids\": [\"28401012\"],\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"\",\n      \"gaps\": [\"Hierarchy between the two pathways not resolved\", \"No direct molecular mechanism of activation\", \"Single lab\"]\n    },\n    {\n      \"year\": 2019,\n      \"claim\": \"Added NF-\\u03baB to the set of NRSN2-activated cascades and confirmed pro-metastatic activity in vivo in breast cancer.\",\n      \"evidence\": \"MTT, in vivo xenograft, PI3K/AKT/mTOR and NF-\\u03baB western blots in breast cancer cells\",\n      \"pmids\": [\"31885716\"],\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"\",\n      \"gaps\": [\"Direct link between NRSN2 and NF-\\u03baB activation not shown\", \"No partner identified\", \"Single lab\"]\n    },\n    {\n      \"year\": 2021,\n      \"claim\": \"Identified the first upstream regulatory layer, showing NRSN2 is a direct microRNA target embedded in extracellular-vesicle-delivered lncRNA ceRNA networks.\",\n      \"evidence\": \"Dual luciferase validation of miR-143 and miR-1913 targeting NRSN2, with EV/exosome delivery and rescue in osteosarcoma\",\n      \"pmids\": [\"33564242\", \"34650689\"],\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"\",\n      \"gaps\": [\"Whether NRSN2 protein output is the sole effector of these axes unclear\", \"Generalizability beyond osteosarcoma untested\", \"Single lab per axis\"]\n    },\n    {\n      \"year\": 2022,\n      \"claim\": \"Defined the ERK1/2-Bcl-xL pathway as an NRSN2 effector and established a complete transcriptional control point via E2F1.\",\n      \"evidence\": \"Dual luciferase and ChIP showing E2F1 transcribes NRSN2 in ESCC; miR-206/EV experiments linking NRSN2 to ERK1/2-Bcl-xL in osteosarcoma\",\n      \"pmids\": [\"35730574\", \"35662041\"],\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"\",\n      \"gaps\": [\"How NRSN2 physically engages ERK1/2 or Bcl-xL unknown\", \"E2F1 regulation shown only in ESCC\", \"Single lab per study\"]\n    },\n    {\n      \"year\": 2024,\n      \"claim\": \"Connected NRSN2 to autophagy regulation through AMPK/ULK1 and reinforced the ESCC ceRNA axis with an additional lncRNA-miRNA pair.\",\n      \"evidence\": \"AMPK/ULK1 pathway inhibition and autophagosome TEM in HPV-laryngeal carcinoma; luciferase and rescue for LUESCC/miR-6785-5p/NRSN2 in ESCC\",\n      \"pmids\": [\"38569536\", \"38457049\"],\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"\",\n      \"gaps\": [\"Direct molecular mechanism linking NRSN2 to AMPK activation absent\", \"Whether autophagy is cause or consequence of malignancy unclear\", \"Single lab per study\"]\n    },\n    {\n      \"year\": 2024,\n      \"claim\": \"Provided the first in vivo evidence of an endogenous neuronal function, implicating NRSN2 in functional brain network connectivity.\",\n      \"evidence\": \"11.7 T multimodal structural and functional MRI in Nrsn2 knockout versus wild-type mice\",\n      \"pmids\": [\"39723223\"],\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"\",\n      \"gaps\": [\"Cellular/molecular basis of connectivity changes not defined\", \"No behavioral correlation established\", \"Relationship to its cancer signaling roles unknown\"]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"The intrinsic biochemical activity of the NRSN2 protein and its direct molecular partners remain undefined, leaving open how it mechanistically engages the many signaling pathways attributed to it.\",\n      \"evidence\": \"No discovery in the corpus reports a direct binding partner, enzymatic activity, or structural mechanism for NRSN2\",\n      \"pmids\": [],\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"\",\n      \"gaps\": [\"No direct physical interactor identified\", \"No structural or biochemical characterization\", \"Context-dependent oncogene-versus-suppressor switch unexplained\"]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [],\n    \"localization\": [],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-162582\", \"supporting_discovery_ids\": [0, 3, 4, 8, 9]},\n      {\"term_id\": \"R-HSA-9612973\", \"supporting_discovery_ids\": [9]},\n      {\"term_id\": \"R-HSA-112316\", \"supporting_discovery_ids\": [10]}\n    ],\n    \"complexes\": [],\n    \"partners\": [],\n    \"other_free_text\": []\n  }\n}","audit_flag":null,"evaluation":{"faith_supported":5,"faith_total":5,"faith_pct":100.0}}