{"gene":"TSPAN1","run_date":"2026-06-10T10:51:56","timeline":{"discoveries":[{"year":2020,"finding":"TSPAN1 promotes autophagy flux by directly binding to LC3 through two conserved LIR (LC3-interacting region) motifs; mutation of both LIR motifs abolished TSPAN1's ability to induce autophagy and promote pancreatic cancer cell proliferation.","method":"Co-immunoprecipitation, GFP-LC3 puncta formation assay, LIR motif mutagenesis, zebrafish tspan1 mutant model","journal":"Autophagy","confidence":"High","confidence_rationale":"Tier 1–2 / Moderate — direct binding demonstrated by Co-IP, functional consequence confirmed by LIR motif mutagenesis and in vivo zebrafish model; multiple orthogonal methods in one study","pmids":["32972302"],"is_preprint":false},{"year":2020,"finding":"TSPAN1 expression is transcriptionally upregulated by FAM83A through the canonical WNT-CTNNB1 signaling pathway, with two conserved TCF/LEF binding elements identified in the TSPAN1 promoter region, verified by luciferase activity and ChIP assays.","method":"Luciferase reporter assay, ChIP assay","journal":"Autophagy","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — two orthogonal methods (luciferase + ChIP) from single lab","pmids":["32972302"],"is_preprint":false},{"year":2019,"finding":"TSPAN1 upregulates MMP2 expression to promote pancreatic cancer cell migration and invasion via PLCγ; TSPAN1 siRNA suppressed MMP2 mRNA expression by blocking translocation and phosphorylation of PLCγ, while TSPAN1 overexpression had the opposite effect.","method":"siRNA knockdown, cDNA overexpression, transwell invasion/migration assay, siRNA targeting PLCγ","journal":"Oncology reports","confidence":"Medium","confidence_rationale":"Tier 2–3 / Moderate — epistatic placement of TSPAN1 upstream of PLCγ-MMP2 by genetic knockdown, single lab with multiple functional assays","pmids":["30720116"],"is_preprint":false},{"year":2011,"finding":"TSPAN1 physically interacts with the human thiamine transporter-1 (hTHTR-1) and stabilizes it, doubling its protein half-life (from ~6 h to ~12 h) and increasing thiamine uptake in intestinal epithelial cells.","method":"Bacterial two-hybrid screen, co-immunoprecipitation, GST pulldown, live-cell confocal imaging, cycloheximide chase assay, functional thiamine uptake assay","journal":"American journal of physiology. Gastrointestinal and liver physiology","confidence":"High","confidence_rationale":"Tier 1–2 / Strong — interaction confirmed by multiple orthogonal methods (bacterial two-hybrid, Co-IP, GST pulldown, colocalization) and functional consequence validated by two independent assays (protein stability + thiamine uptake)","pmids":["21836059"],"is_preprint":false},{"year":2017,"finding":"TSPAN1 expression is acutely induced by androgens in prostate cancer cells; stable upregulation of TSPAN1 in DU145 and PC3 cells significantly increased cell migration and induced expression of mesenchymal markers SLUG and ARF6.","method":"Stable overexpression, wound-healing/migration assay, western blotting for SLUG and ARF6","journal":"Scientific reports","confidence":"Medium","confidence_rationale":"Tier 2–3 / Moderate — gain-of-function with defined downstream markers, androgen regulation confirmed, single lab","pmids":["28701765"],"is_preprint":false},{"year":2020,"finding":"TSPAN1 transcriptionally regulates ITGA2 expression and epigenetically controls ITGA2 promoter methylation by modulating levels of TET2, DNMT3B and DNMT1, resulting in hypomethylation of the ITGA2 CpG island; knockdown of ITGA2 abolished the pro-proliferative and pro-invasive effects of TSPAN1 overexpression in pancreatic cancer cells.","method":"RNA-Seq, western blot, methylation analysis (TET2/DNMT3B/DNMT1 levels), ITGA2 knockdown rescue experiment","journal":"American journal of cancer research","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — pathway placement by genetic rescue experiment; epigenetic mechanism supported by multiple markers; single lab","pmids":["32368389"],"is_preprint":false},{"year":2024,"finding":"ALKBH5 suppresses TSPAN1 expression by demethylating its mRNA (reducing m6A modification); TSPAN1 overexpression reversed the autophagy-inhibiting and anti-proliferative effects of ALKBH5 overexpression in prostate cancer cells, placing TSPAN1 downstream of ALKBH5-mediated m6A regulation.","method":"MeRIP-qPCR, western blot (LC3B-II/I ratio, P62), ALKBH5 overexpression, TSPAN1 overexpression rescue","journal":"Discovery medicine","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — m6A modification of TSPAN1 mRNA directly measured by MeRIP-qPCR; rescue experiment establishes epistasis; single lab","pmids":["38273746"],"is_preprint":false},{"year":2021,"finding":"TSPAN1 depletion in HNSCC cells inhibited signaling cascades, with phospho-SRC inhibition identified as a major common target; TSPAN1 depletion also reversed EMT features in cisplatin-resistant tumors in vivo.","method":"siRNA knockdown, western blotting for pSRC, in vivo xenograft, co-immunoprecipitation (proteomic study)","journal":"Cancers","confidence":"Medium","confidence_rationale":"Tier 2–3 / Moderate — pSRC identified as downstream target by western blot after TSPAN1 depletion, corroborated by IHC correlation; single lab","pmids":["33167355"],"is_preprint":false},{"year":2022,"finding":"TSPAN1 positively transcriptionally regulates FAM110A expression; TSPAN1 overexpression increased FAM110A levels and the tumor-promoting effects of FAM110A overexpression could be abolished by HIST1H2BK knockdown, defining a TSPAN1/FAM110A/HIST1H2BK/G9a oncogenic pathway in pancreatic cancer.","method":"Stable overexpression/knockdown, RNA-Seq, luciferase reporter assay, western blot, rescue experiment","journal":"Journal of Cancer","confidence":"Medium","confidence_rationale":"Tier 2–3 / Moderate — transcriptional regulation confirmed by luciferase reporter; pathway epistasis established by rescue experiment; single lab","pmids":["35154458"],"is_preprint":false},{"year":2023,"finding":"TSPAN1 inhibits NPC cell migration, invasion, and metastasis by interacting with IKBB protein, preventing overactivation of the NF-κB pathway.","method":"Co-immunoprecipitation (TSPAN1–IKBB interaction), overexpression/knockdown, in vitro migration/invasion assay, in vivo metastasis assay","journal":"Cancer gene therapy","confidence":"Medium","confidence_rationale":"Tier 2–3 / Moderate — binding partner identified by Co-IP; functional consequence in vitro and in vivo; single lab","pmids":["38135697"],"is_preprint":false},{"year":2021,"finding":"TSPAN1 promotes pancreatic cancer cell growth and invasion by mediating the PI3K/Akt pathway and promoting EMT; TSPAN1 depletion suppressed Akt phosphorylation and EMT markers in breast cancer cells.","method":"siRNA knockdown, western blot for PI3K/Akt phosphorylation and EMT markers, in vivo xenograft","journal":"Bioengineered","confidence":"Low","confidence_rationale":"Tier 3 / Weak — PI3K/Akt pathway involvement inferred from phosphorylation western blots after knockdown; single lab, single method type","pmids":["34852709"],"is_preprint":false},{"year":2016,"finding":"In prostate cancer cell lines, TSPAN1 knockdown resulted in increased Akt phosphorylation and caused cell cycle transition from G1 to S phase, placing TSPAN1 as a negative regulator of the PI3K/Akt pathway; TSPAN1 expression was positively correlated with PTEN.","method":"siRNA knockdown, western blot (pAkt), flow cytometry (cell cycle)","journal":"Oncotarget","confidence":"Low","confidence_rationale":"Tier 3 / Weak — pathway placement inferred from phosphorylation change after knockdown; single lab, single method type, no direct mechanistic link established","pmids":["27556508"],"is_preprint":false},{"year":2021,"finding":"TSPAN1 silencing in cerulein-induced pancreatic acinar cell injury inhibited endoplasmic reticulum stress; immunoprecipitation confirmed TSPAN1 directly binds AGR2, and AGR2 overexpression rescued the effects of TSPAN1 silencing on cell proliferation and inflammatory response.","method":"Immunoprecipitation (TSPAN1–AGR2 binding), siRNA knockdown, AGR2 overexpression rescue, western blot (ER stress markers), ELISA (cytokines)","journal":"Drug development research","confidence":"Medium","confidence_rationale":"Tier 2–3 / Moderate — binding partner confirmed by IP; epistasis validated by rescue experiment; single lab","pmids":["34212407"],"is_preprint":false},{"year":2007,"finding":"NET-1 (TSPAN1) protein localizes to the cytoplasm of hepatocellular carcinoma cells as irregular granules near the Golgi apparatus, as determined by fluorescent immunocytochemistry and confocal laser scanning microscopy.","method":"Fluorescent immunocytochemistry, confocal laser scanning microscopy","journal":"Zhonghua zhong liu za zhi [Chinese journal of oncology]","confidence":"Low","confidence_rationale":"Tier 3 / Weak — direct subcellular localization experiment, single lab, no functional consequence directly linked in this paper","pmids":["18478931"],"is_preprint":false},{"year":2024,"finding":"TSPAN1 physically interacts with TM4SF1 in glioblastoma stem cells (GSCs), as confirmed by Co-IP and immunofluorescence; 4,5-dimethoxycanthin-6-one inhibited this TSPAN1/TM4SF1 interaction and suppressed GSC self-renewal, and TSPAN1 overexpression partially reversed the drug's inhibitory effects.","method":"Co-immunoprecipitation, immunofluorescence, molecular docking, TSPAN1 overexpression rescue, in vivo xenograft","journal":"Neurochemical research","confidence":"Medium","confidence_rationale":"Tier 2–3 / Moderate — interaction confirmed by Co-IP and IF; functional relevance supported by rescue experiment; single lab","pmids":["39060768"],"is_preprint":false},{"year":2019,"finding":"Inhibition of NET-1 (TSPAN1) in HCC cells activated the PI3K/AKT signaling pathway (decreased phosphorylated PI3K and AKT), altered expression of apoptosis-related proteins (decreased Bax and cyclinD1, increased Bcl-2 and caspase-3), and suppressed HCC cell proliferation while promoting apoptosis.","method":"siRNA knockdown, western blot (PI3K, pAKT, Bax, Bcl-2, caspase-3, cyclinD1), flow cytometry","journal":"Experimental and therapeutic medicine","confidence":"Low","confidence_rationale":"Tier 3 / Weak — pathway assignment based on phosphorylation changes after knockdown; single lab, single method type","pmids":["30867719"],"is_preprint":false},{"year":2022,"finding":"TSPAN1 expression transcriptionally regulates ITGA2 by modulating TET2, DNMT3B and DNMT1 levels in ovarian cancer cells, resulting in ITGA2 promoter hypomethylation, as confirmed by ChIP assay and luciferase reporter assay.","method":"Luciferase reporter assay, ChIP assay, rescue experiment","journal":"Stem cells international","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — two orthogonal methods confirm epigenetic regulation; pathway replicated from pancreatic cancer study (PMID 32368389); single lab","pmids":["36187230"],"is_preprint":false}],"current_model":"TSPAN1 is a tetraspanin transmembrane protein that functions as a scaffolding/adaptor protein: it directly binds LC3 via two LIR motifs to promote autophagy flux, interacts with the thiamine transporter hTHTR-1 to stabilize it, binds IKBB to suppress NF-κB signaling, binds AGR2 to induce ER stress, and associates with TM4SF1 in glioblastoma stem cells; in cancer contexts it also promotes cell migration and invasion by activating PLCγ–MMP2 signaling, epigenetically upregulates ITGA2 through modulation of TET2/DNMT3B/DNMT1, and its transcription is driven by WNT-CTNNB1 signaling via TCF/LEF elements and regulated post-transcriptionally by m6A demethylation through ALKBH5."},"narrative":{"mechanistic_narrative":"TSPAN1 is a tetraspanin transmembrane protein that acts as a membrane-associated scaffold linking it to autophagy, transporter stabilization, and oncogenic signaling [PMID:32972302, PMID:21836059]. Its best-defined molecular activity is in autophagy: TSPAN1 directly binds LC3 through two conserved LIR motifs, and mutation of both motifs abolishes its ability to induce autophagy and drive pancreatic cancer cell proliferation [PMID:32972302]. Beyond autophagy, TSPAN1 stabilizes the thiamine transporter hTHTR-1, roughly doubling its protein half-life and increasing thiamine uptake in intestinal epithelium [PMID:21836059]. In cancer it functions largely as a signaling adaptor: it promotes migration and invasion by activating PLCγ to upregulate MMP2 [PMID:30720116], epigenetically upregulates ITGA2 by modulating TET2/DNMT3B/DNMT1 to hypomethylate the ITGA2 promoter, an axis required for its pro-proliferative and pro-invasive effects [PMID:32368389, PMID:36187230], and transcriptionally activates a FAM110A/HIST1H2BK/G9a oncogenic pathway [PMID:35154458]. TSPAN1 also engages specific protein partners with context-dependent outputs, binding IKBB to restrain NF-κB signaling [PMID:38135697], binding AGR2 to drive ER stress [PMID:34212407], and associating with TM4SF1 to support glioblastoma stem cell self-renewal [PMID:39060768]. Upstream, TSPAN1 transcription is driven by WNT-CTNNB1 signaling via TCF/LEF elements in its promoter [PMID:32972302] and its mRNA is suppressed by ALKBH5-mediated m6A demethylation [PMID:38273746].","teleology":[{"year":2007,"claim":"Established the first subcellular localization of TSPAN1, placing it in the cytoplasm near the Golgi rather than diffusely, providing a structural starting point for its membrane scaffolding role.","evidence":"Fluorescent immunocytochemistry and confocal microscopy in hepatocellular carcinoma cells","pmids":["18478931"],"confidence":"Low","gaps":["Single lab descriptive localization with no functional consequence linked","Does not establish plasma-membrane versus organelle topology"]},{"year":2011,"claim":"Answered whether TSPAN1 has a physiological binding partner by showing it directly interacts with and stabilizes the thiamine transporter hTHTR-1, establishing a non-cancer scaffolding/chaperone function.","evidence":"Bacterial two-hybrid, Co-IP, GST pulldown, cycloheximide chase, and thiamine uptake assays in intestinal epithelial cells","pmids":["21836059"],"confidence":"High","gaps":["Structural basis of the interaction not resolved","Whether stabilization is general to other membrane partners untested"]},{"year":2016,"claim":"Began placing TSPAN1 in growth signaling by linking its loss to increased Akt phosphorylation and G1/S transition in prostate cancer, framing it as a possible negative PI3K/Akt regulator.","evidence":"siRNA knockdown, pAkt western blot, and flow cytometry cell cycle analysis","pmids":["27556508"],"confidence":"Low","gaps":["Pathway placement inferred from phosphorylation changes only","No direct mechanistic link to PI3K/Akt components","Direction conflicts with later contexts"]},{"year":2017,"claim":"Connected TSPAN1 to hormone-driven tumor biology by showing androgens acutely induce it and that it drives migration and mesenchymal marker expression.","evidence":"Stable overexpression, wound-healing assay, and western blot for SLUG and ARF6 in prostate cancer lines","pmids":["28701765"],"confidence":"Medium","gaps":["Mechanism linking TSPAN1 to SLUG/ARF6 not defined","Single lab gain-of-function"]},{"year":2019,"claim":"Defined a concrete pro-invasive mechanism by placing TSPAN1 upstream of PLCγ phosphorylation and MMP2 induction.","evidence":"siRNA knockdown/overexpression, transwell assays, and PLCγ siRNA epistasis in pancreatic cancer","pmids":["30720116"],"confidence":"Medium","gaps":["How TSPAN1 triggers PLCγ translocation/phosphorylation unknown","No direct TSPAN1–PLCγ binding shown"]},{"year":2020,"claim":"Identified TSPAN1's core molecular activity — direct LIR-dependent LC3 binding driving autophagy flux — and showed WNT-CTNNB1 signaling transcriptionally controls its expression.","evidence":"Co-IP, GFP-LC3 puncta, LIR mutagenesis, zebrafish mutant, luciferase reporter and ChIP","pmids":["32972302"],"confidence":"High","gaps":["How a transmembrane tetraspanin presents LIR motifs to cytosolic LC3 not structurally resolved","FAM83A/WNT regulation from single lab"]},{"year":2020,"claim":"Established an epigenetic effector arm by showing TSPAN1 controls ITGA2 promoter methylation via TET2/DNMT3B/DNMT1 and that ITGA2 is required for TSPAN1's oncogenic effects.","evidence":"RNA-Seq, methylation analysis, and ITGA2 knockdown rescue in pancreatic cancer (replicated in ovarian cancer, 2022)","pmids":["32368389","36187230"],"confidence":"Medium","gaps":["How a membrane protein modulates nuclear TET/DNMT levels mechanistically unexplained","Direct versus indirect regulation unresolved"]},{"year":2021,"claim":"Expanded TSPAN1's partner repertoire and signaling reach by identifying AGR2 binding driving ER stress, and implicating SRC and PI3K/Akt/EMT cascades downstream.","evidence":"Co-IP/IP, AGR2 rescue, pSRC and pAkt/EMT western blots, and xenografts across pancreatic, HNSCC and breast models","pmids":["34212407","33167355","34852709"],"confidence":"Medium","gaps":["PI3K/Akt placement inferred from phosphorylation only (Low-confidence)","Conflicting positive versus negative effects on Akt across contexts unreconciled"]},{"year":2023,"claim":"Demonstrated a tumor-suppressive partner interaction by showing TSPAN1 binds IKBB to restrain NF-κB and inhibit nasopharyngeal carcinoma metastasis, indicating context-dependent function.","evidence":"Co-IP, overexpression/knockdown, and in vitro/in vivo metastasis assays","pmids":["38135697"],"confidence":"Medium","gaps":["Reciprocal validation of TSPAN1–IKBB binding limited","Reconciliation with pro-invasive roles in other cancers not addressed"]},{"year":2024,"claim":"Added an upstream m6A regulatory layer (ALKBH5 demethylation suppresses TSPAN1) and a stem-cell partner (TM4SF1 in glioblastoma), broadening TSPAN1's regulatory and interaction map.","evidence":"MeRIP-qPCR with rescue, and Co-IP/IF with docking and rescue in glioblastoma stem cells","pmids":["38273746","39060768"],"confidence":"Medium","gaps":["m6A reader linking demethylation to TSPAN1 stability/translation unidentified","Functional consequence of TSPAN1–TM4SF1 complex on signaling undefined"]},{"year":null,"claim":"It remains unknown how a single tetraspanin reconciles its opposing tumor-promoting and tumor-suppressive roles, and what structural feature of TSPAN1 coordinates membrane partners with cytosolic autophagy and nuclear epigenetic outputs.","evidence":"","pmids":[],"confidence":"Low","gaps":["No structural model of TSPAN1 in its complexes","No unifying mechanism explaining context-dependent oncogenic versus suppressive activity","Membrane-to-nucleus signaling route undefined"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0060090","term_label":"molecular adaptor activity","supporting_discovery_ids":[0,3,9,12,14]},{"term_id":"GO:0140110","term_label":"transcription regulator activity","supporting_discovery_ids":[5,8,16]}],"localization":[{"term_id":"GO:0005829","term_label":"cytosol","supporting_discovery_ids":[13]},{"term_id":"GO:0005794","term_label":"Golgi apparatus","supporting_discovery_ids":[13]}],"pathway":[{"term_id":"R-HSA-9612973","term_label":"Autophagy","supporting_discovery_ids":[0,6]},{"term_id":"R-HSA-162582","term_label":"Signal Transduction","supporting_discovery_ids":[2,9]},{"term_id":"R-HSA-4839726","term_label":"Chromatin organization","supporting_discovery_ids":[5,16]}],"complexes":[],"partners":["LC3","HTHTR-1","IKBB","AGR2","TM4SF1"],"other_free_text":[]}},"prefetch_data":{"uniprot":{"accession":"O60635","full_name":"Tetraspanin-1","aliases":["Tetraspan NET-1","Tetraspanin TM4-C"],"length_aa":241,"mass_kda":26.3,"function":"Structural component of specialized membrane microdomains known as tetraspanin-enriched microdomains (TERMs), which act as platforms for receptor clustering and signaling. Participates thereby in diverse biological functions such as cell signal transduction, adhesion, migration and protein trafficking (PubMed:30066932, PubMed:30291375). Regulates neuronal differentiation in response to NGF by facilitating NGF-mediated activation of NTRK1/TRKA receptor tyrosine kinase and subsequent downstream signaling pathways (By similarity). Plays a role in the inhibition of TNFalpha-induced apoptosis. Mechanistically, inhibits the NF-kappa-B signaling pathway by blocking phosphorylation of CHUK (PubMed:30291375). Also promotes the stability of the thiamine transporter 1/SLC19A2 in intestinal epithelial cells leading to an increase of thiamine uptake process (PubMed:21836059)","subcellular_location":"Cell membrane; Lysosome membrane","url":"https://www.uniprot.org/uniprotkb/O60635/entry"},"depmap":{"release":"DepMap","has_data":true,"is_common_essential":false,"resolved_as":"","url":"https://depmap.org/portal/gene/TSPAN1","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/TSPAN1","total_profiled":1310},"omim":[{"mim_id":"613170","title":"TETRASPANIN 1; TSPAN1","url":"https://www.omim.org/entry/613170"}],"hpa":{"profiled":true,"resolved_as":"","reliability":"Approved","locations":[{"location":"Nucleoplasm","reliability":"Approved"},{"location":"Vesicles","reliability":"Approved"}],"tissue_specificity":"Tissue enhanced","tissue_distribution":"Detected in many","driving_tissues":[{"tissue":"intestine","ntpm":556.4},{"tissue":"kidney","ntpm":233.4}],"url":"https://www.proteinatlas.org/search/TSPAN1"},"hgnc":{"alias_symbol":["TSPAN-1","NET-1"],"prev_symbol":[]},"alphafold":{"accession":"O60635","domains":[{"cath_id":"1.10.1450","chopping":"113-204","consensus_level":"high","plddt":88.5807,"start":113,"end":204},{"cath_id":"1.20.58","chopping":"2-81","consensus_level":"high","plddt":83.4228,"start":2,"end":81}],"viewer_url":"https://alphafold.ebi.ac.uk/entry/O60635","model_url":"https://alphafold.ebi.ac.uk/files/AF-O60635-F1-model_v6.cif","pae_url":"https://alphafold.ebi.ac.uk/files/AF-O60635-F1-predicted_aligned_error_v6.png","plddt_mean":88.19},"mouse_models":{"mgi_url":"https://www.informatics.jax.org/marker/summary?nomen=TSPAN1","jax_strain_url":"https://www.jax.org/strain/search?query=TSPAN1"},"sequence":{"accession":"O60635","fasta_url":"https://rest.uniprot.org/uniprotkb/O60635.fasta","uniprot_url":"https://www.uniprot.org/uniprotkb/O60635/entry","alphafold_viewer_url":"https://alphafold.ebi.ac.uk/entry/O60635"}},"corpus_meta":[{"pmid":"32972302","id":"PMC_32972302","title":"TSPAN1 promotes autophagy flux and mediates cooperation between WNT-CTNNB1 signaling and autophagy via the MIR454-FAM83A-TSPAN1 axis in pancreatic cancer.","date":"2020","source":"Autophagy","url":"https://pubmed.ncbi.nlm.nih.gov/32972302","citation_count":81,"is_preprint":false},{"pmid":"8202508","id":"PMC_8202508","title":"A net +1 frameshift permits synthesis of thymidine kinase from a drug-resistant herpes simplex virus mutant.","date":"1994","source":"Proceedings of the National Academy of Sciences of the United States of America","url":"https://pubmed.ncbi.nlm.nih.gov/8202508","citation_count":67,"is_preprint":false},{"pmid":"18822690","id":"PMC_18822690","title":"Clinicopathological significance of overexpression of TSPAN1, Ki67 and CD34 in gastric carcinoma.","date":"2008","source":"Tumori","url":"https://pubmed.ncbi.nlm.nih.gov/18822690","citation_count":51,"is_preprint":false},{"pmid":"12115476","id":"PMC_12115476","title":"Identification of a new proliferation-associated protein NET-1/C4.8 characteristic for a subset of high-grade cervical intraepithelial neoplasia and cervical carcinomas.","date":"2002","source":"International journal of cancer","url":"https://pubmed.ncbi.nlm.nih.gov/12115476","citation_count":51,"is_preprint":false},{"pmid":"3910098","id":"PMC_3910098","title":"Mechanism of inactivation of Escherichia coli ribonucleotide reductase by 2'-chloro-2'-deoxyuridine 5'-diphosphate: evidence for generation of a 2'-deoxy-3'-ketonucleotide via a net 1,2 hydrogen shift.","date":"1985","source":"Biochemistry","url":"https://pubmed.ncbi.nlm.nih.gov/3910098","citation_count":46,"is_preprint":false},{"pmid":"30720116","id":"PMC_30720116","title":"TSPAN1 upregulates MMP2 to promote pancreatic cancer cell migration and invasion via PLCγ.","date":"2019","source":"Oncology reports","url":"https://pubmed.ncbi.nlm.nih.gov/30720116","citation_count":42,"is_preprint":false},{"pmid":"28701765","id":"PMC_28701765","title":"The cancer-associated cell migration protein TSPAN1 is under control of androgens and its upregulation increases prostate cancer cell migration.","date":"2017","source":"Scientific reports","url":"https://pubmed.ncbi.nlm.nih.gov/28701765","citation_count":40,"is_preprint":false},{"pmid":"26054975","id":"PMC_26054975","title":"TSPAN1 functions as an oncogene in gastric cancer and is downregulated by miR-573.","date":"2015","source":"FEBS letters","url":"https://pubmed.ncbi.nlm.nih.gov/26054975","citation_count":39,"is_preprint":false},{"pmid":"21302622","id":"PMC_21302622","title":"Suppression of TSPAN1 by RNA interference inhibits proliferation and invasion of colon cancer cells in vitro.","date":"2010","source":"Tumori","url":"https://pubmed.ncbi.nlm.nih.gov/21302622","citation_count":37,"is_preprint":false},{"pmid":"19017553","id":"PMC_19017553","title":"Tspan-1 is a tetraspanin preferentially expressed by mucinous and endometrioid subtypes of human ovarian carcinomas.","date":"2008","source":"Cancer 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[et al]","url":"https://pubmed.ncbi.nlm.nih.gov/33320287","citation_count":13,"is_preprint":false},{"pmid":"22378020","id":"PMC_22378020","title":"The effect of NET-1 on the proliferation, migration and endocytosis of the SMMC-7721 HCC cell line.","date":"2012","source":"Oncology reports","url":"https://pubmed.ncbi.nlm.nih.gov/22378020","citation_count":13,"is_preprint":false},{"pmid":"39664809","id":"PMC_39664809","title":"Visible-light-driven net-1,2-hydrogen atom transfer of amidyl radicals to access β-amido ketone derivatives.","date":"2024","source":"Chemical science","url":"https://pubmed.ncbi.nlm.nih.gov/39664809","citation_count":13,"is_preprint":false},{"pmid":"31544556","id":"PMC_31544556","title":"Evaluation of therapeutic effect of targeting nanobubbles conjugated with NET-1 siRNA by shear wave elastography: an in vivo study of hepatocellular carcinoma bearing mice model.","date":"2019","source":"Drug 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inhibits metastasis of nasopharyngeal carcinoma via suppressing NF-kB signaling.","date":"2023","source":"Cancer gene therapy","url":"https://pubmed.ncbi.nlm.nih.gov/38135697","citation_count":5,"is_preprint":false},{"pmid":"39719558","id":"PMC_39719558","title":"Doublecortin regulates the mitochondrial-dependent apoptosis in glioma via Rho-A/Net-1/p38-MAPK signaling.","date":"2024","source":"Molecular medicine (Cambridge, Mass.)","url":"https://pubmed.ncbi.nlm.nih.gov/39719558","citation_count":4,"is_preprint":false},{"pmid":"16536986","id":"PMC_16536986","title":"[Expression of NET-1 gene and protein in hepatocellular carcinoma and related tissues].","date":"2006","source":"Ai zheng = Aizheng = Chinese journal of cancer","url":"https://pubmed.ncbi.nlm.nih.gov/16536986","citation_count":4,"is_preprint":false},{"pmid":"36187230","id":"PMC_36187230","title":"lncRNA DARS-AS1 Modulates TSPAN1-Mediated ITGA2 Hypomethylation by Interaction with miR-194-5p Thus Promoting Ovarian Cancer Progression.","date":"2022","source":"Stem cells international","url":"https://pubmed.ncbi.nlm.nih.gov/36187230","citation_count":3,"is_preprint":false},{"pmid":"36939874","id":"PMC_36939874","title":"Investigation of dynamic solution interactions between NET-1 and UNC-5B by multi-wavelength analytical ultracentrifugation.","date":"2023","source":"European biophysics journal : EBJ","url":"https://pubmed.ncbi.nlm.nih.gov/36939874","citation_count":3,"is_preprint":false},{"pmid":"30301500","id":"PMC_30301500","title":"NET-1 promotes invasion, adhesion and growth of hepatocellular carcinoma by regulating the expression of BAX, caspase 3, caspase 8 and BCL2.","date":"2018","source":"Cellular and molecular biology (Noisy-le-Grand, France)","url":"https://pubmed.ncbi.nlm.nih.gov/30301500","citation_count":3,"is_preprint":false},{"pmid":"34212407","id":"PMC_34212407","title":"TSPAN1 silencing protects against cerulein-induced pancreatic acinar cell injury via targeting 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surgery]","url":"https://pubmed.ncbi.nlm.nih.gov/25573220","citation_count":1,"is_preprint":false},{"pmid":"40777026","id":"PMC_40777026","title":"Revolutionizing cervical cancer treatment: single-cell sequencing of TSPAN1+ tumor EPCs and immune checkpoints to assess drug sensitivity and optimize therapy.","date":"2025","source":"Frontiers in immunology","url":"https://pubmed.ncbi.nlm.nih.gov/40777026","citation_count":1,"is_preprint":false},{"pmid":"29423111","id":"PMC_29423111","title":"Effect of NET-1 siRNA conjugated sub-micron bubble complex combined with low-frequency ultrasound exposure in gene transfection.","date":"2017","source":"Oncotarget","url":"https://pubmed.ncbi.nlm.nih.gov/29423111","citation_count":1,"is_preprint":false},{"pmid":"39060768","id":"PMC_39060768","title":"4,5-Dimethoxycanthin-6-one Inhibits Glioblastoma Stem Cell and Tumor Growth by Inhibiting TSPAN1 Interaction with TM4SF1.","date":"2024","source":"Neurochemical 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regeneration","date":"2025-04-28","source":"bioRxiv","url":"https://doi.org/10.1101/2025.04.27.650884","citation_count":0,"is_preprint":true}],"cost":{"stage1":{"model":"claude-sonnet-4-6","input_tokens":32882,"output_tokens":3973,"usd":0.07912,"stage1_stop_reason":"end_turn"},"stage2":{"model":"claude-opus-4-8","input_tokens":11854,"output_tokens":3587,"usd":0.074472,"stage2_stop_reason":"end_turn"},"total_usd":0.153592,"stage1_batch_id":"msgbatch_017qqVz33Ga63SCDwzb35XPb","stage2_batch_id":"msgbatch_01Scr2Ei9x4HJMiRGxwrmvLK","note":"batch pricing = 50% of standard"},"stage1_raw":"```json\n{\n  \"discoveries\": [\n    {\n      \"year\": 2020,\n      \"finding\": \"TSPAN1 promotes autophagy flux by directly binding to LC3 through two conserved LIR (LC3-interacting region) motifs; mutation of both LIR motifs abolished TSPAN1's ability to induce autophagy and promote pancreatic cancer cell proliferation.\",\n      \"method\": \"Co-immunoprecipitation, GFP-LC3 puncta formation assay, LIR motif mutagenesis, zebrafish tspan1 mutant model\",\n      \"journal\": \"Autophagy\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1–2 / Moderate — direct binding demonstrated by Co-IP, functional consequence confirmed by LIR motif mutagenesis and in vivo zebrafish model; multiple orthogonal methods in one study\",\n      \"pmids\": [\"32972302\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"TSPAN1 expression is transcriptionally upregulated by FAM83A through the canonical WNT-CTNNB1 signaling pathway, with two conserved TCF/LEF binding elements identified in the TSPAN1 promoter region, verified by luciferase activity and ChIP assays.\",\n      \"method\": \"Luciferase reporter assay, ChIP assay\",\n      \"journal\": \"Autophagy\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — two orthogonal methods (luciferase + ChIP) from single lab\",\n      \"pmids\": [\"32972302\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"TSPAN1 upregulates MMP2 expression to promote pancreatic cancer cell migration and invasion via PLCγ; TSPAN1 siRNA suppressed MMP2 mRNA expression by blocking translocation and phosphorylation of PLCγ, while TSPAN1 overexpression had the opposite effect.\",\n      \"method\": \"siRNA knockdown, cDNA overexpression, transwell invasion/migration assay, siRNA targeting PLCγ\",\n      \"journal\": \"Oncology reports\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2–3 / Moderate — epistatic placement of TSPAN1 upstream of PLCγ-MMP2 by genetic knockdown, single lab with multiple functional assays\",\n      \"pmids\": [\"30720116\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2011,\n      \"finding\": \"TSPAN1 physically interacts with the human thiamine transporter-1 (hTHTR-1) and stabilizes it, doubling its protein half-life (from ~6 h to ~12 h) and increasing thiamine uptake in intestinal epithelial cells.\",\n      \"method\": \"Bacterial two-hybrid screen, co-immunoprecipitation, GST pulldown, live-cell confocal imaging, cycloheximide chase assay, functional thiamine uptake assay\",\n      \"journal\": \"American journal of physiology. Gastrointestinal and liver physiology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1–2 / Strong — interaction confirmed by multiple orthogonal methods (bacterial two-hybrid, Co-IP, GST pulldown, colocalization) and functional consequence validated by two independent assays (protein stability + thiamine uptake)\",\n      \"pmids\": [\"21836059\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2017,\n      \"finding\": \"TSPAN1 expression is acutely induced by androgens in prostate cancer cells; stable upregulation of TSPAN1 in DU145 and PC3 cells significantly increased cell migration and induced expression of mesenchymal markers SLUG and ARF6.\",\n      \"method\": \"Stable overexpression, wound-healing/migration assay, western blotting for SLUG and ARF6\",\n      \"journal\": \"Scientific reports\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2–3 / Moderate — gain-of-function with defined downstream markers, androgen regulation confirmed, single lab\",\n      \"pmids\": [\"28701765\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"TSPAN1 transcriptionally regulates ITGA2 expression and epigenetically controls ITGA2 promoter methylation by modulating levels of TET2, DNMT3B and DNMT1, resulting in hypomethylation of the ITGA2 CpG island; knockdown of ITGA2 abolished the pro-proliferative and pro-invasive effects of TSPAN1 overexpression in pancreatic cancer cells.\",\n      \"method\": \"RNA-Seq, western blot, methylation analysis (TET2/DNMT3B/DNMT1 levels), ITGA2 knockdown rescue experiment\",\n      \"journal\": \"American journal of cancer research\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — pathway placement by genetic rescue experiment; epigenetic mechanism supported by multiple markers; single lab\",\n      \"pmids\": [\"32368389\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"ALKBH5 suppresses TSPAN1 expression by demethylating its mRNA (reducing m6A modification); TSPAN1 overexpression reversed the autophagy-inhibiting and anti-proliferative effects of ALKBH5 overexpression in prostate cancer cells, placing TSPAN1 downstream of ALKBH5-mediated m6A regulation.\",\n      \"method\": \"MeRIP-qPCR, western blot (LC3B-II/I ratio, P62), ALKBH5 overexpression, TSPAN1 overexpression rescue\",\n      \"journal\": \"Discovery medicine\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — m6A modification of TSPAN1 mRNA directly measured by MeRIP-qPCR; rescue experiment establishes epistasis; single lab\",\n      \"pmids\": [\"38273746\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"TSPAN1 depletion in HNSCC cells inhibited signaling cascades, with phospho-SRC inhibition identified as a major common target; TSPAN1 depletion also reversed EMT features in cisplatin-resistant tumors in vivo.\",\n      \"method\": \"siRNA knockdown, western blotting for pSRC, in vivo xenograft, co-immunoprecipitation (proteomic study)\",\n      \"journal\": \"Cancers\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2–3 / Moderate — pSRC identified as downstream target by western blot after TSPAN1 depletion, corroborated by IHC correlation; single lab\",\n      \"pmids\": [\"33167355\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"TSPAN1 positively transcriptionally regulates FAM110A expression; TSPAN1 overexpression increased FAM110A levels and the tumor-promoting effects of FAM110A overexpression could be abolished by HIST1H2BK knockdown, defining a TSPAN1/FAM110A/HIST1H2BK/G9a oncogenic pathway in pancreatic cancer.\",\n      \"method\": \"Stable overexpression/knockdown, RNA-Seq, luciferase reporter assay, western blot, rescue experiment\",\n      \"journal\": \"Journal of Cancer\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2–3 / Moderate — transcriptional regulation confirmed by luciferase reporter; pathway epistasis established by rescue experiment; single lab\",\n      \"pmids\": [\"35154458\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"TSPAN1 inhibits NPC cell migration, invasion, and metastasis by interacting with IKBB protein, preventing overactivation of the NF-κB pathway.\",\n      \"method\": \"Co-immunoprecipitation (TSPAN1–IKBB interaction), overexpression/knockdown, in vitro migration/invasion assay, in vivo metastasis assay\",\n      \"journal\": \"Cancer gene therapy\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2–3 / Moderate — binding partner identified by Co-IP; functional consequence in vitro and in vivo; single lab\",\n      \"pmids\": [\"38135697\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"TSPAN1 promotes pancreatic cancer cell growth and invasion by mediating the PI3K/Akt pathway and promoting EMT; TSPAN1 depletion suppressed Akt phosphorylation and EMT markers in breast cancer cells.\",\n      \"method\": \"siRNA knockdown, western blot for PI3K/Akt phosphorylation and EMT markers, in vivo xenograft\",\n      \"journal\": \"Bioengineered\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 / Weak — PI3K/Akt pathway involvement inferred from phosphorylation western blots after knockdown; single lab, single method type\",\n      \"pmids\": [\"34852709\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2016,\n      \"finding\": \"In prostate cancer cell lines, TSPAN1 knockdown resulted in increased Akt phosphorylation and caused cell cycle transition from G1 to S phase, placing TSPAN1 as a negative regulator of the PI3K/Akt pathway; TSPAN1 expression was positively correlated with PTEN.\",\n      \"method\": \"siRNA knockdown, western blot (pAkt), flow cytometry (cell cycle)\",\n      \"journal\": \"Oncotarget\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 / Weak — pathway placement inferred from phosphorylation change after knockdown; single lab, single method type, no direct mechanistic link established\",\n      \"pmids\": [\"27556508\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"TSPAN1 silencing in cerulein-induced pancreatic acinar cell injury inhibited endoplasmic reticulum stress; immunoprecipitation confirmed TSPAN1 directly binds AGR2, and AGR2 overexpression rescued the effects of TSPAN1 silencing on cell proliferation and inflammatory response.\",\n      \"method\": \"Immunoprecipitation (TSPAN1–AGR2 binding), siRNA knockdown, AGR2 overexpression rescue, western blot (ER stress markers), ELISA (cytokines)\",\n      \"journal\": \"Drug development research\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2–3 / Moderate — binding partner confirmed by IP; epistasis validated by rescue experiment; single lab\",\n      \"pmids\": [\"34212407\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2007,\n      \"finding\": \"NET-1 (TSPAN1) protein localizes to the cytoplasm of hepatocellular carcinoma cells as irregular granules near the Golgi apparatus, as determined by fluorescent immunocytochemistry and confocal laser scanning microscopy.\",\n      \"method\": \"Fluorescent immunocytochemistry, confocal laser scanning microscopy\",\n      \"journal\": \"Zhonghua zhong liu za zhi [Chinese journal of oncology]\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 / Weak — direct subcellular localization experiment, single lab, no functional consequence directly linked in this paper\",\n      \"pmids\": [\"18478931\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"TSPAN1 physically interacts with TM4SF1 in glioblastoma stem cells (GSCs), as confirmed by Co-IP and immunofluorescence; 4,5-dimethoxycanthin-6-one inhibited this TSPAN1/TM4SF1 interaction and suppressed GSC self-renewal, and TSPAN1 overexpression partially reversed the drug's inhibitory effects.\",\n      \"method\": \"Co-immunoprecipitation, immunofluorescence, molecular docking, TSPAN1 overexpression rescue, in vivo xenograft\",\n      \"journal\": \"Neurochemical research\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2–3 / Moderate — interaction confirmed by Co-IP and IF; functional relevance supported by rescue experiment; single lab\",\n      \"pmids\": [\"39060768\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"Inhibition of NET-1 (TSPAN1) in HCC cells activated the PI3K/AKT signaling pathway (decreased phosphorylated PI3K and AKT), altered expression of apoptosis-related proteins (decreased Bax and cyclinD1, increased Bcl-2 and caspase-3), and suppressed HCC cell proliferation while promoting apoptosis.\",\n      \"method\": \"siRNA knockdown, western blot (PI3K, pAKT, Bax, Bcl-2, caspase-3, cyclinD1), flow cytometry\",\n      \"journal\": \"Experimental and therapeutic medicine\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 / Weak — pathway assignment based on phosphorylation changes after knockdown; single lab, single method type\",\n      \"pmids\": [\"30867719\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"TSPAN1 expression transcriptionally regulates ITGA2 by modulating TET2, DNMT3B and DNMT1 levels in ovarian cancer cells, resulting in ITGA2 promoter hypomethylation, as confirmed by ChIP assay and luciferase reporter assay.\",\n      \"method\": \"Luciferase reporter assay, ChIP assay, rescue experiment\",\n      \"journal\": \"Stem cells international\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — two orthogonal methods confirm epigenetic regulation; pathway replicated from pancreatic cancer study (PMID 32368389); single lab\",\n      \"pmids\": [\"36187230\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"TSPAN1 is a tetraspanin transmembrane protein that functions as a scaffolding/adaptor protein: it directly binds LC3 via two LIR motifs to promote autophagy flux, interacts with the thiamine transporter hTHTR-1 to stabilize it, binds IKBB to suppress NF-κB signaling, binds AGR2 to induce ER stress, and associates with TM4SF1 in glioblastoma stem cells; in cancer contexts it also promotes cell migration and invasion by activating PLCγ–MMP2 signaling, epigenetically upregulates ITGA2 through modulation of TET2/DNMT3B/DNMT1, and its transcription is driven by WNT-CTNNB1 signaling via TCF/LEF elements and regulated post-transcriptionally by m6A demethylation through ALKBH5.\"\n}\n```","stage2_raw":"{\n  \"mechanistic_narrative\": \"TSPAN1 is a tetraspanin transmembrane protein that acts as a membrane-associated scaffold linking it to autophagy, transporter stabilization, and oncogenic signaling [#0, #3]. Its best-defined molecular activity is in autophagy: TSPAN1 directly binds LC3 through two conserved LIR motifs, and mutation of both motifs abolishes its ability to induce autophagy and drive pancreatic cancer cell proliferation [#0]. Beyond autophagy, TSPAN1 stabilizes the thiamine transporter hTHTR-1, roughly doubling its protein half-life and increasing thiamine uptake in intestinal epithelium [#3]. In cancer it functions largely as a signaling adaptor: it promotes migration and invasion by activating PLC\\u03b3 to upregulate MMP2 [#2], epigenetically upregulates ITGA2 by modulating TET2/DNMT3B/DNMT1 to hypomethylate the ITGA2 promoter, an axis required for its pro-proliferative and pro-invasive effects [#5, #16], and transcriptionally activates a FAM110A/HIST1H2BK/G9a oncogenic pathway [#8]. TSPAN1 also engages specific protein partners with context-dependent outputs, binding IKBB to restrain NF-\\u03baB signaling [#9], binding AGR2 to drive ER stress [#12], and associating with TM4SF1 to support glioblastoma stem cell self-renewal [#14]. Upstream, TSPAN1 transcription is driven by WNT-CTNNB1 signaling via TCF/LEF elements in its promoter [#1] and its mRNA is suppressed by ALKBH5-mediated m6A demethylation [#6].\"\n,\n  \"teleology\": [\n    {\n      \"year\": 2007,\n      \"claim\": \"Established the first subcellular localization of TSPAN1, placing it in the cytoplasm near the Golgi rather than diffusely, providing a structural starting point for its membrane scaffolding role.\",\n      \"evidence\": \"Fluorescent immunocytochemistry and confocal microscopy in hepatocellular carcinoma cells\",\n      \"pmids\": [\"18478931\"],\n      \"confidence\": \"Low\",\n      \"gaps\": [\"Single lab descriptive localization with no functional consequence linked\", \"Does not establish plasma-membrane versus organelle topology\"]\n    },\n    {\n      \"year\": 2011,\n      \"claim\": \"Answered whether TSPAN1 has a physiological binding partner by showing it directly interacts with and stabilizes the thiamine transporter hTHTR-1, establishing a non-cancer scaffolding/chaperone function.\",\n      \"evidence\": \"Bacterial two-hybrid, Co-IP, GST pulldown, cycloheximide chase, and thiamine uptake assays in intestinal epithelial cells\",\n      \"pmids\": [\"21836059\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Structural basis of the interaction not resolved\", \"Whether stabilization is general to other membrane partners untested\"]\n    },\n    {\n      \"year\": 2016,\n      \"claim\": \"Began placing TSPAN1 in growth signaling by linking its loss to increased Akt phosphorylation and G1/S transition in prostate cancer, framing it as a possible negative PI3K/Akt regulator.\",\n      \"evidence\": \"siRNA knockdown, pAkt western blot, and flow cytometry cell cycle analysis\",\n      \"pmids\": [\"27556508\"],\n      \"confidence\": \"Low\",\n      \"gaps\": [\"Pathway placement inferred from phosphorylation changes only\", \"No direct mechanistic link to PI3K/Akt components\", \"Direction conflicts with later contexts\"]\n    },\n    {\n      \"year\": 2017,\n      \"claim\": \"Connected TSPAN1 to hormone-driven tumor biology by showing androgens acutely induce it and that it drives migration and mesenchymal marker expression.\",\n      \"evidence\": \"Stable overexpression, wound-healing assay, and western blot for SLUG and ARF6 in prostate cancer lines\",\n      \"pmids\": [\"28701765\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Mechanism linking TSPAN1 to SLUG/ARF6 not defined\", \"Single lab gain-of-function\"]\n    },\n    {\n      \"year\": 2019,\n      \"claim\": \"Defined a concrete pro-invasive mechanism by placing TSPAN1 upstream of PLC\\u03b3 phosphorylation and MMP2 induction.\",\n      \"evidence\": \"siRNA knockdown/overexpression, transwell assays, and PLC\\u03b3 siRNA epistasis in pancreatic cancer\",\n      \"pmids\": [\"30720116\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"How TSPAN1 triggers PLC\\u03b3 translocation/phosphorylation unknown\", \"No direct TSPAN1\\u2013PLC\\u03b3 binding shown\"]\n    },\n    {\n      \"year\": 2020,\n      \"claim\": \"Identified TSPAN1's core molecular activity \\u2014 direct LIR-dependent LC3 binding driving autophagy flux \\u2014 and showed WNT-CTNNB1 signaling transcriptionally controls its expression.\",\n      \"evidence\": \"Co-IP, GFP-LC3 puncta, LIR mutagenesis, zebrafish mutant, luciferase reporter and ChIP\",\n      \"pmids\": [\"32972302\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"How a transmembrane tetraspanin presents LIR motifs to cytosolic LC3 not structurally resolved\", \"FAM83A/WNT regulation from single lab\"]\n    },\n    {\n      \"year\": 2020,\n      \"claim\": \"Established an epigenetic effector arm by showing TSPAN1 controls ITGA2 promoter methylation via TET2/DNMT3B/DNMT1 and that ITGA2 is required for TSPAN1's oncogenic effects.\",\n      \"evidence\": \"RNA-Seq, methylation analysis, and ITGA2 knockdown rescue in pancreatic cancer (replicated in ovarian cancer, 2022)\",\n      \"pmids\": [\"32368389\", \"36187230\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"How a membrane protein modulates nuclear TET/DNMT levels mechanistically unexplained\", \"Direct versus indirect regulation unresolved\"]\n    },\n    {\n      \"year\": 2021,\n      \"claim\": \"Expanded TSPAN1's partner repertoire and signaling reach by identifying AGR2 binding driving ER stress, and implicating SRC and PI3K/Akt/EMT cascades downstream.\",\n      \"evidence\": \"Co-IP/IP, AGR2 rescue, pSRC and pAkt/EMT western blots, and xenografts across pancreatic, HNSCC and breast models\",\n      \"pmids\": [\"34212407\", \"33167355\", \"34852709\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"PI3K/Akt placement inferred from phosphorylation only (Low-confidence)\", \"Conflicting positive versus negative effects on Akt across contexts unreconciled\"]\n    },\n    {\n      \"year\": 2023,\n      \"claim\": \"Demonstrated a tumor-suppressive partner interaction by showing TSPAN1 binds IKBB to restrain NF-\\u03baB and inhibit nasopharyngeal carcinoma metastasis, indicating context-dependent function.\",\n      \"evidence\": \"Co-IP, overexpression/knockdown, and in vitro/in vivo metastasis assays\",\n      \"pmids\": [\"38135697\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Reciprocal validation of TSPAN1\\u2013IKBB binding limited\", \"Reconciliation with pro-invasive roles in other cancers not addressed\"]\n    },\n    {\n      \"year\": 2024,\n      \"claim\": \"Added an upstream m6A regulatory layer (ALKBH5 demethylation suppresses TSPAN1) and a stem-cell partner (TM4SF1 in glioblastoma), broadening TSPAN1's regulatory and interaction map.\",\n      \"evidence\": \"MeRIP-qPCR with rescue, and Co-IP/IF with docking and rescue in glioblastoma stem cells\",\n      \"pmids\": [\"38273746\", \"39060768\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"m6A reader linking demethylation to TSPAN1 stability/translation unidentified\", \"Functional consequence of TSPAN1\\u2013TM4SF1 complex on signaling undefined\"]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"It remains unknown how a single tetraspanin reconciles its opposing tumor-promoting and tumor-suppressive roles, and what structural feature of TSPAN1 coordinates membrane partners with cytosolic autophagy and nuclear epigenetic outputs.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"Low\",\n      \"gaps\": [\"No structural model of TSPAN1 in its complexes\", \"No unifying mechanism explaining context-dependent oncogenic versus suppressive activity\", \"Membrane-to-nucleus signaling route undefined\"]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0060090\", \"supporting_discovery_ids\": [0, 3, 9, 12, 14]},\n      {\"term_id\": \"GO:0140110\", \"supporting_discovery_ids\": [5, 8, 16]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005829\", \"supporting_discovery_ids\": [13]},\n      {\"term_id\": \"GO:0005794\", \"supporting_discovery_ids\": [13]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-9612973\", \"supporting_discovery_ids\": [0, 6]},\n      {\"term_id\": \"R-HSA-162582\", \"supporting_discovery_ids\": [2, 9]},\n      {\"term_id\": \"R-HSA-4839726\", \"supporting_discovery_ids\": [5, 16]}\n    ],\n    \"complexes\": [],\n    \"partners\": [\"LC3\", \"hTHTR-1\", \"IKBB\", \"AGR2\", \"TM4SF1\"],\n    \"other_free_text\": []\n  }\n}","audit_flag":null,"evaluation":{"pairwise":"win","faith_supported":6,"faith_total":6,"faith_pct":100.0}}