{"gene":"TMED3","run_date":"2026-06-10T10:51:55","timeline":{"discoveries":[{"year":2014,"finding":"TMED3 knockdown in human colon cancer cells reduces endogenous WNT-TCF signaling activity, identifying TMED3 as a positive modulator of the WNT-TCF pathway; in vivo genome-wide RNAi screen showed TMED3 knockdown increases metastatic growth and induces lung metastases from subcutaneous xenografts, establishing TMED3 as a metastatic suppressor functioning through WNT-TCF promotion.","method":"Genome-wide in vivo RNAi screen in primary human tumor cells in mice; WNT-TCF reporter assays; xenograft models","journal":"EMBO molecular medicine","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — in vivo functional screen with multiple cancer cell contexts and pathway reporter assays, single lab","pmids":["24920608"],"is_preprint":false},{"year":2019,"finding":"TMED3 knockdown leads to enhanced expression of TMED9; TMED9 acts downstream of TMED3 and opposes TMED3-WNT-TCF signaling. TMED9 promotes colon cancer metastasis via TGFα secretion and CNIH4 (a TGFα exporter), and TMED9 knockdown compromises TGFα biogenesis. TMED9/TMED3 antagonism modulates both WNT-TCF and GLI signaling, establishing a TMED3→TMED9→CNIH4/TGFα/GLI pro-metastatic axis opposing TMED3-WNT-TCF.","method":"RNAi knockdown, transcriptional rescue assays, gene expression analysis in three colon cancer cell lines, functional migration assays, in vivo xenograft models","journal":"Oncogene","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — epistatic ordering by functional rescue, multiple colon cancer cell contexts, single lab","pmids":["31253868"],"is_preprint":false},{"year":2016,"finding":"TMED3 promotes hepatocellular carcinoma (HCC) metastasis through IL-11/STAT3 signaling; gene microarray of TMED3-knockdown cells revealed decreased IL-11 expression, and TMED3 overexpression promoted cell migration and invasion, which was suppressed by TMED3 knockdown both in vitro and in vivo.","method":"Gene microarray analysis of TMED3-knockdown HCC cells; overexpression and knockdown functional assays; in vivo metastasis models","journal":"Scientific reports","confidence":"Low","confidence_rationale":"Tier 3 / Weak — single lab, IL-11/STAT3 pathway link inferred from microarray without direct mechanistic confirmation in the abstract","pmids":["27901021"],"is_preprint":false},{"year":2022,"finding":"TMED3 recognizes ER core-glycosylated transmembrane protein cargos and, as part of a heteromeric TMED2/3/9/10 complex, mediates ER stress-associated unconventional protein secretion (UPS) of transmembrane proteins including CFTR, pendrin, and SARS-CoV-2 Spike via a Golgi-independent pathway. Co-expression of all four TMEDs improves UPS, while individual silencing reduces it.","method":"Gene silencing (siRNA) of individual and combined TMED family members; functional ion transport assays for ΔF508-CFTR and p.H723R-pendrin; SARS-CoV-2 viral release assay; mechanistic co-expression studies","journal":"Advanced science (Weinheim, Baden-Wurttemberg, Germany)","confidence":"High","confidence_rationale":"Tier 1–2 / Strong — multiple orthogonal functional assays (ion transport, viral release, cargo recognition), heteromeric complex defined, multiple cargo proteins tested, single lab with rigorous controls","pmids":["35748162"],"is_preprint":false},{"year":2019,"finding":"miR-876-3p directly targets TMED3 (confirmed by luciferase reporter and biotin-miRNA pulldown assay); TMED3 siRNA mimics miR-876-3p's inhibitory effects on cisplatin resistance and stem cell-like properties (Sox-2, Oct-4, CD133, CD44 expression) in gastric cancer cells, placing TMED3 downstream of miR-876-3p in regulating drug resistance.","method":"TargetScan prediction; luciferase reporter assay; biotin-miRNA pulldown assay; siRNA knockdown functional rescue","journal":"Journal of gastroenterology and hepatology","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — direct target validated by two orthogonal methods (luciferase + pulldown), functional rescue confirms pathway placement, single lab","pmids":["30843262"],"is_preprint":false},{"year":2021,"finding":"TMED3 promotes lung squamous cell carcinoma (LUSC) progression by regulating EZR (Ezrin); RNA sequencing and IPA identified EZR as a downstream target; EZR inhibition suppresses LUSC progression and reduces the promoting effects of TMED3 overexpression, and TMED3 knockdown inhibits cell migration through regulation of EMT.","method":"RNA sequencing; Ingenuity Pathway Analysis; loss-of-function assays; EZR overexpression rescue experiments; xenograft mouse model","journal":"Cell death & disease","confidence":"Low","confidence_rationale":"Tier 3 / Weak — downstream target identified by transcriptomic screen and partial rescue, single lab, limited direct mechanistic validation","pmids":["34429402"],"is_preprint":false},{"year":2021,"finding":"TMED3 promotes osteosarcoma progression via RPS15A (ribosomal protein S15A) as a downstream target; simultaneous knockdown of both TMED3 and RPS15A intensified inhibitory effects, and RPS15A knockdown reversed the pro-tumorigenic effects of TMED3 overexpression.","method":"Gene expression profile analysis; loss-of-function assays; epistatic rescue by RPS15A knockdown/TMED3 overexpression combination; xenograft tumor model","journal":"Cancer cell international","confidence":"Low","confidence_rationale":"Tier 3 / Weak — epistatic rescue with single downstream target, single lab, pathway mechanism not directly defined","pmids":["34838013"],"is_preprint":false},{"year":2020,"finding":"TMED3 promotes proliferation and migration of breast cancer cells through activation of Wnt/β-catenin signaling; immunofluorescence and Western blot showed TMED3 modulates expression of Wnt/β-catenin pathway proteins and cell cycle/migration-related proteins.","method":"Overexpression and knockdown functional assays; Western blot and immunofluorescence of Wnt/β-catenin pathway components; cell cycle analysis; in vitro proliferation/migration/invasion assays","journal":"OncoTargets and therapy","confidence":"Low","confidence_rationale":"Tier 3 / Weak — single lab, pathway association by protein expression without direct mechanistic dissection","pmids":["32606792"],"is_preprint":false},{"year":2021,"finding":"TMED3 knockdown in chordoma cells reduces expression of Bcl-2, HSP27, IGF-I, IGF-II, IGFBP-2, Livin, Akt, CDK6, and cyclin D1, while upregulating MAPK9; these changes accompany inhibition of cell viability and migration and enhanced apoptosis, linking TMED3 to pro-survival signaling nodes.","method":"Apoptosis antibody array; Western blot; RT-qPCR; in vitro functional assays; xenograft model","journal":"International journal of oncology","confidence":"Low","confidence_rationale":"Tier 3 / Weak — protein array profiling without direct mechanistic pathway validation, single lab","pmids":["33760171"],"is_preprint":false},{"year":2021,"finding":"In breast cancer, the lncRNA RP11-283G6.5 acts upstream of miR-188-3p/TMED3/Wnt/β-catenin signaling; overexpression of TMED3 or inhibition of miR-188-3p rescued the suppressive effects of RP11-283G6.5, placing TMED3 as a direct downstream effector of miR-188-3p and upstream of Wnt/β-catenin signaling in this axis.","method":"Luciferase reporter assay; RNA pulldown; functional rescue experiments (miR-188-3p inhibition, TMED3 overexpression); in vivo xenograft","journal":"RNA biology","confidence":"Low","confidence_rationale":"Tier 3 / Weak — TMED3 mechanistic placement is secondary to lncRNA study; functional rescue validates pathway order but direct TMED3 mechanism not primary focus, single lab","pmids":["34130584"],"is_preprint":false},{"year":2021,"finding":"TMED3 promotes NSCLC progression by enhancing Wnt/β-catenin signaling through modulation of AKT; TMED3 silencing reduced AKT activity and Wnt/β-catenin pathway activity, AKT inhibition blocked TMED3-overexpression-mediated Wnt/β-catenin enhancement, and Wnt/β-catenin inhibition reversed TMED3 overexpression effects on proliferation/invasion.","method":"Knockdown and overexpression; AKT inhibitor treatment; Wnt/β-catenin inhibitor treatment; epistatic rescue experiments; in vivo xenograft","journal":"Toxicology and applied pharmacology","confidence":"Low","confidence_rationale":"Tier 3 / Weak — epistatic ordering via inhibitors, single lab, no direct biochemical interaction demonstrated","pmids":["34758370"],"is_preprint":false},{"year":2023,"finding":"TMED3 interacts with CDCA8 (Cell division cycle associated 8) in melanoma cells; CDCA8 overexpression reversed the inhibitory effects of TMED3 knockdown on cell viability and motility, and TMED3 knockdown decreased P-AKT and P-PI3K levels which were rescued by CDCA8 overexpression, placing TMED3 upstream of CDCA8 and PI3K/AKT signaling in melanoma.","method":"Co-immunoprecipitation or pulldown (interaction); CDCA8 knockdown/overexpression rescue; SC79 (AKT activator) rescue; Western blot for PI3K/AKT phosphorylation; in vitro and in vivo functional assays","journal":"Cell & bioscience","confidence":"Low","confidence_rationale":"Tier 3 / Weak — interaction and epistatic rescue reported, but mechanistic depth limited by abstract description, single lab","pmids":["36991473"],"is_preprint":false},{"year":2024,"finding":"TMED3 promotes prostate cancer progression via phosphorylation of FOXO1a and FOXO3a; TMED3 inhibition decreased FOXO1a and FOXO3a phosphorylation both in vitro and in vivo, linking TMED3 activity to FOXO pathway suppression.","method":"shRNA knockdown; KEGG pathway enrichment analysis; Western blot for FOXO1a/FOXO3a phosphorylation; in vivo prostate cancer mouse model","journal":"Oncology research","confidence":"Low","confidence_rationale":"Tier 3 / Weak — phosphorylation change observed upon TMED3 knockdown but direct biochemical mechanism of how TMED3 regulates FOXO phosphorylation not established, single lab","pmids":["39735675"],"is_preprint":false},{"year":2024,"finding":"TMED3 stabilizes SMAD2 by inhibiting NEDD4 E3 ligase-mediated ubiquitination of SMAD2 in ovarian cancer; SMAD2 knockdown reversed TMED3 overexpression effects on cancer cells; PI3K-AKT pathway was also involved in TMED3-mediated ovarian cancer promotion.","method":"shRNA knockdown; functional rescue experiments (SMAD2 knockdown in TMED3-overexpressing cells); ubiquitination assay (implied); Western blot; subcutaneous xenograft model","journal":"Molecular carcinogenesis","confidence":"Medium","confidence_rationale":"Tier 2 / Weak — mechanistic claim (NEDD4-mediated ubiquitination counteracted by TMED3) with functional rescue validation, single lab, abstract does not detail ubiquitination assay methods explicitly","pmids":["38411267"],"is_preprint":false},{"year":2022,"finding":"TMED3 knockdown in esophageal squamous cell carcinoma cells alters gene expression profile; FAM60A was identified as a functionally related downstream gene, and FAM60A knockdown reversed TMED3's pro-tumorigenic effects, placing FAM60A downstream of TMED3.","method":"Affymetrix microarray; IPA; FAM60A knockdown rescue experiments; in vitro and in vivo functional assays","journal":"Biomedical journal","confidence":"Low","confidence_rationale":"Tier 3 / Weak — downstream target identified by transcriptomics with partial rescue, single lab, limited mechanistic depth","pmids":["35358714"],"is_preprint":false}],"current_model":"TMED3 is a p24 family ER-to-Golgi trafficking protein that functions as part of a heteromeric TMED2/3/9/10 complex to recognize ER core-glycosylated transmembrane cargo proteins and mediate their ER stress-associated unconventional (Golgi-independent) secretion; in cancer contexts, TMED3 promotes WNT-TCF signaling to suppress colon cancer metastasis (while its antagonist TMED9 drives a pro-metastatic TGFα/GLI axis), and in other tumor types TMED3 activates oncogenic signaling through Wnt/β-catenin, PI3K/AKT, IL-11/STAT3, and FOXO pathways, and stabilizes SMAD2 by counteracting NEDD4-mediated ubiquitination."},"narrative":{"mechanistic_narrative":"TMED3 is a p24-family endoplasmic reticulum-to-Golgi cargo protein that recognizes ER core-glycosylated transmembrane cargos and, as a subunit of a heteromeric TMED2/3/9/10 complex, mediates ER stress-associated unconventional (Golgi-independent) secretion of transmembrane proteins including CFTR, pendrin, and SARS-CoV-2 Spike; co-expression of all four TMEDs improves this secretion while individual silencing reduces it [PMID:35748162]. Beyond this trafficking role, TMED3 acts as a context-dependent modulator of oncogenic signaling. In colon cancer it positively modulates WNT-TCF signaling and behaves as a metastatic suppressor, with its knockdown increasing metastatic growth [PMID:24920608]; this WNT-TCF–promoting activity is opposed by TMED9, which is upregulated upon TMED3 loss and drives a pro-metastatic CNIH4/TGFα/GLI axis, defining a TMED3→TMED9 antagonism that balances WNT-TCF and GLI signaling [PMID:31253868]. In ovarian cancer, TMED3 stabilizes SMAD2 by counteracting NEDD4 E3 ligase-mediated ubiquitination [PMID:38411267]. The literature places TMED3 upstream of multiple downstream effectors, but most of these connections rest on lower-confidence transcriptomic and epistatic studies in individual tumor types; the precise biochemical mechanism by which TMED3 engages these signaling pathways has not been resolved in the available corpus.","teleology":[{"year":2014,"claim":"Established that TMED3 is a functional modulator of WNT-TCF signaling and, unexpectedly, a suppressor of metastasis rather than a generic oncogene.","evidence":"Genome-wide in vivo RNAi screen in primary human colon tumor cells with WNT-TCF reporter assays and xenografts","pmids":["24920608"],"confidence":"Medium","gaps":["No direct biochemical mechanism linking TMED3 to WNT-TCF components","Metastatic suppression shown in xenografts but not in autochthonous models"]},{"year":2016,"claim":"Extended TMED3 function to hepatocellular carcinoma, where it appeared to act as a pro-metastatic factor via IL-11/STAT3 signaling, hinting at tissue-context-dependent roles.","evidence":"Gene microarray of TMED3-knockdown HCC cells with overexpression/knockdown functional assays and in vivo metastasis models","pmids":["27901021"],"confidence":"Low","gaps":["IL-11/STAT3 link inferred from microarray without direct mechanistic confirmation","Opposite directionality from colon cancer not reconciled mechanistically"]},{"year":2019,"claim":"Resolved the epistatic relationship between TMED3 and its paralog TMED9, showing antagonistic control of WNT-TCF versus a CNIH4/TGFα/GLI pro-metastatic axis.","evidence":"RNAi knockdown with transcriptional rescue and migration assays in three colon cancer lines plus xenografts","pmids":["31253868"],"confidence":"Medium","gaps":["Molecular basis of TMED3-TMED9 antagonism not defined","How TMED3 loss upregulates TMED9 is unknown"]},{"year":2019,"claim":"Placed TMED3 downstream of miR-876-3p in controlling chemoresistance and stem-like properties, establishing a regulatory input on TMED3 expression.","evidence":"Luciferase reporter and biotin-miRNA pulldown confirming direct targeting, with siRNA functional rescue in gastric cancer cells","pmids":["30843262"],"confidence":"Medium","gaps":["Effector mechanism downstream of TMED3 in this axis not defined","Single cancer-type context"]},{"year":2020,"claim":"Associated TMED3 with Wnt/β-catenin pathway activation in breast cancer, broadening its signaling connections beyond colon WNT-TCF.","evidence":"Overexpression/knockdown assays with Western blot and immunofluorescence of Wnt/β-catenin components","pmids":["32606792"],"confidence":"Low","gaps":["Pathway association by protein expression without direct mechanistic dissection","No demonstrated physical interaction"]},{"year":2021,"claim":"A series of tumor-specific studies positioned TMED3 upstream of diverse downstream effectors (EZR, RPS15A, AKT, pro-survival nodes, lncRNA/miR axes) across multiple cancers.","evidence":"Transcriptomic profiling, apoptosis/protein arrays, and epistatic rescue across LUSC, osteosarcoma, NSCLC, chordoma, and breast cancer with xenografts","pmids":["34429402","34838013","34758370","33760171","34130584"],"confidence":"Low","gaps":["Downstream targets identified largely by transcriptomic screens with partial rescue","No direct biochemical interactions established for most effectors","Pathway mechanisms inferred via inhibitors rather than reconstitution"]},{"year":2022,"claim":"Defined the core molecular function of TMED3: recognition of ER core-glycosylated transmembrane cargos and assembly into a TMED2/3/9/10 complex driving Golgi-independent unconventional secretion.","evidence":"Combinatorial siRNA silencing, ΔF508-CFTR and pendrin ion-transport assays, SARS-CoV-2 release assay, and co-expression studies","pmids":["35748162"],"confidence":"High","gaps":["Structural basis of cargo recognition not resolved","Stoichiometry and assembly order of the TMED2/3/9/10 complex unknown","Link between this trafficking function and cancer signaling roles not established"]},{"year":2022,"claim":"Identified FAM60A as a downstream gene mediating TMED3 pro-tumorigenic effects in esophageal squamous cell carcinoma.","evidence":"Affymetrix microarray and IPA with FAM60A knockdown rescue in vitro and in vivo","pmids":["35358714"],"confidence":"Low","gaps":["Downstream target identified by transcriptomics with partial rescue","No direct mechanistic link from TMED3 to FAM60A"]},{"year":2023,"claim":"Reported a physical interaction between TMED3 and CDCA8 placing TMED3 upstream of PI3K/AKT signaling in melanoma.","evidence":"Interaction assay with CDCA8 and AKT-activator (SC79) rescue plus Western blot for PI3K/AKT phosphorylation","pmids":["36991473"],"confidence":"Low","gaps":["Single Co-IP/pulldown without reciprocal validation","Mechanism by which the interaction modulates PI3K/AKT not defined"]},{"year":2024,"claim":"Provided a more concrete biochemical mechanism: TMED3 stabilizes SMAD2 by antagonizing NEDD4-mediated ubiquitination in ovarian cancer.","evidence":"shRNA knockdown, SMAD2 functional rescue, and ubiquitination assays with xenografts","pmids":["38411267"],"confidence":"Medium","gaps":["Ubiquitination assay methodology not detailed","Whether TMED3 directly binds SMAD2 or NEDD4 not shown"]},{"year":2024,"claim":"Linked TMED3 activity to FOXO1a/FOXO3a phosphorylation in prostate cancer, extending its signaling footprint.","evidence":"shRNA knockdown with KEGG enrichment and Western blot for FOXO phosphorylation in vitro and in vivo","pmids":["39735675"],"confidence":"Low","gaps":["Direct biochemical mechanism linking TMED3 to FOXO phosphorylation not established","No identified kinase intermediary"]},{"year":null,"claim":"How TMED3's core ER-to-Golgi cargo-trafficking and unconventional-secretion function mechanistically connects to its diverse, often opposing, roles in cancer signaling remains unresolved.","evidence":"","pmids":[],"confidence":"Low","gaps":["No unifying model reconciling metastatic-suppressor versus pro-tumorigenic phenotypes across tissues","Whether cancer signaling effects derive from cargo trafficking is untested","Structural and stoichiometric details of the TMED2/3/9/10 complex unknown"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0038024","term_label":"cargo receptor activity","supporting_discovery_ids":[3]}],"localization":[{"term_id":"GO:0005783","term_label":"endoplasmic reticulum","supporting_discovery_ids":[3]},{"term_id":"GO:0005794","term_label":"Golgi apparatus","supporting_discovery_ids":[3]}],"pathway":[{"term_id":"R-HSA-5653656","term_label":"Vesicle-mediated transport","supporting_discovery_ids":[3]},{"term_id":"R-HSA-9609507","term_label":"Protein localization","supporting_discovery_ids":[3]}],"complexes":["TMED2/3/9/10 complex"],"partners":["TMED9","TMED2","TMED10","SMAD2","NEDD4","CDCA8"],"other_free_text":[]}},"prefetch_data":{"uniprot":{"accession":"Q9Y3Q3","full_name":"Transmembrane emp24 domain-containing protein 3","aliases":["Membrane protein p24B","p24 family protein gamma-4","p24gamma4","p26"],"length_aa":217,"mass_kda":24.8,"function":"Potential role in vesicular protein trafficking, mainly in the early secretory pathway. Contributes to the coupled localization of TMED2 and TMED10 in the cis-Golgi network","subcellular_location":"Endoplasmic reticulum-Golgi intermediate compartment membrane; Golgi apparatus, cis-Golgi network membrane; Golgi apparatus, Golgi stack membrane; Endoplasmic reticulum membrane; Cytoplasmic vesicle, COPI-coated vesicle membrane","url":"https://www.uniprot.org/uniprotkb/Q9Y3Q3/entry"},"depmap":{"release":"DepMap","has_data":true,"is_common_essential":false,"resolved_as":"","url":"https://depmap.org/portal/gene/TMED3","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":[{"gene":"MYBBP1A","stoichiometry":4.0},{"gene":"TMED10","stoichiometry":4.0},{"gene":"PGRMC1","stoichiometry":0.2}],"url":"https://opencell.sf.czbiohub.org/search/TMED3","total_profiled":1310},"omim":[{"mim_id":"620437","title":"TRANSMEMBRANE p24 TRAFFICKING PROTEIN 3; TMED3","url":"https://www.omim.org/entry/620437"},{"mim_id":"620436","title":"TRANSMEMBRANE p24 TRAFFICKING PROTEIN 9; TMED9","url":"https://www.omim.org/entry/620436"},{"mim_id":"619642","title":"TRANSMEMBRANE p24 TRAFFICKING PROTEIN 2; TMED2","url":"https://www.omim.org/entry/619642"},{"mim_id":"605406","title":"TRANSMEMBRANE p24 TRAFFICKING PROTEIN 10; TMED10","url":"https://www.omim.org/entry/605406"}],"hpa":{"profiled":true,"resolved_as":"","reliability":"Supported","locations":[{"location":"Golgi apparatus","reliability":"Supported"}],"tissue_specificity":"Tissue enhanced","tissue_distribution":"Detected in all","driving_tissues":[{"tissue":"salivary gland","ntpm":386.0}],"url":"https://www.proteinatlas.org/search/TMED3"},"hgnc":{"alias_symbol":["p24B","p24gamma4","p24g4"],"prev_symbol":["C15orf22"]},"alphafold":{"accession":"Q9Y3Q3","domains":[{"cath_id":"2.60.120.380","chopping":"22-124","consensus_level":"medium","plddt":92.9811,"start":22,"end":124},{"cath_id":"1.20.5","chopping":"142-206","consensus_level":"medium","plddt":94.2348,"start":142,"end":206}],"viewer_url":"https://alphafold.ebi.ac.uk/entry/Q9Y3Q3","model_url":"https://alphafold.ebi.ac.uk/files/AF-Q9Y3Q3-F1-model_v6.cif","pae_url":"https://alphafold.ebi.ac.uk/files/AF-Q9Y3Q3-F1-predicted_aligned_error_v6.png","plddt_mean":88.0},"mouse_models":{"mgi_url":"https://www.informatics.jax.org/marker/summary?nomen=TMED3","jax_strain_url":"https://www.jax.org/strain/search?query=TMED3"},"sequence":{"accession":"Q9Y3Q3","fasta_url":"https://rest.uniprot.org/uniprotkb/Q9Y3Q3.fasta","uniprot_url":"https://www.uniprot.org/uniprotkb/Q9Y3Q3/entry","alphafold_viewer_url":"https://alphafold.ebi.ac.uk/entry/Q9Y3Q3"}},"corpus_meta":[{"pmid":"24920608","id":"PMC_24920608","title":"A novel genome-wide in vivo screen for metastatic suppressors in human colon cancer identifies the positive WNT-TCF pathway modulators TMED3 and SOX12.","date":"2014","source":"EMBO molecular medicine","url":"https://pubmed.ncbi.nlm.nih.gov/24920608","citation_count":73,"is_preprint":false},{"pmid":"27901021","id":"PMC_27901021","title":"TMED3 promotes hepatocellular carcinoma progression via IL-11/STAT3 signaling.","date":"2016","source":"Scientific reports","url":"https://pubmed.ncbi.nlm.nih.gov/27901021","citation_count":58,"is_preprint":false},{"pmid":"22761906","id":"PMC_22761906","title":"High-throughput transcriptomic and RNAi analysis identifies AIM1, ERGIC1, TMED3 and TPX2 as potential drug targets in prostate cancer.","date":"2012","source":"PloS one","url":"https://pubmed.ncbi.nlm.nih.gov/22761906","citation_count":56,"is_preprint":false},{"pmid":"30843262","id":"PMC_30843262","title":"MiR-876-3p regulates cisplatin resistance and stem cell-like properties of gastric cancer cells by targeting TMED3.","date":"2019","source":"Journal of gastroenterology and hepatology","url":"https://pubmed.ncbi.nlm.nih.gov/30843262","citation_count":49,"is_preprint":false},{"pmid":"31253868","id":"PMC_31253868","title":"The protein secretion modulator TMED9 drives CNIH4/TGFα/GLI signaling opposing TMED3-WNT-TCF to promote colon cancer metastases.","date":"2019","source":"Oncogene","url":"https://pubmed.ncbi.nlm.nih.gov/31253868","citation_count":49,"is_preprint":false},{"pmid":"30976199","id":"PMC_30976199","title":"TMED3 promotes cell proliferation and motility in breast cancer and is negatively modulated by miR-188-3p.","date":"2019","source":"Cancer cell international","url":"https://pubmed.ncbi.nlm.nih.gov/30976199","citation_count":39,"is_preprint":false},{"pmid":"32606792","id":"PMC_32606792","title":"TMED3 Promotes Proliferation and Migration in Breast Cancer Cells by Activating Wnt/β-Catenin Signaling.","date":"2020","source":"OncoTargets and therapy","url":"https://pubmed.ncbi.nlm.nih.gov/32606792","citation_count":26,"is_preprint":false},{"pmid":"35748162","id":"PMC_35748162","title":"TMED3 Complex Mediates ER Stress-Associated Secretion of CFTR, Pendrin, and SARS-CoV-2 Spike.","date":"2022","source":"Advanced science (Weinheim, Baden-Wurttemberg, Germany)","url":"https://pubmed.ncbi.nlm.nih.gov/35748162","citation_count":22,"is_preprint":false},{"pmid":"34429402","id":"PMC_34429402","title":"TMED3 promotes the progression and development of lung squamous cell carcinoma by regulating EZR.","date":"2021","source":"Cell death & disease","url":"https://pubmed.ncbi.nlm.nih.gov/34429402","citation_count":17,"is_preprint":false},{"pmid":"33760171","id":"PMC_33760171","title":"Knockdown of TMED3 inhibits cell viability and migration and increases apoptosis in human chordoma cells.","date":"2021","source":"International journal of oncology","url":"https://pubmed.ncbi.nlm.nih.gov/33760171","citation_count":15,"is_preprint":false},{"pmid":"34838013","id":"PMC_34838013","title":"TMED3/RPS15A Axis promotes the development and progression of osteosarcoma.","date":"2021","source":"Cancer cell international","url":"https://pubmed.ncbi.nlm.nih.gov/34838013","citation_count":15,"is_preprint":false},{"pmid":"34130584","id":"PMC_34130584","title":"Long non-coding RNA RP11-283G6.5 confines breast cancer development through modulating miR-188-3p/TMED3/Wnt/β-catenin signalling.","date":"2021","source":"RNA biology","url":"https://pubmed.ncbi.nlm.nih.gov/34130584","citation_count":15,"is_preprint":false},{"pmid":"34758370","id":"PMC_34758370","title":"TMED3 exerts a protumor function in non-small cell lung cancer by enhancing the Wnt/β-catenin pathway via regulation of AKT.","date":"2021","source":"Toxicology and applied pharmacology","url":"https://pubmed.ncbi.nlm.nih.gov/34758370","citation_count":10,"is_preprint":false},{"pmid":"36991473","id":"PMC_36991473","title":"TMED3 promotes the development of malignant melanoma by targeting CDCA8 and regulating PI3K/Akt pathway.","date":"2023","source":"Cell & bioscience","url":"https://pubmed.ncbi.nlm.nih.gov/36991473","citation_count":9,"is_preprint":false},{"pmid":"35358714","id":"PMC_35358714","title":"Trafficking protein TMED3 promotes esophageal squamous cell carcinoma.","date":"2022","source":"Biomedical journal","url":"https://pubmed.ncbi.nlm.nih.gov/35358714","citation_count":5,"is_preprint":false},{"pmid":"36764873","id":"PMC_36764873","title":"Circ_0108942 Regulates the Progression of Breast Cancer by Regulating the MiR-1178-3p/TMED3 Axis.","date":"2022","source":"Clinical breast cancer","url":"https://pubmed.ncbi.nlm.nih.gov/36764873","citation_count":5,"is_preprint":false},{"pmid":"35854294","id":"PMC_35854294","title":"Depleting TMED3 alleviates the development of endometrial carcinoma.","date":"2022","source":"Cancer cell international","url":"https://pubmed.ncbi.nlm.nih.gov/35854294","citation_count":3,"is_preprint":false},{"pmid":"39735675","id":"PMC_39735675","title":"TMED3 promotes prostate cancer via FOXO1a and FOXO3a phosphorylation.","date":"2024","source":"Oncology research","url":"https://pubmed.ncbi.nlm.nih.gov/39735675","citation_count":2,"is_preprint":false},{"pmid":"38411267","id":"PMC_38411267","title":"TMED3 stabilizes SMAD2 by counteracting NEDD4-mediated ubiquitination to promote ovarian cancer.","date":"2024","source":"Molecular carcinogenesis","url":"https://pubmed.ncbi.nlm.nih.gov/38411267","citation_count":1,"is_preprint":false}],"cost":{"stage1":{"model":"claude-sonnet-4-6","input_tokens":12032,"output_tokens":4025,"usd":0.048236,"stage1_stop_reason":"end_turn"},"stage2":{"model":"claude-opus-4-8","input_tokens":11874,"output_tokens":3566,"usd":0.07426,"stage2_stop_reason":"end_turn"},"total_usd":0.122496,"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\": 2014,\n      \"finding\": \"TMED3 knockdown in human colon cancer cells reduces endogenous WNT-TCF signaling activity, identifying TMED3 as a positive modulator of the WNT-TCF pathway; in vivo genome-wide RNAi screen showed TMED3 knockdown increases metastatic growth and induces lung metastases from subcutaneous xenografts, establishing TMED3 as a metastatic suppressor functioning through WNT-TCF promotion.\",\n      \"method\": \"Genome-wide in vivo RNAi screen in primary human tumor cells in mice; WNT-TCF reporter assays; xenograft models\",\n      \"journal\": \"EMBO molecular medicine\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — in vivo functional screen with multiple cancer cell contexts and pathway reporter assays, single lab\",\n      \"pmids\": [\"24920608\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"TMED3 knockdown leads to enhanced expression of TMED9; TMED9 acts downstream of TMED3 and opposes TMED3-WNT-TCF signaling. TMED9 promotes colon cancer metastasis via TGFα secretion and CNIH4 (a TGFα exporter), and TMED9 knockdown compromises TGFα biogenesis. TMED9/TMED3 antagonism modulates both WNT-TCF and GLI signaling, establishing a TMED3→TMED9→CNIH4/TGFα/GLI pro-metastatic axis opposing TMED3-WNT-TCF.\",\n      \"method\": \"RNAi knockdown, transcriptional rescue assays, gene expression analysis in three colon cancer cell lines, functional migration assays, in vivo xenograft models\",\n      \"journal\": \"Oncogene\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — epistatic ordering by functional rescue, multiple colon cancer cell contexts, single lab\",\n      \"pmids\": [\"31253868\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2016,\n      \"finding\": \"TMED3 promotes hepatocellular carcinoma (HCC) metastasis through IL-11/STAT3 signaling; gene microarray of TMED3-knockdown cells revealed decreased IL-11 expression, and TMED3 overexpression promoted cell migration and invasion, which was suppressed by TMED3 knockdown both in vitro and in vivo.\",\n      \"method\": \"Gene microarray analysis of TMED3-knockdown HCC cells; overexpression and knockdown functional assays; in vivo metastasis models\",\n      \"journal\": \"Scientific reports\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 / Weak — single lab, IL-11/STAT3 pathway link inferred from microarray without direct mechanistic confirmation in the abstract\",\n      \"pmids\": [\"27901021\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"TMED3 recognizes ER core-glycosylated transmembrane protein cargos and, as part of a heteromeric TMED2/3/9/10 complex, mediates ER stress-associated unconventional protein secretion (UPS) of transmembrane proteins including CFTR, pendrin, and SARS-CoV-2 Spike via a Golgi-independent pathway. Co-expression of all four TMEDs improves UPS, while individual silencing reduces it.\",\n      \"method\": \"Gene silencing (siRNA) of individual and combined TMED family members; functional ion transport assays for ΔF508-CFTR and p.H723R-pendrin; SARS-CoV-2 viral release assay; mechanistic co-expression studies\",\n      \"journal\": \"Advanced science (Weinheim, Baden-Wurttemberg, Germany)\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1–2 / Strong — multiple orthogonal functional assays (ion transport, viral release, cargo recognition), heteromeric complex defined, multiple cargo proteins tested, single lab with rigorous controls\",\n      \"pmids\": [\"35748162\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"miR-876-3p directly targets TMED3 (confirmed by luciferase reporter and biotin-miRNA pulldown assay); TMED3 siRNA mimics miR-876-3p's inhibitory effects on cisplatin resistance and stem cell-like properties (Sox-2, Oct-4, CD133, CD44 expression) in gastric cancer cells, placing TMED3 downstream of miR-876-3p in regulating drug resistance.\",\n      \"method\": \"TargetScan prediction; luciferase reporter assay; biotin-miRNA pulldown assay; siRNA knockdown functional rescue\",\n      \"journal\": \"Journal of gastroenterology and hepatology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — direct target validated by two orthogonal methods (luciferase + pulldown), functional rescue confirms pathway placement, single lab\",\n      \"pmids\": [\"30843262\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"TMED3 promotes lung squamous cell carcinoma (LUSC) progression by regulating EZR (Ezrin); RNA sequencing and IPA identified EZR as a downstream target; EZR inhibition suppresses LUSC progression and reduces the promoting effects of TMED3 overexpression, and TMED3 knockdown inhibits cell migration through regulation of EMT.\",\n      \"method\": \"RNA sequencing; Ingenuity Pathway Analysis; loss-of-function assays; EZR overexpression rescue experiments; xenograft mouse model\",\n      \"journal\": \"Cell death & disease\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 / Weak — downstream target identified by transcriptomic screen and partial rescue, single lab, limited direct mechanistic validation\",\n      \"pmids\": [\"34429402\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"TMED3 promotes osteosarcoma progression via RPS15A (ribosomal protein S15A) as a downstream target; simultaneous knockdown of both TMED3 and RPS15A intensified inhibitory effects, and RPS15A knockdown reversed the pro-tumorigenic effects of TMED3 overexpression.\",\n      \"method\": \"Gene expression profile analysis; loss-of-function assays; epistatic rescue by RPS15A knockdown/TMED3 overexpression combination; xenograft tumor model\",\n      \"journal\": \"Cancer cell international\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 / Weak — epistatic rescue with single downstream target, single lab, pathway mechanism not directly defined\",\n      \"pmids\": [\"34838013\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"TMED3 promotes proliferation and migration of breast cancer cells through activation of Wnt/β-catenin signaling; immunofluorescence and Western blot showed TMED3 modulates expression of Wnt/β-catenin pathway proteins and cell cycle/migration-related proteins.\",\n      \"method\": \"Overexpression and knockdown functional assays; Western blot and immunofluorescence of Wnt/β-catenin pathway components; cell cycle analysis; in vitro proliferation/migration/invasion assays\",\n      \"journal\": \"OncoTargets and therapy\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 / Weak — single lab, pathway association by protein expression without direct mechanistic dissection\",\n      \"pmids\": [\"32606792\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"TMED3 knockdown in chordoma cells reduces expression of Bcl-2, HSP27, IGF-I, IGF-II, IGFBP-2, Livin, Akt, CDK6, and cyclin D1, while upregulating MAPK9; these changes accompany inhibition of cell viability and migration and enhanced apoptosis, linking TMED3 to pro-survival signaling nodes.\",\n      \"method\": \"Apoptosis antibody array; Western blot; RT-qPCR; in vitro functional assays; xenograft model\",\n      \"journal\": \"International journal of oncology\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 / Weak — protein array profiling without direct mechanistic pathway validation, single lab\",\n      \"pmids\": [\"33760171\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"In breast cancer, the lncRNA RP11-283G6.5 acts upstream of miR-188-3p/TMED3/Wnt/β-catenin signaling; overexpression of TMED3 or inhibition of miR-188-3p rescued the suppressive effects of RP11-283G6.5, placing TMED3 as a direct downstream effector of miR-188-3p and upstream of Wnt/β-catenin signaling in this axis.\",\n      \"method\": \"Luciferase reporter assay; RNA pulldown; functional rescue experiments (miR-188-3p inhibition, TMED3 overexpression); in vivo xenograft\",\n      \"journal\": \"RNA biology\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 / Weak — TMED3 mechanistic placement is secondary to lncRNA study; functional rescue validates pathway order but direct TMED3 mechanism not primary focus, single lab\",\n      \"pmids\": [\"34130584\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"TMED3 promotes NSCLC progression by enhancing Wnt/β-catenin signaling through modulation of AKT; TMED3 silencing reduced AKT activity and Wnt/β-catenin pathway activity, AKT inhibition blocked TMED3-overexpression-mediated Wnt/β-catenin enhancement, and Wnt/β-catenin inhibition reversed TMED3 overexpression effects on proliferation/invasion.\",\n      \"method\": \"Knockdown and overexpression; AKT inhibitor treatment; Wnt/β-catenin inhibitor treatment; epistatic rescue experiments; in vivo xenograft\",\n      \"journal\": \"Toxicology and applied pharmacology\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 / Weak — epistatic ordering via inhibitors, single lab, no direct biochemical interaction demonstrated\",\n      \"pmids\": [\"34758370\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"TMED3 interacts with CDCA8 (Cell division cycle associated 8) in melanoma cells; CDCA8 overexpression reversed the inhibitory effects of TMED3 knockdown on cell viability and motility, and TMED3 knockdown decreased P-AKT and P-PI3K levels which were rescued by CDCA8 overexpression, placing TMED3 upstream of CDCA8 and PI3K/AKT signaling in melanoma.\",\n      \"method\": \"Co-immunoprecipitation or pulldown (interaction); CDCA8 knockdown/overexpression rescue; SC79 (AKT activator) rescue; Western blot for PI3K/AKT phosphorylation; in vitro and in vivo functional assays\",\n      \"journal\": \"Cell & bioscience\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 / Weak — interaction and epistatic rescue reported, but mechanistic depth limited by abstract description, single lab\",\n      \"pmids\": [\"36991473\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"TMED3 promotes prostate cancer progression via phosphorylation of FOXO1a and FOXO3a; TMED3 inhibition decreased FOXO1a and FOXO3a phosphorylation both in vitro and in vivo, linking TMED3 activity to FOXO pathway suppression.\",\n      \"method\": \"shRNA knockdown; KEGG pathway enrichment analysis; Western blot for FOXO1a/FOXO3a phosphorylation; in vivo prostate cancer mouse model\",\n      \"journal\": \"Oncology research\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 / Weak — phosphorylation change observed upon TMED3 knockdown but direct biochemical mechanism of how TMED3 regulates FOXO phosphorylation not established, single lab\",\n      \"pmids\": [\"39735675\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"TMED3 stabilizes SMAD2 by inhibiting NEDD4 E3 ligase-mediated ubiquitination of SMAD2 in ovarian cancer; SMAD2 knockdown reversed TMED3 overexpression effects on cancer cells; PI3K-AKT pathway was also involved in TMED3-mediated ovarian cancer promotion.\",\n      \"method\": \"shRNA knockdown; functional rescue experiments (SMAD2 knockdown in TMED3-overexpressing cells); ubiquitination assay (implied); Western blot; subcutaneous xenograft model\",\n      \"journal\": \"Molecular carcinogenesis\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Weak — mechanistic claim (NEDD4-mediated ubiquitination counteracted by TMED3) with functional rescue validation, single lab, abstract does not detail ubiquitination assay methods explicitly\",\n      \"pmids\": [\"38411267\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"TMED3 knockdown in esophageal squamous cell carcinoma cells alters gene expression profile; FAM60A was identified as a functionally related downstream gene, and FAM60A knockdown reversed TMED3's pro-tumorigenic effects, placing FAM60A downstream of TMED3.\",\n      \"method\": \"Affymetrix microarray; IPA; FAM60A knockdown rescue experiments; in vitro and in vivo functional assays\",\n      \"journal\": \"Biomedical journal\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 / Weak — downstream target identified by transcriptomics with partial rescue, single lab, limited mechanistic depth\",\n      \"pmids\": [\"35358714\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"TMED3 is a p24 family ER-to-Golgi trafficking protein that functions as part of a heteromeric TMED2/3/9/10 complex to recognize ER core-glycosylated transmembrane cargo proteins and mediate their ER stress-associated unconventional (Golgi-independent) secretion; in cancer contexts, TMED3 promotes WNT-TCF signaling to suppress colon cancer metastasis (while its antagonist TMED9 drives a pro-metastatic TGFα/GLI axis), and in other tumor types TMED3 activates oncogenic signaling through Wnt/β-catenin, PI3K/AKT, IL-11/STAT3, and FOXO pathways, and stabilizes SMAD2 by counteracting NEDD4-mediated ubiquitination.\"\n}\n```","stage2_raw":"{\n  \"mechanistic_narrative\": \"TMED3 is a p24-family endoplasmic reticulum-to-Golgi cargo protein that recognizes ER core-glycosylated transmembrane cargos and, as a subunit of a heteromeric TMED2/3/9/10 complex, mediates ER stress-associated unconventional (Golgi-independent) secretion of transmembrane proteins including CFTR, pendrin, and SARS-CoV-2 Spike; co-expression of all four TMEDs improves this secretion while individual silencing reduces it [#3]. Beyond this trafficking role, TMED3 acts as a context-dependent modulator of oncogenic signaling. In colon cancer it positively modulates WNT-TCF signaling and behaves as a metastatic suppressor, with its knockdown increasing metastatic growth [#0]; this WNT-TCF–promoting activity is opposed by TMED9, which is upregulated upon TMED3 loss and drives a pro-metastatic CNIH4/TGFα/GLI axis, defining a TMED3→TMED9 antagonism that balances WNT-TCF and GLI signaling [#1]. In ovarian cancer, TMED3 stabilizes SMAD2 by counteracting NEDD4 E3 ligase-mediated ubiquitination [#13]. The literature places TMED3 upstream of multiple downstream effectors, but most of these connections rest on lower-confidence transcriptomic and epistatic studies in individual tumor types; the precise biochemical mechanism by which TMED3 engages these signaling pathways has not been resolved in the available corpus.\",\n  \"teleology\": [\n    {\n      \"year\": 2014,\n      \"claim\": \"Established that TMED3 is a functional modulator of WNT-TCF signaling and, unexpectedly, a suppressor of metastasis rather than a generic oncogene.\",\n      \"evidence\": \"Genome-wide in vivo RNAi screen in primary human colon tumor cells with WNT-TCF reporter assays and xenografts\",\n      \"pmids\": [\"24920608\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"No direct biochemical mechanism linking TMED3 to WNT-TCF components\", \"Metastatic suppression shown in xenografts but not in autochthonous models\"]\n    },\n    {\n      \"year\": 2016,\n      \"claim\": \"Extended TMED3 function to hepatocellular carcinoma, where it appeared to act as a pro-metastatic factor via IL-11/STAT3 signaling, hinting at tissue-context-dependent roles.\",\n      \"evidence\": \"Gene microarray of TMED3-knockdown HCC cells with overexpression/knockdown functional assays and in vivo metastasis models\",\n      \"pmids\": [\"27901021\"],\n      \"confidence\": \"Low\",\n      \"gaps\": [\"IL-11/STAT3 link inferred from microarray without direct mechanistic confirmation\", \"Opposite directionality from colon cancer not reconciled mechanistically\"]\n    },\n    {\n      \"year\": 2019,\n      \"claim\": \"Resolved the epistatic relationship between TMED3 and its paralog TMED9, showing antagonistic control of WNT-TCF versus a CNIH4/TGFα/GLI pro-metastatic axis.\",\n      \"evidence\": \"RNAi knockdown with transcriptional rescue and migration assays in three colon cancer lines plus xenografts\",\n      \"pmids\": [\"31253868\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Molecular basis of TMED3-TMED9 antagonism not defined\", \"How TMED3 loss upregulates TMED9 is unknown\"]\n    },\n    {\n      \"year\": 2019,\n      \"claim\": \"Placed TMED3 downstream of miR-876-3p in controlling chemoresistance and stem-like properties, establishing a regulatory input on TMED3 expression.\",\n      \"evidence\": \"Luciferase reporter and biotin-miRNA pulldown confirming direct targeting, with siRNA functional rescue in gastric cancer cells\",\n      \"pmids\": [\"30843262\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Effector mechanism downstream of TMED3 in this axis not defined\", \"Single cancer-type context\"]\n    },\n    {\n      \"year\": 2020,\n      \"claim\": \"Associated TMED3 with Wnt/β-catenin pathway activation in breast cancer, broadening its signaling connections beyond colon WNT-TCF.\",\n      \"evidence\": \"Overexpression/knockdown assays with Western blot and immunofluorescence of Wnt/β-catenin components\",\n      \"pmids\": [\"32606792\"],\n      \"confidence\": \"Low\",\n      \"gaps\": [\"Pathway association by protein expression without direct mechanistic dissection\", \"No demonstrated physical interaction\"]\n    },\n    {\n      \"year\": 2021,\n      \"claim\": \"A series of tumor-specific studies positioned TMED3 upstream of diverse downstream effectors (EZR, RPS15A, AKT, pro-survival nodes, lncRNA/miR axes) across multiple cancers.\",\n      \"evidence\": \"Transcriptomic profiling, apoptosis/protein arrays, and epistatic rescue across LUSC, osteosarcoma, NSCLC, chordoma, and breast cancer with xenografts\",\n      \"pmids\": [\"34429402\", \"34838013\", \"34758370\", \"33760171\", \"34130584\"],\n      \"confidence\": \"Low\",\n      \"gaps\": [\"Downstream targets identified largely by transcriptomic screens with partial rescue\", \"No direct biochemical interactions established for most effectors\", \"Pathway mechanisms inferred via inhibitors rather than reconstitution\"]\n    },\n    {\n      \"year\": 2022,\n      \"claim\": \"Defined the core molecular function of TMED3: recognition of ER core-glycosylated transmembrane cargos and assembly into a TMED2/3/9/10 complex driving Golgi-independent unconventional secretion.\",\n      \"evidence\": \"Combinatorial siRNA silencing, ΔF508-CFTR and pendrin ion-transport assays, SARS-CoV-2 release assay, and co-expression studies\",\n      \"pmids\": [\"35748162\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Structural basis of cargo recognition not resolved\", \"Stoichiometry and assembly order of the TMED2/3/9/10 complex unknown\", \"Link between this trafficking function and cancer signaling roles not established\"]\n    },\n    {\n      \"year\": 2022,\n      \"claim\": \"Identified FAM60A as a downstream gene mediating TMED3 pro-tumorigenic effects in esophageal squamous cell carcinoma.\",\n      \"evidence\": \"Affymetrix microarray and IPA with FAM60A knockdown rescue in vitro and in vivo\",\n      \"pmids\": [\"35358714\"],\n      \"confidence\": \"Low\",\n      \"gaps\": [\"Downstream target identified by transcriptomics with partial rescue\", \"No direct mechanistic link from TMED3 to FAM60A\"]\n    },\n    {\n      \"year\": 2023,\n      \"claim\": \"Reported a physical interaction between TMED3 and CDCA8 placing TMED3 upstream of PI3K/AKT signaling in melanoma.\",\n      \"evidence\": \"Interaction assay with CDCA8 and AKT-activator (SC79) rescue plus Western blot for PI3K/AKT phosphorylation\",\n      \"pmids\": [\"36991473\"],\n      \"confidence\": \"Low\",\n      \"gaps\": [\"Single Co-IP/pulldown without reciprocal validation\", \"Mechanism by which the interaction modulates PI3K/AKT not defined\"]\n    },\n    {\n      \"year\": 2024,\n      \"claim\": \"Provided a more concrete biochemical mechanism: TMED3 stabilizes SMAD2 by antagonizing NEDD4-mediated ubiquitination in ovarian cancer.\",\n      \"evidence\": \"shRNA knockdown, SMAD2 functional rescue, and ubiquitination assays with xenografts\",\n      \"pmids\": [\"38411267\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Ubiquitination assay methodology not detailed\", \"Whether TMED3 directly binds SMAD2 or NEDD4 not shown\"]\n    },\n    {\n      \"year\": 2024,\n      \"claim\": \"Linked TMED3 activity to FOXO1a/FOXO3a phosphorylation in prostate cancer, extending its signaling footprint.\",\n      \"evidence\": \"shRNA knockdown with KEGG enrichment and Western blot for FOXO phosphorylation in vitro and in vivo\",\n      \"pmids\": [\"39735675\"],\n      \"confidence\": \"Low\",\n      \"gaps\": [\"Direct biochemical mechanism linking TMED3 to FOXO phosphorylation not established\", \"No identified kinase intermediary\"]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"How TMED3's core ER-to-Golgi cargo-trafficking and unconventional-secretion function mechanistically connects to its diverse, often opposing, roles in cancer signaling remains unresolved.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"Low\",\n      \"gaps\": [\"No unifying model reconciling metastatic-suppressor versus pro-tumorigenic phenotypes across tissues\", \"Whether cancer signaling effects derive from cargo trafficking is untested\", \"Structural and stoichiometric details of the TMED2/3/9/10 complex unknown\"]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0038024\", \"supporting_discovery_ids\": [3]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005783\", \"supporting_discovery_ids\": [3]},\n      {\"term_id\": \"GO:0005794\", \"supporting_discovery_ids\": [3]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-5653656\", \"supporting_discovery_ids\": [3]},\n      {\"term_id\": \"R-HSA-9609507\", \"supporting_discovery_ids\": [3]}\n    ],\n    \"complexes\": [\"TMED2/3/9/10 complex\"],\n    \"partners\": [\"TMED9\", \"TMED2\", \"TMED10\", \"SMAD2\", \"NEDD4\", \"CDCA8\"],\n    \"other_free_text\": []\n  }\n}","audit_flag":null,"evaluation":{"pairwise":"win","faith_supported":3,"faith_total":3,"faith_pct":100.0}}