{"gene":"AMIGO2","run_date":"2026-06-09T22:02:43","timeline":{"discoveries":[{"year":2015,"finding":"AMIGO2 directly binds the pleckstrin homology (PH) domain of PDK1 via amino acid residues 465–474 in AMIGO2, acting as a membrane scaffold that recruits PDK1 to the plasma membrane and promotes Akt activation. Loss of AMIGO2 in endothelial cells leads to apoptosis and inhibition of angiogenesis with Akt inactivation. A synthetic peptide containing AMIGO2 residues 465–474 abrogated the AMIGO2–PDK1 interaction, Akt activation, and pathological angiogenesis in vivo.","method":"Co-immunoprecipitation, direct binding assay with synthetic peptides, loss-of-function in endothelial cells, in vivo tumor/retinopathy angiogenesis models","journal":"The Journal of cell biology","confidence":"High","confidence_rationale":"Tier 2 / Strong — reciprocal Co-IP, direct peptide competition assay, in vitro apoptosis phenotype, and two in vivo models; multiple orthogonal methods in one study","pmids":["26553931"],"is_preprint":false},{"year":2017,"finding":"AMIGO2 interacts with the pseudokinase PTK7, and AMIGO2 regulates PTK7 proteolytic processing. BET bromodomain proteins BRD2/4 occupy a melanoma-specific promoter and super-enhancer at the AMIGO2 locus; BET inhibitor treatment evicts BRD2/4, silences AMIGO2, and alters PTK7 processing. AMIGO2 silencing in melanoma cells induces G1/S cell-cycle arrest followed by apoptosis.","method":"Co-immunoprecipitation (AMIGO2–PTK7 interaction), epigenomic profiling (ChIP-seq for BRD2/4), CRISPR genome editing of regulatory elements, BETi treatment, loss-of-function RNA silencing with cell-cycle/apoptosis readouts","journal":"Molecular cell","confidence":"High","confidence_rationale":"Tier 2 / Strong — reciprocal Co-IP for PTK7 interaction, ChIP-seq + CRISPR for regulatory mechanism, multiple orthogonal methods in one rigorous study","pmids":["29149598"],"is_preprint":false},{"year":2004,"finding":"DEGA/AMIGO2 is a type I transmembrane protein that localizes to the cell surface (confirmed by DEGA-GFP fusion construct in 293 cells). Stable antisense-mediated knockdown in gastric adenocarcinoma AGS cells caused altered morphology, increased ploidy, chromosomal instability, decreased cell adhesion and migration, and near-complete abrogation of tumorigenicity in nude mice.","method":"GFP fusion localization, stable antisense transfection, nude mouse xenograft, cell adhesion/migration assays, chromosomal analysis","journal":"Oncogene","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — direct localization experiment plus multiple functional readouts in a single lab; no reciprocal validation","pmids":["15107827"],"is_preprint":false},{"year":2017,"finding":"AMIGO2 expression in tumor cells mediates their preferential adhesion to liver endothelial cells (but not lung endothelial cells), promoting liver metastasis. siRNA-mediated knockdown reduced liver endothelial cell adhesion in vitro, and forced expression of AMIGO2 in low-AMIGO2 cells increased liver endothelial cell adhesion and liver metastasis in vivo.","method":"siRNA knockdown, forced expression, ex vivo imaging, intrasplenic injection metastasis model","journal":"Scientific reports","confidence":"High","confidence_rationale":"Tier 2 / Strong — bidirectional manipulation (KD and overexpression), in vitro adhesion assay plus in vivo metastasis model, replicated in later studies","pmids":["28272394"],"is_preprint":false},{"year":2019,"finding":"AMIGO2 functions as a homophilic cell-surface protein in the retina; starburst amacrine cells (SACs) and rod bipolar cells (RBCs) express AMIGO2, and Amigo2 knockout mice display expanded SAC and RBC dendrite arbors while other retinal neuron arbors remain unchanged. Increased SAC dendrite coverage was accompanied by increased direction selectivity of downstream direction-selective ganglion cells (DSGCs), identifying AMIGO2 as a cell-type-specific dendritic scaling factor.","method":"Amigo2 knockout mouse, morphological analysis of dendrite arbors, electrophysiology of DSGCs, immunostaining","journal":"Cell reports","confidence":"High","confidence_rationale":"Tier 2 / Strong — clean KO with specific cellular phenotype, multiple neuron types analyzed as controls, functional electrophysiology readout, rigorous loss-of-function study","pmids":["31693896"],"is_preprint":false},{"year":2017,"finding":"AMIGO2 modulates T-cell functions: Amigo2 knockout impairs T-cell infiltration into secondary lymphoid organs and dampens Th-cell activation, while promoting splenic Th-cell proliferation. Mechanistically, AMIGO2 overexpression in 293T cells dampens NF-κB transcriptional activity, and AMIGO2 deficiency enhances Akt but suppresses GSK-3β phosphorylation and promotes nuclear translocation of NF-κB and NFAT1 in Th cells. Amigo2 KO mice exhibit ameliorated experimental autoimmune encephalomyelitis (EAE).","method":"Amigo2 knockout mice, adoptive T-cell transfer, NF-κB reporter assay, Western blot for Akt/GSK-3β phosphorylation, nuclear fractionation for NF-κB/NFAT1","journal":"Brain, behavior, and immunity","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — KO mouse with defined cellular and molecular phenotypes, NF-κB reporter assay, multiple signaling readouts; single lab","pmids":["28119027"],"is_preprint":false},{"year":2016,"finding":"IL-17A and TNF-α synergistically upregulate AMIGO2 expression in RA synoviocytes via an ERK-dependent mechanism (JNK inhibits AMIGO2 induction). Elevated AMIGO2 promotes cell survival and reduces apoptosis (cadmium-induced toxicity). HMGB1 in inflammatory conditions further increases AMIGO2 expression and reduces cell toxicity.","method":"Cytokine stimulation of synoviocytes, ERK/JNK pharmacological inhibition, apoptosis assay with cadmium, co-culture experiments","journal":"Frontiers in immunology","confidence":"Medium","confidence_rationale":"Tier 3 / Moderate — pharmacological pathway dissection plus functional apoptosis readout, multiple conditions tested; single lab","pmids":["27446084"],"is_preprint":false},{"year":2022,"finding":"AMIGO2 contained in cancer cell-derived extracellular vesicles (EVs) is internalized by human hepatic sinusoidal endothelial cells (HHSECs) and, once inside, significantly enhances adhesion of HHSECs to cancer cells (including cells that themselves lack AMIGO2 expression), identifying a paracrine mechanism for liver metastasis initiation.","method":"Western blot of EVs, EV internalization assay, cell adhesion assay with AMIGO2-overexpressing vs. control EVs","journal":"Scientific reports","confidence":"Medium","confidence_rationale":"Tier 3 / Moderate — functional EV adhesion assay with gain-of-function EV treatment; multiple cancer cell lines tested; single lab","pmids":["35039535"],"is_preprint":false},{"year":2024,"finding":"METTL3-mediated m6A methylation in the 5'-UTR of AMIGO2 mRNA recruits the reader YTHDC2, stabilizing/upregulating AMIGO2 expression in RA fibroblast-like synoviocytes. METTL3 knockdown reduces m6A on AMIGO2 5'-UTR, decreases YTHDC2–AMIGO2 mRNA interaction, and lowers AMIGO2 protein, suppressing RA-FLS proliferation and migration; AMIGO2 overexpression rescues these phenotypes.","method":"m6A-seq/MeRIP for modification site mapping, RIP (YTHDC2–AMIGO2 mRNA), siRNA knockdown of METTL3/YTHDC2, rescue overexpression of AMIGO2","journal":"Biochimica et biophysica acta. Molecular basis of disease","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — MeRIP for m6A site, RIP for reader–mRNA interaction, rescue experiment; single lab","pmids":["38432455"],"is_preprint":false},{"year":2024,"finding":"In esophageal squamous cell carcinoma, METTL3-mediated m6A modification in the 5'-UTR of AMIGO2 pre-mRNA is read by YTHDC1, which regulates AMIGO2 splicing. METTL3 knockdown reduces m6A on AMIGO2 pre-mRNA, impairs YTHDC1-mediated splicing, and decreases AMIGO2 expression, suppressing ESCC proliferation and migration; AMIGO2 overexpression rescues these effects.","method":"MeRIP (m6A site mapping), RIP (YTHDC1–AMIGO2 pre-mRNA), splicing analysis, siRNA knockdown/rescue overexpression, in vitro and in vivo tumor models","journal":"Gene","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — MeRIP, RIP, splicing assay, rescue experiment; single lab; distinct reader (YTHDC1) from the RA study (YTHDC2)","pmids":["38360124"],"is_preprint":false},{"year":2018,"finding":"AMIGO2 deficiency in mice (KO) causes increased cardiomyocyte apoptosis, reduced proliferation, and reduced angiogenesis after myocardial infarction, with weaker cardiac function and larger infarct scar. Molecularly, AMIGO2 KO increases active caspase-3 and decreases PDK1, p-Akt, Bcl-2/Bax, and VEGF expression, consistent with inactivation of the PDK1/Akt survival pathway in cardiomyocytes.","method":"AMIGO2 KO mouse MI model, echocardiography, TUNEL/Ki67/immunohistochemistry, Western blot for PDK1/Akt/Bcl-2/Bax/VEGF","journal":"Cardiology journal","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — clean KO mouse with defined cardiac phenotype and molecular pathway readouts; single lab","pmids":["29718531"],"is_preprint":false},{"year":2023,"finding":"AMIGO2 suppresses innate cisplatin sensitivity in NSCLC by inhibiting cisplatin-induced activation of caspase-8, caspase-9, and caspase-3, thereby preventing GSDME cleavage and pyroptosis. AMIGO2 acts by stimulating the PDK1/Akt (T308) signaling axis. Alteration of AMIGO2 expression changed cisplatin-induced pyroptosis both in vitro and in vivo.","method":"AMIGO2 knockdown/overexpression, caspase activity assays, GSDME cleavage Western blot, PDK1/Akt phosphorylation assays, in vivo xenograft cisplatin treatment","journal":"Journal of cellular and molecular medicine","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — bidirectional manipulation, multiple pathway readouts (caspase cascade, GSDME, PDK1/Akt), in vitro and in vivo; single lab","pmids":["37438979"],"is_preprint":false},{"year":2022,"finding":"AMIGO2 expression in human gastric and colorectal cancer cells is directly associated with their adhesion to human hepatic sinusoidal endothelial cells (HHSECs) in vitro. Constitutive AMIGO2 knockdown clones showed significantly attenuated adhesion to HHSECs and suppressed liver metastasis in nude mice following intrasplenic inoculation, validating the mouse findings in human cancer cells.","method":"AMIGO2 knockdown (constitutive clones), cell adhesion assay to HHSECs, intrasplenic injection nude mouse liver metastasis model","journal":"Pathology, research and practice","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — loss-of-function with defined in vitro and in vivo readouts; single lab; extends prior mouse data to human cells","pmids":["35843033"],"is_preprint":false},{"year":2024,"finding":"AMIGO2 promotes EMT in colorectal cancer cells via activation of the TGFβ/Smad signaling pathway. Treatment with TGFβ receptor inhibitor LY2109761 suppressed AMIGO2-induced EMT. In CRC tissue samples, AMIGO2 was found at the invasive front where it localizes to the nucleus and associates with EMT marker expression, suggesting nuclear translocation of AMIGO2 as a mechanism for inducing EMT.","method":"AMIGO2 overexpression/knockdown, TGFβ/Smad pathway inhibitor treatment, immunohistochemistry/nuclear localization analysis in CRC clinical samples, EMT marker Western blot","journal":"Cancer gene therapy","confidence":"Medium","confidence_rationale":"Tier 3 / Moderate — pharmacological pathway inhibition plus clinical sample localization data; nuclear localization is observational without mechanistic reconstitution; single lab","pmids":["39379686"],"is_preprint":false},{"year":2025,"finding":"AMIGO2-containing small extracellular vesicles (sEVs) from gastric cancer cells activate hepatic stellate cells (HSCs) via NF-κB nuclear translocation, inducing IL-8 secretion. The resulting IL-8-rich conditioned medium significantly enhances gastric cancer cell migration; neutralizing IL-8 with antibodies suppresses this migration, establishing an AMIGO2 sEV→HSC→IL-8→cancer cell migration axis in liver metastasis.","method":"sEV isolation, AMIGO2 overexpression, NF-κB nuclear translocation assay, chemokine array, IL-8 neutralizing antibody, migration assay","journal":"Anticancer research","confidence":"Medium","confidence_rationale":"Tier 3 / Moderate — functional rescue with neutralizing antibody confirms IL-8 role; NF-κB translocation assay; single lab","pmids":["40155010"],"is_preprint":false},{"year":2022,"finding":"AMIGO2 expression in cancer-associated fibroblasts (CAFs) of colorectal cancer is induced by TGF-β. Conditioned media from NAFs with AMIGO2 overexpression enhanced CRC tumor cell proliferation and migration; siRNA-mediated inhibition of AMIGO2 in CAFs attenuated these effects, showing AMIGO2 regulates paracrine tumorigenic secretomes in CAFs.","method":"TGF-β stimulation, AMIGO2 overexpression in NAFs, siRNA knockdown in CAFs, conditioned media transfer, proliferation/migration assays","journal":"The Journal of pathology","confidence":"Medium","confidence_rationale":"Tier 3 / Moderate — bidirectional manipulation (OE and KD) with conditioned media functional readout; single lab","pmids":["39523830"],"is_preprint":false},{"year":2024,"finding":"AMIGO2 knockdown in bladder cancer cells suppresses PPAR-γ expression; PPAR-γ overexpression rescues the proliferation and migration suppressed by AMIGO2 knockdown, placing PPAR-γ downstream of AMIGO2 in a signaling axis controlling bladder cancer cell growth.","method":"siRNA/lentiviral AMIGO2 knockdown, RNA-seq, PPAR-γ Western blot, PPAR-γ rescue overexpression, cell proliferation/migration/xenograft assays","journal":"Hereditas","confidence":"Low","confidence_rationale":"Tier 3 / Weak — rescue experiment placing PPAR-γ downstream, but pathway connection is inferred from RNA-seq without direct mechanistic assay; single lab","pmids":["38978149"],"is_preprint":false},{"year":2021,"finding":"AMIGO2 knockdown via CRISPR/Cas9 in high-grade serous ovarian cancer cells reduces sphere-forming potential, adhesion, and invasion in vitro, and significantly attenuates intraperitoneal metastasis in vivo.","method":"CRISPR/Cas9 knockdown, sphere formation assay, cell adhesion/invasion assay, in vivo IP metastasis model","journal":"Cancer letters","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — CRISPR-based loss-of-function with multiple orthogonal in vitro and in vivo readouts; single lab","pmids":["33524500"],"is_preprint":false},{"year":2022,"finding":"Adhesion molecule Amigo2 is selectively expressed in medial habenula neurons and mediates fasciculation of the fasciculus retroflexus. Amigo2 loss-of-function in mice causes defasciculation of medial habenular axons; gain-of-function generates a more condensed tract and rescues the KO phenotype. AMIGO2 does not alter the course of habenular fibers, only their fasciculation.","method":"Amigo2 KO mouse, gain-of-function overexpression rescue experiment, tract tracing/morphological analysis","journal":"Developmental dynamics","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — bidirectional manipulation (KO and rescue OE) with specific morphological phenotype; single lab","pmids":["35727300"],"is_preprint":false}],"current_model":"AMIGO2 is a type I transmembrane leucine-rich repeat/Ig-domain cell-surface scaffold protein that promotes cell survival and adhesion through multiple mechanisms: it directly binds the PH domain of PDK1 (via residues 465–474) to recruit PDK1 to the plasma membrane and activate Akt, thereby suppressing apoptosis in endothelial and cancer cells; it interacts with and regulates proteolytic processing of the pseudokinase PTK7 in melanoma; it mediates homophilic adhesion to scale dendrite arbors in specific retinal neuron types; it promotes tumor cell adhesion to hepatic endothelial cells (and, in EVs, activates hepatic stellate cells to secrete IL-8) to drive liver metastasis; it induces EMT via TGFβ/Smad signaling at the invasive front; and its expression is post-transcriptionally regulated by METTL3-mediated m6A modification read by YTHDC1 (splicing) or YTHDC2 (stability) in a context-dependent manner."},"narrative":{"mechanistic_narrative":"AMIGO2 is a type I transmembrane cell-surface protein that functions as a membrane scaffold and homophilic adhesion molecule to promote cell survival, adhesion, and tissue patterning [PMID:26553931, PMID:15107827, PMID:31693896]. Its central survival mechanism is direct binding of the PH domain of PDK1 through AMIGO2 residues 465–474, which recruits PDK1 to the plasma membrane and activates Akt; disrupting this interaction with a competing peptide abrogates Akt activation, triggers apoptosis, and blocks pathological angiogenesis [PMID:26553931]. This PDK1/Akt axis underlies AMIGO2's protective roles in cardiomyocytes after myocardial infarction [PMID:29718531] and its suppression of cisplatin-induced caspase activation, GSDME cleavage, and pyroptosis in lung cancer [PMID:37438979]. As a homophilic surface adhesion factor, AMIGO2 scales dendrite arbors of specific retinal neuron types and tunes downstream direction selectivity [PMID:31693896], and mediates fasciculation of medial habenular axons in the fasciculus retroflexus [PMID:35727300]. In cancer, AMIGO2 promotes tumor cell adhesion to hepatic sinusoidal endothelial cells to drive liver metastasis [PMID:28272394, PMID:35843033], a process that also proceeds paracrine: AMIGO2 carried in tumor-derived extracellular vesicles is internalized by hepatic endothelial cells to enhance their adhesiveness [PMID:35039535] and activates hepatic stellate cells via NF-κB to secrete IL-8 that boosts cancer cell migration [PMID:40155010]. AMIGO2 additionally interacts with the pseudokinase PTK7 and regulates its proteolytic processing in melanoma, where it is driven by a BET-dependent super-enhancer [PMID:29149598], and it induces EMT through TGFβ/Smad signaling at the invasive front [PMID:39379686]. AMIGO2 expression is post-transcriptionally controlled by METTL3-deposited m6A in its 5′-UTR, read context-dependently by YTHDC1 to regulate splicing or YTHDC2 to regulate stability [PMID:38360124, PMID:38432455].","teleology":[{"year":2004,"claim":"Established AMIGO2 (DEGA) as a cell-surface type I transmembrane protein required for tumorigenicity, the first functional characterization linking it to adhesion, genomic stability, and cancer cell behavior.","evidence":"GFP-fusion localization and stable antisense knockdown with xenograft, adhesion, and chromosomal readouts in gastric adenocarcinoma cells","pmids":["15107827"],"confidence":"Medium","gaps":["No molecular mechanism for adhesion or ploidy effects","No reciprocal validation of localization","Binding partners unidentified"]},{"year":2015,"claim":"Defined the molecular survival mechanism: AMIGO2 directly binds the PDK1 PH domain via residues 465–474 to recruit PDK1 to the membrane and activate Akt, explaining its anti-apoptotic and pro-angiogenic role.","evidence":"Reciprocal Co-IP, synthetic peptide competition, endothelial loss-of-function, and in vivo angiogenesis models","pmids":["26553931"],"confidence":"High","gaps":["Structural basis of the AMIGO2–PDK1 interface not resolved","How surface AMIGO2 spatially organizes cytoplasmic PDK1 unclear"]},{"year":2016,"claim":"Showed AMIGO2 is an inducible cytoprotective gene downstream of inflammatory cytokines, linking IL-17A/TNF-α/ERK signaling to AMIGO2-mediated survival in synoviocytes.","evidence":"Cytokine stimulation with ERK/JNK pharmacological inhibition and cadmium-induced apoptosis assay in RA synoviocytes","pmids":["27446084"],"confidence":"Medium","gaps":["Transcription factors mediating ERK-dependent induction not identified","Downstream survival effector in synoviocytes not tied to PDK1/Akt"]},{"year":2017,"claim":"Connected AMIGO2 to a second client (PTK7) and revealed its epigenetic regulation, showing a melanoma super-enhancer drives AMIGO2 to sustain proliferation and survival.","evidence":"Co-IP for AMIGO2–PTK7, ChIP-seq for BRD2/4, CRISPR editing of regulatory elements, and BETi with cell-cycle/apoptosis readouts","pmids":["29149598"],"confidence":"High","gaps":["How AMIGO2 controls PTK7 proteolytic processing mechanistically unknown","Whether PTK7 regulation requires the PDK1/Akt axis unclear"]},{"year":2017,"claim":"Identified organ-tropic adhesion as a metastasis mechanism: AMIGO2 mediates selective tumor cell adhesion to liver (not lung) endothelium to drive liver metastasis.","evidence":"siRNA knockdown, forced expression, and intrasplenic injection metastasis model","pmids":["28272394"],"confidence":"High","gaps":["Endothelial counter-receptor for AMIGO2 not identified","Molecular basis of liver-specific tropism unresolved"]},{"year":2017,"claim":"Extended AMIGO2's role to adaptive immunity, showing it constrains T-cell signaling through Akt/GSK-3β and NF-κB/NFAT1 to shape autoimmune responses.","evidence":"Amigo2 KO mice, adoptive transfer, NF-κB reporter, signaling Westerns, nuclear fractionation, and EAE model","pmids":["28119027"],"confidence":"Medium","gaps":["Whether AMIGO2 acts cell-autonomously in T cells via PDK1 unclear","Direct partners in T cells not defined"]},{"year":2018,"claim":"Demonstrated AMIGO2's PDK1/Akt survival axis operates in vivo in cardiomyocytes, linking AMIGO2 loss to worse cardiac outcomes after infarction.","evidence":"Amigo2 KO mouse MI model with echocardiography, TUNEL/Ki67, and PDK1/Akt/Bcl-2/Bax/VEGF Westerns","pmids":["29718531"],"confidence":"Medium","gaps":["Cell-autonomous vs. paracrine contribution not dissected","Direct membrane scaffolding in cardiomyocytes not shown"]},{"year":2019,"claim":"Revealed AMIGO2 as a cell-type-specific homophilic dendritic scaling factor in the retina, with functional consequences for neural circuit output.","evidence":"Amigo2 KO mouse, dendrite arbor morphometry across neuron types, and DSGC electrophysiology","pmids":["31693896"],"confidence":"High","gaps":["Molecular pathway downstream of homophilic adhesion in neurons unknown","Whether PDK1/Akt is involved in dendrite scaling untested"]},{"year":2021,"claim":"Generalized AMIGO2's pro-metastatic adhesion/invasion role to ovarian cancer using CRISPR loss-of-function.","evidence":"CRISPR/Cas9 knockdown with sphere formation, adhesion/invasion, and intraperitoneal metastasis assays","pmids":["33524500"],"confidence":"Medium","gaps":["Molecular effectors of sphere formation and invasion not mapped","No link to specific signaling partner in this context"]},{"year":2022,"claim":"Defined a paracrine, EV-borne mechanism of metastasis, showing AMIGO2 transferred in tumor EVs reprograms recipient hepatic endothelium independent of tumor-cell AMIGO2.","evidence":"EV Western blot, internalization assay, and adhesion assay with AMIGO2-overexpressing vs. control EVs","pmids":["35039535"],"confidence":"Medium","gaps":["Intracellular pathway activated in recipient endothelium unknown","Whether internalized AMIGO2 engages endothelial PDK1 untested"]},{"year":2022,"claim":"Validated AMIGO2-dependent hepatic endothelial adhesion and liver metastasis in human gastric and colorectal cancer cells.","evidence":"Constitutive AMIGO2 knockdown clones with HHSEC adhesion assays and intrasplenic metastasis model","pmids":["35843033"],"confidence":"Medium","gaps":["Endothelial binding partner still unidentified","Single-lab confirmation"]},{"year":2022,"claim":"Placed AMIGO2 in the stromal compartment, showing TGF-β-induced AMIGO2 in cancer-associated fibroblasts drives a pro-tumorigenic paracrine secretome.","evidence":"TGF-β stimulation, AMIGO2 overexpression/knockdown in fibroblasts, and conditioned-media proliferation/migration assays","pmids":["39523830"],"confidence":"Medium","gaps":["Secreted factors mediating the effect not identified","Whether CAF AMIGO2 signals via the same PDK1 axis unknown"]},{"year":2022,"claim":"Established AMIGO2 as a developmental axon fasciculation factor in medial habenula, distinguishing fasciculation from axon guidance.","evidence":"Amigo2 KO and gain-of-function rescue with tract tracing in mice","pmids":["35727300"],"confidence":"Medium","gaps":["Whether fasciculation is homophilic adhesion-dependent not directly shown","Downstream signaling unknown"]},{"year":2023,"claim":"Showed AMIGO2 suppresses chemotherapy-induced pyroptosis, linking its PDK1/Akt activity to evasion of caspase/GSDME-mediated death.","evidence":"Bidirectional manipulation with caspase activity, GSDME cleavage, PDK1/Akt phosphorylation, and xenograft cisplatin readouts in NSCLC","pmids":["37438979"],"confidence":"Medium","gaps":["Direct mechanistic step from Akt to caspase-8/9 suppression not resolved","Single lab"]},{"year":2024,"claim":"Resolved post-transcriptional control of AMIGO2, showing METTL3-deposited 5′-UTR m6A is read by YTHDC2 (stability) or YTHDC1 (splicing) in a context-dependent manner.","evidence":"MeRIP m6A mapping, RIP for reader–mRNA interaction, and METTL3/reader knockdown with AMIGO2 rescue in RA synoviocytes and esophageal carcinoma","pmids":["38432455","38360124"],"confidence":"Medium","gaps":["What determines YTHDC1 vs. YTHDC2 reader choice unknown","Upstream signals controlling METTL3 activity on AMIGO2 not defined"]},{"year":2024,"claim":"Identified an EMT-promoting role via TGFβ/Smad signaling and an unexpected nuclear localization of AMIGO2 at the tumor invasive front.","evidence":"AMIGO2 overexpression/knockdown, TGFβ receptor inhibitor LY2109761, and IHC of CRC clinical samples","pmids":["39379686"],"confidence":"Medium","gaps":["Mechanism by which a transmembrane protein reaches the nucleus not established","Direct role of nuclear AMIGO2 in transcription not reconstituted"]},{"year":2025,"claim":"Completed a paracrine liver-metastasis circuit, defining an AMIGO2 sEV → hepatic stellate cell NF-κB → IL-8 → cancer cell migration axis.","evidence":"sEV isolation, NF-κB translocation assay, chemokine array, and IL-8 neutralization with migration assays","pmids":["40155010"],"confidence":"Medium","gaps":["How EV AMIGO2 activates NF-κB in stellate cells unknown","Single lab"]},{"year":null,"claim":"The identity of AMIGO2's extracellular binding partner(s)—including its endothelial counter-receptor in metastasis and the basis of homophilic adhesion—and the mechanism linking surface or EV AMIGO2 to nuclear/EMT functions remain unresolved.","evidence":"","pmids":[],"confidence":"Medium","gaps":["No extracellular ligand/counter-receptor identified","No structure of AMIGO2 ectodomain or its homophilic interface","Mechanism of nuclear translocation of a transmembrane protein unexplained"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0098631","term_label":"cell adhesion mediator activity","supporting_discovery_ids":[4,18]},{"term_id":"GO:0060089","term_label":"molecular transducer activity","supporting_discovery_ids":[0]},{"term_id":"GO:0060090","term_label":"molecular adaptor activity","supporting_discovery_ids":[0]}],"localization":[{"term_id":"GO:0005886","term_label":"plasma membrane","supporting_discovery_ids":[0,2]},{"term_id":"GO:0031410","term_label":"cytoplasmic vesicle","supporting_discovery_ids":[7,14]},{"term_id":"GO:0005634","term_label":"nucleus","supporting_discovery_ids":[13]}],"pathway":[{"term_id":"R-HSA-162582","term_label":"Signal Transduction","supporting_discovery_ids":[0,11]},{"term_id":"R-HSA-1643685","term_label":"Disease","supporting_discovery_ids":[3,12]},{"term_id":"R-HSA-5357801","term_label":"Programmed Cell Death","supporting_discovery_ids":[0,11]},{"term_id":"R-HSA-1266738","term_label":"Developmental Biology","supporting_discovery_ids":[4,18]}],"complexes":[],"partners":["PDK1","PTK7"],"other_free_text":[]}},"prefetch_data":{"uniprot":{"accession":"Q86SJ2","full_name":"Amphoterin-induced protein 2","aliases":["AMIGO-2","Alivin-1","Differentially expressed in gastric adenocarcinomas","DEGA"],"length_aa":522,"mass_kda":57.9,"function":"Required for depolarization-dependent survival of cultured cerebellar granule neurons. May mediate homophilic as well as heterophilic cell-cell interaction with AMIGO1 or AMIGO3. May contribute to signal transduction through its intracellular domain. May be required for tumorigenesis of a subset of gastric adenocarcinomas","subcellular_location":"Cell membrane; Nucleus","url":"https://www.uniprot.org/uniprotkb/Q86SJ2/entry"},"depmap":{"release":"DepMap","has_data":true,"is_common_essential":false,"resolved_as":"","url":"https://depmap.org/portal/gene/AMIGO2","classification":"Not Classified","n_dependent_lines":1,"n_total_lines":1208,"dependency_fraction":0.0008278145695364238},"opencell":{"profiled":false,"resolved_as":"","ensg_id":"","cell_line_id":"","localizations":[],"interactors":[],"url":"https://opencell.sf.czbiohub.org/search/AMIGO2","total_profiled":1310},"omim":[{"mim_id":"615691","title":"ADHESION MOLECULE WITH Ig-LIKE DOMAIN 3; AMIGO3","url":"https://www.omim.org/entry/615691"},{"mim_id":"615690","title":"ADHESION MOLECULE WITH Ig-LIKE DOMAIN 2; AMIGO2","url":"https://www.omim.org/entry/615690"},{"mim_id":"615689","title":"ADHESION MOLECULE WITH Ig-LIKE DOMAIN 1; AMIGO1","url":"https://www.omim.org/entry/615689"}],"hpa":{"profiled":true,"resolved_as":"","reliability":"Approved","locations":[{"location":"Golgi apparatus","reliability":"Approved"},{"location":"Plasma membrane","reliability":"Approved"},{"location":"Cytosol","reliability":"Additional"}],"tissue_specificity":"Tissue enhanced","tissue_distribution":"Detected in many","driving_tissues":[{"tissue":"blood vessel","ntpm":68.2}],"url":"https://www.proteinatlas.org/search/AMIGO2"},"hgnc":{"alias_symbol":["ALI1","DEGA"],"prev_symbol":[]},"alphafold":{"accession":"Q86SJ2","domains":[{"cath_id":"3.80.10.10","chopping":"45-289","consensus_level":"medium","plddt":90.2519,"start":45,"end":289},{"cath_id":"2.60.40.10","chopping":"296-382","consensus_level":"high","plddt":86.4114,"start":296,"end":382}],"viewer_url":"https://alphafold.ebi.ac.uk/entry/Q86SJ2","model_url":"https://alphafold.ebi.ac.uk/files/AF-Q86SJ2-F1-model_v6.cif","pae_url":"https://alphafold.ebi.ac.uk/files/AF-Q86SJ2-F1-predicted_aligned_error_v6.png","plddt_mean":75.06},"mouse_models":{"mgi_url":"https://www.informatics.jax.org/marker/summary?nomen=AMIGO2","jax_strain_url":"https://www.jax.org/strain/search?query=AMIGO2"},"sequence":{"accession":"Q86SJ2","fasta_url":"https://rest.uniprot.org/uniprotkb/Q86SJ2.fasta","uniprot_url":"https://www.uniprot.org/uniprotkb/Q86SJ2/entry","alphafold_viewer_url":"https://alphafold.ebi.ac.uk/entry/Q86SJ2"}},"corpus_meta":[{"pmid":"29149598","id":"PMC_29149598","title":"Harnessing BET Inhibitor Sensitivity Reveals AMIGO2 as a Melanoma Survival Gene.","date":"2017","source":"Molecular cell","url":"https://pubmed.ncbi.nlm.nih.gov/29149598","citation_count":102,"is_preprint":false},{"pmid":"26553931","id":"PMC_26553931","title":"AMIGO2, a novel membrane anchor of PDK1, controls cell survival and angiogenesis via Akt activation.","date":"2015","source":"The Journal of cell biology","url":"https://pubmed.ncbi.nlm.nih.gov/26553931","citation_count":60,"is_preprint":false},{"pmid":"15107827","id":"PMC_15107827","title":"DEGA/AMIGO-2, a leucine-rich repeat family member, differentially expressed in human gastric adenocarcinoma: effects on ploidy, chromosomal stability, cell adhesion/migration and tumorigenicity.","date":"2004","source":"Oncogene","url":"https://pubmed.ncbi.nlm.nih.gov/15107827","citation_count":54,"is_preprint":false},{"pmid":"22314660","id":"PMC_22314660","title":"AMIGO2 mRNA expression in hippocampal CA2 and CA3a.","date":"2013","source":"Brain structure & function","url":"https://pubmed.ncbi.nlm.nih.gov/22314660","citation_count":40,"is_preprint":false},{"pmid":"28272394","id":"PMC_28272394","title":"Amigo2-upregulation in Tumour Cells Facilitates Their Attachment to Liver Endothelial Cells Resulting in Liver Metastases.","date":"2017","source":"Scientific reports","url":"https://pubmed.ncbi.nlm.nih.gov/28272394","citation_count":35,"is_preprint":false},{"pmid":"31628879","id":"PMC_31628879","title":"Natural variations in the promoter of Awn Length Inhibitor 1 (ALI-1) are associated with awn elongation and grain length in common wheat.","date":"2019","source":"The Plant journal : for cell and molecular biology","url":"https://pubmed.ncbi.nlm.nih.gov/31628879","citation_count":35,"is_preprint":false},{"pmid":"31074142","id":"PMC_31074142","title":"A novel nonsense substitution identified in the AMIGO2 gene in an Occulo-Auriculo-Vertebral spectrum patient.","date":"2019","source":"Orthodontics & craniofacial research","url":"https://pubmed.ncbi.nlm.nih.gov/31074142","citation_count":23,"is_preprint":false},{"pmid":"33732354","id":"PMC_33732354","title":"AMIGO2 as a novel indicator of liver metastasis in patients with colorectal cancer.","date":"2021","source":"Oncology letters","url":"https://pubmed.ncbi.nlm.nih.gov/33732354","citation_count":20,"is_preprint":false},{"pmid":"33524500","id":"PMC_33524500","title":"In vivo selection of highly metastatic human ovarian cancer sublines reveals role for AMIGO2 in intra-peritoneal metastatic regulation.","date":"2021","source":"Cancer letters","url":"https://pubmed.ncbi.nlm.nih.gov/33524500","citation_count":20,"is_preprint":false},{"pmid":"31693896","id":"PMC_31693896","title":"AMIGO2 Scales Dendrite Arbors in the Retina.","date":"2019","source":"Cell reports","url":"https://pubmed.ncbi.nlm.nih.gov/31693896","citation_count":19,"is_preprint":false},{"pmid":"35039535","id":"PMC_35039535","title":"AMIGO2 contained in cancer cell-derived extracellular vesicles enhances the adhesion of liver endothelial cells to cancer cells.","date":"2022","source":"Scientific reports","url":"https://pubmed.ncbi.nlm.nih.gov/35039535","citation_count":17,"is_preprint":false},{"pmid":"27446084","id":"PMC_27446084","title":"IL-17A and TNF-α Increase the Expression of the Antiapoptotic Adhesion Molecule Amigo-2 in Arthritis Synoviocytes.","date":"2016","source":"Frontiers in immunology","url":"https://pubmed.ncbi.nlm.nih.gov/27446084","citation_count":16,"is_preprint":false},{"pmid":"38432455","id":"PMC_38432455","title":"METTL3 knockdown suppresses RA-FLS activation through m6A-YTHDC2-mediated regulation of AMIGO2.","date":"2024","source":"Biochimica et biophysica acta. Molecular basis of disease","url":"https://pubmed.ncbi.nlm.nih.gov/38432455","citation_count":13,"is_preprint":false},{"pmid":"35094710","id":"PMC_35094710","title":"Establishment of an antibody specific for AMIGO2 improves immunohistochemical evaluation of liver metastases and clinical outcomes in patients with colorectal cancer.","date":"2022","source":"Diagnostic pathology","url":"https://pubmed.ncbi.nlm.nih.gov/35094710","citation_count":13,"is_preprint":false},{"pmid":"37438979","id":"PMC_37438979","title":"AMIGO2 attenuates innate cisplatin sensitivity by suppression of GSDME-conferred pyroptosis in non-small cell lung cancer.","date":"2023","source":"Journal of cellular and molecular medicine","url":"https://pubmed.ncbi.nlm.nih.gov/37438979","citation_count":12,"is_preprint":false},{"pmid":"35843033","id":"PMC_35843033","title":"Liver Metastasis Formation Is Defined by AMIGO2 Expression via Adhesion to Hepatic Endothelial Cells in Human Gastric and Colorectal Cancer Cells.","date":"2022","source":"Pathology, research and practice","url":"https://pubmed.ncbi.nlm.nih.gov/35843033","citation_count":12,"is_preprint":false},{"pmid":"38360124","id":"PMC_38360124","title":"The METTL3-m6A-YTHDC1-AMIGO2 axis contributes to cell proliferation and migration in esophageal squamous cell carcinoma.","date":"2024","source":"Gene","url":"https://pubmed.ncbi.nlm.nih.gov/38360124","citation_count":11,"is_preprint":false},{"pmid":"8407808","id":"PMC_8407808","title":"The degA gene product accelerates degradation of Bacillus subtilis phosphoribosylpyrophosphate amidotransferase in Escherichia coli.","date":"1993","source":"Journal of bacteriology","url":"https://pubmed.ncbi.nlm.nih.gov/8407808","citation_count":10,"is_preprint":false},{"pmid":"28119027","id":"PMC_28119027","title":"AMIGO2 modulates T cell functions and its deficiency in mice ameliorates experimental autoimmune encephalomyelitis.","date":"2017","source":"Brain, behavior, and immunity","url":"https://pubmed.ncbi.nlm.nih.gov/28119027","citation_count":8,"is_preprint":false},{"pmid":"28693424","id":"PMC_28693424","title":"Bacillus subtilis IolQ (DegA) is a transcriptional repressor of iolX encoding NAD+-dependent scyllo-inositol dehydrogenase.","date":"2017","source":"BMC microbiology","url":"https://pubmed.ncbi.nlm.nih.gov/28693424","citation_count":7,"is_preprint":false},{"pmid":"38189855","id":"PMC_38189855","title":"AMIGO2 is a pivotal therapeutic target related to M2 polarization of macrophages in pancreatic ductal adenocarcinoma.","date":"2024","source":"Aging","url":"https://pubmed.ncbi.nlm.nih.gov/38189855","citation_count":6,"is_preprint":false},{"pmid":"37323247","id":"PMC_37323247","title":"AMIGO2 expression as a predictor of recurrence in cervical cancer with intermediate risk.","date":"2023","source":"Molecular and clinical oncology","url":"https://pubmed.ncbi.nlm.nih.gov/37323247","citation_count":5,"is_preprint":false},{"pmid":"25362569","id":"PMC_25362569","title":"Properties and distribution of a metallo-β-lactamase (ALI-1) from the fish pathogen Aliivibrio salmonicida LFI1238.","date":"2014","source":"The Journal of antimicrobial chemotherapy","url":"https://pubmed.ncbi.nlm.nih.gov/25362569","citation_count":5,"is_preprint":false},{"pmid":"39049987","id":"PMC_39049987","title":"Role of AMIGO2 in cancer progression: Novel insights (Review).","date":"2024","source":"Oncology letters","url":"https://pubmed.ncbi.nlm.nih.gov/39049987","citation_count":4,"is_preprint":false},{"pmid":"39379686","id":"PMC_39379686","title":"AMIGO2 enhances the invasive potential of colorectal cancer by inducing EMT.","date":"2024","source":"Cancer gene therapy","url":"https://pubmed.ncbi.nlm.nih.gov/39379686","citation_count":4,"is_preprint":false},{"pmid":"40155010","id":"PMC_40155010","title":"Hepatic Stellate Cells Activated by Cancer Cell-derived AMIGO2-containing Small Extracellular Vesicles Promote Cancer Cell Migration by Producing IL-8.","date":"2025","source":"Anticancer research","url":"https://pubmed.ncbi.nlm.nih.gov/40155010","citation_count":4,"is_preprint":false},{"pmid":"38978149","id":"PMC_38978149","title":"Knockdown of AMIGO2 suppresses proliferation and migration through regulating PPAR-γ in bladder cancer.","date":"2024","source":"Hereditas","url":"https://pubmed.ncbi.nlm.nih.gov/38978149","citation_count":3,"is_preprint":false},{"pmid":"29718531","id":"PMC_29718531","title":"Loss of AMIGO2 causes dramatic damage to cardiac preservation after ischemic injury.","date":"2018","source":"Cardiology journal","url":"https://pubmed.ncbi.nlm.nih.gov/29718531","citation_count":3,"is_preprint":false},{"pmid":"35727300","id":"PMC_35727300","title":"Adhesion molecule Amigo2 is involved in the fasciculation process of the fasciculus retroflexus.","date":"2022","source":"Developmental dynamics : an official publication of the American Association of Anatomists","url":"https://pubmed.ncbi.nlm.nih.gov/35727300","citation_count":3,"is_preprint":false},{"pmid":"39523830","id":"PMC_39523830","title":"AMIGO2 characterizes cancer-associated fibroblasts in metastatic colon cancer and induces the release of paracrine active tumorigenic secretomes.","date":"2024","source":"The Journal of pathology","url":"https://pubmed.ncbi.nlm.nih.gov/39523830","citation_count":2,"is_preprint":false},{"pmid":"39548119","id":"PMC_39548119","title":"Prevention of liver metastasis via the pharmacological suppression of AMIGO2 expression in tumor cells.","date":"2024","source":"Scientific reports","url":"https://pubmed.ncbi.nlm.nih.gov/39548119","citation_count":2,"is_preprint":false},{"pmid":"38811439","id":"PMC_38811439","title":"AMIGO2 is involved in the spread of peritoneal metastasis in serous ovarian cancer via promoting adhesion to the peritoneal mesothelial cells.","date":"2024","source":"International journal of clinical oncology","url":"https://pubmed.ncbi.nlm.nih.gov/38811439","citation_count":2,"is_preprint":false},{"pmid":"41254143","id":"PMC_41254143","title":"Metabolomic and transcriptomic profiling of HNSCC identifies AMIGO2 as a therapeutic target modulating tumor microenvironment.","date":"2025","source":"NPJ precision oncology","url":"https://pubmed.ncbi.nlm.nih.gov/41254143","citation_count":1,"is_preprint":false},{"pmid":"40998908","id":"PMC_40998908","title":"AMIGO2 accelerates tumor progression by inducing a cancer stem cell-like phenotype.","date":"2025","source":"Scientific reports","url":"https://pubmed.ncbi.nlm.nih.gov/40998908","citation_count":0,"is_preprint":false},{"pmid":"8299970","id":"PMC_8299970","title":"The alkane-inducible Candida maltosa ALI1 gene product is an NADH:ubiquinone oxidoreductase subunit homologue.","date":"1993","source":"Gene","url":"https://pubmed.ncbi.nlm.nih.gov/8299970","citation_count":0,"is_preprint":false},{"pmid":"31233685","id":"PMC_31233685","title":"[In vivo study of plant protein products from soybeans and non-alkaloid lupine Dega variety].","date":"2019","source":"Voprosy pitaniia","url":"https://pubmed.ncbi.nlm.nih.gov/31233685","citation_count":0,"is_preprint":false},{"pmid":"41732354","id":"PMC_41732354","title":"An open image dataset of Indonesian soybean seed varieties (Anjasmoro, Grobogan, DEGA-1) for agricultural research and machine learning applications.","date":"2026","source":"Data in brief","url":"https://pubmed.ncbi.nlm.nih.gov/41732354","citation_count":0,"is_preprint":false},{"pmid":null,"id":"bio_10.1101_2025.06.11.659103","title":"Hippocampal Commissural Circuitry Shows Asymmetric cAMP-Dependent Synaptic Plasticity","date":"2025-06-11","source":"bioRxiv","url":"https://doi.org/10.1101/2025.06.11.659103","citation_count":0,"is_preprint":true},{"pmid":null,"id":"bio_10.1101_2024.11.07.622463","title":"Perineuronal nets on CA2 pyramidal cells and parvalbumin-expressing cells differentially regulate hippocampal dependent memory","date":"2024-11-08","source":"bioRxiv","url":"https://doi.org/10.1101/2024.11.07.622463","citation_count":0,"is_preprint":true},{"pmid":null,"id":"bio_10.1101_2024.08.12.607550","title":"Exploring adaptive introgression in modern human circadian rhythm genes","date":"2024-08-12","source":"bioRxiv","url":"https://doi.org/10.1101/2024.08.12.607550","citation_count":0,"is_preprint":true}],"cost":{"stage1":{"model":"claude-sonnet-4-6","input_tokens":22055,"output_tokens":5391,"usd":0.073515,"stage1_stop_reason":"end_turn"},"stage2":{"model":"claude-opus-4-8","input_tokens":13996,"output_tokens":5290,"usd":0.101115,"stage2_stop_reason":"end_turn"},"total_usd":0.17463,"stage1_batch_id":"msgbatch_0179Z2pxRYBsXi2xADneafRw","stage2_batch_id":"msgbatch_01LaY9ZaVdgjWArcZo9eqQE5","note":"batch pricing = 50% of standard"},"stage1_raw":"```json\n{\n  \"discoveries\": [\n    {\n      \"year\": 2015,\n      \"finding\": \"AMIGO2 directly binds the pleckstrin homology (PH) domain of PDK1 via amino acid residues 465–474 in AMIGO2, acting as a membrane scaffold that recruits PDK1 to the plasma membrane and promotes Akt activation. Loss of AMIGO2 in endothelial cells leads to apoptosis and inhibition of angiogenesis with Akt inactivation. A synthetic peptide containing AMIGO2 residues 465–474 abrogated the AMIGO2–PDK1 interaction, Akt activation, and pathological angiogenesis in vivo.\",\n      \"method\": \"Co-immunoprecipitation, direct binding assay with synthetic peptides, loss-of-function in endothelial cells, in vivo tumor/retinopathy angiogenesis models\",\n      \"journal\": \"The Journal of cell biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — reciprocal Co-IP, direct peptide competition assay, in vitro apoptosis phenotype, and two in vivo models; multiple orthogonal methods in one study\",\n      \"pmids\": [\"26553931\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2017,\n      \"finding\": \"AMIGO2 interacts with the pseudokinase PTK7, and AMIGO2 regulates PTK7 proteolytic processing. BET bromodomain proteins BRD2/4 occupy a melanoma-specific promoter and super-enhancer at the AMIGO2 locus; BET inhibitor treatment evicts BRD2/4, silences AMIGO2, and alters PTK7 processing. AMIGO2 silencing in melanoma cells induces G1/S cell-cycle arrest followed by apoptosis.\",\n      \"method\": \"Co-immunoprecipitation (AMIGO2–PTK7 interaction), epigenomic profiling (ChIP-seq for BRD2/4), CRISPR genome editing of regulatory elements, BETi treatment, loss-of-function RNA silencing with cell-cycle/apoptosis readouts\",\n      \"journal\": \"Molecular cell\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — reciprocal Co-IP for PTK7 interaction, ChIP-seq + CRISPR for regulatory mechanism, multiple orthogonal methods in one rigorous study\",\n      \"pmids\": [\"29149598\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2004,\n      \"finding\": \"DEGA/AMIGO2 is a type I transmembrane protein that localizes to the cell surface (confirmed by DEGA-GFP fusion construct in 293 cells). Stable antisense-mediated knockdown in gastric adenocarcinoma AGS cells caused altered morphology, increased ploidy, chromosomal instability, decreased cell adhesion and migration, and near-complete abrogation of tumorigenicity in nude mice.\",\n      \"method\": \"GFP fusion localization, stable antisense transfection, nude mouse xenograft, cell adhesion/migration assays, chromosomal analysis\",\n      \"journal\": \"Oncogene\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — direct localization experiment plus multiple functional readouts in a single lab; no reciprocal validation\",\n      \"pmids\": [\"15107827\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2017,\n      \"finding\": \"AMIGO2 expression in tumor cells mediates their preferential adhesion to liver endothelial cells (but not lung endothelial cells), promoting liver metastasis. siRNA-mediated knockdown reduced liver endothelial cell adhesion in vitro, and forced expression of AMIGO2 in low-AMIGO2 cells increased liver endothelial cell adhesion and liver metastasis in vivo.\",\n      \"method\": \"siRNA knockdown, forced expression, ex vivo imaging, intrasplenic injection metastasis model\",\n      \"journal\": \"Scientific reports\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — bidirectional manipulation (KD and overexpression), in vitro adhesion assay plus in vivo metastasis model, replicated in later studies\",\n      \"pmids\": [\"28272394\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"AMIGO2 functions as a homophilic cell-surface protein in the retina; starburst amacrine cells (SACs) and rod bipolar cells (RBCs) express AMIGO2, and Amigo2 knockout mice display expanded SAC and RBC dendrite arbors while other retinal neuron arbors remain unchanged. Increased SAC dendrite coverage was accompanied by increased direction selectivity of downstream direction-selective ganglion cells (DSGCs), identifying AMIGO2 as a cell-type-specific dendritic scaling factor.\",\n      \"method\": \"Amigo2 knockout mouse, morphological analysis of dendrite arbors, electrophysiology of DSGCs, immunostaining\",\n      \"journal\": \"Cell reports\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — clean KO with specific cellular phenotype, multiple neuron types analyzed as controls, functional electrophysiology readout, rigorous loss-of-function study\",\n      \"pmids\": [\"31693896\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2017,\n      \"finding\": \"AMIGO2 modulates T-cell functions: Amigo2 knockout impairs T-cell infiltration into secondary lymphoid organs and dampens Th-cell activation, while promoting splenic Th-cell proliferation. Mechanistically, AMIGO2 overexpression in 293T cells dampens NF-κB transcriptional activity, and AMIGO2 deficiency enhances Akt but suppresses GSK-3β phosphorylation and promotes nuclear translocation of NF-κB and NFAT1 in Th cells. Amigo2 KO mice exhibit ameliorated experimental autoimmune encephalomyelitis (EAE).\",\n      \"method\": \"Amigo2 knockout mice, adoptive T-cell transfer, NF-κB reporter assay, Western blot for Akt/GSK-3β phosphorylation, nuclear fractionation for NF-κB/NFAT1\",\n      \"journal\": \"Brain, behavior, and immunity\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — KO mouse with defined cellular and molecular phenotypes, NF-κB reporter assay, multiple signaling readouts; single lab\",\n      \"pmids\": [\"28119027\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2016,\n      \"finding\": \"IL-17A and TNF-α synergistically upregulate AMIGO2 expression in RA synoviocytes via an ERK-dependent mechanism (JNK inhibits AMIGO2 induction). Elevated AMIGO2 promotes cell survival and reduces apoptosis (cadmium-induced toxicity). HMGB1 in inflammatory conditions further increases AMIGO2 expression and reduces cell toxicity.\",\n      \"method\": \"Cytokine stimulation of synoviocytes, ERK/JNK pharmacological inhibition, apoptosis assay with cadmium, co-culture experiments\",\n      \"journal\": \"Frontiers in immunology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 3 / Moderate — pharmacological pathway dissection plus functional apoptosis readout, multiple conditions tested; single lab\",\n      \"pmids\": [\"27446084\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"AMIGO2 contained in cancer cell-derived extracellular vesicles (EVs) is internalized by human hepatic sinusoidal endothelial cells (HHSECs) and, once inside, significantly enhances adhesion of HHSECs to cancer cells (including cells that themselves lack AMIGO2 expression), identifying a paracrine mechanism for liver metastasis initiation.\",\n      \"method\": \"Western blot of EVs, EV internalization assay, cell adhesion assay with AMIGO2-overexpressing vs. control EVs\",\n      \"journal\": \"Scientific reports\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 3 / Moderate — functional EV adhesion assay with gain-of-function EV treatment; multiple cancer cell lines tested; single lab\",\n      \"pmids\": [\"35039535\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"METTL3-mediated m6A methylation in the 5'-UTR of AMIGO2 mRNA recruits the reader YTHDC2, stabilizing/upregulating AMIGO2 expression in RA fibroblast-like synoviocytes. METTL3 knockdown reduces m6A on AMIGO2 5'-UTR, decreases YTHDC2–AMIGO2 mRNA interaction, and lowers AMIGO2 protein, suppressing RA-FLS proliferation and migration; AMIGO2 overexpression rescues these phenotypes.\",\n      \"method\": \"m6A-seq/MeRIP for modification site mapping, RIP (YTHDC2–AMIGO2 mRNA), siRNA knockdown of METTL3/YTHDC2, rescue overexpression of AMIGO2\",\n      \"journal\": \"Biochimica et biophysica acta. Molecular basis of disease\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — MeRIP for m6A site, RIP for reader–mRNA interaction, rescue experiment; single lab\",\n      \"pmids\": [\"38432455\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"In esophageal squamous cell carcinoma, METTL3-mediated m6A modification in the 5'-UTR of AMIGO2 pre-mRNA is read by YTHDC1, which regulates AMIGO2 splicing. METTL3 knockdown reduces m6A on AMIGO2 pre-mRNA, impairs YTHDC1-mediated splicing, and decreases AMIGO2 expression, suppressing ESCC proliferation and migration; AMIGO2 overexpression rescues these effects.\",\n      \"method\": \"MeRIP (m6A site mapping), RIP (YTHDC1–AMIGO2 pre-mRNA), splicing analysis, siRNA knockdown/rescue overexpression, in vitro and in vivo tumor models\",\n      \"journal\": \"Gene\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — MeRIP, RIP, splicing assay, rescue experiment; single lab; distinct reader (YTHDC1) from the RA study (YTHDC2)\",\n      \"pmids\": [\"38360124\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2018,\n      \"finding\": \"AMIGO2 deficiency in mice (KO) causes increased cardiomyocyte apoptosis, reduced proliferation, and reduced angiogenesis after myocardial infarction, with weaker cardiac function and larger infarct scar. Molecularly, AMIGO2 KO increases active caspase-3 and decreases PDK1, p-Akt, Bcl-2/Bax, and VEGF expression, consistent with inactivation of the PDK1/Akt survival pathway in cardiomyocytes.\",\n      \"method\": \"AMIGO2 KO mouse MI model, echocardiography, TUNEL/Ki67/immunohistochemistry, Western blot for PDK1/Akt/Bcl-2/Bax/VEGF\",\n      \"journal\": \"Cardiology journal\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — clean KO mouse with defined cardiac phenotype and molecular pathway readouts; single lab\",\n      \"pmids\": [\"29718531\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"AMIGO2 suppresses innate cisplatin sensitivity in NSCLC by inhibiting cisplatin-induced activation of caspase-8, caspase-9, and caspase-3, thereby preventing GSDME cleavage and pyroptosis. AMIGO2 acts by stimulating the PDK1/Akt (T308) signaling axis. Alteration of AMIGO2 expression changed cisplatin-induced pyroptosis both in vitro and in vivo.\",\n      \"method\": \"AMIGO2 knockdown/overexpression, caspase activity assays, GSDME cleavage Western blot, PDK1/Akt phosphorylation assays, in vivo xenograft cisplatin treatment\",\n      \"journal\": \"Journal of cellular and molecular medicine\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — bidirectional manipulation, multiple pathway readouts (caspase cascade, GSDME, PDK1/Akt), in vitro and in vivo; single lab\",\n      \"pmids\": [\"37438979\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"AMIGO2 expression in human gastric and colorectal cancer cells is directly associated with their adhesion to human hepatic sinusoidal endothelial cells (HHSECs) in vitro. Constitutive AMIGO2 knockdown clones showed significantly attenuated adhesion to HHSECs and suppressed liver metastasis in nude mice following intrasplenic inoculation, validating the mouse findings in human cancer cells.\",\n      \"method\": \"AMIGO2 knockdown (constitutive clones), cell adhesion assay to HHSECs, intrasplenic injection nude mouse liver metastasis model\",\n      \"journal\": \"Pathology, research and practice\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — loss-of-function with defined in vitro and in vivo readouts; single lab; extends prior mouse data to human cells\",\n      \"pmids\": [\"35843033\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"AMIGO2 promotes EMT in colorectal cancer cells via activation of the TGFβ/Smad signaling pathway. Treatment with TGFβ receptor inhibitor LY2109761 suppressed AMIGO2-induced EMT. In CRC tissue samples, AMIGO2 was found at the invasive front where it localizes to the nucleus and associates with EMT marker expression, suggesting nuclear translocation of AMIGO2 as a mechanism for inducing EMT.\",\n      \"method\": \"AMIGO2 overexpression/knockdown, TGFβ/Smad pathway inhibitor treatment, immunohistochemistry/nuclear localization analysis in CRC clinical samples, EMT marker Western blot\",\n      \"journal\": \"Cancer gene therapy\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 3 / Moderate — pharmacological pathway inhibition plus clinical sample localization data; nuclear localization is observational without mechanistic reconstitution; single lab\",\n      \"pmids\": [\"39379686\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"AMIGO2-containing small extracellular vesicles (sEVs) from gastric cancer cells activate hepatic stellate cells (HSCs) via NF-κB nuclear translocation, inducing IL-8 secretion. The resulting IL-8-rich conditioned medium significantly enhances gastric cancer cell migration; neutralizing IL-8 with antibodies suppresses this migration, establishing an AMIGO2 sEV→HSC→IL-8→cancer cell migration axis in liver metastasis.\",\n      \"method\": \"sEV isolation, AMIGO2 overexpression, NF-κB nuclear translocation assay, chemokine array, IL-8 neutralizing antibody, migration assay\",\n      \"journal\": \"Anticancer research\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 3 / Moderate — functional rescue with neutralizing antibody confirms IL-8 role; NF-κB translocation assay; single lab\",\n      \"pmids\": [\"40155010\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"AMIGO2 expression in cancer-associated fibroblasts (CAFs) of colorectal cancer is induced by TGF-β. Conditioned media from NAFs with AMIGO2 overexpression enhanced CRC tumor cell proliferation and migration; siRNA-mediated inhibition of AMIGO2 in CAFs attenuated these effects, showing AMIGO2 regulates paracrine tumorigenic secretomes in CAFs.\",\n      \"method\": \"TGF-β stimulation, AMIGO2 overexpression in NAFs, siRNA knockdown in CAFs, conditioned media transfer, proliferation/migration assays\",\n      \"journal\": \"The Journal of pathology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 3 / Moderate — bidirectional manipulation (OE and KD) with conditioned media functional readout; single lab\",\n      \"pmids\": [\"39523830\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"AMIGO2 knockdown in bladder cancer cells suppresses PPAR-γ expression; PPAR-γ overexpression rescues the proliferation and migration suppressed by AMIGO2 knockdown, placing PPAR-γ downstream of AMIGO2 in a signaling axis controlling bladder cancer cell growth.\",\n      \"method\": \"siRNA/lentiviral AMIGO2 knockdown, RNA-seq, PPAR-γ Western blot, PPAR-γ rescue overexpression, cell proliferation/migration/xenograft assays\",\n      \"journal\": \"Hereditas\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 / Weak — rescue experiment placing PPAR-γ downstream, but pathway connection is inferred from RNA-seq without direct mechanistic assay; single lab\",\n      \"pmids\": [\"38978149\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"AMIGO2 knockdown via CRISPR/Cas9 in high-grade serous ovarian cancer cells reduces sphere-forming potential, adhesion, and invasion in vitro, and significantly attenuates intraperitoneal metastasis in vivo.\",\n      \"method\": \"CRISPR/Cas9 knockdown, sphere formation assay, cell adhesion/invasion assay, in vivo IP metastasis model\",\n      \"journal\": \"Cancer letters\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — CRISPR-based loss-of-function with multiple orthogonal in vitro and in vivo readouts; single lab\",\n      \"pmids\": [\"33524500\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"Adhesion molecule Amigo2 is selectively expressed in medial habenula neurons and mediates fasciculation of the fasciculus retroflexus. Amigo2 loss-of-function in mice causes defasciculation of medial habenular axons; gain-of-function generates a more condensed tract and rescues the KO phenotype. AMIGO2 does not alter the course of habenular fibers, only their fasciculation.\",\n      \"method\": \"Amigo2 KO mouse, gain-of-function overexpression rescue experiment, tract tracing/morphological analysis\",\n      \"journal\": \"Developmental dynamics\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — bidirectional manipulation (KO and rescue OE) with specific morphological phenotype; single lab\",\n      \"pmids\": [\"35727300\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"AMIGO2 is a type I transmembrane leucine-rich repeat/Ig-domain cell-surface scaffold protein that promotes cell survival and adhesion through multiple mechanisms: it directly binds the PH domain of PDK1 (via residues 465–474) to recruit PDK1 to the plasma membrane and activate Akt, thereby suppressing apoptosis in endothelial and cancer cells; it interacts with and regulates proteolytic processing of the pseudokinase PTK7 in melanoma; it mediates homophilic adhesion to scale dendrite arbors in specific retinal neuron types; it promotes tumor cell adhesion to hepatic endothelial cells (and, in EVs, activates hepatic stellate cells to secrete IL-8) to drive liver metastasis; it induces EMT via TGFβ/Smad signaling at the invasive front; and its expression is post-transcriptionally regulated by METTL3-mediated m6A modification read by YTHDC1 (splicing) or YTHDC2 (stability) in a context-dependent manner.\"\n}\n```","stage2_raw":"{\n  \"mechanistic_narrative\": \"AMIGO2 is a type I transmembrane cell-surface protein that functions as a membrane scaffold and homophilic adhesion molecule to promote cell survival, adhesion, and tissue patterning [#0, #2, #4]. Its central survival mechanism is direct binding of the PH domain of PDK1 through AMIGO2 residues 465–474, which recruits PDK1 to the plasma membrane and activates Akt; disrupting this interaction with a competing peptide abrogates Akt activation, triggers apoptosis, and blocks pathological angiogenesis [#0]. This PDK1/Akt axis underlies AMIGO2's protective roles in cardiomyocytes after myocardial infarction [#10] and its suppression of cisplatin-induced caspase activation, GSDME cleavage, and pyroptosis in lung cancer [#11]. As a homophilic surface adhesion factor, AMIGO2 scales dendrite arbors of specific retinal neuron types and tunes downstream direction selectivity [#4], and mediates fasciculation of medial habenular axons in the fasciculus retroflexus [#18]. In cancer, AMIGO2 promotes tumor cell adhesion to hepatic sinusoidal endothelial cells to drive liver metastasis [#3, #12], a process that also proceeds paracrine: AMIGO2 carried in tumor-derived extracellular vesicles is internalized by hepatic endothelial cells to enhance their adhesiveness [#7] and activates hepatic stellate cells via NF-κB to secrete IL-8 that boosts cancer cell migration [#14]. AMIGO2 additionally interacts with the pseudokinase PTK7 and regulates its proteolytic processing in melanoma, where it is driven by a BET-dependent super-enhancer [#1], and it induces EMT through TGFβ/Smad signaling at the invasive front [#13]. AMIGO2 expression is post-transcriptionally controlled by METTL3-deposited m6A in its 5′-UTR, read context-dependently by YTHDC1 to regulate splicing or YTHDC2 to regulate stability [#9, #8].\",\n  \"teleology\": [\n    {\n      \"year\": 2004,\n      \"claim\": \"Established AMIGO2 (DEGA) as a cell-surface type I transmembrane protein required for tumorigenicity, the first functional characterization linking it to adhesion, genomic stability, and cancer cell behavior.\",\n      \"evidence\": \"GFP-fusion localization and stable antisense knockdown with xenograft, adhesion, and chromosomal readouts in gastric adenocarcinoma cells\",\n      \"pmids\": [\"15107827\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"No molecular mechanism for adhesion or ploidy effects\", \"No reciprocal validation of localization\", \"Binding partners unidentified\"]\n    },\n    {\n      \"year\": 2015,\n      \"claim\": \"Defined the molecular survival mechanism: AMIGO2 directly binds the PDK1 PH domain via residues 465–474 to recruit PDK1 to the membrane and activate Akt, explaining its anti-apoptotic and pro-angiogenic role.\",\n      \"evidence\": \"Reciprocal Co-IP, synthetic peptide competition, endothelial loss-of-function, and in vivo angiogenesis models\",\n      \"pmids\": [\"26553931\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Structural basis of the AMIGO2–PDK1 interface not resolved\", \"How surface AMIGO2 spatially organizes cytoplasmic PDK1 unclear\"]\n    },\n    {\n      \"year\": 2016,\n      \"claim\": \"Showed AMIGO2 is an inducible cytoprotective gene downstream of inflammatory cytokines, linking IL-17A/TNF-α/ERK signaling to AMIGO2-mediated survival in synoviocytes.\",\n      \"evidence\": \"Cytokine stimulation with ERK/JNK pharmacological inhibition and cadmium-induced apoptosis assay in RA synoviocytes\",\n      \"pmids\": [\"27446084\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Transcription factors mediating ERK-dependent induction not identified\", \"Downstream survival effector in synoviocytes not tied to PDK1/Akt\"]\n    },\n    {\n      \"year\": 2017,\n      \"claim\": \"Connected AMIGO2 to a second client (PTK7) and revealed its epigenetic regulation, showing a melanoma super-enhancer drives AMIGO2 to sustain proliferation and survival.\",\n      \"evidence\": \"Co-IP for AMIGO2–PTK7, ChIP-seq for BRD2/4, CRISPR editing of regulatory elements, and BETi with cell-cycle/apoptosis readouts\",\n      \"pmids\": [\"29149598\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"How AMIGO2 controls PTK7 proteolytic processing mechanistically unknown\", \"Whether PTK7 regulation requires the PDK1/Akt axis unclear\"]\n    },\n    {\n      \"year\": 2017,\n      \"claim\": \"Identified organ-tropic adhesion as a metastasis mechanism: AMIGO2 mediates selective tumor cell adhesion to liver (not lung) endothelium to drive liver metastasis.\",\n      \"evidence\": \"siRNA knockdown, forced expression, and intrasplenic injection metastasis model\",\n      \"pmids\": [\"28272394\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Endothelial counter-receptor for AMIGO2 not identified\", \"Molecular basis of liver-specific tropism unresolved\"]\n    },\n    {\n      \"year\": 2017,\n      \"claim\": \"Extended AMIGO2's role to adaptive immunity, showing it constrains T-cell signaling through Akt/GSK-3β and NF-κB/NFAT1 to shape autoimmune responses.\",\n      \"evidence\": \"Amigo2 KO mice, adoptive transfer, NF-κB reporter, signaling Westerns, nuclear fractionation, and EAE model\",\n      \"pmids\": [\"28119027\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Whether AMIGO2 acts cell-autonomously in T cells via PDK1 unclear\", \"Direct partners in T cells not defined\"]\n    },\n    {\n      \"year\": 2018,\n      \"claim\": \"Demonstrated AMIGO2's PDK1/Akt survival axis operates in vivo in cardiomyocytes, linking AMIGO2 loss to worse cardiac outcomes after infarction.\",\n      \"evidence\": \"Amigo2 KO mouse MI model with echocardiography, TUNEL/Ki67, and PDK1/Akt/Bcl-2/Bax/VEGF Westerns\",\n      \"pmids\": [\"29718531\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Cell-autonomous vs. paracrine contribution not dissected\", \"Direct membrane scaffolding in cardiomyocytes not shown\"]\n    },\n    {\n      \"year\": 2019,\n      \"claim\": \"Revealed AMIGO2 as a cell-type-specific homophilic dendritic scaling factor in the retina, with functional consequences for neural circuit output.\",\n      \"evidence\": \"Amigo2 KO mouse, dendrite arbor morphometry across neuron types, and DSGC electrophysiology\",\n      \"pmids\": [\"31693896\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Molecular pathway downstream of homophilic adhesion in neurons unknown\", \"Whether PDK1/Akt is involved in dendrite scaling untested\"]\n    },\n    {\n      \"year\": 2021,\n      \"claim\": \"Generalized AMIGO2's pro-metastatic adhesion/invasion role to ovarian cancer using CRISPR loss-of-function.\",\n      \"evidence\": \"CRISPR/Cas9 knockdown with sphere formation, adhesion/invasion, and intraperitoneal metastasis assays\",\n      \"pmids\": [\"33524500\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Molecular effectors of sphere formation and invasion not mapped\", \"No link to specific signaling partner in this context\"]\n    },\n    {\n      \"year\": 2022,\n      \"claim\": \"Defined a paracrine, EV-borne mechanism of metastasis, showing AMIGO2 transferred in tumor EVs reprograms recipient hepatic endothelium independent of tumor-cell AMIGO2.\",\n      \"evidence\": \"EV Western blot, internalization assay, and adhesion assay with AMIGO2-overexpressing vs. control EVs\",\n      \"pmids\": [\"35039535\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Intracellular pathway activated in recipient endothelium unknown\", \"Whether internalized AMIGO2 engages endothelial PDK1 untested\"]\n    },\n    {\n      \"year\": 2022,\n      \"claim\": \"Validated AMIGO2-dependent hepatic endothelial adhesion and liver metastasis in human gastric and colorectal cancer cells.\",\n      \"evidence\": \"Constitutive AMIGO2 knockdown clones with HHSEC adhesion assays and intrasplenic metastasis model\",\n      \"pmids\": [\"35843033\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Endothelial binding partner still unidentified\", \"Single-lab confirmation\"]\n    },\n    {\n      \"year\": 2022,\n      \"claim\": \"Placed AMIGO2 in the stromal compartment, showing TGF-β-induced AMIGO2 in cancer-associated fibroblasts drives a pro-tumorigenic paracrine secretome.\",\n      \"evidence\": \"TGF-β stimulation, AMIGO2 overexpression/knockdown in fibroblasts, and conditioned-media proliferation/migration assays\",\n      \"pmids\": [\"39523830\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Secreted factors mediating the effect not identified\", \"Whether CAF AMIGO2 signals via the same PDK1 axis unknown\"]\n    },\n    {\n      \"year\": 2022,\n      \"claim\": \"Established AMIGO2 as a developmental axon fasciculation factor in medial habenula, distinguishing fasciculation from axon guidance.\",\n      \"evidence\": \"Amigo2 KO and gain-of-function rescue with tract tracing in mice\",\n      \"pmids\": [\"35727300\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Whether fasciculation is homophilic adhesion-dependent not directly shown\", \"Downstream signaling unknown\"]\n    },\n    {\n      \"year\": 2023,\n      \"claim\": \"Showed AMIGO2 suppresses chemotherapy-induced pyroptosis, linking its PDK1/Akt activity to evasion of caspase/GSDME-mediated death.\",\n      \"evidence\": \"Bidirectional manipulation with caspase activity, GSDME cleavage, PDK1/Akt phosphorylation, and xenograft cisplatin readouts in NSCLC\",\n      \"pmids\": [\"37438979\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Direct mechanistic step from Akt to caspase-8/9 suppression not resolved\", \"Single lab\"]\n    },\n    {\n      \"year\": 2024,\n      \"claim\": \"Resolved post-transcriptional control of AMIGO2, showing METTL3-deposited 5′-UTR m6A is read by YTHDC2 (stability) or YTHDC1 (splicing) in a context-dependent manner.\",\n      \"evidence\": \"MeRIP m6A mapping, RIP for reader–mRNA interaction, and METTL3/reader knockdown with AMIGO2 rescue in RA synoviocytes and esophageal carcinoma\",\n      \"pmids\": [\"38432455\", \"38360124\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"What determines YTHDC1 vs. YTHDC2 reader choice unknown\", \"Upstream signals controlling METTL3 activity on AMIGO2 not defined\"]\n    },\n    {\n      \"year\": 2024,\n      \"claim\": \"Identified an EMT-promoting role via TGFβ/Smad signaling and an unexpected nuclear localization of AMIGO2 at the tumor invasive front.\",\n      \"evidence\": \"AMIGO2 overexpression/knockdown, TGFβ receptor inhibitor LY2109761, and IHC of CRC clinical samples\",\n      \"pmids\": [\"39379686\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Mechanism by which a transmembrane protein reaches the nucleus not established\", \"Direct role of nuclear AMIGO2 in transcription not reconstituted\"]\n    },\n    {\n      \"year\": 2025,\n      \"claim\": \"Completed a paracrine liver-metastasis circuit, defining an AMIGO2 sEV → hepatic stellate cell NF-κB → IL-8 → cancer cell migration axis.\",\n      \"evidence\": \"sEV isolation, NF-κB translocation assay, chemokine array, and IL-8 neutralization with migration assays\",\n      \"pmids\": [\"40155010\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"How EV AMIGO2 activates NF-κB in stellate cells unknown\", \"Single lab\"]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"The identity of AMIGO2's extracellular binding partner(s)—including its endothelial counter-receptor in metastasis and the basis of homophilic adhesion—and the mechanism linking surface or EV AMIGO2 to nuclear/EMT functions remain unresolved.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"No extracellular ligand/counter-receptor identified\", \"No structure of AMIGO2 ectodomain or its homophilic interface\", \"Mechanism of nuclear translocation of a transmembrane protein unexplained\"]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0098631\", \"supporting_discovery_ids\": [4, 18]},\n      {\"term_id\": \"GO:0060089\", \"supporting_discovery_ids\": [0]},\n      {\"term_id\": \"GO:0060090\", \"supporting_discovery_ids\": [0]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005886\", \"supporting_discovery_ids\": [0, 2]},\n      {\"term_id\": \"GO:0031410\", \"supporting_discovery_ids\": [7, 14]},\n      {\"term_id\": \"GO:0005634\", \"supporting_discovery_ids\": [13]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-162582\", \"supporting_discovery_ids\": [0, 11]},\n      {\"term_id\": \"R-HSA-1643685\", \"supporting_discovery_ids\": [3, 12]},\n      {\"term_id\": \"R-HSA-5357801\", \"supporting_discovery_ids\": [0, 11]},\n      {\"term_id\": \"R-HSA-1266738\", \"supporting_discovery_ids\": [4, 18]}\n    ],\n    \"complexes\": [],\n    \"partners\": [\"PDK1\", \"PTK7\"],\n    \"other_free_text\": []\n  }\n}","audit_flag":null,"evaluation":{"pairwise":"win","faith_supported":7,"faith_total":7,"faith_pct":100.0}}