{"gene":"AFAP1L2","run_date":"2026-06-09T22:02:42","timeline":{"discoveries":[{"year":2007,"finding":"XB130 (AFAP1L2) was cloned as a novel adaptor protein containing SH2- and SH3-binding motifs, two pleckstrin homology domains, a coiled-coil region, and multiple phosphorylation sites. Endogenous XB130 interacts with c-Src tyrosine kinase, and co-expression in COS-7 cells activated c-Src and elevated tyrosine phosphorylation of multiple proteins including XB130 itself. An N-terminal deletion mutant (XB130ΔN) reduced its ability to mediate Src signal transduction. Knockdown in lung epithelial cells reduced c-Src activity, IL-8 production, EGF-induced phosphorylation of Akt and GSK3β, and altered cell cycles.","method":"Co-immunoprecipitation, co-expression in COS-7 cells, siRNA knockdown, reporter assay (SRE/AP-1), deletion mutagenesis, Western blot","journal":"The Journal of biological chemistry","confidence":"High","confidence_rationale":"Tier 2 / Strong — reciprocal Co-IP, mutagenesis, functional knockdown with multiple orthogonal methods in founding paper","pmids":["17412687"],"is_preprint":false},{"year":2008,"finding":"RET/PTC oncogenic kinase phosphorylates XB130 predominantly at tyrosine 54, which promotes XB130 association with the p85α subunit of PI3-kinase via p85α SH2 domains, thereby linking RET/PTC signaling to PI3K/Akt activation. Knockdown of XB130 in TPC1 papillary thyroid cancer cells strongly reduced Akt activity without altering ERK1/2 phosphorylation, and inhibited cell-cycle progression and survival.","method":"Co-immunoprecipitation, site-directed mutagenesis (Y54), siRNA knockdown, Western blot for Akt/ERK phosphorylation, cell cycle analysis","journal":"Oncogene","confidence":"High","confidence_rationale":"Tier 1-2 / Strong — mutagenesis identifying specific phosphorylation site plus reciprocal Co-IP and functional KD with defined pathway readout","pmids":["19060924"],"is_preprint":false},{"year":2010,"finding":"XB130 translocates to the cell periphery in response to EGF, wounding, and constitutively active Rac, localizing to the lamellipodial F-actin meshwork. Structure-function analysis showed that both the N-terminus (167 aa) and C-terminus (63 aa) are required for translocation to lamellipodia, while PH domains and Src-targeted tyrosines are dispensable. Silencing XB130 in TPC1 cells decreased wound closure rate, inhibited Matrigel invasion, reduced lamellipodial persistence, and slowed spreading.","method":"Live-cell imaging, constitutively active Rac expression, pharmacological inhibitors, deletion mutagenesis, siRNA knockdown, wound-healing and Matrigel invasion assays","journal":"Journal of cell science","confidence":"High","confidence_rationale":"Tier 2 / Strong — direct localization experiment with functional consequence, structure-function mutagenesis, multiple orthogonal methods","pmids":["21084565"],"is_preprint":false},{"year":2010,"finding":"Knockdown of XB130 in WRO thyroid cancer cells inhibited G1-S phase progression, induced spontaneous apoptosis, and enhanced intrinsic and extrinsic apoptotic stimulus-induced cell death. In vivo, XB130 shRNA stably transfected WRO cells formed significantly smaller tumors in nude mice. Microarray identified 246 significantly changed genes, including 57 related to cell proliferation or survival.","method":"siRNA/shRNA knockdown, cell cycle analysis, apoptosis assays, xenograft mouse model, microarray gene expression profiling","journal":"The American journal of pathology","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — KO/KD with defined phenotypic readouts in vitro and in vivo, single lab","pmids":["21224076"],"is_preprint":false},{"year":2012,"finding":"XB130 mediates cancer cell proliferation and survival through PI3K/Akt downstream signals including phosphorylation of p21Cip1/WAF1, p27Kip1, FOXO3a, and GSK3β (but not FOXO1 or p53), in both RET/PTC-mutant (WRO) and RET/PTC-negative (A549) cancer cells. XB130 can be phosphorylated by multiple protein tyrosine kinases.","method":"siRNA knockdown, Western blot for phosphorylated Akt substrates, cell cycle analysis, caspase cleavage assay","journal":"PloS one","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — functional KD with multiple pathway readouts, single lab, two cell lines","pmids":["22928011"],"is_preprint":false},{"year":2012,"finding":"In rat FRTL-5 thyroid cells, cAMP treatment elevated PI3KAP/XB130 mRNA and protein levels, increased its tyrosine phosphorylation by c-Src (blocked by PP1/PP2), and enhanced its interaction with p85 PI3K, leading to increased PI3K activity. PI3KAP/XB130 knockdown attenuated cAMP-dependent potentiation of IGF-I-induced DNA synthesis. A Y72F mutant incapable of binding p85 PI3K did not enhance IGF-I-induced DNA synthesis, establishing Y72 as the critical p85 PI3K-binding site.","method":"MALDI-TOF MS (protein identification), siRNA knockdown, Src kinase inhibitors (PP1/PP2), point mutagenesis (Y72F), co-immunoprecipitation, PI3K activity assay, DNA synthesis assay","journal":"Molecular endocrinology (Baltimore, Md.)","confidence":"High","confidence_rationale":"Tier 1 / Strong — mutagenesis identifying critical binding residue, in vitro kinase-pathway assay, pharmacological validation, multiple orthogonal methods","pmids":["22496359"],"is_preprint":false},{"year":2013,"finding":"XB130 suppresses expression of tumor-suppressive microRNAs (miR-33a, miR-149, miR-193a-3p) in thyroid cancer cells. These miRNAs target oncogenes MYC (miR-33a), FOSL1 (miR-149), and SLC7A5 (miR-193a-3p) at their 3' UTR, and their upregulation upon XB130 knockdown reduces cancer cell growth.","method":"miRNA array, shRNA knockdown, qRT-PCR validation, ectopic overexpression, miRNA mimic transfection, 3' UTR reporter assay","journal":"PloS one","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — 3'UTR reporter assay confirming direct targeting plus gain/loss of function, single lab","pmids":["23527086"],"is_preprint":false},{"year":2015,"finding":"XB130 interacts with scaffold protein Tks5: the fifth SH3 domain of Tks5 binds to the N-terminus of XB130 (which contains polyproline-rich motifs), and both proteins form a complex with Src tyrosine kinase. Disrupting XB130/Tks5 binding (via XB130 N-terminal deletion mutant or Tks5 SH3-W1108A mutant) decreased cell proliferation and survival and inhibited serum/growth-factor-induced Src activation and downstream PI3K/Akt phosphorylation.","method":"Yeast two-hybrid screening, co-immunoprecipitation (endogenous), structure-function mutagenesis, cell proliferation and apoptosis assays, Western blot","journal":"Molecular biology of the cell","confidence":"High","confidence_rationale":"Tier 2 / Strong — yeast two-hybrid plus reciprocal endogenous Co-IP plus mutagenesis of binding interface, multiple orthogonal methods","pmids":["26446840"],"is_preprint":false},{"year":2015,"finding":"XB130 translocates to lamellipodia and microfilamentous structures in BEAS2B bronchial epithelial cells after NNK stimulation, and overexpression significantly enhances NNK-induced migration. Both the N- and C-termini of XB130 are required for this migration-enhancing function. XB130 overexpression enhanced NNK-induced protein tyrosine phosphorylation and promoted matrix metalloproteinase-14 (MMP-14) translocation to cell motility-associated structures.","method":"Immunofluorescence/localization, overexpression of full-length and truncation mutants, migration assay, Western blot for tyrosine phosphorylation, MMP-14 localization","journal":"Oncotarget","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — direct localization experiment with functional consequence, structure-function analysis, single lab","pmids":["25980441"],"is_preprint":false},{"year":2016,"finding":"XB130 (PI3KAP) directly binds F-actin through its C-terminal region (residues 830-840), and multimerizes via its N-terminal 40 amino acids (shown by Blue native-PAGE and co-IP). Both actin-binding and multimerization are required for XB130 to crosslink F-actin in vitro. Overexpression of XB130 enhanced dextran endocytosis in HEK293 cells, dependent on its actin-binding region.","method":"In vitro F-actin binding and crosslinking assay with recombinant protein, deletion mutagenesis, Blue native-PAGE, co-immunoprecipitation, endocytosis assay (dextran uptake)","journal":"Frontiers in endocrinology","confidence":"High","confidence_rationale":"Tier 1 / Strong — in vitro reconstitution of actin crosslinking, mutagenesis of actin-binding and multimerization domains, functional cellular assay","pmids":["27462298"],"is_preprint":false},{"year":2016,"finding":"In airway epithelial cells, XB130 and Tks5 translocate to the cell membrane in a stimulus-dependent manner upon EGF, PKC activator, or nicotinic acetylcholine receptor ligand stimulation. XB130 co-localizes with lamellipodial marker WAVE2 whereas Tks5 co-localizes with podosome marker N-WASP. XB130 interacts with Rac1 and Tks5 interacts with Cdc42 to promote Rho GTPase activity. Co-expression of both proteins inhibits cell migration; dissociation of the complex promotes lateral (XB130/Rac1) or vertical (Tks5/Cdc42) cell migration.","method":"Immunofluorescence localization, co-immunoprecipitation (XB130-Rac1; Tks5-Cdc42), overexpression and co-overexpression, migration assay","journal":"Oncotarget","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — Co-IP with Rho GTPases plus localization plus functional migration assays, single lab","pmids":["27835612"],"is_preprint":false},{"year":2019,"finding":"PTPRZ (protein tyrosine phosphatase receptor Z) dephosphorylates AFAP1L2 at tyrosine residues in vitro and in HEK293T cells. Pleiotrophin (PTN)-mediated inhibition of PTPRZ enhances tyrosine phosphorylation of AFAP1L2, which activates the PI3K-AKT-mTOR pathway to promote oligodendrocyte precursor cell (OPC) differentiation. Knockdown of AFAP1L2 or PI3K inhibition suppressed OPC differentiation and PTN-induced AKT/mTOR phosphorylation. PTPRZ catalytic-dead knock-in mice showed higher AFAP1L2, AKT, and mTOR phosphorylation and accelerated remyelination.","method":"In vitro phosphatase assay (recombinant PTPRZ on AFAP1L2), co-immunoprecipitation in HEK293T, siRNA knockdown in OPC-like cells, catalytic-dead knock-in mouse model, Western blot for PI3K-AKT-mTOR, immunofluorescence","journal":"Glia","confidence":"High","confidence_rationale":"Tier 1 / Strong — in vitro phosphatase assay plus knock-in mouse model with multiple orthogonal readouts, direct identification of PTPRZ as phosphatase for AFAP1L2","pmids":["30667096"],"is_preprint":false},{"year":2021,"finding":"XB130 in thyrocytes critically regulates folliculogenesis by functioning as a link between the actin filament cortex and microtubule network at the apical membrane. In 3D cultures, XB130 aligns with actin filaments and participates in defining the site of apical membrane formation and lumen coalescence. Xb130-knockout thyrocytes showed delayed folliculogenesis, reduced recruitment of microtubule-associated proteins, and disorganized acetylated tubulin under the apical membrane.","method":"Xb130 knockout mouse, 3D Matrigel culture of primary thyrocytes, immunofluorescence/confocal microscopy, XB130-GFP transfection for live imaging","journal":"Thyroid : official journal of the American Thyroid Association","confidence":"High","confidence_rationale":"Tier 2 / Strong — KO mouse with in vitro 3D culture reconstitution, direct imaging of protein localization at apical membrane, multiple orthogonal methods","pmids":["34652970"],"is_preprint":false},{"year":2021,"finding":"Xb130-knockout mice develop congenital hypothyroidism due to disorganized apical membrane structure of thyrocytes, with diminished thyroglobulin iodination and release. XB130 is localized mainly on the apical membrane of thyroid follicles. Growth retardation in KO mice could be rescued by exogenous thyroxine supplementation, demonstrating that the phenotype is caused by insufficient thyroid hormone production.","method":"Xb130 knockout mouse model, histology, immunohistochemistry, immunofluorescence, Western blot, quantitative RT-PCR, levothyroxine rescue experiment","journal":"Thyroid : official journal of the American Thyroid Association","confidence":"High","confidence_rationale":"Tier 2 / Strong — KO mouse with rescue experiment, multiple orthogonal methods establishing mechanism","pmids":["34470464"],"is_preprint":false},{"year":2023,"finding":"AFAP1L2 activates a SRC-FUNDC1 signaling axis in HCC cells to block FUNDC1 recruitment of LC3B to mitochondria, thereby inhibiting mitophagy. AFAP1L2 overexpression in sorafenib-resistant cells leads to elevated phosphorylation of SRC and FUNDC1 and suppressed mitophagy. Artesunate reduces AFAP1L2 protein expression, suppresses SRC/FUNDC1 phosphorylation, and promotes massive FUNDC1-LC3B co-recruitment to mitochondria, overactivating mitophagy and inducing apoptosis of resistant cells.","method":"Co-immunoprecipitation (AFAP1L2-SRC interaction), CETSA (artesunate-AFAP1L2 target engagement), shRNA knockdown, overexpression, surface plasmon resonance, in vitro and xenograft in vivo models, immunofluorescence, TEM for mitophagy","journal":"Autophagy","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — Co-IP and CETSA establishing AFAP1L2-SRC interaction, SPR for direct binding, functional KD/OE with pathway readout, single lab","pmids":["37733919"],"is_preprint":false},{"year":2017,"finding":"AFAP1L2/RET is a fusion oncogene identified in papillary thyroid cancers of young patients in Fukushima; functional analysis confirmed its transforming ability through activation of the MAPK pathway.","method":"5' RACE identification, functional transformation assay, MAPK pathway activation analysis","journal":"Thyroid : official journal of the American Thyroid Association","confidence":"Medium","confidence_rationale":"Tier 2 / Weak — functional transformation assay confirming MAPK activation, single study, limited mechanistic detail in abstract","pmids":["28351223"],"is_preprint":false},{"year":2022,"finding":"In HER2+ gastric cancer cells resistant to trastuzumab, XB130 expression is increased and p-SRC (Tyr416) facilitates binding of XB130 to PI3K p85α. XB130 knockdown reverses resistance by downregulating p-AKT. XB130 negatively regulates PTEN gene transcription, forming a positive feedback loop (SRC-XB130-PTEN) that sustains PI3K/AKT activation.","method":"Western blot, qRT-PCR, CCK8, co-immunoprecipitation (XB130-PI3K p85α), shRNA knockdown, xenograft mouse model","journal":"Clinical & translational oncology","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — Co-IP confirming XB130-p85α interaction, functional KD reversing drug resistance, single lab","pmids":["36284062"],"is_preprint":false},{"year":2024,"finding":"In bone marrow stromal cells on TiO2 nanotube surfaces, XB130 interacts with Src and activates the downstream PI3K/Akt/GSK-3β/β-catenin pathway. Filamentous actin depolymerization changes XB130 expression and distribution, thereby affecting osteogenic differentiation. XB130 overexpression promotes osteogenic differentiation while knockdown inhibits it.","method":"Co-immunoprecipitation (XB130-Src), siRNA knockdown, overexpression, Western blot for PI3K/Akt/GSK-3β/β-catenin, immunofluorescence, in vitro and in vivo osteogenesis assays","journal":"Acta biomaterialia","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — Co-IP for XB130-Src interaction, functional KD/OE with pathway readout, single lab","pmids":["38360291"],"is_preprint":false},{"year":2025,"finding":"hnRNPC binds to specific regions of the XB130 3'UTR, enhancing XB130 mRNA stability by inhibiting recruitment of nucleases XRN1 and DIS3L2, and simultaneously interacts with eIF4E (component of eIF4F complex) to facilitate XB130 mRNA circularization and increase translation efficiency, resulting in elevated XB130/PI3K/Akt signaling and promotion of NSCLC cell proliferation and EMT.","method":"RNA pull-down assay, RNA immunoprecipitation, dual-luciferase reporter assay, co-immunoprecipitation (hnRNPC-eIF4E), Western blot, qRT-PCR, overexpression/knockdown functional assays","journal":"Cancer cell international","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — RNA pull-down plus RIP plus Co-IP establishing hnRNPC-XB130 mRNA and hnRNPC-eIF4E interactions, single lab","pmids":["39800708"],"is_preprint":false},{"year":2025,"finding":"hnRNPR protects XB130 mRNA from XRN1- and DIS3L2-mediated degradation by binding to specific regions within the XB130 3'UTR, thereby elevating XB130 expression, activating the Akt signaling pathway, and promoting NSCLC cell proliferation and EMT.","method":"RNA pull-down assay, RNA immunoprecipitation, dual-luciferase reporter assay, Western blot, qRT-PCR, overexpression/knockdown, tissue microarray, in vivo experiments","journal":"Cellular signalling","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — RNA pull-down plus RIP confirming hnRNPR-XB130 3'UTR interaction, functional KD/OE, single lab","pmids":["40268079"],"is_preprint":false}],"current_model":"AFAP1L2/XB130 is a cytosolic adaptor/scaffold protein that acts as a substrate and regulator of multiple tyrosine kinases (c-Src, RET/PTC, PTPRZ) and, upon phosphorylation at key tyrosines (Y54 by RET/PTC; Y72 in rodent ortholog), recruits the p85α subunit of PI3K to activate PI3K/Akt signaling; it also directly crosslinks F-actin through a C-terminal actin-binding domain and N-terminal multimerization region, localizes to lamellipodia via Rac1 and branched F-actin to regulate cell motility and endocytosis, forms a complex with Tks5 scaffold protein to sustain Src-mediated proliferation and survival, links filamentous actin to the microtubule network at thyrocyte apical membranes to support folliculogenesis, drives FUNDC1-mediated mitophagy via an AFAP1L2-SRC axis, and has its mRNA stability and translation regulated by RNA-binding proteins hnRNPC and hnRNPR acting on its 3'UTR."},"narrative":{"mechanistic_narrative":"AFAP1L2 (XB130) is a cytosolic adaptor/scaffold protein that couples tyrosine-kinase signaling to PI3K/Akt activation and to actin-based remodeling of the cell cortex [PMID:17412687, PMID:19060924]. It was identified as a c-Src-interacting adaptor bearing SH2/SH3-binding motifs, two PH domains, and a coiled-coil region that itself becomes tyrosine-phosphorylated and amplifies Src signal transduction [PMID:17412687]. Upon phosphorylation at defined tyrosines (Y54 by the RET/PTC oncokinase; Y72 in the rodent ortholog), it recruits the p85α regulatory subunit of PI3K through p85α SH2 domains, channeling kinase input into Akt activation and downstream control of cell-cycle and survival effectors including p21, p27, FOXO3a, and GSK3β [PMID:19060924, PMID:22928011, PMID:22496359]. Its phosphorylation state is set bidirectionally: receptor tyrosine phosphatase PTPRZ dephosphorylates AFAP1L2, and pleiotrophin-mediated PTPRZ inhibition raises AFAP1L2 phosphorylation to drive PI3K-AKT-mTOR signaling and oligodendrocyte precursor differentiation and remyelination [PMID:30667096]. Independently of phosphorylation, AFAP1L2 directly binds and crosslinks F-actin via a C-terminal actin-binding region and N-terminal multimerization domain, translocating to lamellipodia in a Rac1-dependent manner to govern cell spreading, migration, invasion, and endocytosis [PMID:21084565, PMID:27462298]. It nucleates a complex with the Tks5 scaffold and Src that sustains proliferation and survival and partitions migratory output between Rac1- and Cdc42-driven structures [PMID:26446840, PMID:27835612]. In thyrocytes it links the actin cortex to the microtubule network at the apical membrane to direct folliculogenesis, and its loss causes congenital hypothyroidism rescuable by thyroxine [PMID:34652970, PMID:34470464]. AFAP1L2 is broadly oncogenic and drug-resistance-associated: it drives a SRC-FUNDC1 axis that suppresses mitophagy in hepatocellular carcinoma [PMID:37733919], sustains PI3K/AKT in trastuzumab-resistant HER2+ gastric cancer [PMID:36284062], and its mRNA stability and translation are controlled by 3'UTR-binding proteins hnRNPC and hnRNPR [PMID:39800708, PMID:40268079].","teleology":[{"year":2007,"claim":"Established AFAP1L2/XB130 as a bona fide adaptor protein physically and functionally coupled to c-Src, answering whether it participates in tyrosine-kinase signaling.","evidence":"Cloning, reciprocal Co-IP, COS-7 co-expression, siRNA knockdown and deletion mutagenesis in lung epithelial cells","pmids":["17412687"],"confidence":"High","gaps":["Direct substrate/kinase relationship not fully resolved","Which domains mediate Src binding versus scaffolding not delineated"]},{"year":2008,"claim":"Identified Y54 as the kinase-targeted residue that links RET/PTC signaling to PI3K via p85α, defining the molecular route from kinase input to Akt.","evidence":"Site-directed mutagenesis (Y54), Co-IP with p85α, siRNA knockdown with Akt/ERK readout in TPC1 thyroid cancer cells","pmids":["19060924"],"confidence":"High","gaps":["Whether other kinases phosphorylate Y54 not determined","Stoichiometry of the XB130-p85α-PI3K complex unknown"]},{"year":2010,"claim":"Showed AFAP1L2 translocates to the lamellipodial F-actin meshwork under Rac and growth-factor cues and is required for motility, separating its actin/migration role from its kinase-substrate role.","evidence":"Live-cell imaging, constitutively active Rac, deletion mutagenesis, wound-healing and Matrigel invasion assays in TPC1 cells","pmids":["21084565"],"confidence":"High","gaps":["Direct actin binding not yet demonstrated at this stage","Mechanism of Rac-dependent recruitment unresolved"]},{"year":2010,"claim":"Demonstrated AFAP1L2 is required for cancer cell proliferation, survival, and tumor growth in vivo, establishing functional importance beyond signaling biochemistry.","evidence":"shRNA knockdown, cell-cycle/apoptosis assays, xenografts in nude mice, microarray profiling in WRO thyroid cancer cells","pmids":["21224076"],"confidence":"Medium","gaps":["Single-lab, single-tumor-type evidence","Causal effectors among 246 changed genes not validated"]},{"year":2012,"claim":"Mapped the PI3K/Akt-dependent downstream effectors (p21, p27, FOXO3a, GSK3β) controlled by AFAP1L2 in both RET/PTC-positive and -negative cells, generalizing its proliferation/survival mechanism.","evidence":"siRNA knockdown, Western blot of Akt substrates, cell-cycle and caspase assays in WRO and A549 cells","pmids":["22928011"],"confidence":"Medium","gaps":["Direct versus indirect effector regulation not separated","Single lab"]},{"year":2012,"claim":"Defined Y72 in the rodent ortholog as the critical p85-binding residue and placed AFAP1L2 in a cAMP/Src-driven mitogenic loop, confirming the phospho-tyrosine–p85 mechanism in a non-cancer thyroid system.","evidence":"MALDI-TOF identification, Y72F mutagenesis, PP1/PP2 inhibitors, Co-IP, PI3K activity and DNA synthesis assays in rat FRTL-5 cells","pmids":["22496359"],"confidence":"High","gaps":["Correspondence between rodent Y72 and human Y54 not formally reconciled","Upstream kinase identity in this context limited to Src inference"]},{"year":2015,"claim":"Identified the Tks5 scaffold as a direct N-terminal partner forming a Tks5-XB130-Src complex required for Src/PI3K/Akt activation, revealing a higher-order scaffolding mechanism for sustained signaling.","evidence":"Yeast two-hybrid, endogenous Co-IP, interface mutagenesis (XB130 ΔN, Tks5 SH3 W1108A), proliferation/apoptosis assays","pmids":["26446840"],"confidence":"High","gaps":["Structural basis of the SH3-polyproline interface not solved","Whether complex assembles constitutively or stimulus-dependently unclear"]},{"year":2015,"claim":"Extended the migration role to carcinogen (NNK)-induced motility in bronchial epithelium and linked AFAP1L2 to MMP-14 relocalization, connecting it to invasive machinery.","evidence":"Immunofluorescence, full-length/truncation overexpression, migration assays, MMP-14 localization in BEAS2B cells","pmids":["25980441"],"confidence":"Medium","gaps":["Direct AFAP1L2-MMP-14 link not established","Single lab"]},{"year":2016,"claim":"Reconstituted AFAP1L2 as a direct F-actin binder and crosslinker, mapping the C-terminal actin-binding region and N-terminal multimerization domain and tying these to endocytosis, providing the biochemical basis for its cortical functions.","evidence":"In vitro F-actin binding/crosslinking with recombinant protein, deletion mutagenesis, Blue native-PAGE, dextran uptake in HEK293","pmids":["27462298"],"confidence":"High","gaps":["In vivo contribution of crosslinking to motility not quantified","Regulation of actin binding by phosphorylation unknown"]},{"year":2016,"claim":"Showed AFAP1L2/Rac1 and Tks5/Cdc42 modules direct distinct migration modes from a shared complex, defining how the scaffold partitions cytoskeletal output.","evidence":"Immunofluorescence with WAVE2/N-WASP markers, Co-IP (XB130-Rac1; Tks5-Cdc42), co-overexpression and migration assays in airway epithelial cells","pmids":["27835612"],"confidence":"Medium","gaps":["Direct versus indirect Rac1 binding not dissected","Single lab"]},{"year":2019,"claim":"Identified PTPRZ as the phosphatase counteracting AFAP1L2 tyrosine phosphorylation and placed AFAP1L2 in pleiotrophin-controlled PI3K-AKT-mTOR signaling for OPC differentiation and remyelination, establishing a physiological, non-cancer role and a bidirectional phospho-control axis.","evidence":"In vitro phosphatase assay, Co-IP, siRNA knockdown, PTPRZ catalytic-dead knock-in mice, pathway Western blots","pmids":["30667096"],"confidence":"High","gaps":["Specific dephosphorylated tyrosines not mapped","Kinase responsible in OPCs not identified"]},{"year":2021,"claim":"Defined a developmental, kinase-independent function: AFAP1L2 links the apical actin cortex to microtubules in thyrocytes to direct folliculogenesis, and its loss causes congenital hypothyroidism rescuable by thyroxine.","evidence":"Xb130 knockout mice, 3D thyrocyte cultures, confocal imaging, histology, levothyroxine rescue","pmids":["34652970","34470464"],"confidence":"High","gaps":["Molecular bridge to microtubule-associated proteins not identified","Relationship to Src/PI3K signaling in this context unclear"]},{"year":2023,"claim":"Linked AFAP1L2 to mitophagy control via a SRC-FUNDC1 axis driving sorafenib resistance in HCC, expanding its effector repertoire beyond PI3K/Akt.","evidence":"Co-IP, CETSA, SPR, shRNA/overexpression, TEM mitophagy readout, xenografts in HCC cells","pmids":["37733919"],"confidence":"Medium","gaps":["Whether AFAP1L2 directly modulates SRC kinase activity or scaffolds FUNDC1 unclear","Single lab"]},{"year":2022,"claim":"Showed AFAP1L2 sustains PI3K/AKT in trastuzumab-resistant HER2+ gastric cancer and proposed a SRC-XB130-PTEN feedback loop, connecting it to therapy resistance.","evidence":"Co-IP (XB130-p85α), shRNA knockdown, qRT-PCR, xenografts","pmids":["36284062"],"confidence":"Medium","gaps":["Mechanism of PTEN transcriptional repression not defined","Single lab"]},{"year":2025,"claim":"Identified post-transcriptional control of AFAP1L2 by 3'UTR-binding proteins hnRNPC and hnRNPR, which stabilize its mRNA against XRN1/DIS3L2 (and, for hnRNPC, enhance translation via eIF4E), explaining how AFAP1L2 levels are elevated in NSCLC.","evidence":"RNA pull-down, RIP, dual-luciferase 3'UTR reporters, Co-IP (hnRNPC-eIF4E), knockdown/overexpression, in vivo NSCLC models","pmids":["39800708","40268079"],"confidence":"Medium","gaps":["Whether hnRNPC and hnRNPR act cooperatively or redundantly unclear","Single lab for each"]},{"year":null,"claim":"How AFAP1L2's two activities — phospho-dependent PI3K recruitment and phospho-independent actin crosslinking — are coordinated within a single cell and whether they share or compete for the same molecular pool remains unresolved.","evidence":"","pmids":[],"confidence":"Medium","gaps":["No structural model integrating PH domains, actin-binding, and scaffolding regions","No high-resolution structure of any AFAP1L2 complex","Regulation switching between signaling and cytoskeletal modes uncharacterized"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0060090","term_label":"molecular adaptor activity","supporting_discovery_ids":[0,1,7]},{"term_id":"GO:0008092","term_label":"cytoskeletal protein binding","supporting_discovery_ids":[2,9]},{"term_id":"GO:0098772","term_label":"molecular function regulator activity","supporting_discovery_ids":[0,1,5]}],"localization":[{"term_id":"GO:0005829","term_label":"cytosol","supporting_discovery_ids":[0]},{"term_id":"GO:0005856","term_label":"cytoskeleton","supporting_discovery_ids":[2,9,12]},{"term_id":"GO:0005886","term_label":"plasma membrane","supporting_discovery_ids":[10,13]}],"pathway":[{"term_id":"R-HSA-162582","term_label":"Signal Transduction","supporting_discovery_ids":[0,1,5,11]},{"term_id":"R-HSA-1643685","term_label":"Disease","supporting_discovery_ids":[1,14,16]},{"term_id":"R-HSA-1266738","term_label":"Developmental Biology","supporting_discovery_ids":[11,12,13]},{"term_id":"R-HSA-8953854","term_label":"Metabolism of RNA","supporting_discovery_ids":[18,19]},{"term_id":"R-HSA-9612973","term_label":"Autophagy","supporting_discovery_ids":[14]}],"complexes":["XB130-Tks5-Src complex"],"partners":["SRC","PIK3R1","TKS5","RET","PTPRZ1","RAC1","FUNDC1","HNRNPC"],"other_free_text":[]}},"prefetch_data":{"uniprot":{"accession":"Q8N4X5","full_name":"Actin filament-associated protein 1-like 2","aliases":[],"length_aa":818,"mass_kda":91.3,"function":"May play a role in a signaling cascade by enhancing the kinase activity of SRC. Contributes to SRC-regulated transcription activation","subcellular_location":"Cytoplasm","url":"https://www.uniprot.org/uniprotkb/Q8N4X5/entry"},"depmap":{"release":"DepMap","has_data":true,"is_common_essential":false,"resolved_as":"","url":"https://depmap.org/portal/gene/AFAP1L2","classification":"Not Classified","n_dependent_lines":0,"n_total_lines":1208,"dependency_fraction":0.0},"opencell":{"profiled":false,"resolved_as":"","ensg_id":"","cell_line_id":"","localizations":[],"interactors":[],"url":"https://opencell.sf.czbiohub.org/search/AFAP1L2","total_profiled":1310},"omim":[{"mim_id":"612420","title":"ACTIN FILAMENT-ASSOCIATED PROTEIN 1-LIKE PROTEIN 2; AFAP1L2","url":"https://www.omim.org/entry/612420"}],"hpa":{"profiled":true,"resolved_as":"","reliability":"Supported","locations":[{"location":"Cytosol","reliability":"Supported"},{"location":"Plasma membrane","reliability":"Additional"}],"tissue_specificity":"Tissue enhanced","tissue_distribution":"Detected in many","driving_tissues":[{"tissue":"lymphoid tissue","ntpm":60.8},{"tissue":"parathyroid gland","ntpm":64.6},{"tissue":"thyroid gland","ntpm":77.0}],"url":"https://www.proteinatlas.org/search/AFAP1L2"},"hgnc":{"alias_symbol":["FLJ14564","Em:AC005383.4","XB130"],"prev_symbol":["KIAA1914"]},"alphafold":{"accession":"Q8N4X5","domains":[{"cath_id":"2.30.29.30","chopping":"175-271","consensus_level":"high","plddt":89.5118,"start":175,"end":271},{"cath_id":"2.30.29.30","chopping":"358-484","consensus_level":"high","plddt":91.1,"start":358,"end":484},{"cath_id":"4.10.860","chopping":"2-47","consensus_level":"high","plddt":81.2811,"start":2,"end":47},{"cath_id":"1.10.287","chopping":"660-750","consensus_level":"high","plddt":93.034,"start":660,"end":750}],"viewer_url":"https://alphafold.ebi.ac.uk/entry/Q8N4X5","model_url":"https://alphafold.ebi.ac.uk/files/AF-Q8N4X5-F1-model_v6.cif","pae_url":"https://alphafold.ebi.ac.uk/files/AF-Q8N4X5-F1-predicted_aligned_error_v6.png","plddt_mean":63.59},"mouse_models":{"mgi_url":"https://www.informatics.jax.org/marker/summary?nomen=AFAP1L2","jax_strain_url":"https://www.jax.org/strain/search?query=AFAP1L2"},"sequence":{"accession":"Q8N4X5","fasta_url":"https://rest.uniprot.org/uniprotkb/Q8N4X5.fasta","uniprot_url":"https://www.uniprot.org/uniprotkb/Q8N4X5/entry","alphafold_viewer_url":"https://alphafold.ebi.ac.uk/entry/Q8N4X5"}},"corpus_meta":[{"pmid":"24387290","id":"PMC_24387290","title":"XB130 promotes proliferation and invasion of gastric cancer cells.","date":"2014","source":"Journal of translational medicine","url":"https://pubmed.ncbi.nlm.nih.gov/24387290","citation_count":86,"is_preprint":false},{"pmid":"17412687","id":"PMC_17412687","title":"XB130, a novel adaptor protein for signal transduction.","date":"2007","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/17412687","citation_count":74,"is_preprint":false},{"pmid":"37733919","id":"PMC_37733919","title":"Artesunate Sensitizes human hepatocellular carcinoma to sorafenib via exacerbating AFAP1L2-SRC-FUNDC1 axis-dependent mitophagy.","date":"2023","source":"Autophagy","url":"https://pubmed.ncbi.nlm.nih.gov/37733919","citation_count":69,"is_preprint":false},{"pmid":"19060924","id":"PMC_19060924","title":"XB130, a tissue-specific adaptor protein that couples the RET/PTC oncogenic kinase to PI 3-kinase pathway.","date":"2008","source":"Oncogene","url":"https://pubmed.ncbi.nlm.nih.gov/19060924","citation_count":57,"is_preprint":false},{"pmid":"28351223","id":"PMC_28351223","title":"Identification of Three Novel Fusion Oncogenes, SQSTM1/NTRK3, AFAP1L2/RET, and PPFIBP2/RET, in Thyroid Cancers of Young Patients in Fukushima.","date":"2017","source":"Thyroid : official journal of the American Thyroid Association","url":"https://pubmed.ncbi.nlm.nih.gov/28351223","citation_count":44,"is_preprint":false},{"pmid":"21224076","id":"PMC_21224076","title":"XB130, a novel adaptor protein, promotes thyroid tumor growth.","date":"2010","source":"The American journal of pathology","url":"https://pubmed.ncbi.nlm.nih.gov/21224076","citation_count":41,"is_preprint":false},{"pmid":"21084565","id":"PMC_21084565","title":"Adaptor protein XB130 is a Rac-controlled component of lamellipodia that regulates cell motility and invasion.","date":"2010","source":"Journal of cell science","url":"https://pubmed.ncbi.nlm.nih.gov/21084565","citation_count":41,"is_preprint":false},{"pmid":"22928011","id":"PMC_22928011","title":"XB130 mediates cancer cell proliferation and survival through multiple signaling events downstream of Akt.","date":"2012","source":"PloS one","url":"https://pubmed.ncbi.nlm.nih.gov/22928011","citation_count":37,"is_preprint":false},{"pmid":"30667096","id":"PMC_30667096","title":"The PTN-PTPRZ signal activates the AFAP1L2-dependent PI3K-AKT pathway for oligodendrocyte differentiation: Targeted inactivation of PTPRZ activity in mice.","date":"2019","source":"Glia","url":"https://pubmed.ncbi.nlm.nih.gov/30667096","citation_count":37,"is_preprint":false},{"pmid":"23527086","id":"PMC_23527086","title":"XB130, a new adaptor protein, regulates expression of tumor suppressive microRNAs in cancer cells.","date":"2013","source":"PloS one","url":"https://pubmed.ncbi.nlm.nih.gov/23527086","citation_count":32,"is_preprint":false},{"pmid":"21884627","id":"PMC_21884627","title":"Roles of XB130, a novel adaptor protein, in cancer.","date":"2011","source":"Journal of clinical bioinformatics","url":"https://pubmed.ncbi.nlm.nih.gov/21884627","citation_count":27,"is_preprint":false},{"pmid":"24995146","id":"PMC_24995146","title":"XB130-A Novel Adaptor Protein: Gene, Function, and Roles in Tumorigenesis.","date":"2014","source":"Scientifica","url":"https://pubmed.ncbi.nlm.nih.gov/24995146","citation_count":26,"is_preprint":false},{"pmid":"22496359","id":"PMC_22496359","title":"Phosphatidylinositol 3-kinase-binding protein, PI3KAP/XB130, is required for cAMP-induced amplification of IGF mitogenic activity in FRTL-5 thyroid cells.","date":"2012","source":"Molecular endocrinology (Baltimore, Md.)","url":"https://pubmed.ncbi.nlm.nih.gov/22496359","citation_count":21,"is_preprint":false},{"pmid":"26998266","id":"PMC_26998266","title":"XB130: A novel adaptor protein in cancer signal transduction.","date":"2016","source":"Biomedical reports","url":"https://pubmed.ncbi.nlm.nih.gov/26998266","citation_count":21,"is_preprint":false},{"pmid":"26446840","id":"PMC_26446840","title":"XB130/Tks5 scaffold protein interaction regulates Src-mediated cell proliferation and survival.","date":"2015","source":"Molecular biology of the cell","url":"https://pubmed.ncbi.nlm.nih.gov/26446840","citation_count":18,"is_preprint":false},{"pmid":"32159887","id":"PMC_32159887","title":"XB130, regulated by miR-203, miR-219, and miR-4782-3p, mediates the proliferation and metastasis of non-small-cell lung cancer cells.","date":"2020","source":"Molecular carcinogenesis","url":"https://pubmed.ncbi.nlm.nih.gov/32159887","citation_count":16,"is_preprint":false},{"pmid":"31219645","id":"PMC_31219645","title":"Knockdown of XB130 restrains cancer stem cell-like phenotype through inhibition of Wnt/β-Catenin signaling in breast cancer.","date":"2019","source":"Molecular carcinogenesis","url":"https://pubmed.ncbi.nlm.nih.gov/31219645","citation_count":16,"is_preprint":false},{"pmid":"25980441","id":"PMC_25980441","title":"XB130 translocation to microfilamentous structures mediates NNK-induced migration of human bronchial epithelial cells.","date":"2015","source":"Oncotarget","url":"https://pubmed.ncbi.nlm.nih.gov/25980441","citation_count":16,"is_preprint":false},{"pmid":"27509056","id":"PMC_27509056","title":"XB130 is overexpressed in prostate cancer and involved in cell growth and invasion.","date":"2016","source":"Oncotarget","url":"https://pubmed.ncbi.nlm.nih.gov/27509056","citation_count":15,"is_preprint":false},{"pmid":"30820526","id":"PMC_30820526","title":"XB130 deficiency enhances carcinogen-induced skin tumorigenesis.","date":"2019","source":"Carcinogenesis","url":"https://pubmed.ncbi.nlm.nih.gov/30820526","citation_count":13,"is_preprint":false},{"pmid":"25892537","id":"PMC_25892537","title":"Integrated pathway and epistasis analysis reveals interactive effect of genetic variants at TERF1 and AFAP1L2 loci on melanoma risk.","date":"2015","source":"International journal of cancer","url":"https://pubmed.ncbi.nlm.nih.gov/25892537","citation_count":13,"is_preprint":false},{"pmid":"38360291","id":"PMC_38360291","title":"TiO2 nanotube topography enhances osteogenesis through filamentous actin and XB130-protein-mediated mechanotransduction.","date":"2024","source":"Acta biomaterialia","url":"https://pubmed.ncbi.nlm.nih.gov/38360291","citation_count":12,"is_preprint":false},{"pmid":"25272040","id":"PMC_25272040","title":"XB130 deficiency affects tracheal epithelial differentiation during airway repair.","date":"2014","source":"PloS one","url":"https://pubmed.ncbi.nlm.nih.gov/25272040","citation_count":12,"is_preprint":false},{"pmid":"26360608","id":"PMC_26360608","title":"XB130 promotes bronchioalveolar stem cell and Club cell proliferation in airway epithelial repair and regeneration.","date":"2015","source":"Oncotarget","url":"https://pubmed.ncbi.nlm.nih.gov/26360608","citation_count":10,"is_preprint":false},{"pmid":"34652970","id":"PMC_34652970","title":"XB130 Plays an Essential Role in Folliculogenesis Through Mediating Interactions Between Microfilament and Microtubule Systems in Thyrocytes.","date":"2021","source":"Thyroid : official journal of the American Thyroid Association","url":"https://pubmed.ncbi.nlm.nih.gov/34652970","citation_count":9,"is_preprint":false},{"pmid":"26191231","id":"PMC_26191231","title":"Expression of XB130 in human ductal breast cancer.","date":"2015","source":"International journal of clinical and experimental pathology","url":"https://pubmed.ncbi.nlm.nih.gov/26191231","citation_count":9,"is_preprint":false},{"pmid":"30246718","id":"PMC_30246718","title":"XB130 Knockdown Inhibits the Proliferation, Invasiveness, and Metastasis of Hepatocellular Carcinoma Cells and Sensitizes them to TRAIL-Induced Apoptosis.","date":"2018","source":"Chinese medical journal","url":"https://pubmed.ncbi.nlm.nih.gov/30246718","citation_count":9,"is_preprint":false},{"pmid":"27462298","id":"PMC_27462298","title":"Phosphatidylinositol 3-Kinase-Associated Protein (PI3KAP)/XB130 Crosslinks Actin Filaments through Its Actin Binding and Multimerization Properties In Vitro and Enhances Endocytosis in HEK293 Cells.","date":"2016","source":"Frontiers in endocrinology","url":"https://pubmed.ncbi.nlm.nih.gov/27462298","citation_count":9,"is_preprint":false},{"pmid":"34780466","id":"PMC_34780466","title":"Adaptor protein XB130 regulates the aggressiveness of cholangiocarcinoma.","date":"2021","source":"PloS one","url":"https://pubmed.ncbi.nlm.nih.gov/34780466","citation_count":8,"is_preprint":false},{"pmid":"26045762","id":"PMC_26045762","title":"XB130 expression in human osteosarcoma: a clinical and experimental study.","date":"2015","source":"International journal of clinical and experimental pathology","url":"https://pubmed.ncbi.nlm.nih.gov/26045762","citation_count":8,"is_preprint":false},{"pmid":"27835612","id":"PMC_27835612","title":"Stimulus-dependent dissociation between XB130 and Tks5 scaffold proteins promotes airway epithelial cell migration.","date":"2016","source":"Oncotarget","url":"https://pubmed.ncbi.nlm.nih.gov/27835612","citation_count":8,"is_preprint":false},{"pmid":"34470464","id":"PMC_34470464","title":"XB130 Deficiency Causes Congenital Hypothyroidism in Mice due to Disorganized Apical Membrane Structure and Function of Thyrocytes.","date":"2021","source":"Thyroid : official journal of the American Thyroid Association","url":"https://pubmed.ncbi.nlm.nih.gov/34470464","citation_count":8,"is_preprint":false},{"pmid":"28849225","id":"PMC_28849225","title":"XB130 enhances invasion and migration of human colorectal cancer cells by promoting epithelial‑mesenchymal transition.","date":"2017","source":"Molecular medicine reports","url":"https://pubmed.ncbi.nlm.nih.gov/28849225","citation_count":7,"is_preprint":false},{"pmid":"27029000","id":"PMC_27029000","title":"XB130 deficiency enhances lipopolysaccharide-induced septic response and acute lung injury.","date":"2016","source":"Oncotarget","url":"https://pubmed.ncbi.nlm.nih.gov/27029000","citation_count":7,"is_preprint":false},{"pmid":"35326426","id":"PMC_35326426","title":"Thyroidal Transcriptomic Profiles of Pathoadaptive Responses to Congenital Hypothyroidism in XB130 Knockout Mice.","date":"2022","source":"Cells","url":"https://pubmed.ncbi.nlm.nih.gov/35326426","citation_count":5,"is_preprint":false},{"pmid":"36284062","id":"PMC_36284062","title":"Molecular mechanism of XB130 adaptor protein mediates trastuzumab resistance in gastric cancer.","date":"2022","source":"Clinical & translational oncology : official publication of the Federation of Spanish Oncology Societies and of the National Cancer Institute of Mexico","url":"https://pubmed.ncbi.nlm.nih.gov/36284062","citation_count":2,"is_preprint":false},{"pmid":"37771334","id":"PMC_37771334","title":"XB130 inhibits healing of diabetic skin ulcers through the PI3K/Akt signalling pathway.","date":"2023","source":"World journal of diabetes","url":"https://pubmed.ncbi.nlm.nih.gov/37771334","citation_count":2,"is_preprint":false},{"pmid":"37720672","id":"PMC_37720672","title":"The 3'‑untranslated region of XB130 regulates its mRNA stability and translational efficiency in non‑small cell lung cancer cells.","date":"2023","source":"Oncology letters","url":"https://pubmed.ncbi.nlm.nih.gov/37720672","citation_count":2,"is_preprint":false},{"pmid":"39800708","id":"PMC_39800708","title":"Heterogeneous nuclear ribonucleoprotein C promotes non-small cell lung cancer progression by enhancing XB130 mRNA stability and translation.","date":"2025","source":"Cancer cell international","url":"https://pubmed.ncbi.nlm.nih.gov/39800708","citation_count":2,"is_preprint":false},{"pmid":"34915750","id":"PMC_34915750","title":"Pathogenesis of Multinodular Goiter in Elderly XB130-Deficient Mice: Alteration of Thyroperoxidase Affinity with Iodide and Hydrogen Peroxide.","date":"2022","source":"Thyroid : official journal of the American Thyroid Association","url":"https://pubmed.ncbi.nlm.nih.gov/34915750","citation_count":2,"is_preprint":false},{"pmid":"40268079","id":"PMC_40268079","title":"HnRNPR promotes non-small cell lung cancer progression by protecting XB130 mRNA from XRN1- and DIS3L2-mediated degradation.","date":"2025","source":"Cellular signalling","url":"https://pubmed.ncbi.nlm.nih.gov/40268079","citation_count":0,"is_preprint":false},{"pmid":"25997546","id":"PMC_25997546","title":"WITHDRAWN: XB130 expression in human osteosarcoma: a clinical and experimental study.","date":"2015","source":"International journal of clinical pharmacology and therapeutics","url":"https://pubmed.ncbi.nlm.nih.gov/25997546","citation_count":0,"is_preprint":false}],"cost":{"stage1":{"model":"claude-sonnet-4-6","input_tokens":25399,"output_tokens":6085,"usd":0.083736,"stage1_stop_reason":"end_turn"},"stage2":{"model":"claude-opus-4-8","input_tokens":14650,"output_tokens":5310,"usd":0.103,"stage2_stop_reason":"end_turn"},"total_usd":0.186736,"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\": 2007,\n      \"finding\": \"XB130 (AFAP1L2) was cloned as a novel adaptor protein containing SH2- and SH3-binding motifs, two pleckstrin homology domains, a coiled-coil region, and multiple phosphorylation sites. Endogenous XB130 interacts with c-Src tyrosine kinase, and co-expression in COS-7 cells activated c-Src and elevated tyrosine phosphorylation of multiple proteins including XB130 itself. An N-terminal deletion mutant (XB130ΔN) reduced its ability to mediate Src signal transduction. Knockdown in lung epithelial cells reduced c-Src activity, IL-8 production, EGF-induced phosphorylation of Akt and GSK3β, and altered cell cycles.\",\n      \"method\": \"Co-immunoprecipitation, co-expression in COS-7 cells, siRNA knockdown, reporter assay (SRE/AP-1), deletion mutagenesis, Western blot\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — reciprocal Co-IP, mutagenesis, functional knockdown with multiple orthogonal methods in founding paper\",\n      \"pmids\": [\"17412687\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2008,\n      \"finding\": \"RET/PTC oncogenic kinase phosphorylates XB130 predominantly at tyrosine 54, which promotes XB130 association with the p85α subunit of PI3-kinase via p85α SH2 domains, thereby linking RET/PTC signaling to PI3K/Akt activation. Knockdown of XB130 in TPC1 papillary thyroid cancer cells strongly reduced Akt activity without altering ERK1/2 phosphorylation, and inhibited cell-cycle progression and survival.\",\n      \"method\": \"Co-immunoprecipitation, site-directed mutagenesis (Y54), siRNA knockdown, Western blot for Akt/ERK phosphorylation, cell cycle analysis\",\n      \"journal\": \"Oncogene\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1-2 / Strong — mutagenesis identifying specific phosphorylation site plus reciprocal Co-IP and functional KD with defined pathway readout\",\n      \"pmids\": [\"19060924\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2010,\n      \"finding\": \"XB130 translocates to the cell periphery in response to EGF, wounding, and constitutively active Rac, localizing to the lamellipodial F-actin meshwork. Structure-function analysis showed that both the N-terminus (167 aa) and C-terminus (63 aa) are required for translocation to lamellipodia, while PH domains and Src-targeted tyrosines are dispensable. Silencing XB130 in TPC1 cells decreased wound closure rate, inhibited Matrigel invasion, reduced lamellipodial persistence, and slowed spreading.\",\n      \"method\": \"Live-cell imaging, constitutively active Rac expression, pharmacological inhibitors, deletion mutagenesis, siRNA knockdown, wound-healing and Matrigel invasion assays\",\n      \"journal\": \"Journal of cell science\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — direct localization experiment with functional consequence, structure-function mutagenesis, multiple orthogonal methods\",\n      \"pmids\": [\"21084565\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2010,\n      \"finding\": \"Knockdown of XB130 in WRO thyroid cancer cells inhibited G1-S phase progression, induced spontaneous apoptosis, and enhanced intrinsic and extrinsic apoptotic stimulus-induced cell death. In vivo, XB130 shRNA stably transfected WRO cells formed significantly smaller tumors in nude mice. Microarray identified 246 significantly changed genes, including 57 related to cell proliferation or survival.\",\n      \"method\": \"siRNA/shRNA knockdown, cell cycle analysis, apoptosis assays, xenograft mouse model, microarray gene expression profiling\",\n      \"journal\": \"The American journal of pathology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — KO/KD with defined phenotypic readouts in vitro and in vivo, single lab\",\n      \"pmids\": [\"21224076\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2012,\n      \"finding\": \"XB130 mediates cancer cell proliferation and survival through PI3K/Akt downstream signals including phosphorylation of p21Cip1/WAF1, p27Kip1, FOXO3a, and GSK3β (but not FOXO1 or p53), in both RET/PTC-mutant (WRO) and RET/PTC-negative (A549) cancer cells. XB130 can be phosphorylated by multiple protein tyrosine kinases.\",\n      \"method\": \"siRNA knockdown, Western blot for phosphorylated Akt substrates, cell cycle analysis, caspase cleavage assay\",\n      \"journal\": \"PloS one\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — functional KD with multiple pathway readouts, single lab, two cell lines\",\n      \"pmids\": [\"22928011\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2012,\n      \"finding\": \"In rat FRTL-5 thyroid cells, cAMP treatment elevated PI3KAP/XB130 mRNA and protein levels, increased its tyrosine phosphorylation by c-Src (blocked by PP1/PP2), and enhanced its interaction with p85 PI3K, leading to increased PI3K activity. PI3KAP/XB130 knockdown attenuated cAMP-dependent potentiation of IGF-I-induced DNA synthesis. A Y72F mutant incapable of binding p85 PI3K did not enhance IGF-I-induced DNA synthesis, establishing Y72 as the critical p85 PI3K-binding site.\",\n      \"method\": \"MALDI-TOF MS (protein identification), siRNA knockdown, Src kinase inhibitors (PP1/PP2), point mutagenesis (Y72F), co-immunoprecipitation, PI3K activity assay, DNA synthesis assay\",\n      \"journal\": \"Molecular endocrinology (Baltimore, Md.)\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Strong — mutagenesis identifying critical binding residue, in vitro kinase-pathway assay, pharmacological validation, multiple orthogonal methods\",\n      \"pmids\": [\"22496359\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2013,\n      \"finding\": \"XB130 suppresses expression of tumor-suppressive microRNAs (miR-33a, miR-149, miR-193a-3p) in thyroid cancer cells. These miRNAs target oncogenes MYC (miR-33a), FOSL1 (miR-149), and SLC7A5 (miR-193a-3p) at their 3' UTR, and their upregulation upon XB130 knockdown reduces cancer cell growth.\",\n      \"method\": \"miRNA array, shRNA knockdown, qRT-PCR validation, ectopic overexpression, miRNA mimic transfection, 3' UTR reporter assay\",\n      \"journal\": \"PloS one\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — 3'UTR reporter assay confirming direct targeting plus gain/loss of function, single lab\",\n      \"pmids\": [\"23527086\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2015,\n      \"finding\": \"XB130 interacts with scaffold protein Tks5: the fifth SH3 domain of Tks5 binds to the N-terminus of XB130 (which contains polyproline-rich motifs), and both proteins form a complex with Src tyrosine kinase. Disrupting XB130/Tks5 binding (via XB130 N-terminal deletion mutant or Tks5 SH3-W1108A mutant) decreased cell proliferation and survival and inhibited serum/growth-factor-induced Src activation and downstream PI3K/Akt phosphorylation.\",\n      \"method\": \"Yeast two-hybrid screening, co-immunoprecipitation (endogenous), structure-function mutagenesis, cell proliferation and apoptosis assays, Western blot\",\n      \"journal\": \"Molecular biology of the cell\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — yeast two-hybrid plus reciprocal endogenous Co-IP plus mutagenesis of binding interface, multiple orthogonal methods\",\n      \"pmids\": [\"26446840\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2015,\n      \"finding\": \"XB130 translocates to lamellipodia and microfilamentous structures in BEAS2B bronchial epithelial cells after NNK stimulation, and overexpression significantly enhances NNK-induced migration. Both the N- and C-termini of XB130 are required for this migration-enhancing function. XB130 overexpression enhanced NNK-induced protein tyrosine phosphorylation and promoted matrix metalloproteinase-14 (MMP-14) translocation to cell motility-associated structures.\",\n      \"method\": \"Immunofluorescence/localization, overexpression of full-length and truncation mutants, migration assay, Western blot for tyrosine phosphorylation, MMP-14 localization\",\n      \"journal\": \"Oncotarget\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — direct localization experiment with functional consequence, structure-function analysis, single lab\",\n      \"pmids\": [\"25980441\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2016,\n      \"finding\": \"XB130 (PI3KAP) directly binds F-actin through its C-terminal region (residues 830-840), and multimerizes via its N-terminal 40 amino acids (shown by Blue native-PAGE and co-IP). Both actin-binding and multimerization are required for XB130 to crosslink F-actin in vitro. Overexpression of XB130 enhanced dextran endocytosis in HEK293 cells, dependent on its actin-binding region.\",\n      \"method\": \"In vitro F-actin binding and crosslinking assay with recombinant protein, deletion mutagenesis, Blue native-PAGE, co-immunoprecipitation, endocytosis assay (dextran uptake)\",\n      \"journal\": \"Frontiers in endocrinology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Strong — in vitro reconstitution of actin crosslinking, mutagenesis of actin-binding and multimerization domains, functional cellular assay\",\n      \"pmids\": [\"27462298\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2016,\n      \"finding\": \"In airway epithelial cells, XB130 and Tks5 translocate to the cell membrane in a stimulus-dependent manner upon EGF, PKC activator, or nicotinic acetylcholine receptor ligand stimulation. XB130 co-localizes with lamellipodial marker WAVE2 whereas Tks5 co-localizes with podosome marker N-WASP. XB130 interacts with Rac1 and Tks5 interacts with Cdc42 to promote Rho GTPase activity. Co-expression of both proteins inhibits cell migration; dissociation of the complex promotes lateral (XB130/Rac1) or vertical (Tks5/Cdc42) cell migration.\",\n      \"method\": \"Immunofluorescence localization, co-immunoprecipitation (XB130-Rac1; Tks5-Cdc42), overexpression and co-overexpression, migration assay\",\n      \"journal\": \"Oncotarget\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — Co-IP with Rho GTPases plus localization plus functional migration assays, single lab\",\n      \"pmids\": [\"27835612\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"PTPRZ (protein tyrosine phosphatase receptor Z) dephosphorylates AFAP1L2 at tyrosine residues in vitro and in HEK293T cells. Pleiotrophin (PTN)-mediated inhibition of PTPRZ enhances tyrosine phosphorylation of AFAP1L2, which activates the PI3K-AKT-mTOR pathway to promote oligodendrocyte precursor cell (OPC) differentiation. Knockdown of AFAP1L2 or PI3K inhibition suppressed OPC differentiation and PTN-induced AKT/mTOR phosphorylation. PTPRZ catalytic-dead knock-in mice showed higher AFAP1L2, AKT, and mTOR phosphorylation and accelerated remyelination.\",\n      \"method\": \"In vitro phosphatase assay (recombinant PTPRZ on AFAP1L2), co-immunoprecipitation in HEK293T, siRNA knockdown in OPC-like cells, catalytic-dead knock-in mouse model, Western blot for PI3K-AKT-mTOR, immunofluorescence\",\n      \"journal\": \"Glia\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Strong — in vitro phosphatase assay plus knock-in mouse model with multiple orthogonal readouts, direct identification of PTPRZ as phosphatase for AFAP1L2\",\n      \"pmids\": [\"30667096\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"XB130 in thyrocytes critically regulates folliculogenesis by functioning as a link between the actin filament cortex and microtubule network at the apical membrane. In 3D cultures, XB130 aligns with actin filaments and participates in defining the site of apical membrane formation and lumen coalescence. Xb130-knockout thyrocytes showed delayed folliculogenesis, reduced recruitment of microtubule-associated proteins, and disorganized acetylated tubulin under the apical membrane.\",\n      \"method\": \"Xb130 knockout mouse, 3D Matrigel culture of primary thyrocytes, immunofluorescence/confocal microscopy, XB130-GFP transfection for live imaging\",\n      \"journal\": \"Thyroid : official journal of the American Thyroid Association\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — KO mouse with in vitro 3D culture reconstitution, direct imaging of protein localization at apical membrane, multiple orthogonal methods\",\n      \"pmids\": [\"34652970\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"Xb130-knockout mice develop congenital hypothyroidism due to disorganized apical membrane structure of thyrocytes, with diminished thyroglobulin iodination and release. XB130 is localized mainly on the apical membrane of thyroid follicles. Growth retardation in KO mice could be rescued by exogenous thyroxine supplementation, demonstrating that the phenotype is caused by insufficient thyroid hormone production.\",\n      \"method\": \"Xb130 knockout mouse model, histology, immunohistochemistry, immunofluorescence, Western blot, quantitative RT-PCR, levothyroxine rescue experiment\",\n      \"journal\": \"Thyroid : official journal of the American Thyroid Association\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — KO mouse with rescue experiment, multiple orthogonal methods establishing mechanism\",\n      \"pmids\": [\"34470464\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"AFAP1L2 activates a SRC-FUNDC1 signaling axis in HCC cells to block FUNDC1 recruitment of LC3B to mitochondria, thereby inhibiting mitophagy. AFAP1L2 overexpression in sorafenib-resistant cells leads to elevated phosphorylation of SRC and FUNDC1 and suppressed mitophagy. Artesunate reduces AFAP1L2 protein expression, suppresses SRC/FUNDC1 phosphorylation, and promotes massive FUNDC1-LC3B co-recruitment to mitochondria, overactivating mitophagy and inducing apoptosis of resistant cells.\",\n      \"method\": \"Co-immunoprecipitation (AFAP1L2-SRC interaction), CETSA (artesunate-AFAP1L2 target engagement), shRNA knockdown, overexpression, surface plasmon resonance, in vitro and xenograft in vivo models, immunofluorescence, TEM for mitophagy\",\n      \"journal\": \"Autophagy\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — Co-IP and CETSA establishing AFAP1L2-SRC interaction, SPR for direct binding, functional KD/OE with pathway readout, single lab\",\n      \"pmids\": [\"37733919\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2017,\n      \"finding\": \"AFAP1L2/RET is a fusion oncogene identified in papillary thyroid cancers of young patients in Fukushima; functional analysis confirmed its transforming ability through activation of the MAPK pathway.\",\n      \"method\": \"5' RACE identification, functional transformation assay, MAPK pathway activation analysis\",\n      \"journal\": \"Thyroid : official journal of the American Thyroid Association\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Weak — functional transformation assay confirming MAPK activation, single study, limited mechanistic detail in abstract\",\n      \"pmids\": [\"28351223\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"In HER2+ gastric cancer cells resistant to trastuzumab, XB130 expression is increased and p-SRC (Tyr416) facilitates binding of XB130 to PI3K p85α. XB130 knockdown reverses resistance by downregulating p-AKT. XB130 negatively regulates PTEN gene transcription, forming a positive feedback loop (SRC-XB130-PTEN) that sustains PI3K/AKT activation.\",\n      \"method\": \"Western blot, qRT-PCR, CCK8, co-immunoprecipitation (XB130-PI3K p85α), shRNA knockdown, xenograft mouse model\",\n      \"journal\": \"Clinical & translational oncology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — Co-IP confirming XB130-p85α interaction, functional KD reversing drug resistance, single lab\",\n      \"pmids\": [\"36284062\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"In bone marrow stromal cells on TiO2 nanotube surfaces, XB130 interacts with Src and activates the downstream PI3K/Akt/GSK-3β/β-catenin pathway. Filamentous actin depolymerization changes XB130 expression and distribution, thereby affecting osteogenic differentiation. XB130 overexpression promotes osteogenic differentiation while knockdown inhibits it.\",\n      \"method\": \"Co-immunoprecipitation (XB130-Src), siRNA knockdown, overexpression, Western blot for PI3K/Akt/GSK-3β/β-catenin, immunofluorescence, in vitro and in vivo osteogenesis assays\",\n      \"journal\": \"Acta biomaterialia\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — Co-IP for XB130-Src interaction, functional KD/OE with pathway readout, single lab\",\n      \"pmids\": [\"38360291\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"hnRNPC binds to specific regions of the XB130 3'UTR, enhancing XB130 mRNA stability by inhibiting recruitment of nucleases XRN1 and DIS3L2, and simultaneously interacts with eIF4E (component of eIF4F complex) to facilitate XB130 mRNA circularization and increase translation efficiency, resulting in elevated XB130/PI3K/Akt signaling and promotion of NSCLC cell proliferation and EMT.\",\n      \"method\": \"RNA pull-down assay, RNA immunoprecipitation, dual-luciferase reporter assay, co-immunoprecipitation (hnRNPC-eIF4E), Western blot, qRT-PCR, overexpression/knockdown functional assays\",\n      \"journal\": \"Cancer cell international\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — RNA pull-down plus RIP plus Co-IP establishing hnRNPC-XB130 mRNA and hnRNPC-eIF4E interactions, single lab\",\n      \"pmids\": [\"39800708\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"hnRNPR protects XB130 mRNA from XRN1- and DIS3L2-mediated degradation by binding to specific regions within the XB130 3'UTR, thereby elevating XB130 expression, activating the Akt signaling pathway, and promoting NSCLC cell proliferation and EMT.\",\n      \"method\": \"RNA pull-down assay, RNA immunoprecipitation, dual-luciferase reporter assay, Western blot, qRT-PCR, overexpression/knockdown, tissue microarray, in vivo experiments\",\n      \"journal\": \"Cellular signalling\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — RNA pull-down plus RIP confirming hnRNPR-XB130 3'UTR interaction, functional KD/OE, single lab\",\n      \"pmids\": [\"40268079\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"AFAP1L2/XB130 is a cytosolic adaptor/scaffold protein that acts as a substrate and regulator of multiple tyrosine kinases (c-Src, RET/PTC, PTPRZ) and, upon phosphorylation at key tyrosines (Y54 by RET/PTC; Y72 in rodent ortholog), recruits the p85α subunit of PI3K to activate PI3K/Akt signaling; it also directly crosslinks F-actin through a C-terminal actin-binding domain and N-terminal multimerization region, localizes to lamellipodia via Rac1 and branched F-actin to regulate cell motility and endocytosis, forms a complex with Tks5 scaffold protein to sustain Src-mediated proliferation and survival, links filamentous actin to the microtubule network at thyrocyte apical membranes to support folliculogenesis, drives FUNDC1-mediated mitophagy via an AFAP1L2-SRC axis, and has its mRNA stability and translation regulated by RNA-binding proteins hnRNPC and hnRNPR acting on its 3'UTR.\"\n}\n```","stage2_raw":"{\n  \"mechanistic_narrative\": \"AFAP1L2 (XB130) is a cytosolic adaptor/scaffold protein that couples tyrosine-kinase signaling to PI3K/Akt activation and to actin-based remodeling of the cell cortex [#0, #1]. It was identified as a c-Src-interacting adaptor bearing SH2/SH3-binding motifs, two PH domains, and a coiled-coil region that itself becomes tyrosine-phosphorylated and amplifies Src signal transduction [#0]. Upon phosphorylation at defined tyrosines (Y54 by the RET/PTC oncokinase; Y72 in the rodent ortholog), it recruits the p85\\u03b1 regulatory subunit of PI3K through p85\\u03b1 SH2 domains, channeling kinase input into Akt activation and downstream control of cell-cycle and survival effectors including p21, p27, FOXO3a, and GSK3\\u03b2 [#1, #4, #5]. Its phosphorylation state is set bidirectionally: receptor tyrosine phosphatase PTPRZ dephosphorylates AFAP1L2, and pleiotrophin-mediated PTPRZ inhibition raises AFAP1L2 phosphorylation to drive PI3K-AKT-mTOR signaling and oligodendrocyte precursor differentiation and remyelination [#11]. Independently of phosphorylation, AFAP1L2 directly binds and crosslinks F-actin via a C-terminal actin-binding region and N-terminal multimerization domain, translocating to lamellipodia in a Rac1-dependent manner to govern cell spreading, migration, invasion, and endocytosis [#2, #9]. It nucleates a complex with the Tks5 scaffold and Src that sustains proliferation and survival and partitions migratory output between Rac1- and Cdc42-driven structures [#7, #10]. In thyrocytes it links the actin cortex to the microtubule network at the apical membrane to direct folliculogenesis, and its loss causes congenital hypothyroidism rescuable by thyroxine [#12, #13]. AFAP1L2 is broadly oncogenic and drug-resistance-associated: it drives a SRC-FUNDC1 axis that suppresses mitophagy in hepatocellular carcinoma [#14], sustains PI3K/AKT in trastuzumab-resistant HER2+ gastric cancer [#16], and its mRNA stability and translation are controlled by 3'UTR-binding proteins hnRNPC and hnRNPR [#18, #19].\",\n  \"teleology\": [\n    {\n      \"year\": 2007,\n      \"claim\": \"Established AFAP1L2/XB130 as a bona fide adaptor protein physically and functionally coupled to c-Src, answering whether it participates in tyrosine-kinase signaling.\",\n      \"evidence\": \"Cloning, reciprocal Co-IP, COS-7 co-expression, siRNA knockdown and deletion mutagenesis in lung epithelial cells\",\n      \"pmids\": [\"17412687\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Direct substrate/kinase relationship not fully resolved\", \"Which domains mediate Src binding versus scaffolding not delineated\"]\n    },\n    {\n      \"year\": 2008,\n      \"claim\": \"Identified Y54 as the kinase-targeted residue that links RET/PTC signaling to PI3K via p85\\u03b1, defining the molecular route from kinase input to Akt.\",\n      \"evidence\": \"Site-directed mutagenesis (Y54), Co-IP with p85\\u03b1, siRNA knockdown with Akt/ERK readout in TPC1 thyroid cancer cells\",\n      \"pmids\": [\"19060924\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Whether other kinases phosphorylate Y54 not determined\", \"Stoichiometry of the XB130-p85\\u03b1-PI3K complex unknown\"]\n    },\n    {\n      \"year\": 2010,\n      \"claim\": \"Showed AFAP1L2 translocates to the lamellipodial F-actin meshwork under Rac and growth-factor cues and is required for motility, separating its actin/migration role from its kinase-substrate role.\",\n      \"evidence\": \"Live-cell imaging, constitutively active Rac, deletion mutagenesis, wound-healing and Matrigel invasion assays in TPC1 cells\",\n      \"pmids\": [\"21084565\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Direct actin binding not yet demonstrated at this stage\", \"Mechanism of Rac-dependent recruitment unresolved\"]\n    },\n    {\n      \"year\": 2010,\n      \"claim\": \"Demonstrated AFAP1L2 is required for cancer cell proliferation, survival, and tumor growth in vivo, establishing functional importance beyond signaling biochemistry.\",\n      \"evidence\": \"shRNA knockdown, cell-cycle/apoptosis assays, xenografts in nude mice, microarray profiling in WRO thyroid cancer cells\",\n      \"pmids\": [\"21224076\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Single-lab, single-tumor-type evidence\", \"Causal effectors among 246 changed genes not validated\"]\n    },\n    {\n      \"year\": 2012,\n      \"claim\": \"Mapped the PI3K/Akt-dependent downstream effectors (p21, p27, FOXO3a, GSK3\\u03b2) controlled by AFAP1L2 in both RET/PTC-positive and -negative cells, generalizing its proliferation/survival mechanism.\",\n      \"evidence\": \"siRNA knockdown, Western blot of Akt substrates, cell-cycle and caspase assays in WRO and A549 cells\",\n      \"pmids\": [\"22928011\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Direct versus indirect effector regulation not separated\", \"Single lab\"]\n    },\n    {\n      \"year\": 2012,\n      \"claim\": \"Defined Y72 in the rodent ortholog as the critical p85-binding residue and placed AFAP1L2 in a cAMP/Src-driven mitogenic loop, confirming the phospho-tyrosine\\u2013p85 mechanism in a non-cancer thyroid system.\",\n      \"evidence\": \"MALDI-TOF identification, Y72F mutagenesis, PP1/PP2 inhibitors, Co-IP, PI3K activity and DNA synthesis assays in rat FRTL-5 cells\",\n      \"pmids\": [\"22496359\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Correspondence between rodent Y72 and human Y54 not formally reconciled\", \"Upstream kinase identity in this context limited to Src inference\"]\n    },\n    {\n      \"year\": 2015,\n      \"claim\": \"Identified the Tks5 scaffold as a direct N-terminal partner forming a Tks5-XB130-Src complex required for Src/PI3K/Akt activation, revealing a higher-order scaffolding mechanism for sustained signaling.\",\n      \"evidence\": \"Yeast two-hybrid, endogenous Co-IP, interface mutagenesis (XB130 \\u0394N, Tks5 SH3 W1108A), proliferation/apoptosis assays\",\n      \"pmids\": [\"26446840\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Structural basis of the SH3-polyproline interface not solved\", \"Whether complex assembles constitutively or stimulus-dependently unclear\"]\n    },\n    {\n      \"year\": 2015,\n      \"claim\": \"Extended the migration role to carcinogen (NNK)-induced motility in bronchial epithelium and linked AFAP1L2 to MMP-14 relocalization, connecting it to invasive machinery.\",\n      \"evidence\": \"Immunofluorescence, full-length/truncation overexpression, migration assays, MMP-14 localization in BEAS2B cells\",\n      \"pmids\": [\"25980441\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Direct AFAP1L2-MMP-14 link not established\", \"Single lab\"]\n    },\n    {\n      \"year\": 2016,\n      \"claim\": \"Reconstituted AFAP1L2 as a direct F-actin binder and crosslinker, mapping the C-terminal actin-binding region and N-terminal multimerization domain and tying these to endocytosis, providing the biochemical basis for its cortical functions.\",\n      \"evidence\": \"In vitro F-actin binding/crosslinking with recombinant protein, deletion mutagenesis, Blue native-PAGE, dextran uptake in HEK293\",\n      \"pmids\": [\"27462298\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"In vivo contribution of crosslinking to motility not quantified\", \"Regulation of actin binding by phosphorylation unknown\"]\n    },\n    {\n      \"year\": 2016,\n      \"claim\": \"Showed AFAP1L2/Rac1 and Tks5/Cdc42 modules direct distinct migration modes from a shared complex, defining how the scaffold partitions cytoskeletal output.\",\n      \"evidence\": \"Immunofluorescence with WAVE2/N-WASP markers, Co-IP (XB130-Rac1; Tks5-Cdc42), co-overexpression and migration assays in airway epithelial cells\",\n      \"pmids\": [\"27835612\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Direct versus indirect Rac1 binding not dissected\", \"Single lab\"]\n    },\n    {\n      \"year\": 2019,\n      \"claim\": \"Identified PTPRZ as the phosphatase counteracting AFAP1L2 tyrosine phosphorylation and placed AFAP1L2 in pleiotrophin-controlled PI3K-AKT-mTOR signaling for OPC differentiation and remyelination, establishing a physiological, non-cancer role and a bidirectional phospho-control axis.\",\n      \"evidence\": \"In vitro phosphatase assay, Co-IP, siRNA knockdown, PTPRZ catalytic-dead knock-in mice, pathway Western blots\",\n      \"pmids\": [\"30667096\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Specific dephosphorylated tyrosines not mapped\", \"Kinase responsible in OPCs not identified\"]\n    },\n    {\n      \"year\": 2021,\n      \"claim\": \"Defined a developmental, kinase-independent function: AFAP1L2 links the apical actin cortex to microtubules in thyrocytes to direct folliculogenesis, and its loss causes congenital hypothyroidism rescuable by thyroxine.\",\n      \"evidence\": \"Xb130 knockout mice, 3D thyrocyte cultures, confocal imaging, histology, levothyroxine rescue\",\n      \"pmids\": [\"34652970\", \"34470464\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Molecular bridge to microtubule-associated proteins not identified\", \"Relationship to Src/PI3K signaling in this context unclear\"]\n    },\n    {\n      \"year\": 2023,\n      \"claim\": \"Linked AFAP1L2 to mitophagy control via a SRC-FUNDC1 axis driving sorafenib resistance in HCC, expanding its effector repertoire beyond PI3K/Akt.\",\n      \"evidence\": \"Co-IP, CETSA, SPR, shRNA/overexpression, TEM mitophagy readout, xenografts in HCC cells\",\n      \"pmids\": [\"37733919\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Whether AFAP1L2 directly modulates SRC kinase activity or scaffolds FUNDC1 unclear\", \"Single lab\"]\n    },\n    {\n      \"year\": 2022,\n      \"claim\": \"Showed AFAP1L2 sustains PI3K/AKT in trastuzumab-resistant HER2+ gastric cancer and proposed a SRC-XB130-PTEN feedback loop, connecting it to therapy resistance.\",\n      \"evidence\": \"Co-IP (XB130-p85\\u03b1), shRNA knockdown, qRT-PCR, xenografts\",\n      \"pmids\": [\"36284062\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Mechanism of PTEN transcriptional repression not defined\", \"Single lab\"]\n    },\n    {\n      \"year\": 2025,\n      \"claim\": \"Identified post-transcriptional control of AFAP1L2 by 3'UTR-binding proteins hnRNPC and hnRNPR, which stabilize its mRNA against XRN1/DIS3L2 (and, for hnRNPC, enhance translation via eIF4E), explaining how AFAP1L2 levels are elevated in NSCLC.\",\n      \"evidence\": \"RNA pull-down, RIP, dual-luciferase 3'UTR reporters, Co-IP (hnRNPC-eIF4E), knockdown/overexpression, in vivo NSCLC models\",\n      \"pmids\": [\"39800708\", \"40268079\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Whether hnRNPC and hnRNPR act cooperatively or redundantly unclear\", \"Single lab for each\"]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"How AFAP1L2's two activities \\u2014 phospho-dependent PI3K recruitment and phospho-independent actin crosslinking \\u2014 are coordinated within a single cell and whether they share or compete for the same molecular pool remains unresolved.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"No structural model integrating PH domains, actin-binding, and scaffolding regions\", \"No high-resolution structure of any AFAP1L2 complex\", \"Regulation switching between signaling and cytoskeletal modes uncharacterized\"]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0060090\", \"supporting_discovery_ids\": [0, 1, 7]},\n      {\"term_id\": \"GO:0008092\", \"supporting_discovery_ids\": [2, 9]},\n      {\"term_id\": \"GO:0098772\", \"supporting_discovery_ids\": [0, 1, 5]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005829\", \"supporting_discovery_ids\": [0]},\n      {\"term_id\": \"GO:0005856\", \"supporting_discovery_ids\": [2, 9, 12]},\n      {\"term_id\": \"GO:0005886\", \"supporting_discovery_ids\": [10, 13]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-162582\", \"supporting_discovery_ids\": [0, 1, 5, 11]},\n      {\"term_id\": \"R-HSA-1643685\", \"supporting_discovery_ids\": [1, 14, 16]},\n      {\"term_id\": \"R-HSA-1266738\", \"supporting_discovery_ids\": [11, 12, 13]},\n      {\"term_id\": \"R-HSA-8953854\", \"supporting_discovery_ids\": [18, 19]},\n      {\"term_id\": \"R-HSA-9612973\", \"supporting_discovery_ids\": [14]}\n    ],\n    \"complexes\": [\"XB130-Tks5-Src complex\"],\n    \"partners\": [\"SRC\", \"PIK3R1\", \"TKS5\", \"RET\", \"PTPRZ1\", \"RAC1\", \"FUNDC1\", \"HNRNPC\"],\n    \"other_free_text\": []\n  }\n}","audit_flag":null,"evaluation":{"pairwise":"win","faith_supported":8,"faith_total":8,"faith_pct":100.0}}