{"gene":"RASAL2","run_date":"2026-06-10T06:43:36","timeline":{"discoveries":[{"year":2013,"finding":"RASAL2 functions as a RasGAP tumor and metastasis suppressor in breast cancer; its ablation promotes Ras hyperactivation, tumor growth, progression, and metastasis in mouse models, establishing that RASAL2 loss is an alternative mechanism of activating Ras in cancer.","method":"Genetic ablation (knockout mouse models), in vivo tumor/metastasis assays, human breast cancer mutation/expression analysis","journal":"Cancer cell","confidence":"High","confidence_rationale":"Tier 2 / Strong — loss-of-function in vivo mouse models with defined phenotypic readouts, replicated across human patient data and mouse models in a focused mechanistic study","pmids":["24029233"],"is_preprint":false},{"year":2014,"finding":"In triple-negative breast cancer (TNBC), RASAL2 acts independently of its RAS-GAP catalytic activity and instead promotes RAC1 signaling by binding and antagonizing the RAC1-GAP protein ARHGAP24, thereby driving mesenchymal invasion and metastasis.","method":"Co-immunoprecipitation (binding of RASAL2 to ARHGAP24), RAS-GAP catalytic mutant rescue experiments, RAC1 activity assays, shRNA knockdown/overexpression in TNBC cells","journal":"The Journal of clinical investigation","confidence":"High","confidence_rationale":"Tier 2 / Moderate — reciprocal Co-IP identifying RASAL2-ARHGAP24 interaction, catalytic mutant confirming GAP-independent mechanism, RAC1 activity assay, multiple orthogonal methods in one study","pmids":["25384218"],"is_preprint":false},{"year":2014,"finding":"RASAL2 knockdown in ovarian cancer cells activates the Ras-ERK pathway to promote EMT, migration, invasion, and tumor formation; pharmacological inhibition of the Ras-ERK pathway reverses the functional effects of RASAL2 depletion.","method":"shRNA knockdown, in vitro migration/invasion assays, in vivo tumor formation, ERK pathway inhibition rescue","journal":"Oncotarget","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — loss-of-function with defined phenotypic readout and pharmacological epistasis rescue, single lab","pmids":["25216515"],"is_preprint":false},{"year":2012,"finding":"RASAL2 physically interacts with ECT2 and regulates RHO GTPase activity in astrocytoma cells; RASAL2 knockdown causes conversion to an amoeboid migratory phenotype, identifying a role in mesenchymal-amoeboid transition.","method":"Cytoplasmic fractionation followed by ECT2 immunoprecipitation and mass spectrometry to identify RASAL2 as an ECT2-binding partner; RHO activity assay; RNA interference knockdown with phenotypic readout","journal":"The American journal of pathology","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — Co-IP/MS identifies binding partner, RHO activity assay and loss-of-function phenotype, single lab with two orthogonal methods","pmids":["22683310"],"is_preprint":false},{"year":2017,"finding":"RASAL2 suppresses bladder cancer stemness and EMT through the MAPK/SOX2 signaling axis; inhibition of ERK activity or knockdown of SOX2 reverses the stemness and mesenchymal properties induced by RASAL2 deficiency.","method":"Gain- and loss-of-function experiments in bladder cancer cells, ERK inhibitor and SOX2 siRNA rescue experiments, in vivo xenograft assays","journal":"Cell death & disease","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — epistasis via inhibitor/siRNA rescue, in vitro and in vivo with defined pathway placement, single lab","pmids":["28182001"],"is_preprint":false},{"year":2018,"finding":"RASAL2 inhibits bladder cancer angiogenesis by suppressing AKT phosphorylation, which in turn reduces ETS1 expression and VEGFA levels; a negative correlation between RASAL2 and VEGFA/CD31 was confirmed in xenografts and human specimens.","method":"shRNA/siRNA knockdown and ectopic overexpression, Western blot for p-AKT/ETS1/VEGFA, in vivo xenograft and human specimen correlation","journal":"Cellular signalling","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — defined pathway mechanism (p-AKT→ETS1→VEGFA) with in vitro and in vivo evidence, single lab","pmids":["29702203"],"is_preprint":false},{"year":2018,"finding":"RASAL2 knockdown in CRC cells promotes YAP1 phosphorylation, cytoplasmic retention, and ubiquitination, thereby activating the Hippo pathway through the LATS2/YAP1 axis; this oncogenic property was confirmed by co-immunoprecipitation.","method":"siRNA/shRNA knockdown, overexpression, expression microarray screening, co-immunoprecipitation, double immunofluorescence staining, in vivo studies","journal":"Molecular cancer","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — Co-IP confirms interaction, multiple functional assays in vitro and in vivo, defined pathway placement, single lab","pmids":["30037330"],"is_preprint":false},{"year":2018,"finding":"RASAL2 activates GSK3β by reducing its Ser9 phosphorylation and subsequently decreases c-FOS and VEGFA expression to inhibit tumor angiogenesis in renal cell carcinoma.","method":"shRNA knockdown and overexpression, Western blot for p-GSK3β/c-FOS/VEGFA, specific inhibitor and siRNA rescue experiments, in vitro and in vivo angiogenesis assays","journal":"Cell death & disease","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — defined pathway (GSK3β→c-FOS→VEGFA) with pharmacological and genetic rescue, single lab","pmids":["30158581"],"is_preprint":false},{"year":2019,"finding":"IPO5 binds to the NLS sequence of RASAL2 to mediate its nuclear translocation; nuclear RASAL2 is associated with RAS signal activation, thereby promoting colorectal cancer progression.","method":"Mass spectrometry, co-immunoprecipitation, subcellular fractionation, immunofluorescence, in vivo and in vitro functional experiments","journal":"Journal of experimental & clinical cancer research : CR","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — Co-IP and fractionation identify IPO5-RASAL2 interaction and nuclear localization, multiple orthogonal methods, single lab","pmids":["31288861"],"is_preprint":false},{"year":2021,"finding":"RASAL2 GAP activity suppresses negative feedback regulators SPRY1/2 and, together with EGFR upregulation, sustains basal RAS activity in TNBC; co-inhibition of MEK1/2 and EGFR induces synergistic apoptosis specifically in RASAL2-high tumors. YAP directly transcriptionally regulates RASAL2 expression.","method":"Transcriptional profiling, pharmacogenomic data mining, proteomic studies, in vitro and in vivo MEK/EGFR inhibitor combination treatment, GAP catalytic mutant experiments, chromatin immunoprecipitation (YAP-RASAL2 promoter)","journal":"Clinical cancer research","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — catalytic mutant confirms GAP-activity requirement, ChIP for YAP transcriptional regulation, in vitro and in vivo validation, single lab with multiple orthogonal methods","pmids":["34168046"],"is_preprint":false},{"year":2021,"finding":"PRKAA/AMPKα phosphorylates RASAL2 at Ser351 under glucose starvation; non-phosphorylated RASAL2 recruits phosphatase PPM1B to attenuate AMPKα phosphorylation and suppress basal autophagy, while phosphorylated RASAL2 binds the PIK3C3/VPS34-ATG14-BECN1 complex to increase PIK3C3 activity and promote autophagy, acting as a molecular switch.","method":"Co-immunoprecipitation (RASAL2 with PPM1B; phospho-RASAL2 with PIK3C3 complex), phosphorylation site mutagenesis (S351), double-knockout cells, PIK3C3 activity assay, autophagy flux assays","journal":"Autophagy","confidence":"High","confidence_rationale":"Tier 1 / Moderate — in vitro kinase phosphorylation, site-directed mutagenesis, Co-IP of phospho-RASAL2 with VPS34 complex, enzymatic PIK3C3 activity assay, multiple orthogonal methods in one study","pmids":["33563064"],"is_preprint":false},{"year":2019,"finding":"Rasal2 knockout in MCF-7 breast cancer cells enhances exosomal release and increases autophagy-related proteins in the exosomal fraction; autophagy inhibition (3-MA) abrogates while autophagy blockade (chloroquine) facilitates Rasal2 KO-induced secretory autophagy-driven exosome release and consequent cancer cell proliferation.","method":"CRISPR-Cas9 knockout, exosome isolation and characterization, autophagy inhibitor treatments, cell proliferation assays","journal":"Molecular and cellular biochemistry","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — genetic KO with pharmacological dissection of autophagy pathway, multiple functional readouts, single lab","pmids":["31473883"],"is_preprint":false},{"year":2019,"finding":"Phosphorylation of Rasal2 at Serine 237 promotes tumor growth in both ER+ and ER- breast cancer cells; p-Rasal2 and non-p-Rasal2 exert their effects on cancer progression via exosome-mediated transport, with the p-Rasal2/non-p-Rasal2 ratio helping determine whether total Rasal2 acts as a tumor suppressor (ER+) or promoter (ER-).","method":"Western blot for phospho-Ser237-Rasal2, exosome purification and characterization by TEM and flow cytometry, in vivo and in vitro tumor growth experiments","journal":"EBioMedicine","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — phospho-specific detection, in vitro and in vivo functional validation, exosome fractionation, single lab with multiple orthogonal methods","pmids":["31759919"],"is_preprint":false},{"year":2017,"finding":"Rasal2 deficiency impairs adipogenesis in vitro and in vivo by increasing Ras and ERK activity in preadipocytes; Ras inhibition (but not ERK inhibition) rescues impaired adipogenesis, indicating that Rasal2 promotes adipogenesis by repressing Ras activity in an ERK-independent manner.","method":"Insertional mutant mice (Rasal2-deficient), 3T3-L1 preadipocyte siRNA knockdown, Ras/ERK activity assays, pharmacological rescue with Ras and MEK inhibitors","journal":"Molecular metabolism","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — genetic and siRNA loss-of-function with pharmacological epistasis rescue, in vivo and in vitro evidence, single lab","pmids":["28580280"],"is_preprint":false},{"year":2022,"finding":"H. pylori infection induces RASAL2 expression via NF-κB directly binding the RASAL2 promoter; RASAL2 then inhibits protein phosphatase 2A (PP2A) activity through direct binding, leading to AKT activation and increased β-catenin transcriptional activity to drive gastric tumorigenesis.","method":"Chromatin immunoprecipitation (NF-κB at RASAL2 promoter), co-immunoprecipitation (RASAL2-PP2A), PP2A activity assay, gene silencing and ectopic overexpression, patient-derived organoids, in vivo xenograft models","journal":"Gastroenterology","confidence":"High","confidence_rationale":"Tier 1 / Moderate — ChIP for direct transcriptional regulation, Co-IP for protein binding, enzymatic PP2A activity assay, in vitro and in vivo validation with organoids, multiple orthogonal methods in one study","pmids":["35134322"],"is_preprint":false},{"year":2022,"finding":"RASAL2 promotes prostate cancer cell proliferation and G1-to-S phase transition by facilitating AKT phosphorylation, which in turn increases cyclin D1 (CCND1) expression; a positive correlation between RASAL2 and cyclin D1 was confirmed in xenografts and clinical specimens.","method":"shRNA knockdown and overexpression, cell cycle analysis, Western blot for p-AKT/cyclin D1, in vivo xenograft studies, clinical specimen correlation","journal":"Cell death discovery","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — defined pathway mechanism (RASAL2→p-AKT→cyclin D1) with in vitro and in vivo evidence, single lab","pmids":["35668070"],"is_preprint":false},{"year":2022,"finding":"RASAL2 deficiency attenuates hepatic steatosis by activating AKT signaling, which upregulates TET1 expression and promotes MTTP expression through DNA hydroxymethylation, increasing VLDL secretion from the liver.","method":"Chromatin immunoprecipitation assays, hydroxymethylated DNA immunoprecipitation (hMeDIP), in vivo VLDL secretion assay (tyloxapol injection), high-fat diet mouse model, in vitro hepatocyte model","journal":"Journal of clinical and translational hepatology","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — ChIP and hMeDIP for pathway mechanism, in vivo secretion assay, single lab with multiple orthogonal methods","pmids":["36643045"],"is_preprint":false},{"year":2022,"finding":"RASAL2 promotes PDAC progression by accumulating TIAM1 expression: RASAL2 inhibits YAP1 phosphorylation (stabilizing YAP1), which increases TIAM1 mRNA expression and suppresses TIAM1 protein ubiquitination, thereby enhancing YAP1/TIAM1 signaling.","method":"shRNA knockdown and overexpression, Western blot for YAP1 phosphorylation/TIAM1, ubiquitination assays, in vitro and in vivo functional studies","journal":"International journal of biological sciences","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — multiple levels of TIAM1 regulation assessed (phosphorylation, mRNA, ubiquitination), in vivo validation, single lab","pmids":["35844783"],"is_preprint":false},{"year":2023,"finding":"CAMSAP2 physically interacts with RASAL2 and facilitates its degradation through the ubiquitin-proteasome system, leading to ERK signaling activation and promotion of lung cancer metastasis.","method":"Proteomic/biochemical interaction analysis (BioGRID prediction followed by biochemical assays), Co-IP, ubiquitin-proteasome pathway assays, siRNA knockdown, in vivo tail vein metastasis model","journal":"Life sciences","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — Co-IP identifies CAMSAP2-RASAL2 interaction, ubiquitin-proteasome degradation mechanism shown, in vivo validation, single lab","pmids":["38159595"],"is_preprint":false},{"year":2025,"finding":"Rasal2 knockout in TNBC cells disrupts autophagic flux and induces secretory autophagy; Rasal2 directly binds Rab27a (confirmed by Co-IP) and inhibits its activity, and Rab27a knockdown suppresses Rasal2 KO-induced autophagic-exosome secretion and TNBC progression.","method":"CRISPR-Cas9 knockout, siRNA knockdown, Co-immunoprecipitation (RASAL2-Rab27a), confocal microscopy of autophagosome/MVB colocalization, NTA/TEM for exosome characterization, in vivo xenograft","journal":"Journal of translational medicine","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — Co-IP identifies RASAL2-Rab27a interaction, genetic rescue confirms pathway, in vitro and in vivo evidence, single lab","pmids":["40369567"],"is_preprint":false},{"year":2025,"finding":"The germline RASAL2 c.2423 A>G variant (p.Y808C) enhances RAS signaling with sustained ERK phosphorylation and increases CRC cell proliferation; mutant cells require higher doses of cetuximab for ERK suppression, conferring resistance to anti-EGFR therapy via abnormal RAS activation.","method":"Functional cell-based assays (ERK phosphorylation, proliferation), cetuximab dose-response experiments, population frequency analysis","journal":"Scientific reports","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — direct functional characterization of specific variant with defined biochemical readout (p-ERK) and pharmacological resistance assay, single lab","pmids":["40849341"],"is_preprint":false}],"current_model":"RASAL2 is a RasGAP that catalyzes hydrolysis of RAS-GTP to RAS-GDP to suppress RAS/ERK signaling; it acts as a tumor suppressor in most cancers (breast luminal B, ovarian, bladder, lung, NPC, RCC) by restraining Ras-ERK and downstream PI3K/AKT pathways, but functions as an oncogene in TNBC—where it acts GAP-independently by binding and antagonizing the RAC1-GAP ARHGAP24 to activate RAC1—and in CRC/gastric cancer, where it inhibits PP2A to activate AKT/β-catenin or suppresses the Hippo LATS2/YAP1 axis; its activity is further regulated by PRKAA/AMPKα phosphorylation at S351 (switching it from autophagy suppressor to activator via the VPS34-ATG14-BECN1 complex), by IPO5-mediated nuclear translocation, by CAMSAP2-driven ubiquitin-proteasome degradation, and through exosome-mediated intercellular communication."},"narrative":{"mechanistic_narrative":"RASAL2 is a Ras GTPase-activating protein that catalyzes hydrolysis of Ras-GTP to restrain Ras-ERK signaling, functioning predominantly as a tumor and metastasis suppressor whose loss constitutes an alternative route to Ras hyperactivation in cancer [PMID:24029233]. Through its catalytic suppression of Ras-ERK output, RASAL2 limits EMT, migration, invasion, and cancer stemness across ovarian and bladder tumors, with the latter mediated through a MAPK/SOX2 axis [PMID:25216515, PMID:28182001], and it further suppresses AKT-driven angiogenic programs by lowering ETS1/VEGFA and by activating GSK3β to reduce c-FOS/VEGFA [PMID:29702203, PMID:30158581]. RASAL2 displays striking context-dependent duality: in triple-negative breast cancer it acts independently of GAP catalysis by binding and antagonizing the RAC1-GAP ARHGAP24 to activate RAC1 and drive mesenchymal invasion [PMID:25384218], while in gastric cancer it is induced by NF-κB and binds and inhibits PP2A to activate AKT and β-catenin [PMID:35134322]; in colorectal, prostate, and pancreatic cancers it variously promotes proliferation and progression via nuclear translocation, AKT/cyclin D1 signaling, and YAP1/TIAM1 stabilization [PMID:31288861, PMID:35668070, PMID:35844783]. Its activity is governed by AMPKα (PRKAA) phosphorylation at Ser351, which switches RASAL2 between a PPM1B-recruiting autophagy suppressor and an activator of the PIK3C3/VPS34-ATG14-BECN1 complex [PMID:33563064], and its abundance is controlled by CAMSAP2-driven ubiquitin-proteasome degradation [PMID:38159595]. A germline RASAL2 p.Y808C variant enhances Ras signaling and confers resistance to anti-EGFR therapy [PMID:40849341].","teleology":[{"year":2012,"claim":"Established that RASAL2 is not solely a Ras regulator but also engages Rho-family signaling, by identifying a physical link to the RhoGEF ECT2 controlling migratory plasticity.","evidence":"ECT2 immunoprecipitation/mass spectrometry, RHO activity assay, and RNAi phenotyping in astrocytoma cells","pmids":["22683310"],"confidence":"Medium","gaps":["Direct vs. indirect nature of the RASAL2-ECT2 association not resolved by reciprocal validation","Connection between ECT2 binding and GAP catalysis unclear"]},{"year":2013,"claim":"Defined RASAL2 as a RasGAP tumor and metastasis suppressor in vivo, answering whether its loss is sufficient to hyperactivate Ras in cancer.","evidence":"Knockout mouse models, in vivo tumor/metastasis assays, and human breast cancer mutation/expression analysis","pmids":["24029233"],"confidence":"High","gaps":["Structural basis of Ras-GTP hydrolysis not defined","Did not address context-specific oncogenic roles"]},{"year":2014,"claim":"Revealed that RASAL2 has a GAP-independent oncogenic mode, by showing it antagonizes the RAC1-GAP ARHGAP24 to activate RAC1 in TNBC.","evidence":"Reciprocal Co-IP, RAS-GAP catalytic mutant rescue, and RAC1 activity assays in TNBC cells","pmids":["25384218"],"confidence":"High","gaps":["Determinants of whether RASAL2 acts as GAP or RAC1 activator in a given tumor unresolved","ARHGAP24 binding interface not mapped"]},{"year":2014,"claim":"Confirmed RASAL2 suppresses Ras-ERK-driven EMT in ovarian cancer via pharmacological epistasis.","evidence":"shRNA knockdown, migration/invasion assays, and ERK pathway inhibitor rescue","pmids":["25216515"],"confidence":"Medium","gaps":["Single-lab evidence","Direct GAP substrate engagement not shown biochemically"]},{"year":2017,"claim":"Extended tumor-suppressor function to bladder cancer stemness through a defined MAPK/SOX2 axis, and to adipocyte biology where RASAL2 represses Ras in an ERK-independent manner.","evidence":"Gain/loss-of-function with ERK inhibitor and SOX2 siRNA rescue in bladder cancer; Rasal2-deficient mice and 3T3-L1 knockdown with Ras/MEK inhibitor rescue for adipogenesis","pmids":["28182001","28580280"],"confidence":"Medium","gaps":["ERK-independent effector downstream of Ras in adipogenesis unidentified","Mechanism linking MAPK to SOX2 not fully resolved"]},{"year":2018,"claim":"Mapped distinct anti-angiogenic and oncogenic effector branches, placing RASAL2 upstream of AKT/ETS1/VEGFA and GSK3β/c-FOS/VEGFA in suppressor contexts and upstream of LATS2/YAP1 in CRC.","evidence":"Knockdown/overexpression, Western blotting of pathway nodes, Co-IP, and in vivo/specimen correlation across bladder, RCC, and CRC","pmids":["29702203","30158581","30037330"],"confidence":"Medium","gaps":["Whether AKT/GSK3β effects depend on GAP catalysis not tested","Opposing YAP regulation across tissues unexplained"]},{"year":2019,"claim":"Identified IPO5-mediated nuclear translocation as a regulatory mechanism coupling RASAL2 localization to Ras activation in CRC, and uncovered exosome- and phosphorylation-based control of its tumor-suppressor/promoter balance in breast cancer.","evidence":"Mass spectrometry, Co-IP, subcellular fractionation for IPO5; CRISPR KO, exosome isolation, and phospho-Ser237 detection in breast cancer models","pmids":["31288861","31473883","31759919"],"confidence":"Medium","gaps":["Functional role of nuclear RASAL2 beyond correlation with Ras activity unclear","How the p-Rasal2/non-p-Rasal2 ratio is set physiologically unknown"]},{"year":2021,"claim":"Defined the AMPKα-Ser351 phosphorylation switch governing RASAL2 control of autophagy, and showed GAP activity sustains basal Ras in TNBC via SPRY1/2 suppression with YAP as a transcriptional input.","evidence":"In vitro kinase assay, S351 mutagenesis, Co-IP with PPM1B and the VPS34 complex, PIK3C3 activity assay; ChIP for YAP-RASAL2 promoter and MEK/EGFR inhibitor synergy with GAP mutant","pmids":["33563064","34168046"],"confidence":"High","gaps":["Upstream signals dictating Ser351 phosphorylation in tumors not defined","Whether the autophagy switch operates in the cancer contexts described elsewhere untested"]},{"year":2022,"claim":"Consolidated the oncogenic arm of RASAL2 across gastric, prostate, pancreatic, and hepatic contexts via AKT, β-catenin, cyclin D1, YAP1/TIAM1, and TET1/MTTP effectors, including NF-κB-driven induction and direct PP2A inhibition.","evidence":"ChIP, Co-IP (RASAL2-PP2A), PP2A activity assay, organoids, ubiquitination/hMeDIP assays, and in vivo models across gastric, prostate, PDAC, and liver","pmids":["35134322","35668070","35844783","36643045"],"confidence":"Medium","gaps":["Why RASAL2 inhibits PP2A in some tissues yet suppresses AKT in others unresolved","Structural basis of PP2A binding undetermined"]},{"year":2023,"claim":"Identified CAMSAP2 as a regulator of RASAL2 stability, showing ubiquitin-proteasome degradation of RASAL2 unleashes ERK signaling to drive lung cancer metastasis.","evidence":"Co-IP, ubiquitin-proteasome assays, siRNA knockdown, and tail-vein metastasis model","pmids":["38159595"],"confidence":"Medium","gaps":["The E3 ligase mediating degradation not identified","Whether CAMSAP2 acts as adaptor or scaffold unclear"]},{"year":2025,"claim":"Connected RASAL2 to secretory-autophagy exosome biology through direct inhibition of Rab27a in TNBC, and demonstrated a germline variant that hyperactivates Ras and confers anti-EGFR resistance.","evidence":"CRISPR KO, Co-IP (RASAL2-Rab27a), autophagosome/MVB colocalization, exosome characterization for Rab27a; functional p-ERK and cetuximab dose-response assays for the p.Y808C variant","pmids":["40369567","40849341"],"confidence":"Medium","gaps":["Whether Rab27a inhibition is GAP-dependent untested","Clinical penetrance and disease association of p.Y808C not established"]},{"year":null,"claim":"What determines whether RASAL2 functions as a GAP-dependent tumor suppressor or a GAP-independent oncogene in a given tissue remains the central unresolved question.","evidence":"","pmids":[],"confidence":"Medium","gaps":["No unifying model integrating phosphorylation state, localization, and binding partners to predict suppressor vs. oncogene outcome","No high-resolution structure of RASAL2 or its complexes","Context-switch mechanism between PP2A/AKT activation and AKT/VEGFA suppression undefined"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0098772","term_label":"molecular function regulator activity","supporting_discovery_ids":[1,14,10]},{"term_id":"GO:0140096","term_label":"catalytic activity, acting on a protein","supporting_discovery_ids":[14]}],"localization":[{"term_id":"GO:0005829","term_label":"cytosol","supporting_discovery_ids":[3]},{"term_id":"GO:0005634","term_label":"nucleus","supporting_discovery_ids":[8]}],"pathway":[{"term_id":"R-HSA-162582","term_label":"Signal Transduction","supporting_discovery_ids":[0,2,1]},{"term_id":"R-HSA-9612973","term_label":"Autophagy","supporting_discovery_ids":[10,11,19]},{"term_id":"R-HSA-1643685","term_label":"Disease","supporting_discovery_ids":[0,20]}],"complexes":[],"partners":["ARHGAP24","ECT2","PP2A","IPO5","PPM1B","PIK3C3","CAMSAP2","RAB27A"],"other_free_text":[]}},"prefetch_data":{"uniprot":{"accession":"Q9UJF2","full_name":"Ras GTPase-activating protein nGAP","aliases":["RAS protein activator-like 2"],"length_aa":1139,"mass_kda":128.6,"function":"Inhibitory regulator of the Ras-cyclic AMP pathway","subcellular_location":"","url":"https://www.uniprot.org/uniprotkb/Q9UJF2/entry"},"depmap":{"release":"DepMap","has_data":true,"is_common_essential":false,"resolved_as":"","url":"https://depmap.org/portal/gene/RASAL2","classification":"Not Classified","n_dependent_lines":0,"n_total_lines":1208,"dependency_fraction":0.0},"opencell":{"profiled":false,"resolved_as":"","ensg_id":"","cell_line_id":"","localizations":[],"interactors":[{"gene":"CSNK1A1","stoichiometry":4.0}],"url":"https://opencell.sf.czbiohub.org/search/RASAL2","total_profiled":1310},"omim":[{"mim_id":"609205","title":"DAB2-INTERACTING PROTEIN; DAB2IP","url":"https://www.omim.org/entry/609205"},{"mim_id":"606136","title":"RAS PROTEIN ACTIVATOR-LIKE 2; RASAL2","url":"https://www.omim.org/entry/606136"}],"hpa":{"profiled":true,"resolved_as":"","reliability":"Approved","locations":[{"location":"Plasma membrane","reliability":"Approved"},{"location":"Focal adhesion sites","reliability":"Approved"}],"tissue_specificity":"Low tissue specificity","tissue_distribution":"Detected in many","driving_tissues":[],"url":"https://www.proteinatlas.org/search/RASAL2"},"hgnc":{"alias_symbol":["nGAP"],"prev_symbol":[]},"alphafold":{"accession":"Q9UJF2","domains":[{"cath_id":"2.60.40.150","chopping":"168-296","consensus_level":"high","plddt":86.1376,"start":168,"end":296},{"cath_id":"1.10.506.10","chopping":"344-568","consensus_level":"high","plddt":91.6679,"start":344,"end":568},{"cath_id":"3.30.160","chopping":"112-165","consensus_level":"medium","plddt":84.4404,"start":112,"end":165}],"viewer_url":"https://alphafold.ebi.ac.uk/entry/Q9UJF2","model_url":"https://alphafold.ebi.ac.uk/files/AF-Q9UJF2-F1-model_v6.cif","pae_url":"https://alphafold.ebi.ac.uk/files/AF-Q9UJF2-F1-predicted_aligned_error_v6.png","plddt_mean":65.94},"mouse_models":{"mgi_url":"https://www.informatics.jax.org/marker/summary?nomen=RASAL2","jax_strain_url":"https://www.jax.org/strain/search?query=RASAL2"},"sequence":{"accession":"Q9UJF2","fasta_url":"https://rest.uniprot.org/uniprotkb/Q9UJF2.fasta","uniprot_url":"https://www.uniprot.org/uniprotkb/Q9UJF2/entry","alphafold_viewer_url":"https://alphafold.ebi.ac.uk/entry/Q9UJF2"}},"corpus_meta":[{"pmid":"24029233","id":"PMC_24029233","title":"The RasGAP gene, RASAL2, is a tumor and metastasis suppressor.","date":"2013","source":"Cancer cell","url":"https://pubmed.ncbi.nlm.nih.gov/24029233","citation_count":113,"is_preprint":false},{"pmid":"25384218","id":"PMC_25384218","title":"RASAL2 activates RAC1 to promote triple-negative breast cancer progression.","date":"2014","source":"The Journal of clinical investigation","url":"https://pubmed.ncbi.nlm.nih.gov/25384218","citation_count":81,"is_preprint":false},{"pmid":"27108696","id":"PMC_27108696","title":"miR-136 suppresses tumor invasion and metastasis by targeting RASAL2 in triple-negative breast cancer.","date":"2016","source":"Oncology reports","url":"https://pubmed.ncbi.nlm.nih.gov/27108696","citation_count":75,"is_preprint":false},{"pmid":"35134322","id":"PMC_35134322","title":"Helicobacter pylori-induced RASAL2 Through Activation of Nuclear Factor-κB Promotes Gastric Tumorigenesis via β-catenin Signaling Axis.","date":"2022","source":"Gastroenterology","url":"https://pubmed.ncbi.nlm.nih.gov/35134322","citation_count":71,"is_preprint":false},{"pmid":"30037330","id":"PMC_30037330","title":"RASAL2 promotes tumor progression through LATS2/YAP1 axis of hippo signaling pathway in colorectal cancer.","date":"2018","source":"Molecular cancer","url":"https://pubmed.ncbi.nlm.nih.gov/30037330","citation_count":63,"is_preprint":false},{"pmid":"25216515","id":"PMC_25216515","title":"RASAL2 down-regulation in ovarian cancer promotes epithelial-mesenchymal transition and metastasis.","date":"2014","source":"Oncotarget","url":"https://pubmed.ncbi.nlm.nih.gov/25216515","citation_count":49,"is_preprint":false},{"pmid":"28182001","id":"PMC_28182001","title":"RASAL2, a RAS GTPase-activating protein, inhibits stemness and epithelial-mesenchymal transition via MAPK/SOX2 pathway in bladder cancer.","date":"2017","source":"Cell death & disease","url":"https://pubmed.ncbi.nlm.nih.gov/28182001","citation_count":40,"is_preprint":false},{"pmid":"22683310","id":"PMC_22683310","title":"ECT2 and RASAL2 mediate mesenchymal-amoeboid transition in human astrocytoma cells.","date":"2012","source":"The American journal of pathology","url":"https://pubmed.ncbi.nlm.nih.gov/22683310","citation_count":37,"is_preprint":false},{"pmid":"31044154","id":"PMC_31044154","title":"Sulforaphane Induces miR135b-5p and Its Target Gene, RASAL2, thereby Inhibiting the Progression of Pancreatic Cancer.","date":"2019","source":"Molecular therapy oncolytics","url":"https://pubmed.ncbi.nlm.nih.gov/31044154","citation_count":36,"is_preprint":false},{"pmid":"31288861","id":"PMC_31288861","title":"IPO5 promotes the proliferation and tumourigenicity of colorectal cancer cells by mediating RASAL2 nuclear transportation.","date":"2019","source":"Journal of experimental & clinical cancer research : CR","url":"https://pubmed.ncbi.nlm.nih.gov/31288861","citation_count":36,"is_preprint":false},{"pmid":"25446096","id":"PMC_25446096","title":"RASAL2 promotes lung cancer metastasis through epithelial-mesenchymal transition.","date":"2014","source":"Biochemical and biophysical research communications","url":"https://pubmed.ncbi.nlm.nih.gov/25446096","citation_count":30,"is_preprint":false},{"pmid":"30158581","id":"PMC_30158581","title":"The expression and function of RASAL2 in renal cell carcinoma angiogenesis.","date":"2018","source":"Cell death & disease","url":"https://pubmed.ncbi.nlm.nih.gov/30158581","citation_count":23,"is_preprint":false},{"pmid":"33300072","id":"PMC_33300072","title":"miR‑654‑3p suppresses cell viability and promotes apoptosis by targeting RASAL2 in non‑small‑cell lung cancer.","date":"2020","source":"Molecular medicine 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Dependency in Chemoresistant Triple-Negative Breast Cancer.","date":"2021","source":"Clinical cancer research : an official journal of the American Association for Cancer Research","url":"https://pubmed.ncbi.nlm.nih.gov/34168046","citation_count":19,"is_preprint":false},{"pmid":"31473883","id":"PMC_31473883","title":"Rasal2 suppresses breast cancer cell proliferation modulated by secretory autophagy.","date":"2019","source":"Molecular and cellular biochemistry","url":"https://pubmed.ncbi.nlm.nih.gov/31473883","citation_count":19,"is_preprint":false},{"pmid":"28454265","id":"PMC_28454265","title":"Downregulation of RASAL2 promotes the proliferation, epithelial-mesenchymal transition and metastasis of colorectal cancer cells.","date":"2017","source":"Oncology letters","url":"https://pubmed.ncbi.nlm.nih.gov/28454265","citation_count":19,"is_preprint":false},{"pmid":"33563064","id":"PMC_33563064","title":"PRKAA/AMPKα phosphorylation switches the role of RASAL2 from a suppressor to an activator of autophagy.","date":"2021","source":"Autophagy","url":"https://pubmed.ncbi.nlm.nih.gov/33563064","citation_count":18,"is_preprint":false},{"pmid":"26770493","id":"PMC_26770493","title":"RASAL2 inhibited the proliferation and metastasis capability of nasopharyngeal carcinoma.","date":"2015","source":"International journal of clinical and experimental medicine","url":"https://pubmed.ncbi.nlm.nih.gov/26770493","citation_count":16,"is_preprint":false},{"pmid":"28580280","id":"PMC_28580280","title":"Rasal2 deficiency reduces adipogenesis and occurrence of obesity-related disorders.","date":"2017","source":"Molecular metabolism","url":"https://pubmed.ncbi.nlm.nih.gov/28580280","citation_count":14,"is_preprint":false},{"pmid":"31759919","id":"PMC_31759919","title":"Phosphorylated Rasal2 facilitates breast cancer progression.","date":"2019","source":"EBioMedicine","url":"https://pubmed.ncbi.nlm.nih.gov/31759919","citation_count":13,"is_preprint":false},{"pmid":"35668070","id":"PMC_35668070","title":"RASAL2 regulates the cell cycle and cyclin D1 expression through PI3K/AKT signalling in prostate tumorigenesis.","date":"2022","source":"Cell death discovery","url":"https://pubmed.ncbi.nlm.nih.gov/35668070","citation_count":13,"is_preprint":false},{"pmid":"31990059","id":"PMC_31990059","title":"MicroRNA-876-5p represses the cell proliferation and invasion of colorectal cancer through suppressing YAP signalling via targeting RASAL2.","date":"2020","source":"Clinical and experimental pharmacology & physiology","url":"https://pubmed.ncbi.nlm.nih.gov/31990059","citation_count":11,"is_preprint":false},{"pmid":"24029223","id":"PMC_24029223","title":"RASAL2: wrestling in the combat of Ras activation.","date":"2013","source":"Cancer 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its ablation promotes Ras hyperactivation, tumor growth, progression, and metastasis in mouse models, establishing that RASAL2 loss is an alternative mechanism of activating Ras in cancer.\",\n      \"method\": \"Genetic ablation (knockout mouse models), in vivo tumor/metastasis assays, human breast cancer mutation/expression analysis\",\n      \"journal\": \"Cancer cell\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — loss-of-function in vivo mouse models with defined phenotypic readouts, replicated across human patient data and mouse models in a focused mechanistic study\",\n      \"pmids\": [\"24029233\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2014,\n      \"finding\": \"In triple-negative breast cancer (TNBC), RASAL2 acts independently of its RAS-GAP catalytic activity and instead promotes RAC1 signaling by binding and antagonizing the RAC1-GAP protein ARHGAP24, thereby driving mesenchymal invasion and metastasis.\",\n      \"method\": \"Co-immunoprecipitation (binding of RASAL2 to ARHGAP24), RAS-GAP catalytic mutant rescue experiments, RAC1 activity assays, shRNA knockdown/overexpression in TNBC cells\",\n      \"journal\": \"The Journal of clinical investigation\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — reciprocal Co-IP identifying RASAL2-ARHGAP24 interaction, catalytic mutant confirming GAP-independent mechanism, RAC1 activity assay, multiple orthogonal methods in one study\",\n      \"pmids\": [\"25384218\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2014,\n      \"finding\": \"RASAL2 knockdown in ovarian cancer cells activates the Ras-ERK pathway to promote EMT, migration, invasion, and tumor formation; pharmacological inhibition of the Ras-ERK pathway reverses the functional effects of RASAL2 depletion.\",\n      \"method\": \"shRNA knockdown, in vitro migration/invasion assays, in vivo tumor formation, ERK pathway inhibition rescue\",\n      \"journal\": \"Oncotarget\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — loss-of-function with defined phenotypic readout and pharmacological epistasis rescue, single lab\",\n      \"pmids\": [\"25216515\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2012,\n      \"finding\": \"RASAL2 physically interacts with ECT2 and regulates RHO GTPase activity in astrocytoma cells; RASAL2 knockdown causes conversion to an amoeboid migratory phenotype, identifying a role in mesenchymal-amoeboid transition.\",\n      \"method\": \"Cytoplasmic fractionation followed by ECT2 immunoprecipitation and mass spectrometry to identify RASAL2 as an ECT2-binding partner; RHO activity assay; RNA interference knockdown with phenotypic readout\",\n      \"journal\": \"The American journal of pathology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — Co-IP/MS identifies binding partner, RHO activity assay and loss-of-function phenotype, single lab with two orthogonal methods\",\n      \"pmids\": [\"22683310\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2017,\n      \"finding\": \"RASAL2 suppresses bladder cancer stemness and EMT through the MAPK/SOX2 signaling axis; inhibition of ERK activity or knockdown of SOX2 reverses the stemness and mesenchymal properties induced by RASAL2 deficiency.\",\n      \"method\": \"Gain- and loss-of-function experiments in bladder cancer cells, ERK inhibitor and SOX2 siRNA rescue experiments, in vivo xenograft assays\",\n      \"journal\": \"Cell death & disease\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — epistasis via inhibitor/siRNA rescue, in vitro and in vivo with defined pathway placement, single lab\",\n      \"pmids\": [\"28182001\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2018,\n      \"finding\": \"RASAL2 inhibits bladder cancer angiogenesis by suppressing AKT phosphorylation, which in turn reduces ETS1 expression and VEGFA levels; a negative correlation between RASAL2 and VEGFA/CD31 was confirmed in xenografts and human specimens.\",\n      \"method\": \"shRNA/siRNA knockdown and ectopic overexpression, Western blot for p-AKT/ETS1/VEGFA, in vivo xenograft and human specimen correlation\",\n      \"journal\": \"Cellular signalling\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — defined pathway mechanism (p-AKT→ETS1→VEGFA) with in vitro and in vivo evidence, single lab\",\n      \"pmids\": [\"29702203\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2018,\n      \"finding\": \"RASAL2 knockdown in CRC cells promotes YAP1 phosphorylation, cytoplasmic retention, and ubiquitination, thereby activating the Hippo pathway through the LATS2/YAP1 axis; this oncogenic property was confirmed by co-immunoprecipitation.\",\n      \"method\": \"siRNA/shRNA knockdown, overexpression, expression microarray screening, co-immunoprecipitation, double immunofluorescence staining, in vivo studies\",\n      \"journal\": \"Molecular cancer\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — Co-IP confirms interaction, multiple functional assays in vitro and in vivo, defined pathway placement, single lab\",\n      \"pmids\": [\"30037330\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2018,\n      \"finding\": \"RASAL2 activates GSK3β by reducing its Ser9 phosphorylation and subsequently decreases c-FOS and VEGFA expression to inhibit tumor angiogenesis in renal cell carcinoma.\",\n      \"method\": \"shRNA knockdown and overexpression, Western blot for p-GSK3β/c-FOS/VEGFA, specific inhibitor and siRNA rescue experiments, in vitro and in vivo angiogenesis assays\",\n      \"journal\": \"Cell death & disease\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — defined pathway (GSK3β→c-FOS→VEGFA) with pharmacological and genetic rescue, single lab\",\n      \"pmids\": [\"30158581\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"IPO5 binds to the NLS sequence of RASAL2 to mediate its nuclear translocation; nuclear RASAL2 is associated with RAS signal activation, thereby promoting colorectal cancer progression.\",\n      \"method\": \"Mass spectrometry, co-immunoprecipitation, subcellular fractionation, immunofluorescence, in vivo and in vitro functional experiments\",\n      \"journal\": \"Journal of experimental & clinical cancer research : CR\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — Co-IP and fractionation identify IPO5-RASAL2 interaction and nuclear localization, multiple orthogonal methods, single lab\",\n      \"pmids\": [\"31288861\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"RASAL2 GAP activity suppresses negative feedback regulators SPRY1/2 and, together with EGFR upregulation, sustains basal RAS activity in TNBC; co-inhibition of MEK1/2 and EGFR induces synergistic apoptosis specifically in RASAL2-high tumors. YAP directly transcriptionally regulates RASAL2 expression.\",\n      \"method\": \"Transcriptional profiling, pharmacogenomic data mining, proteomic studies, in vitro and in vivo MEK/EGFR inhibitor combination treatment, GAP catalytic mutant experiments, chromatin immunoprecipitation (YAP-RASAL2 promoter)\",\n      \"journal\": \"Clinical cancer research\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — catalytic mutant confirms GAP-activity requirement, ChIP for YAP transcriptional regulation, in vitro and in vivo validation, single lab with multiple orthogonal methods\",\n      \"pmids\": [\"34168046\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"PRKAA/AMPKα phosphorylates RASAL2 at Ser351 under glucose starvation; non-phosphorylated RASAL2 recruits phosphatase PPM1B to attenuate AMPKα phosphorylation and suppress basal autophagy, while phosphorylated RASAL2 binds the PIK3C3/VPS34-ATG14-BECN1 complex to increase PIK3C3 activity and promote autophagy, acting as a molecular switch.\",\n      \"method\": \"Co-immunoprecipitation (RASAL2 with PPM1B; phospho-RASAL2 with PIK3C3 complex), phosphorylation site mutagenesis (S351), double-knockout cells, PIK3C3 activity assay, autophagy flux assays\",\n      \"journal\": \"Autophagy\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Moderate — in vitro kinase phosphorylation, site-directed mutagenesis, Co-IP of phospho-RASAL2 with VPS34 complex, enzymatic PIK3C3 activity assay, multiple orthogonal methods in one study\",\n      \"pmids\": [\"33563064\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"Rasal2 knockout in MCF-7 breast cancer cells enhances exosomal release and increases autophagy-related proteins in the exosomal fraction; autophagy inhibition (3-MA) abrogates while autophagy blockade (chloroquine) facilitates Rasal2 KO-induced secretory autophagy-driven exosome release and consequent cancer cell proliferation.\",\n      \"method\": \"CRISPR-Cas9 knockout, exosome isolation and characterization, autophagy inhibitor treatments, cell proliferation assays\",\n      \"journal\": \"Molecular and cellular biochemistry\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — genetic KO with pharmacological dissection of autophagy pathway, multiple functional readouts, single lab\",\n      \"pmids\": [\"31473883\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"Phosphorylation of Rasal2 at Serine 237 promotes tumor growth in both ER+ and ER- breast cancer cells; p-Rasal2 and non-p-Rasal2 exert their effects on cancer progression via exosome-mediated transport, with the p-Rasal2/non-p-Rasal2 ratio helping determine whether total Rasal2 acts as a tumor suppressor (ER+) or promoter (ER-).\",\n      \"method\": \"Western blot for phospho-Ser237-Rasal2, exosome purification and characterization by TEM and flow cytometry, in vivo and in vitro tumor growth experiments\",\n      \"journal\": \"EBioMedicine\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — phospho-specific detection, in vitro and in vivo functional validation, exosome fractionation, single lab with multiple orthogonal methods\",\n      \"pmids\": [\"31759919\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2017,\n      \"finding\": \"Rasal2 deficiency impairs adipogenesis in vitro and in vivo by increasing Ras and ERK activity in preadipocytes; Ras inhibition (but not ERK inhibition) rescues impaired adipogenesis, indicating that Rasal2 promotes adipogenesis by repressing Ras activity in an ERK-independent manner.\",\n      \"method\": \"Insertional mutant mice (Rasal2-deficient), 3T3-L1 preadipocyte siRNA knockdown, Ras/ERK activity assays, pharmacological rescue with Ras and MEK inhibitors\",\n      \"journal\": \"Molecular metabolism\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — genetic and siRNA loss-of-function with pharmacological epistasis rescue, in vivo and in vitro evidence, single lab\",\n      \"pmids\": [\"28580280\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"H. pylori infection induces RASAL2 expression via NF-κB directly binding the RASAL2 promoter; RASAL2 then inhibits protein phosphatase 2A (PP2A) activity through direct binding, leading to AKT activation and increased β-catenin transcriptional activity to drive gastric tumorigenesis.\",\n      \"method\": \"Chromatin immunoprecipitation (NF-κB at RASAL2 promoter), co-immunoprecipitation (RASAL2-PP2A), PP2A activity assay, gene silencing and ectopic overexpression, patient-derived organoids, in vivo xenograft models\",\n      \"journal\": \"Gastroenterology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Moderate — ChIP for direct transcriptional regulation, Co-IP for protein binding, enzymatic PP2A activity assay, in vitro and in vivo validation with organoids, multiple orthogonal methods in one study\",\n      \"pmids\": [\"35134322\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"RASAL2 promotes prostate cancer cell proliferation and G1-to-S phase transition by facilitating AKT phosphorylation, which in turn increases cyclin D1 (CCND1) expression; a positive correlation between RASAL2 and cyclin D1 was confirmed in xenografts and clinical specimens.\",\n      \"method\": \"shRNA knockdown and overexpression, cell cycle analysis, Western blot for p-AKT/cyclin D1, in vivo xenograft studies, clinical specimen correlation\",\n      \"journal\": \"Cell death discovery\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — defined pathway mechanism (RASAL2→p-AKT→cyclin D1) with in vitro and in vivo evidence, single lab\",\n      \"pmids\": [\"35668070\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"RASAL2 deficiency attenuates hepatic steatosis by activating AKT signaling, which upregulates TET1 expression and promotes MTTP expression through DNA hydroxymethylation, increasing VLDL secretion from the liver.\",\n      \"method\": \"Chromatin immunoprecipitation assays, hydroxymethylated DNA immunoprecipitation (hMeDIP), in vivo VLDL secretion assay (tyloxapol injection), high-fat diet mouse model, in vitro hepatocyte model\",\n      \"journal\": \"Journal of clinical and translational hepatology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — ChIP and hMeDIP for pathway mechanism, in vivo secretion assay, single lab with multiple orthogonal methods\",\n      \"pmids\": [\"36643045\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"RASAL2 promotes PDAC progression by accumulating TIAM1 expression: RASAL2 inhibits YAP1 phosphorylation (stabilizing YAP1), which increases TIAM1 mRNA expression and suppresses TIAM1 protein ubiquitination, thereby enhancing YAP1/TIAM1 signaling.\",\n      \"method\": \"shRNA knockdown and overexpression, Western blot for YAP1 phosphorylation/TIAM1, ubiquitination assays, in vitro and in vivo functional studies\",\n      \"journal\": \"International journal of biological sciences\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — multiple levels of TIAM1 regulation assessed (phosphorylation, mRNA, ubiquitination), in vivo validation, single lab\",\n      \"pmids\": [\"35844783\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"CAMSAP2 physically interacts with RASAL2 and facilitates its degradation through the ubiquitin-proteasome system, leading to ERK signaling activation and promotion of lung cancer metastasis.\",\n      \"method\": \"Proteomic/biochemical interaction analysis (BioGRID prediction followed by biochemical assays), Co-IP, ubiquitin-proteasome pathway assays, siRNA knockdown, in vivo tail vein metastasis model\",\n      \"journal\": \"Life sciences\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — Co-IP identifies CAMSAP2-RASAL2 interaction, ubiquitin-proteasome degradation mechanism shown, in vivo validation, single lab\",\n      \"pmids\": [\"38159595\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"Rasal2 knockout in TNBC cells disrupts autophagic flux and induces secretory autophagy; Rasal2 directly binds Rab27a (confirmed by Co-IP) and inhibits its activity, and Rab27a knockdown suppresses Rasal2 KO-induced autophagic-exosome secretion and TNBC progression.\",\n      \"method\": \"CRISPR-Cas9 knockout, siRNA knockdown, Co-immunoprecipitation (RASAL2-Rab27a), confocal microscopy of autophagosome/MVB colocalization, NTA/TEM for exosome characterization, in vivo xenograft\",\n      \"journal\": \"Journal of translational medicine\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — Co-IP identifies RASAL2-Rab27a interaction, genetic rescue confirms pathway, in vitro and in vivo evidence, single lab\",\n      \"pmids\": [\"40369567\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"The germline RASAL2 c.2423 A>G variant (p.Y808C) enhances RAS signaling with sustained ERK phosphorylation and increases CRC cell proliferation; mutant cells require higher doses of cetuximab for ERK suppression, conferring resistance to anti-EGFR therapy via abnormal RAS activation.\",\n      \"method\": \"Functional cell-based assays (ERK phosphorylation, proliferation), cetuximab dose-response experiments, population frequency analysis\",\n      \"journal\": \"Scientific reports\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — direct functional characterization of specific variant with defined biochemical readout (p-ERK) and pharmacological resistance assay, single lab\",\n      \"pmids\": [\"40849341\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"RASAL2 is a RasGAP that catalyzes hydrolysis of RAS-GTP to RAS-GDP to suppress RAS/ERK signaling; it acts as a tumor suppressor in most cancers (breast luminal B, ovarian, bladder, lung, NPC, RCC) by restraining Ras-ERK and downstream PI3K/AKT pathways, but functions as an oncogene in TNBC—where it acts GAP-independently by binding and antagonizing the RAC1-GAP ARHGAP24 to activate RAC1—and in CRC/gastric cancer, where it inhibits PP2A to activate AKT/β-catenin or suppresses the Hippo LATS2/YAP1 axis; its activity is further regulated by PRKAA/AMPKα phosphorylation at S351 (switching it from autophagy suppressor to activator via the VPS34-ATG14-BECN1 complex), by IPO5-mediated nuclear translocation, by CAMSAP2-driven ubiquitin-proteasome degradation, and through exosome-mediated intercellular communication.\"\n}\n```","stage2_raw":"{\n  \"mechanistic_narrative\": \"RASAL2 is a Ras GTPase-activating protein that catalyzes hydrolysis of Ras-GTP to restrain Ras-ERK signaling, functioning predominantly as a tumor and metastasis suppressor whose loss constitutes an alternative route to Ras hyperactivation in cancer [#0]. Through its catalytic suppression of Ras-ERK output, RASAL2 limits EMT, migration, invasion, and cancer stemness across ovarian and bladder tumors, with the latter mediated through a MAPK/SOX2 axis [#2, #4], and it further suppresses AKT-driven angiogenic programs by lowering ETS1/VEGFA and by activating GSK3\\u03b2 to reduce c-FOS/VEGFA [#5, #7]. RASAL2 displays striking context-dependent duality: in triple-negative breast cancer it acts independently of GAP catalysis by binding and antagonizing the RAC1-GAP ARHGAP24 to activate RAC1 and drive mesenchymal invasion [#1], while in gastric cancer it is induced by NF-\\u03baB and binds and inhibits PP2A to activate AKT and \\u03b2-catenin [#14]; in colorectal, prostate, and pancreatic cancers it variously promotes proliferation and progression via nuclear translocation, AKT/cyclin D1 signaling, and YAP1/TIAM1 stabilization [#8, #15, #17]. Its activity is governed by AMPK\\u03b1 (PRKAA) phosphorylation at Ser351, which switches RASAL2 between a PPM1B-recruiting autophagy suppressor and an activator of the PIK3C3/VPS34-ATG14-BECN1 complex [#10], and its abundance is controlled by CAMSAP2-driven ubiquitin-proteasome degradation [#18]. A germline RASAL2 p.Y808C variant enhances Ras signaling and confers resistance to anti-EGFR therapy [#20].\",\n  \"teleology\": [\n    {\n      \"year\": 2012,\n      \"claim\": \"Established that RASAL2 is not solely a Ras regulator but also engages Rho-family signaling, by identifying a physical link to the RhoGEF ECT2 controlling migratory plasticity.\",\n      \"evidence\": \"ECT2 immunoprecipitation/mass spectrometry, RHO activity assay, and RNAi phenotyping in astrocytoma cells\",\n      \"pmids\": [\"22683310\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Direct vs. indirect nature of the RASAL2-ECT2 association not resolved by reciprocal validation\", \"Connection between ECT2 binding and GAP catalysis unclear\"]\n    },\n    {\n      \"year\": 2013,\n      \"claim\": \"Defined RASAL2 as a RasGAP tumor and metastasis suppressor in vivo, answering whether its loss is sufficient to hyperactivate Ras in cancer.\",\n      \"evidence\": \"Knockout mouse models, in vivo tumor/metastasis assays, and human breast cancer mutation/expression analysis\",\n      \"pmids\": [\"24029233\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Structural basis of Ras-GTP hydrolysis not defined\", \"Did not address context-specific oncogenic roles\"]\n    },\n    {\n      \"year\": 2014,\n      \"claim\": \"Revealed that RASAL2 has a GAP-independent oncogenic mode, by showing it antagonizes the RAC1-GAP ARHGAP24 to activate RAC1 in TNBC.\",\n      \"evidence\": \"Reciprocal Co-IP, RAS-GAP catalytic mutant rescue, and RAC1 activity assays in TNBC cells\",\n      \"pmids\": [\"25384218\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Determinants of whether RASAL2 acts as GAP or RAC1 activator in a given tumor unresolved\", \"ARHGAP24 binding interface not mapped\"]\n    },\n    {\n      \"year\": 2014,\n      \"claim\": \"Confirmed RASAL2 suppresses Ras-ERK-driven EMT in ovarian cancer via pharmacological epistasis.\",\n      \"evidence\": \"shRNA knockdown, migration/invasion assays, and ERK pathway inhibitor rescue\",\n      \"pmids\": [\"25216515\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Single-lab evidence\", \"Direct GAP substrate engagement not shown biochemically\"]\n    },\n    {\n      \"year\": 2017,\n      \"claim\": \"Extended tumor-suppressor function to bladder cancer stemness through a defined MAPK/SOX2 axis, and to adipocyte biology where RASAL2 represses Ras in an ERK-independent manner.\",\n      \"evidence\": \"Gain/loss-of-function with ERK inhibitor and SOX2 siRNA rescue in bladder cancer; Rasal2-deficient mice and 3T3-L1 knockdown with Ras/MEK inhibitor rescue for adipogenesis\",\n      \"pmids\": [\"28182001\", \"28580280\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"ERK-independent effector downstream of Ras in adipogenesis unidentified\", \"Mechanism linking MAPK to SOX2 not fully resolved\"]\n    },\n    {\n      \"year\": 2018,\n      \"claim\": \"Mapped distinct anti-angiogenic and oncogenic effector branches, placing RASAL2 upstream of AKT/ETS1/VEGFA and GSK3\\u03b2/c-FOS/VEGFA in suppressor contexts and upstream of LATS2/YAP1 in CRC.\",\n      \"evidence\": \"Knockdown/overexpression, Western blotting of pathway nodes, Co-IP, and in vivo/specimen correlation across bladder, RCC, and CRC\",\n      \"pmids\": [\"29702203\", \"30158581\", \"30037330\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Whether AKT/GSK3\\u03b2 effects depend on GAP catalysis not tested\", \"Opposing YAP regulation across tissues unexplained\"]\n    },\n    {\n      \"year\": 2019,\n      \"claim\": \"Identified IPO5-mediated nuclear translocation as a regulatory mechanism coupling RASAL2 localization to Ras activation in CRC, and uncovered exosome- and phosphorylation-based control of its tumor-suppressor/promoter balance in breast cancer.\",\n      \"evidence\": \"Mass spectrometry, Co-IP, subcellular fractionation for IPO5; CRISPR KO, exosome isolation, and phospho-Ser237 detection in breast cancer models\",\n      \"pmids\": [\"31288861\", \"31473883\", \"31759919\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Functional role of nuclear RASAL2 beyond correlation with Ras activity unclear\", \"How the p-Rasal2/non-p-Rasal2 ratio is set physiologically unknown\"]\n    },\n    {\n      \"year\": 2021,\n      \"claim\": \"Defined the AMPK\\u03b1-Ser351 phosphorylation switch governing RASAL2 control of autophagy, and showed GAP activity sustains basal Ras in TNBC via SPRY1/2 suppression with YAP as a transcriptional input.\",\n      \"evidence\": \"In vitro kinase assay, S351 mutagenesis, Co-IP with PPM1B and the VPS34 complex, PIK3C3 activity assay; ChIP for YAP-RASAL2 promoter and MEK/EGFR inhibitor synergy with GAP mutant\",\n      \"pmids\": [\"33563064\", \"34168046\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Upstream signals dictating Ser351 phosphorylation in tumors not defined\", \"Whether the autophagy switch operates in the cancer contexts described elsewhere untested\"]\n    },\n    {\n      \"year\": 2022,\n      \"claim\": \"Consolidated the oncogenic arm of RASAL2 across gastric, prostate, pancreatic, and hepatic contexts via AKT, \\u03b2-catenin, cyclin D1, YAP1/TIAM1, and TET1/MTTP effectors, including NF-\\u03baB-driven induction and direct PP2A inhibition.\",\n      \"evidence\": \"ChIP, Co-IP (RASAL2-PP2A), PP2A activity assay, organoids, ubiquitination/hMeDIP assays, and in vivo models across gastric, prostate, PDAC, and liver\",\n      \"pmids\": [\"35134322\", \"35668070\", \"35844783\", \"36643045\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Why RASAL2 inhibits PP2A in some tissues yet suppresses AKT in others unresolved\", \"Structural basis of PP2A binding undetermined\"]\n    },\n    {\n      \"year\": 2023,\n      \"claim\": \"Identified CAMSAP2 as a regulator of RASAL2 stability, showing ubiquitin-proteasome degradation of RASAL2 unleashes ERK signaling to drive lung cancer metastasis.\",\n      \"evidence\": \"Co-IP, ubiquitin-proteasome assays, siRNA knockdown, and tail-vein metastasis model\",\n      \"pmids\": [\"38159595\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"The E3 ligase mediating degradation not identified\", \"Whether CAMSAP2 acts as adaptor or scaffold unclear\"]\n    },\n    {\n      \"year\": 2025,\n      \"claim\": \"Connected RASAL2 to secretory-autophagy exosome biology through direct inhibition of Rab27a in TNBC, and demonstrated a germline variant that hyperactivates Ras and confers anti-EGFR resistance.\",\n      \"evidence\": \"CRISPR KO, Co-IP (RASAL2-Rab27a), autophagosome/MVB colocalization, exosome characterization for Rab27a; functional p-ERK and cetuximab dose-response assays for the p.Y808C variant\",\n      \"pmids\": [\"40369567\", \"40849341\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Whether Rab27a inhibition is GAP-dependent untested\", \"Clinical penetrance and disease association of p.Y808C not established\"]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"What determines whether RASAL2 functions as a GAP-dependent tumor suppressor or a GAP-independent oncogene in a given tissue remains the central unresolved question.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"No unifying model integrating phosphorylation state, localization, and binding partners to predict suppressor vs. oncogene outcome\", \"No high-resolution structure of RASAL2 or its complexes\", \"Context-switch mechanism between PP2A/AKT activation and AKT/VEGFA suppression undefined\"]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0005096\", \"supporting_discovery_ids\": [0]},\n      {\"term_id\": \"GO:0098772\", \"supporting_discovery_ids\": [1, 14, 10]},\n      {\"term_id\": \"GO:0140096\", \"supporting_discovery_ids\": [14]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005829\", \"supporting_discovery_ids\": [3]},\n      {\"term_id\": \"GO:0005634\", \"supporting_discovery_ids\": [8]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-162582\", \"supporting_discovery_ids\": [0, 2, 1]},\n      {\"term_id\": \"R-HSA-9612973\", \"supporting_discovery_ids\": [10, 11, 19]},\n      {\"term_id\": \"R-HSA-1643685\", \"supporting_discovery_ids\": [0, 20]}\n    ],\n    \"complexes\": [],\n    \"partners\": [\"ARHGAP24\", \"ECT2\", \"PP2A\", \"IPO5\", \"PPM1B\", \"PIK3C3\", \"CAMSAP2\", \"RAB27A\"],\n    \"other_free_text\": []\n  }\n}","audit_flag":null,"evaluation":{"faith_supported":5,"faith_total":5,"faith_pct":100.0}}