{"gene":"KRAS","run_date":"2026-04-28T18:30:27","timeline":{"discoveries":[{"year":1988,"finding":"Oncogenic mutations at codon 12 of c-K-ras (KRAS) are present in 21 of 22 human pancreatic carcinomas, establishing KRAS mutation as a near-universal and early critical event in exocrine pancreatic oncogenesis.","method":"PCR amplification and RNase A mismatch cleavage mutation detection on primary tumor and metastasis specimens","journal":"Cell","confidence":"High","confidence_rationale":"Tier 1 — direct sequencing of primary tumors and matched metastases, replicated across >100 subsequent studies","pmids":["2453289"],"is_preprint":false},{"year":1989,"finding":"KRAS2 codon 12 mutations occur early in human colon carcinoma development, preceding ploidy changes and existing in diploid cells from which aneuploid subpopulations arise; mutations can be present in histologically normal mucosa adjacent to carcinoma.","method":"Histological enrichment, cell sorting, PCR amplification, and direct DNA sequencing of colon carcinomas, adenomas, and adjacent normal tissue","journal":"Proceedings of the National Academy of Sciences of the United States of America","confidence":"High","confidence_rationale":"Tier 2 — multiple orthogonal methods, staged tissue analysis establishing temporal order of mutation","pmids":["2648401"],"is_preprint":false},{"year":1993,"finding":"Human SOS1 (hSos1) functions as a guanine nucleotide exchange factor (GEF) for RAS, with its CDC25-related domain specifically stimulating guanine nucleotide exchange on mammalian Ras proteins in vitro; hSos1 binds GRB2 via SH3 domain interactions, coupling receptor tyrosine kinases to RAS/KRAS signaling.","method":"In vitro GEF assay, yeast complementation, co-immunoprecipitation, overexpression in mammalian cells","journal":"Science","confidence":"High","confidence_rationale":"Tier 1 — in vitro reconstitution of GEF activity plus yeast complementation and co-IP, foundational paper","pmids":["8493579"],"is_preprint":false},{"year":1995,"finding":"K-Ras4B processing is more sensitive to inhibition by geranylgeranyltransferase I (GGTase I) inhibitor GGTI-286 than to farnesyltransferase inhibitor FTI-277, demonstrating that K-Ras4B can be alternatively geranylgeranylated and that its oncogenic signaling can be disrupted by GGTase I inhibition.","method":"In vitro prenylation assays, whole-cell processing assays, MAP kinase activity assays in intact cells","journal":"The Journal of biological chemistry","confidence":"High","confidence_rationale":"Tier 1 — in vitro enzyme assays with cell-based validation, first demonstration of K-Ras4B alternative prenylation","pmids":["7592913"],"is_preprint":false},{"year":2000,"finding":"Crystal structures of rat farnesyltransferase (FTase) ternary complexes with farnesyl diphosphate analogs and K-Ras4B peptide substrates revealed that the K-Ras4B polybasic region forms a type I beta turn and binds along the rim of the hydrophobic cavity, conferring the highest affinity of any natural FTase substrate; zinc is essential for productive Ca1a2X peptide binding.","method":"X-ray crystallography of ternary complexes at 2 Å resolution","journal":"Structure","confidence":"High","confidence_rationale":"Tier 1 — crystal structure at 2 Å with four independent complexes, defines substrate binding mechanism","pmids":["10673434"],"is_preprint":false},{"year":2003,"finding":"K-Ras4B resistance to farnesyltransferase inhibitors (FTIs) arises from two independent mechanisms: (1) its polybasic domain increases affinity for FTase, and (2) its CAAX motif can be alternatively geranylgeranylated. Either the polybasic domain alone or an alternatively prenylated CAAX alone renders K-Ras4B FTI-resistant, and K-Ras4B function is independent of the identity of the prenyl group.","method":"Chimeric Ras protein constructs, Elk-1 activation assays, anchorage-independent colony formation, microarray analysis","journal":"The Journal of biological chemistry","confidence":"High","confidence_rationale":"Tier 2 — multiple orthogonal methods with structure-function chimeras, mechanistically dissects two independent resistance mechanisms","pmids":["12882980"],"is_preprint":false},{"year":2009,"finding":"The C-terminal hypervariable region (HVR) of K-Ras4B, specifically its polybasic farnesylated tail, is responsible for isoform-specific interaction with calmodulin (CaM); the HVR binds the C-terminal domain of Ca2+-loaded CaM with micromolar affinity, while the GTP-loaded catalytic domain may additionally interact with the N-terminal CaM domain, linking nucleotide state to CaM binding.","method":"NMR spectroscopy, isothermal titration calorimetry (ITC)","journal":"Biochemistry","confidence":"High","confidence_rationale":"Tier 1 — NMR and ITC provide quantitative binding data with domain-level resolution","pmids":["19583261"],"is_preprint":false},{"year":2009,"finding":"TBK1 is a synthetic lethal partner of oncogenic KRAS; TBK1 selectively activates NF-κB anti-apoptotic signals (via c-Rel and BCL-XL) in KRAS-mutant cancer cells, and its suppression induces apoptosis specifically in KRAS-dependent cells.","method":"Genome-wide RNA interference screen, mechanistic validation by epistasis and apoptosis assays in human cancer cell lines","journal":"Nature","confidence":"High","confidence_rationale":"Tier 2 — genome-wide RNAi screen followed by mechanistic pathway epistasis, replicated across multiple KRAS-mutant lines","pmids":["19847166"],"is_preprint":false},{"year":2012,"finding":"KRAS is enriched for rare codons relative to HRAS, limiting KRAS protein expression. Converting rare to common codons increases K-Ras expression and tumorigenicity to mirror that of H-Ras, demonstrating that synonymous nucleotide differences affecting codon usage underlie differences in HRas vs KRas expression and oncogenic function.","method":"Synonymous codon-optimized transgenes expressed from identical loci, protein expression quantification, transformation assays","journal":"Current biology","confidence":"High","confidence_rationale":"Tier 2 — isogenic codon-substituted alleles with functional readouts, identifies hardwired translational regulatory mechanism","pmids":["23246410"],"is_preprint":false},{"year":2013,"finding":"Oncogenic KRAS reprograms glutamine metabolism in pancreatic ductal adenocarcinoma (PDAC) via transcriptional upregulation of RREBP1 and MYC and downregulation of GLUD1: PDAC cells use a non-canonical pathway converting glutamine-derived aspartate to oxaloacetate (via GOT1), then to malate and pyruvate, maintaining NADPH/NADP+ ratio and redox balance. Knockdown of any enzyme in this pathway suppresses PDAC growth in vitro and in vivo.","method":"Isotope tracing with 13C-glutamine, siRNA knockdown, xenograft tumor models, gene expression analysis","journal":"Nature","confidence":"High","confidence_rationale":"Tier 1 — metabolic flux tracing plus genetic validation with multiple enzymes, in vivo confirmation","pmids":["23535601"],"is_preprint":false},{"year":2014,"finding":"YAP1 can substitute for oncogenic KRAS to rescue cell viability in KRAS-dependent cancer cells; KRAS and YAP1 converge on transcription factor FOS to activate a transcriptional program regulating the epithelial-mesenchymal transition (EMT). YAP1 is required for KRAS-induced cell transformation, and acquired resistance to Kras suppression in a murine lung cancer model involves increased YAP1 signaling.","method":"ORFeome rescue screen (15,294 ORFs) in KRAS-dependent cells with inducible KRAS shRNA, epistasis analysis, murine lung cancer model","journal":"Cell","confidence":"High","confidence_rationale":"Tier 2 — genome-scale functional screen plus mechanistic in vitro and in vivo validation","pmids":["24954536"],"is_preprint":false},{"year":2014,"finding":"In mice, codon-optimized Kras alleles (Kras(ex3op)) producing more K-Ras protein from the endogenous locus lead to fewer carcinogen-induced tumors and induce growth arrest when oncogenically mutated, demonstrating that the rare codon bias of KRAS is a tumor-suppressive mechanism that limits oncogenic K-Ras protein levels in vivo.","method":"Knock-in mice with synonymous codon-optimized Kras exon 3, urethane carcinogenesis, tumor burden quantification","journal":"The Journal of clinical investigation","confidence":"High","confidence_rationale":"Tier 2 — in vivo genetic model with clean isogenic comparison, validates translational regulation of KRAS oncogenesis","pmids":["25437878"],"is_preprint":false},{"year":2015,"finding":"The C-terminal HVR of K-Ras4B directly interacts with the active site/effector-binding region of the catalytic domain with ~100-fold higher affinity in the GDP-bound vs. GTP-bound state; HVR binding interferes with Ras-Raf interaction, modulates phospholipid binding, and slightly slows nucleotide exchange, establishing an autoinhibitory mechanism.","method":"NMR spectroscopy, surface plasmon resonance, isothermal titration calorimetry","journal":"Biophysical journal","confidence":"High","confidence_rationale":"Tier 1 — multiple biophysical methods (NMR, SPR, ITC) demonstrating direct intramolecular interaction with quantitative affinities","pmids":["26682817"],"is_preprint":false},{"year":2015,"finding":"GTP-bound K-Ras4B forms stable homodimers; two major dimer interfaces were identified: a β-sheet interface overlapping effector binding sites (potentially inhibitory) and a helical interface that may promote Raf dimerization and activation. Ras self-association can regulate effector binding and activity.","method":"Analytical ultracentrifugation, molecular dynamics simulations, small-angle X-ray scattering","journal":"Structure","confidence":"Medium","confidence_rationale":"Tier 2 — multiple biophysical methods but single lab, without cell-based validation of the dimer interfaces","pmids":["26051715"],"is_preprint":false},{"year":2015,"finding":"K-Ras4B membrane binding is driven by farnesyl group insertion into disordered lipid microdomains; phosphorylation of Ser-181 prohibits spontaneous farnesyl membrane insertion; the polybasic polylysine sequence modulates specific binding to anionic phospholipids and farnesyl membrane orientation.","method":"Confocal microscopy, surface plasmon resonance, molecular dynamics simulations","journal":"The Journal of biological chemistry","confidence":"Medium","confidence_rationale":"Tier 2 — orthogonal experimental and computational methods, single lab","pmids":["25713064"],"is_preprint":false},{"year":2016,"finding":"Molecular dynamics simulations reveal that oncogenic mutations G12C/G12D/G12V/G13D/Q61H differentially drive inactive-to-active conformational transitions in K-Ras4B-GTP; GAP not only donates its R789 arginine finger but stabilizes the catalytically competent conformation and pre-organizes Q61; oncogenic mutations disrupt R789/Q61 organization, impairing GAP-mediated GTP hydrolysis.","method":"6.4 μs cumulative molecular dynamics simulations of WT and mutant K-Ras4B with and without GAP","journal":"Scientific reports","confidence":"Medium","confidence_rationale":"Tier 4 for computational, but extensive simulation coverage; mechanistic model consistent with established biochemistry","pmids":["26902995"],"is_preprint":false},{"year":2016,"finding":"Kras is required for B cell lymphopoiesis: hematopoietic-specific deletion of Kras impairs early B cell development at the pre-B cell stage and late B cell maturation. Kras deficiency specifically impairs pre-BCR- and BCR-induced activation of the Raf-1/MEK/ERK pathway, while T cell development is unaffected, demonstrating Kras as the unique Ras family member critical for the Raf-1/MEK/ERK axis in B cells.","method":"Conditional knockout mice (hematopoietic-specific and B cell-specific Cre), bone marrow chimeras, flow cytometry, proliferation and signaling assays","journal":"Journal of Immunology","confidence":"High","confidence_rationale":"Tier 2 — clean conditional KO in two cell-type-specific models with defined signaling pathway readout","pmids":["26773157"],"is_preprint":false},{"year":2016,"finding":"Phosphorylation at Ser-181 of K-Ras4B reduces but does not fully abolish membrane binding and clustering; phosphorylated K-Ras4B maintains association with cytosolic shuttle PDEδ; phosphorylation does not alter localization to liquid-disordered lipid subdomains but facilitates dissociation from the plasma membrane.","method":"Semisynthesis of triply modified K-Ras4B (phosphate + farnesyl + methyl), supported lipid bilayer studies, fluorescence spectroscopy, cell microinjection","journal":"ACS Chemical Biology","confidence":"High","confidence_rationale":"Tier 1 — chemically defined post-translationally modified protein studied by multiple spectroscopic methods plus cell experiments","pmids":["28448716"],"is_preprint":false},{"year":2016,"finding":"Ca2+/calmodulin (CaM) extracts K-Ras4B from negatively charged membranes in a nucleotide-independent manner; the CaM/K-Ras4B complex is stable in the presence of anionic membranes and shows no membrane binding. PDEδ and CaM affect K-Ras4B membrane interaction through different mechanisms.","method":"Surface plasmon resonance, fluorescence spectroscopy (FCS, FRET), model membrane studies, FRAP","journal":"Biophysical journal","confidence":"High","confidence_rationale":"Tier 1 — multiple spectroscopic and biophysical techniques with chemically defined proteins and membranes","pmids":["27410739"],"is_preprint":false},{"year":2017,"finding":"Oncogenic KRAS drives a non-canonical glutamine-to-aspartate metabolic pathway through transcriptional regulation of key enzymes; the KRAS-regulated kinome in PDAC includes WEE1 among kinases downregulated upon KRAS loss, and combined WEE1 + ERK inhibition causes enhanced PDAC growth suppression and apoptosis.","method":"Multiplexed inhibitor bead/MS kinomics, siRNA knockdown, pharmacological inhibition, synergy studies in PDAC cell lines","journal":"The Journal of biological chemistry","confidence":"Medium","confidence_rationale":"Tier 2 — system-wide kinome profiling across six cell lines with pharmacological validation","pmids":["34688654"],"is_preprint":false},{"year":2017,"finding":"CaM preferentially binds unfolded K-Ras4B HVR (not α-helical HVR) using all three CaM domains; interaction is stabilized by docking of farnesyl to hydrophobic pockets in both CaM lobes; CaM wraps around the polybasic anchor region of HVR, enabling membrane extraction of K-Ras4B to form a K-Ras4B–CaM–PI3Kα ternary complex that activates PI3Kα.","method":"Molecular dynamics simulations, fluorescence binding experiments","journal":"The Journal of biological chemistry","confidence":"Medium","confidence_rationale":"Tier 3 — computational modeling supported by fluorescence experiments; ternary complex model not directly confirmed by structure","pmids":["28623230"],"is_preprint":false},{"year":2017,"finding":"USP18 deubiquitinase stabilizes the KRAS oncoprotein; USP18 loss reduces KRAS protein half-life and mislocalizes KRAS from the plasma membrane; USP18 gain increases KRAS stability; Usp18 loss in Kras-mutant mice significantly reduces lung tumor burden.","method":"Cycloheximide chase, subcellular fractionation, conditional KO mice, immunohistochemistry","journal":"Molecular Cancer Research","confidence":"Medium","confidence_rationale":"Tier 2 — protein stability and localization experiments with in vivo genetic confirmation","pmids":["28242811"],"is_preprint":false},{"year":2018,"finding":"KRAS dimerization at the α4-α5 interface mediates wild-type KRAS-dependent fitness of KRAS-mutant lung adenocarcinoma cells and underlies resistance to MEK inhibition; KRASD154Q (dimerization-deficient mutant) abrogates these effects both in vitro and in vivo; dimerization also has a critical role in the oncogenic activity of mutant KRAS.","method":"Genetically inducible KRAS LOH model, dimerization-disrupting mutant KRASD154Q, MEK inhibitor response assays, in vitro and in vivo tumor models","journal":"Cell","confidence":"High","confidence_rationale":"Tier 2 — clean genetic models with structure-guided mutant, multiple in vitro and in vivo readouts, replicated across human and murine systems","pmids":["29336889"],"is_preprint":false},{"year":2018,"finding":"Full-length, fully processed (farnesylated + methylated) K-Ras4B lacks intrinsic dimerization capability on supported lipid bilayers across a wide range of surface densities and lipid compositions including cholesterol-containing membranes.","method":"Fluorescence correlation spectroscopy (FCS), single-molecule tracking on supported lipid bilayers with natively processed K-Ras4B","journal":"Biophysical journal","confidence":"High","confidence_rationale":"Tier 1 — single-molecule imaging with natively processed protein under multiple lipid conditions, directly contradicts other reports","pmids":["29320680"],"is_preprint":false},{"year":2018,"finding":"K-Ras4B mutant (G12C/G12D) HVR shows preferential interaction with phosphatidic acid (PA) over other phospholipids; in the GDP-bound state the HVR shields the effector-binding site (autoinhibition); GTP binding and oncogenic mutations release HVR, enabling calmodulin interaction.","method":"Molecular dynamics simulations, NMR, phospholipid binding assays","journal":"Current opinion in structural biology","confidence":"Medium","confidence_rationale":"Tier 2 — combines computational and NMR data; mutation-specific phospholipid binding specificity established","pmids":["26709496"],"is_preprint":false},{"year":2018,"finding":"A compound (Cmpd2) inhibits K-RAS4B by stabilizing membrane-dependent occlusion of the effector-binding site: it simultaneously engages a shallow pocket on KRAS and the lipid bilayer, orienting membrane-associated prenylated KRAS so the membrane sterically occludes the effector-binding site, reducing RAF binding and impairing RAF activation.","method":"NMR, lipid bilayer binding assays, cell-based RAF activation assays, structure-based mechanism elucidation","journal":"Cell Chemical Biology","confidence":"High","confidence_rationale":"Tier 1 — NMR structural characterization of mechanism with cell-based functional validation","pmids":["30122370"],"is_preprint":false},{"year":2019,"finding":"BI-2852 binds with nanomolar affinity to the switch I/II pocket on KRAS (present in both active and inactive forms), blocking all GEF, GAP, and effector interactions simultaneously, demonstrating that this pocket is druggable; binding inhibits downstream signaling and has antiproliferative effects in KRAS-mutant cells.","method":"Structure-based drug design, X-ray crystallography, biochemical GEF/GAP/effector competition assays, cell-based signaling assays","journal":"Proceedings of the National Academy of Sciences of the United States of America","confidence":"High","confidence_rationale":"Tier 1 — crystal structure plus biochemical assays showing blockade of multiple interaction partners","pmids":["31332011"],"is_preprint":false},{"year":2019,"finding":"K-Ras4B allosterically activates PI3Kα by binding-induced conformational changes that disrupt p110/p85 (nSH2) interactions, exposing the kinase domain for membrane association and substrate phosphorylation; allosteric signaling is rewired from helical to kinase domain in the K-Ras4B/PI3Kα complex.","method":"Accelerated molecular dynamics simulations, allosteric pathway analysis, community network analysis","journal":"International journal of biological macromolecules","confidence":"Low","confidence_rationale":"Tier 4 — computational only, no in vitro or cell-based validation","pmids":["31816384"],"is_preprint":false},{"year":2019,"finding":"A small molecule KRAS agonist (KRA-533) binds the GTP/GDP-binding pocket of KRAS and prevents GTP cleavage, accumulating active GTP-KRAS; K117A mutation in KRAS abolishes KRA-533 binding and blocks its activity; KRA-533-mediated KRAS hyperactivation promotes apoptosis and autophagic cell death preferentially in KRAS-mutant cancer cells.","method":"GDP/GTP exchange assay, site-directed mutagenesis, cell viability assays, xenograft and GEMM models","journal":"Molecular cancer","confidence":"Medium","confidence_rationale":"Tier 2 — biochemical assay plus mutagenesis plus in vivo models from single lab","pmids":["30971271"],"is_preprint":false},{"year":2020,"finding":"Urethane carcinogenesis specificity is determined by the sequence specificity of urethane mutagenesis coupled with transcription bias and isoform locus: the initiating Kras Q61L/R mutation was captured days after urethane exposure using error-corrected high-throughput sequencing, demonstrating that transcription rate and isoform-specific context drive RAS mutation tropism.","method":"Error-corrected high-throughput sequencing of mouse Ras genes at multiple time points post-carcinogen exposure","journal":"Nature communications","confidence":"High","confidence_rationale":"Tier 2 — direct sequencing capturing the initiating mutation in vivo with mechanistic dissection of specificity determinants","pmids":["32286309"],"is_preprint":false},{"year":2021,"finding":"KRAS-dependent transcription is driven predominantly through the ERK/MAPK cascade; KRAS-regulated ERK signaling deregulates the anaphase promoting complex/cyclosome (APC/C) and cell cycle machinery as key drivers of PDAC growth; the KRAS-dependent gene signature diverges substantially from the Hallmark KRAS signaling gene signature.","method":"Inducible KRAS knockdown, transcriptomics, phosphoproteomics, total proteomics, patient tumor data integration","journal":"Science","confidence":"High","confidence_rationale":"Tier 2 — multiple omics layers integrated with genetic perturbation and patient data validation","pmids":["38843331"],"is_preprint":false},{"year":2022,"finding":"Resistance to KRASG12C inhibitors is driven primarily by upstream feedback activation of wild-type RAS (rather than reactivation of KRASG12C to its GTP-bound state); multiple RTKs can independently drive this KRASG12C-independent RAS-MAPK reactivation; convergent upstream or downstream blockade can overcome resistance.","method":"RAS activity assays, RTK inhibitor combinations, cell signaling studies in KRASG12C-mutant cancer cells treated with G12C inhibitors","journal":"Cell reports","confidence":"High","confidence_rationale":"Tier 2 — mechanistic dissection with multiple inhibitors and RAS activity measurements identifying WT RAS as the resistance driver","pmids":["35732135"],"is_preprint":false},{"year":2023,"finding":"Cancer-associated fibroblast (CAF)-derived NRG1 activates cancer cell ERBB2/ERBB3 receptor tyrosine kinases to support KRAS*-independent pancreatic cancer growth; genetic extinction of KRAS* upregulates ERBB2/ERBB3 in cancer cells, which then utilize paracrine CAF-NRG1 as a survival factor; ERBB2/3 or NRG1 depletion abolishes KRAS* bypass.","method":"Genetic KRAS extinction, pharmacological inhibition, paracrine co-culture models, mouse and human PDAC models","journal":"Genes & development","confidence":"High","confidence_rationale":"Tier 2 — genetic and pharmacological evidence in multiple model systems with defined paracrine mechanism","pmids":["37775182"],"is_preprint":false},{"year":2023,"finding":"A comprehensive deep mutational scan quantified >26,000 mutations' effects on KRAS folding and binding to six interaction partners, mapping >22,000 causal free energy changes; allosteric propagation is particularly effective across the central β-sheet; multiple surface pockets are validated as allosterically active including a distal C-terminal lobe pocket; most allosteric mutations inhibit all effectors but some can alter binding specificity.","method":"Deep mutational scanning, double-mutant genetic interaction analysis, free energy change inference","journal":"Nature","confidence":"High","confidence_rationale":"Tier 1 — genome-scale biophysical measurements with double-mutant epistasis providing causal energy landscapes","pmids":["38109937"],"is_preprint":false},{"year":2021,"finding":"Acquired resistance to adagrasib (KRASG12C inhibitor) involves diverse mechanisms including secondary KRAS mutations (G12D/R/V/W, G13D, Q61H, R68S, H95D/Q/R, Y96C), KRAS amplification, and bypass alterations (MET amplification, NRAS/BRAF/MAP2K1/RET mutations, oncogenic fusions involving ALK/RET/BRAF/RAF1/FGFR3, NF1/PTEN loss); deep mutational scanning systematically defined the landscape of KRAS mutations conferring inhibitor resistance.","method":"Genomic sequencing of paired pre-/post-treatment biopsies, in vitro deep mutational scanning screen","journal":"The New England journal of medicine","confidence":"High","confidence_rationale":"Tier 2 — clinical genomics plus systematic in vitro resistance landscape, multiple orthogonal evidence types","pmids":["34161704"],"is_preprint":false},{"year":2017,"finding":"KRAS can bind numerous effector proteins (RAF, PI3K, RalGDS families, and others); combinatorial siRNA knockdown of 41 KRAS effector nodes in 92 cell lines identified two major subtypes of KRAS-mutant cancers with distinct effector dependencies, demonstrating that each cell line has a unique effector engagement pattern.","method":"Arrayed combinatorial siRNA screen of 41 effector nodes across 92 cancer cell lines, quantitative phenotype assessment","journal":"Cell reports","confidence":"High","confidence_rationale":"Tier 2 — large-scale systematic knockdown across diverse cell lines with quantitative phenotypic readouts","pmids":["29444439"],"is_preprint":false}],"current_model":"KRAS encodes a small GTPase that cycles between GDP-bound (inactive) and GTP-bound (active) states regulated by GEFs (e.g., SOS1) and GAPs; oncogenic mutations at codons 12, 13, or 61 impair GAP-mediated GTP hydrolysis, locking KRAS in its active state to constitutively signal through multiple effector pathways (RAF-MEK-ERK, PI3K-AKT, RalGDS); membrane association via farnesylation of the C-terminal CAAX motif and the polybasic HVR is essential for oncogenic signaling; the HVR can autoinhibit the effector-binding site in the GDP-bound state; KRAS dimerization at the α4-α5 interface modulates MEK inhibitor sensitivity and oncogenic activity; calmodulin specifically extracts K-Ras4B from the membrane via HVR interaction to promote PI3Kα activation; rare codon bias in KRAS limits protein expression as a built-in tumor-suppressive mechanism; oncogenic KRAS reprograms glutamine metabolism through a non-canonical GOT1-dependent pathway to maintain redox balance; KRAS signals predominantly through ERK/MAPK to regulate APC/C and cell cycle machinery driving tumor growth; and resistance to covalent KRASG12C inhibitors occurs via secondary KRAS mutations, KRAS amplification, and bypass RTK/RAS pathway reactivation."},"narrative":{"teleology":[{"year":1988,"claim":"Establishing that KRAS mutations are a near-universal initiating event in pancreatic cancer answered whether a single oncogene could account for virtually all cases of this malignancy and defined KRAS as the central driver.","evidence":"PCR-based mutation detection in 22 primary pancreatic carcinomas and colon tumors with staged tissue analysis","pmids":["2453289","2648401"],"confidence":"High","gaps":["Mechanism by which mutant KRAS drives tumorigenesis was undefined","No effector pathway specificity was determined","Functional contribution versus passenger role not formally tested"]},{"year":1993,"claim":"Identification of SOS1 as the GEF coupling RTKs to RAS via GRB2 resolved how extracellular signals activate KRAS, placing it downstream of growth factor receptors.","evidence":"In vitro GEF reconstitution, yeast complementation, co-immunoprecipitation","pmids":["8493579"],"confidence":"High","gaps":["How GAP-mediated inactivation was specifically impaired by oncogenic mutations remained structural speculation","Isoform-specific GEF regulation was not addressed"]},{"year":1995,"claim":"Discovery that K-Ras4B can be alternatively geranylgeranylated explained the clinical failure of farnesyltransferase inhibitors against KRAS-driven cancers, revealing redundancy in prenylation.","evidence":"In vitro prenylation assays and cell-based processing/MAPK assays with FTI and GGTase I inhibitors","pmids":["7592913","12882980"],"confidence":"High","gaps":["Structural basis for K-Ras4B's dual prenylation susceptibility was not resolved","In vivo pharmacological confirmation was lacking"]},{"year":2000,"claim":"Crystal structures of K-Ras4B peptide–FTase complexes revealed how the polybasic HVR confers the highest FTase affinity among Ras isoforms, providing structural understanding of C-terminal processing.","evidence":"X-ray crystallography at 2 Å resolution of ternary FTase complexes","pmids":["10673434"],"confidence":"High","gaps":["Full-length K-Ras4B structure with lipid modifications was unavailable","Mechanism of alternative geranylgeranylation was not structurally addressed"]},{"year":2009,"claim":"NMR and calorimetry defined the calmodulin–K-Ras4B HVR interaction and a genome-wide RNAi screen identified TBK1 as a synthetic lethal partner, jointly revealing non-canonical signaling outputs and vulnerabilities of mutant KRAS.","evidence":"NMR/ITC for CaM binding; genome-wide RNAi screen with mechanistic epistasis in KRAS-mutant cell lines","pmids":["19583261","19847166"],"confidence":"High","gaps":["Whether CaM-dependent extraction operates in vivo was not shown","TBK1 synthetic lethality was not validated in clinical settings"]},{"year":2012,"claim":"Demonstrating that KRAS rare codon bias limits protein expression and that codon optimization increases tumorigenicity uncovered a hard-wired translational tumor suppressor mechanism embedded in the coding sequence.","evidence":"Isogenic codon-optimized transgenes with transformation assays in vitro; knock-in mice with codon-optimized Kras exon 3 and carcinogenesis assays","pmids":["23246410","25437878"],"confidence":"High","gaps":["The ribosomal or tRNA mechanisms mediating rare-codon suppression were not identified","Whether codon bias affects mutant vs. wild-type KRAS differentially was not resolved"]},{"year":2013,"claim":"Isotope tracing revealed that oncogenic KRAS reprograms glutamine metabolism through a non-canonical GOT1-dependent pathway to maintain NADPH and redox balance, establishing metabolic rewiring as a core function of mutant KRAS in PDAC.","evidence":"13C-glutamine flux analysis, siRNA knockdown of pathway enzymes, xenograft validation","pmids":["23535601"],"confidence":"High","gaps":["Whether this pathway operates in KRAS-mutant cancers beyond PDAC was untested","Direct transcriptional mechanism linking KRAS to enzyme expression was incompletely defined"]},{"year":2015,"claim":"Biophysical studies established that the K-Ras4B HVR autoinhibits the effector-binding site in the GDP state with ~100-fold selectivity, and that GTP-bound K-Ras4B forms homodimers at two distinct interfaces, resolving how intramolecular and intermolecular interactions regulate effector access.","evidence":"NMR/SPR/ITC for autoinhibition; analytical ultracentrifugation, SAXS, and MD for dimerization","pmids":["26682817","26051715"],"confidence":"High","gaps":["Dimerization interfaces lacked cell-based validation","Whether autoinhibition is modulated by specific lipid environments was unknown"]},{"year":2016,"claim":"Conditional knockout of Kras in hematopoietic cells demonstrated a non-redundant requirement for KRAS in B cell lymphopoiesis through the Raf-1/MEK/ERK axis, establishing a physiological developmental role distinct from oncogenesis.","evidence":"Hematopoietic- and B cell-specific conditional KO mice, bone marrow chimeras, signaling assays","pmids":["26773157"],"confidence":"High","gaps":["Whether KRAS has analogous non-redundant roles in other non-hematopoietic lineages was not tested","Mechanism for KRAS isoform specificity in B cells was unclear"]},{"year":2016,"claim":"Studies of Ser-181 phosphorylation, PDEδ shuttling, and calmodulin-mediated membrane extraction defined three orthogonal mechanisms controlling K-Ras4B plasma membrane residence, clarifying how localization is dynamically regulated.","evidence":"Semisynthetic phosphorylated/farnesylated K-Ras4B on supported bilayers; SPR/FCS/FRET with CaM and PDEδ","pmids":["28448716","27410739"],"confidence":"High","gaps":["Relative contributions of CaM, PDEδ, and phosphorylation in living cells were not quantified","Whether these mechanisms are differentially engaged in mutant vs. WT KRAS was not resolved"]},{"year":2018,"claim":"Genetic evidence that KRAS dimerization at the α4-α5 interface mediates fitness of KRAS-mutant cells and MEK inhibitor resistance provided a structural rationale for wild-type KRAS dependency in mutant KRAS tumors, though single-molecule studies challenged whether full-length processed K-Ras4B dimerizes on membranes.","evidence":"KRASD154Q dimerization-deficient mutant in LOH model and tumor assays; single-molecule FCS on supported lipid bilayers","pmids":["29336889","29320680"],"confidence":"High","gaps":["Discrepancy between cell-based genetic dimerization data and biophysical monomeric behavior on bilayers is unresolved","Whether scaffold proteins mediate dimerization in cells was not tested"]},{"year":2019,"claim":"Structure-based drug design yielded compounds targeting two distinct KRAS pockets—BI-2852 at the switch I/II interface blocking all GEF/GAP/effector interactions, and Cmpd2 stabilizing membrane-occluded KRAS—proving KRAS is druggable at multiple sites.","evidence":"X-ray crystallography and NMR with biochemical competition and cell-based signaling assays","pmids":["31332011","30122370"],"confidence":"High","gaps":["In vivo efficacy and selectivity of these compounds were not demonstrated","Whether these sites are accessible in all KRAS mutant contexts was untested"]},{"year":2021,"claim":"Clinical genomics and deep mutational scanning systematically mapped the resistance landscape to KRASG12C inhibitors, revealing that secondary KRAS mutations, gene amplification, and RTK-driven wild-type RAS reactivation represent convergent escape mechanisms.","evidence":"Paired pre-/post-treatment biopsy sequencing; in vitro deep mutational scanning; RAS activity assays with RTK inhibitor combinations","pmids":["34161704","35732135"],"confidence":"High","gaps":["Therapeutic strategies to preempt multi-mechanism resistance were not clinically validated","Whether resistance mechanisms differ across tissue types was not systematically tested"]},{"year":2021,"claim":"Multi-omic profiling showed that KRAS-dependent transcription in PDAC is driven predominantly through ERK/MAPK, with APC/C and cell-cycle machinery as key downstream effectors, revising the canonical KRAS transcriptional signature.","evidence":"Inducible KRAS knockdown with integrated transcriptomics, phosphoproteomics, proteomics, and patient tumor data","pmids":["38843331"],"confidence":"High","gaps":["Whether APC/C deregulation is a direct or indirect effect of ERK signaling was not distinguished","Contribution of non-ERK effectors to KRAS-dependent transcription was not fully quantified"]},{"year":2023,"claim":"A comprehensive deep mutational scan of >26,000 KRAS mutations mapped the allosteric network across the central β-sheet and identified multiple allosterically active surface pockets, providing a near-complete biophysical atlas of KRAS function.","evidence":"Deep mutational scanning with double-mutant epistasis and free energy inference across six binding partners","pmids":["38109937"],"confidence":"High","gaps":["Structural validation of newly identified allosteric pockets by crystallography or cryo-EM is pending","Whether allosteric mutations differentially affect KRAS signaling in cellular contexts was not tested"]},{"year":2023,"claim":"CAF-derived NRG1 activating ERBB2/3 was identified as a microenvironment-driven bypass mechanism enabling KRAS-independent PDAC survival, demonstrating that KRAS-targeted therapy must account for paracrine escape.","evidence":"Genetic KRAS extinction, co-culture paracrine models, pharmacological ERBB inhibition in mouse and human PDAC","pmids":["37775182"],"confidence":"High","gaps":["Whether NRG1-ERBB bypass operates in non-pancreatic KRAS-mutant cancers is unknown","The full repertoire of microenvironment-mediated escape routes has not been mapped"]},{"year":null,"claim":"Key unresolved questions include whether KRAS dimerization occurs on native plasma membranes under physiological conditions, the complete structural basis for isoform-specific effector engagement in vivo, and how to therapeutically target the full spectrum of resistance mechanisms arising during KRAS inhibitor treatment.","evidence":"","pmids":[],"confidence":"High","gaps":["No consensus on in vivo KRAS dimerization stoichiometry or scaffold dependency","Full-length, lipid-modified KRAS structure in a membrane environment is unavailable","Clinical strategies to preempt convergent resistance remain undefined"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0003924","term_label":"GTPase activity","supporting_discovery_ids":[15,26,28,33]},{"term_id":"GO:0098772","term_label":"molecular function regulator activity","supporting_discovery_ids":[30,35]},{"term_id":"GO:0008289","term_label":"lipid binding","supporting_discovery_ids":[14,24,25]}],"localization":[{"term_id":"GO:0005886","term_label":"plasma membrane","supporting_discovery_ids":[14,17,18,21,23,25]},{"term_id":"GO:0005829","term_label":"cytosol","supporting_discovery_ids":[17,18]}],"pathway":[{"term_id":"R-HSA-162582","term_label":"Signal Transduction","supporting_discovery_ids":[2,7,10,20,27,30,35]},{"term_id":"R-HSA-1640170","term_label":"Cell Cycle","supporting_discovery_ids":[30]},{"term_id":"R-HSA-1430728","term_label":"Metabolism","supporting_discovery_ids":[9,19]},{"term_id":"R-HSA-1643685","term_label":"Disease","supporting_discovery_ids":[0,1,31,34]}],"complexes":[],"partners":["SOS1","BRAF","RAF1","PIK3CA","CALM1","YAP1","USP18","PDED"],"other_free_text":[]},"mechanistic_narrative":"KRAS encodes a small GTPase that cycles between GDP-bound (inactive) and GTP-bound (active) states, transducing signals from receptor tyrosine kinases through effector pathways including RAF-MEK-ERK, PI3K-AKT, and RalGDS to control cell proliferation, survival, and metabolism [PMID:8493579, PMID:29444439]. Oncogenic mutations at codons 12, 13, and 61 impair GAP-mediated GTP hydrolysis, locking KRAS in the active state and constitutively driving downstream signaling; these mutations are near-universal early events in pancreatic and colorectal carcinogenesis [PMID:2453289, PMID:2648401, PMID:26902995]. Membrane association via C-terminal farnesylation and the polybasic hypervariable region is essential for signaling, and this region also mediates nucleotide-state-dependent autoinhibition of the effector-binding site, calmodulin-dependent membrane extraction feeding PI3Kα activation, and regulation by phosphorylation at Ser-181 [PMID:26682817, PMID:19583261, PMID:27410739, PMID:28448716]. Oncogenic KRAS reprograms glutamine metabolism through a non-canonical GOT1-dependent pathway to maintain redox balance, signals predominantly through ERK to deregulate APC/C and cell-cycle machinery, and its protein output is constrained by rare codon usage that acts as a built-in tumor-suppressive mechanism [PMID:23535601, PMID:38843331, PMID:23246410, PMID:25437878]."},"prefetch_data":{"uniprot":{"accession":"P01116","full_name":"GTPase KRas","aliases":["K-Ras 2","Ki-Ras","c-K-ras","c-Ki-ras"],"length_aa":189,"mass_kda":21.7,"function":"Ras proteins bind GDP/GTP and possess intrinsic GTPase activity (PubMed:20949621, PubMed:39809765). Plays an important role in the regulation of cell proliferation (PubMed:22711838, PubMed:23698361). Activates MAPK1/MAPK3 resulting in phosphorylation and ultimately degradation of GJA1 (By similarity). Plays a role in promoting oncogenic events by inducing transcriptional silencing of tumor suppressor genes (TSGs) in colorectal cancer (CRC) cells in a ZNF304-dependent manner (PubMed:24623306)","subcellular_location":"Cell membrane","url":"https://www.uniprot.org/uniprotkb/P01116/entry"},"depmap":{"release":"DepMap","has_data":true,"is_common_essential":true,"resolved_as":"","url":"https://depmap.org/portal/gene/KRAS","classification":"Common Essential","n_dependent_lines":696,"n_total_lines":1208,"dependency_fraction":0.5761589403973509},"opencell":{"profiled":true,"resolved_as":"","ensg_id":"ENSG00000133703","cell_line_id":"CID000479","localizations":[{"compartment":"membrane","grade":3},{"compartment":"cytoplasmic","grade":1}],"interactors":[{"gene":"YWHAH","stoichiometry":10.0},{"gene":"YWHAQ","stoichiometry":10.0},{"gene":"YWHAB","stoichiometry":10.0},{"gene":"YWHAG","stoichiometry":4.0},{"gene":"ACTR2","stoichiometry":4.0},{"gene":"YWHAZ","stoichiometry":0.2},{"gene":"YWHAE","stoichiometry":0.2},{"gene":"KIAA0430","stoichiometry":0.2},{"gene":"APPL1","stoichiometry":0.2},{"gene":"TRAPPC4","stoichiometry":0.2}],"url":"https://opencell.sf.czbiohub.org/target/CID000479","total_profiled":1310},"omim":[{"mim_id":"621467","title":"NCBP2 ANTISENSE RNA 2 (HEAD TO HEAD); NCBP2AS2","url":"https://www.omim.org/entry/621467"},{"mim_id":"621092","title":"IQ MOTIF-CONTAINING GTPase-ACTIVATING PROTEIN 3; IQGAP3","url":"https://www.omim.org/entry/621092"},{"mim_id":"620654","title":"THROMBOCYTOPENIA 11 WITH MULTIPLE CONGENITAL ANOMALIES AND DYSMORPHIC FACIES; THC11","url":"https://www.omim.org/entry/620654"},{"mim_id":"620302","title":"WD REPEAT-CONTAINING PROTEIN 76; WDR76","url":"https://www.omim.org/entry/620302"},{"mim_id":"618712","title":"ANKYRIN REPEAT DOMAIN-CONTAINING PROTEIN 45; ANKRD45","url":"https://www.omim.org/entry/618712"}],"hpa":{"profiled":true,"resolved_as":"","reliability":"Supported","locations":[{"location":"Cytosol","reliability":"Supported"}],"tissue_specificity":"Low tissue specificity","tissue_distribution":"Detected in all","driving_tissues":[],"url":"https://www.proteinatlas.org/search/KRAS"},"hgnc":{"alias_symbol":["KRAS1","K-Ras4B"],"prev_symbol":["KRAS2"]},"alphafold":{"accession":"P01116","domains":[{"cath_id":"3.40.50.300","chopping":"1-175","consensus_level":"high","plddt":95.1591,"start":1,"end":175}],"viewer_url":"https://alphafold.ebi.ac.uk/entry/P01116","model_url":"https://alphafold.ebi.ac.uk/files/AF-P01116-F1-model_v6.cif","pae_url":"https://alphafold.ebi.ac.uk/files/AF-P01116-F1-predicted_aligned_error_v6.png","plddt_mean":91.5},"mouse_models":{"mgi_url":"https://www.informatics.jax.org/marker/summary?nomen=KRAS","jax_strain_url":"https://www.jax.org/strain/search?query=KRAS"},"sequence":{"accession":"P01116","fasta_url":"https://rest.uniprot.org/uniprotkb/P01116.fasta","uniprot_url":"https://www.uniprot.org/uniprotkb/P01116/entry","alphafold_viewer_url":"https://alphafold.ebi.ac.uk/entry/P01116"}},"corpus_meta":[{"pmid":"34776511","id":"PMC_34776511","title":"KRAS mutation: from undruggable to druggable in cancer.","date":"2021","source":"Signal transduction and targeted therapy","url":"https://pubmed.ncbi.nlm.nih.gov/34776511","citation_count":796,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"24954536","id":"PMC_24954536","title":"KRAS and YAP1 converge to regulate EMT and tumor survival.","date":"2014","source":"Cell","url":"https://pubmed.ncbi.nlm.nih.gov/24954536","citation_count":650,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"29364867","id":"PMC_29364867","title":"Evolutionary routes and KRAS dosage define pancreatic cancer phenotypes.","date":"2018","source":"Nature","url":"https://pubmed.ncbi.nlm.nih.gov/29364867","citation_count":334,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"31332011","id":"PMC_31332011","title":"Drugging an undruggable pocket on KRAS.","date":"2019","source":"Proceedings of the National Academy of Sciences of the United States of America","url":"https://pubmed.ncbi.nlm.nih.gov/31332011","citation_count":328,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"20617134","id":"PMC_20617134","title":"Clinical relevance of KRAS in human cancers.","date":"2010","source":"Journal of biomedicine & biotechnology","url":"https://pubmed.ncbi.nlm.nih.gov/20617134","citation_count":289,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"33676749","id":"PMC_33676749","title":"KRAS mutation in pancreatic cancer.","date":"2021","source":"Seminars in oncology","url":"https://pubmed.ncbi.nlm.nih.gov/33676749","citation_count":276,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"31649840","id":"PMC_31649840","title":"Targeting the untargetable KRAS in cancer therapy.","date":"2019","source":"Acta pharmaceutica Sinica. B","url":"https://pubmed.ncbi.nlm.nih.gov/31649840","citation_count":269,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"25878360","id":"PMC_25878360","title":"KRAS as a Therapeutic Target.","date":"2015","source":"Clinical cancer research : an official journal of the American Association for Cancer Research","url":"https://pubmed.ncbi.nlm.nih.gov/25878360","citation_count":254,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"35046095","id":"PMC_35046095","title":"Expanding the Reach of Precision Oncology by Drugging All KRAS Mutants.","date":"2022","source":"Cancer discovery","url":"https://pubmed.ncbi.nlm.nih.gov/35046095","citation_count":247,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"38637634","id":"PMC_38637634","title":"Targeting KRAS in cancer.","date":"2024","source":"Nature medicine","url":"https://pubmed.ncbi.nlm.nih.gov/38637634","citation_count":213,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"29336889","id":"PMC_29336889","title":"KRAS Dimerization Impacts MEK Inhibitor Sensitivity and Oncogenic Activity of Mutant KRAS.","date":"2018","source":"Cell","url":"https://pubmed.ncbi.nlm.nih.gov/29336889","citation_count":212,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"23401440","id":"PMC_23401440","title":"KRAS mutation: should we test for it, and does it matter?","date":"2013","source":"Journal of clinical oncology : official journal of the American Society of Clinical Oncology","url":"https://pubmed.ncbi.nlm.nih.gov/23401440","citation_count":202,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"2648401","id":"PMC_2648401","title":"Mutations in the KRAS2 oncogene during progressive stages of human colon carcinoma.","date":"1989","source":"Proceedings of the National Academy of Sciences of the United States of America","url":"https://pubmed.ncbi.nlm.nih.gov/2648401","citation_count":199,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"26902995","id":"PMC_26902995","title":"The Structural Basis of Oncogenic Mutations G12, G13 and Q61 in Small GTPase K-Ras4B.","date":"2016","source":"Scientific reports","url":"https://pubmed.ncbi.nlm.nih.gov/26902995","citation_count":181,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"26051715","id":"PMC_26051715","title":"GTP-Dependent K-Ras Dimerization.","date":"2015","source":"Structure (London, England : 1993)","url":"https://pubmed.ncbi.nlm.nih.gov/26051715","citation_count":174,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"7592913","id":"PMC_7592913","title":"Disruption of oncogenic K-Ras4B processing and signaling by a potent geranylgeranyltransferase I inhibitor.","date":"1995","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/7592913","citation_count":168,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"34471232","id":"PMC_34471232","title":"The KRAS-G12C inhibitor: activity and resistance.","date":"2021","source":"Cancer gene therapy","url":"https://pubmed.ncbi.nlm.nih.gov/34471232","citation_count":157,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"32014824","id":"PMC_32014824","title":"KRAS G12C Game of Thrones, which direct KRAS inhibitor will claim the iron throne?","date":"2020","source":"Cancer treatment reviews","url":"https://pubmed.ncbi.nlm.nih.gov/32014824","citation_count":155,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"29061961","id":"PMC_29061961","title":"Survival of pancreatic cancer cells lacking KRAS function.","date":"2017","source":"Nature communications","url":"https://pubmed.ncbi.nlm.nih.gov/29061961","citation_count":152,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"35732135","id":"PMC_35732135","title":"KRASG12C-independent feedback activation of wild-type RAS constrains KRASG12C inhibitor efficacy.","date":"2022","source":"Cell reports","url":"https://pubmed.ncbi.nlm.nih.gov/35732135","citation_count":128,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"10989276","id":"PMC_10989276","title":"The importance of being K-Ras.","date":"2000","source":"Cellular signalling","url":"https://pubmed.ncbi.nlm.nih.gov/10989276","citation_count":127,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"29444439","id":"PMC_29444439","title":"Differential Effector Engagement by Oncogenic KRAS.","date":"2018","source":"Cell reports","url":"https://pubmed.ncbi.nlm.nih.gov/29444439","citation_count":120,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"23246410","id":"PMC_23246410","title":"Rare codons regulate KRas oncogenesis.","date":"2012","source":"Current biology : CB","url":"https://pubmed.ncbi.nlm.nih.gov/23246410","citation_count":118,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"21125676","id":"PMC_21125676","title":"KRAS and BRAF: drug targets and predictive biomarkers.","date":"2010","source":"The Journal of pathology","url":"https://pubmed.ncbi.nlm.nih.gov/21125676","citation_count":118,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"12189555","id":"PMC_12189555","title":"Differential diagnosis between chronic pancreatitis and pancreatic cancer: value of the detection of KRAS2 mutations in circulating DNA.","date":"2002","source":"British journal of cancer","url":"https://pubmed.ncbi.nlm.nih.gov/12189555","citation_count":112,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"10673434","id":"PMC_10673434","title":"The basis for K-Ras4B binding specificity to protein farnesyltransferase revealed by 2 A resolution ternary complex structures.","date":"2000","source":"Structure (London, England : 1993)","url":"https://pubmed.ncbi.nlm.nih.gov/10673434","citation_count":112,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"20857619","id":"PMC_20857619","title":"EGFR and KRAS in colorectal cancer.","date":"2010","source":"Advances in clinical chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/20857619","citation_count":101,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"38109937","id":"PMC_38109937","title":"The energetic and allosteric landscape for KRAS inhibition.","date":"2023","source":"Nature","url":"https://pubmed.ncbi.nlm.nih.gov/38109937","citation_count":99,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"32591521","id":"PMC_32591521","title":"A potent KRAS macromolecule degrader specifically targeting tumours with mutant KRAS.","date":"2020","source":"Nature communications","url":"https://pubmed.ncbi.nlm.nih.gov/32591521","citation_count":97,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"25713064","id":"PMC_25713064","title":"Mechanisms of membrane binding of small GTPase K-Ras4B farnesylated hypervariable region.","date":"2015","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/25713064","citation_count":95,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"35319967","id":"PMC_35319967","title":"Landscape of KRASG12C, Associated Genomic Alterations, and Interrelation With Immuno-Oncology Biomarkers in KRAS-Mutated Cancers.","date":"2022","source":"JCO precision oncology","url":"https://pubmed.ncbi.nlm.nih.gov/35319967","citation_count":94,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"27193833","id":"PMC_27193833","title":"Suppression of KRas-mutant cancer through the combined inhibition of KRAS with PLK1 and ROCK.","date":"2016","source":"Nature communications","url":"https://pubmed.ncbi.nlm.nih.gov/27193833","citation_count":83,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"29326299","id":"PMC_29326299","title":"Targeting mutant KRAS with CRISPR-Cas9 controls tumor growth.","date":"2018","source":"Genome research","url":"https://pubmed.ncbi.nlm.nih.gov/29326299","citation_count":82,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"38843331","id":"PMC_38843331","title":"Defining the KRAS- and ERK-dependent transcriptome in KRAS-mutant cancers.","date":"2024","source":"Science (New York, N.Y.)","url":"https://pubmed.ncbi.nlm.nih.gov/38843331","citation_count":79,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"12882980","id":"PMC_12882980","title":"High affinity for farnesyltransferase and alternative prenylation contribute individually to K-Ras4B resistance to farnesyltransferase inhibitors.","date":"2003","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/12882980","citation_count":77,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"30122370","id":"PMC_30122370","title":"Inhibition of K-RAS4B by a Unique Mechanism of Action: Stabilizing Membrane-Dependent Occlusion of the Effector-Binding Site.","date":"2018","source":"Cell chemical biology","url":"https://pubmed.ncbi.nlm.nih.gov/30122370","citation_count":77,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"26682817","id":"PMC_26682817","title":"High-Affinity Interaction of the K-Ras4B Hypervariable Region with the Ras Active Site.","date":"2015","source":"Biophysical journal","url":"https://pubmed.ncbi.nlm.nih.gov/26682817","citation_count":72,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"34830757","id":"PMC_34830757","title":"Oncogenic KRAS: Signaling and Drug Resistance.","date":"2021","source":"Cancers","url":"https://pubmed.ncbi.nlm.nih.gov/34830757","citation_count":69,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"28462395","id":"PMC_28462395","title":"Calmodulin and PI3K Signaling in KRAS Cancers.","date":"2017","source":"Trends in cancer","url":"https://pubmed.ncbi.nlm.nih.gov/28462395","citation_count":68,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"24368337","id":"PMC_24368337","title":"The proto-oncogene KRAS is targeted by miR-200c.","date":"2014","source":"Oncotarget","url":"https://pubmed.ncbi.nlm.nih.gov/24368337","citation_count":67,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"25437878","id":"PMC_25437878","title":"Rare codons capacitate Kras-driven de novo tumorigenesis.","date":"2014","source":"The Journal of clinical investigation","url":"https://pubmed.ncbi.nlm.nih.gov/25437878","citation_count":66,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"34607583","id":"PMC_34607583","title":"Oncogenic KRAS blockade therapy: renewed enthusiasm and persistent challenges.","date":"2021","source":"Molecular cancer","url":"https://pubmed.ncbi.nlm.nih.gov/34607583","citation_count":65,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"29320680","id":"PMC_29320680","title":"K-Ras4B Remains Monomeric on Membranes over a Wide Range of Surface Densities and Lipid Compositions.","date":"2018","source":"Biophysical journal","url":"https://pubmed.ncbi.nlm.nih.gov/29320680","citation_count":63,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"30711927","id":"PMC_30711927","title":"The Role of KRAS in Endometrial Cancer: A Mini-Review.","date":"2019","source":"Anticancer research","url":"https://pubmed.ncbi.nlm.nih.gov/30711927","citation_count":59,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"25245423","id":"PMC_25245423","title":"MUC1-C confers EMT and KRAS independence in mutant KRAS lung cancer cells.","date":"2014","source":"Oncotarget","url":"https://pubmed.ncbi.nlm.nih.gov/25245423","citation_count":59,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"19583261","id":"PMC_19583261","title":"The hypervariable region of K-Ras4B is responsible for its specific interactions with calmodulin.","date":"2009","source":"Biochemistry","url":"https://pubmed.ncbi.nlm.nih.gov/19583261","citation_count":57,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"30971271","id":"PMC_30971271","title":"Small Molecule KRAS Agonist for Mutant KRAS Cancer Therapy.","date":"2019","source":"Molecular cancer","url":"https://pubmed.ncbi.nlm.nih.gov/30971271","citation_count":57,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"14724583","id":"PMC_14724583","title":"Mutations of BRAF and KRAS2 in the development of Barrett's adenocarcinoma.","date":"2004","source":"Oncogene","url":"https://pubmed.ncbi.nlm.nih.gov/14724583","citation_count":57,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"38666907","id":"PMC_38666907","title":"Significance of TP53, CDKN2A, SMAD4 and KRAS in Pancreatic Cancer.","date":"2024","source":"Current issues in molecular biology","url":"https://pubmed.ncbi.nlm.nih.gov/38666907","citation_count":54,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"33995590","id":"PMC_33995590","title":"KRAS/LKB1 and KRAS/TP53 co-mutations create divergent immune signatures in lung adenocarcinomas.","date":"2021","source":"Therapeutic advances in medical oncology","url":"https://pubmed.ncbi.nlm.nih.gov/33995590","citation_count":52,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"18075308","id":"PMC_18075308","title":"Sensitive and quantitative detection of KRAS2 gene mutations in pancreatic duct juice differentiates patients with pancreatic cancer from chronic pancreatitis, potential for early detection.","date":"2008","source":"Cancer biology & therapy","url":"https://pubmed.ncbi.nlm.nih.gov/18075308","citation_count":51,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"21745016","id":"PMC_21745016","title":"Laboratory methods for KRAS mutation analysis.","date":"2011","source":"Expert review of molecular diagnostics","url":"https://pubmed.ncbi.nlm.nih.gov/21745016","citation_count":50,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"38686056","id":"PMC_38686056","title":"Targeting KRAS in pancreatic cancer.","date":"2024","source":"Oncology research","url":"https://pubmed.ncbi.nlm.nih.gov/38686056","citation_count":49,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"28597297","id":"PMC_28597297","title":"Intrinsic protein disorder in oncogenic KRAS signaling.","date":"2017","source":"Cellular and molecular life sciences : CMLS","url":"https://pubmed.ncbi.nlm.nih.gov/28597297","citation_count":48,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"29120102","id":"PMC_29120102","title":"Deciphering lipid codes: K-Ras as a paradigm.","date":"2017","source":"Traffic (Copenhagen, Denmark)","url":"https://pubmed.ncbi.nlm.nih.gov/29120102","citation_count":47,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"36414681","id":"PMC_36414681","title":"Glimmers of hope for targeting oncogenic KRAS-G12D.","date":"2022","source":"Cancer gene therapy","url":"https://pubmed.ncbi.nlm.nih.gov/36414681","citation_count":46,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"34303932","id":"PMC_34303932","title":"Stopping the beating heart of cancer: KRAS reviewed.","date":"2021","source":"Current opinion in structural biology","url":"https://pubmed.ncbi.nlm.nih.gov/34303932","citation_count":46,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"35014625","id":"PMC_35014625","title":"Targeting the undruggable oncogenic KRAS: the dawn of hope.","date":"2022","source":"JCI insight","url":"https://pubmed.ncbi.nlm.nih.gov/35014625","citation_count":45,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"23208496","id":"PMC_23208496","title":"CD44 promotes Kras-dependent lung adenocarcinoma.","date":"2012","source":"Oncogene","url":"https://pubmed.ncbi.nlm.nih.gov/23208496","citation_count":45,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"27410739","id":"PMC_27410739","title":"Regulation of K-Ras4B Membrane Binding by Calmodulin.","date":"2016","source":"Biophysical journal","url":"https://pubmed.ncbi.nlm.nih.gov/27410739","citation_count":43,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"39799325","id":"PMC_39799325","title":"Targeting KRAS: from metabolic regulation to cancer treatment.","date":"2025","source":"Molecular cancer","url":"https://pubmed.ncbi.nlm.nih.gov/39799325","citation_count":42,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"26709496","id":"PMC_26709496","title":"The disordered hypervariable region and the folded catalytic domain of oncogenic K-Ras4B partner in phospholipid binding.","date":"2015","source":"Current opinion in structural biology","url":"https://pubmed.ncbi.nlm.nih.gov/26709496","citation_count":41,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"28623230","id":"PMC_28623230","title":"Flexible-body motions of calmodulin and the farnesylated hypervariable region yield a high-affinity interaction enabling K-Ras4B membrane extraction.","date":"2017","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/28623230","citation_count":39,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"35561303","id":"PMC_35561303","title":"More to the RAS Story: KRASG12C Inhibition, Resistance Mechanisms, and Moving Beyond KRASG12C.","date":"2022","source":"American Society of Clinical Oncology educational book. American Society of Clinical Oncology. Annual Meeting","url":"https://pubmed.ncbi.nlm.nih.gov/35561303","citation_count":39,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"28242811","id":"PMC_28242811","title":"Deubiquitinase USP18 Loss Mislocalizes and Destabilizes KRAS in Lung Cancer.","date":"2017","source":"Molecular cancer research : MCR","url":"https://pubmed.ncbi.nlm.nih.gov/28242811","citation_count":38,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"34064352","id":"PMC_34064352","title":"Targeting KRAS in Solid Tumors: Current Challenges and Future Opportunities of Novel KRAS Inhibitors.","date":"2021","source":"Pharmaceutics","url":"https://pubmed.ncbi.nlm.nih.gov/34064352","citation_count":37,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"9921983","id":"PMC_9921983","title":"Differences in patterns of TP53 and KRAS2 mutations in a large series of endometrial carcinomas with or without microsatellite instability.","date":"1999","source":"Cancer","url":"https://pubmed.ncbi.nlm.nih.gov/9921983","citation_count":36,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"39047616","id":"PMC_39047616","title":"The next-generation KRAS inhibitors…What comes after sotorasib and adagrasib?","date":"2024","source":"Lung cancer (Amsterdam, Netherlands)","url":"https://pubmed.ncbi.nlm.nih.gov/39047616","citation_count":34,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"37190303","id":"PMC_37190303","title":"The Therapeutic Landscape for KRAS-Mutated Colorectal Cancers.","date":"2023","source":"Cancers","url":"https://pubmed.ncbi.nlm.nih.gov/37190303","citation_count":34,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"31816384","id":"PMC_31816384","title":"Insight into the mechanism of allosteric activation of PI3Kα by oncoprotein K-Ras4B.","date":"2019","source":"International journal of biological macromolecules","url":"https://pubmed.ncbi.nlm.nih.gov/31816384","citation_count":34,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"28448716","id":"PMC_28448716","title":"Phosphorylation Weakens but Does Not Inhibit Membrane Binding and Clustering of K-Ras4B.","date":"2017","source":"ACS chemical biology","url":"https://pubmed.ncbi.nlm.nih.gov/28448716","citation_count":33,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"38668053","id":"PMC_38668053","title":"KRAS: Biology, Inhibition, and Mechanisms of Inhibitor Resistance.","date":"2024","source":"Current oncology (Toronto, Ont.)","url":"https://pubmed.ncbi.nlm.nih.gov/38668053","citation_count":32,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"30089720","id":"PMC_30089720","title":"Requirement for MUC5AC in KRAS-dependent lung carcinogenesis.","date":"2018","source":"JCI insight","url":"https://pubmed.ncbi.nlm.nih.gov/30089720","citation_count":32,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"25371176","id":"PMC_25371176","title":"Wild-type KRAS inhibits oncogenic KRAS-induced T-ALL in mice.","date":"2014","source":"Leukemia","url":"https://pubmed.ncbi.nlm.nih.gov/25371176","citation_count":32,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"35994329","id":"PMC_35994329","title":"Markov State Models and Molecular Dynamics Simulations Reveal the Conformational Transition of the Intrinsically Disordered Hypervariable Region of K-Ras4B to the Ordered Conformation.","date":"2022","source":"Journal of chemical information and modeling","url":"https://pubmed.ncbi.nlm.nih.gov/35994329","citation_count":30,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"39638567","id":"PMC_39638567","title":"\"Undruggable KRAS\": druggable after all.","date":"2025","source":"Genes & development","url":"https://pubmed.ncbi.nlm.nih.gov/39638567","citation_count":29,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"34520956","id":"PMC_34520956","title":"Targeting mutated GTPase KRAS in tumor therapies.","date":"2021","source":"European journal of medicinal chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/34520956","citation_count":29,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"32286309","id":"PMC_32286309","title":"Capturing the primordial Kras mutation initiating urethane carcinogenesis.","date":"2020","source":"Nature communications","url":"https://pubmed.ncbi.nlm.nih.gov/32286309","citation_count":29,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"32796566","id":"PMC_32796566","title":"Targeting Mutant KRAS in Pancreatic Cancer: Futile or Promising?","date":"2020","source":"Biomedicines","url":"https://pubmed.ncbi.nlm.nih.gov/32796566","citation_count":28,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"26873344","id":"PMC_26873344","title":"K-Ras4B/calmodulin/PI3Kα: A promising new adenocarcinoma-specific drug target?","date":"2016","source":"Expert opinion on therapeutic targets","url":"https://pubmed.ncbi.nlm.nih.gov/26873344","citation_count":28,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"29358307","id":"PMC_29358307","title":"miR-422a inhibits osteosarcoma proliferation by targeting BCL2L2 and KRAS.","date":"2018","source":"Bioscience reports","url":"https://pubmed.ncbi.nlm.nih.gov/29358307","citation_count":27,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"34688654","id":"PMC_34688654","title":"The KRAS-regulated kinome identifies WEE1 and ERK coinhibition as a potential therapeutic strategy in KRAS-mutant pancreatic cancer.","date":"2021","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/34688654","citation_count":26,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"14511407","id":"PMC_14511407","title":"RASSF1A promoter methylation and Kras2 mutations in non small cell lung cancer.","date":"2003","source":"Neoplasia (New York, N.Y.)","url":"https://pubmed.ncbi.nlm.nih.gov/14511407","citation_count":25,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"39167893","id":"PMC_39167893","title":"Structural insights into small-molecule KRAS inhibitors for targeting KRAS mutant cancers.","date":"2024","source":"European journal of medicinal chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/39167893","citation_count":24,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"26773157","id":"PMC_26773157","title":"Kras Is Critical for B Cell Lymphopoiesis.","date":"2016","source":"Journal of immunology (Baltimore, Md. : 1950)","url":"https://pubmed.ncbi.nlm.nih.gov/26773157","citation_count":24,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"38116743","id":"PMC_38116743","title":"Decrypting Allostery in Membrane-Bound K-Ras4B Using Complementary In Silico Approaches Based on Unbiased Molecular Dynamics Simulations.","date":"2023","source":"Journal of the American Chemical Society","url":"https://pubmed.ncbi.nlm.nih.gov/38116743","citation_count":23,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"35093302","id":"PMC_35093302","title":"Interplay between K-RAS and miRNAs.","date":"2022","source":"Trends in cancer","url":"https://pubmed.ncbi.nlm.nih.gov/35093302","citation_count":22,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"34040167","id":"PMC_34040167","title":"AMPKα loss promotes KRAS-mediated lung tumorigenesis.","date":"2021","source":"Cell death and differentiation","url":"https://pubmed.ncbi.nlm.nih.gov/34040167","citation_count":22,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"29499269","id":"PMC_29499269","title":"Oncogenic KRas mobility in the membrane and signaling response.","date":"2018","source":"Seminars in cancer biology","url":"https://pubmed.ncbi.nlm.nih.gov/29499269","citation_count":22,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"37775182","id":"PMC_37775182","title":"Stromal-derived NRG1 enables oncogenic KRAS bypass in pancreas cancer.","date":"2023","source":"Genes & development","url":"https://pubmed.ncbi.nlm.nih.gov/37775182","citation_count":21,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"20232877","id":"PMC_20232877","title":"Imaging human pancreatic cancer xenografts by targeting mutant KRAS2 mRNA with [(111)In]DOTA(n)-poly(diamidopropanoyl)(m)-KRAS2 PNA-D(Cys-Ser-Lys-Cys) nanoparticles.","date":"2010","source":"Bioconjugate chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/20232877","citation_count":21,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"38937452","id":"PMC_38937452","title":"A pan-KRAS degrader for the treatment of KRAS-mutant cancers.","date":"2024","source":"Cell discovery","url":"https://pubmed.ncbi.nlm.nih.gov/38937452","citation_count":20,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"36946612","id":"PMC_36946612","title":"KRAS Hijacks the miRNA Regulatory Pathway in Cancer.","date":"2023","source":"Cancer research","url":"https://pubmed.ncbi.nlm.nih.gov/36946612","citation_count":20,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"31327761","id":"PMC_31327761","title":"miR-548d-3p inhibits osteosarcoma by downregulating KRAS.","date":"2019","source":"Aging","url":"https://pubmed.ncbi.nlm.nih.gov/31327761","citation_count":20,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"31277422","id":"PMC_31277422","title":"Activating Mutations in PTPN11 and KRAS in Canine Histiocytic Sarcomas.","date":"2019","source":"Genes","url":"https://pubmed.ncbi.nlm.nih.gov/31277422","citation_count":20,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"33838397","id":"PMC_33838397","title":"Targeting mutant KRAS.","date":"2021","source":"Current opinion in chemical biology","url":"https://pubmed.ncbi.nlm.nih.gov/33838397","citation_count":19,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"37696812","id":"PMC_37696812","title":"CBX4 deletion promotes tumorigenesis under KrasG12D background by inducing genomic instability.","date":"2023","source":"Signal transduction and targeted therapy","url":"https://pubmed.ncbi.nlm.nih.gov/37696812","citation_count":19,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"22569482","id":"PMC_22569482","title":"K-Ras4B lipoprotein synthesis: biochemical characterization, functional properties, and dimer formation.","date":"2012","source":"Protein expression and purification","url":"https://pubmed.ncbi.nlm.nih.gov/22569482","citation_count":18,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"30976068","id":"PMC_30976068","title":"Detection of KRAS mutation via ligation-initiated LAMP reaction.","date":"2019","source":"Scientific reports","url":"https://pubmed.ncbi.nlm.nih.gov/30976068","citation_count":18,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"33202255","id":"PMC_33202255","title":"A CRISPR-Cas9 repressor for epigenetic silencing of KRAS.","date":"2020","source":"Pharmacological research","url":"https://pubmed.ncbi.nlm.nih.gov/33202255","citation_count":18,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"19339720","id":"PMC_19339720","title":"Cetuximab and chemotherapy as initial treatment for metastatic colorectal cancer.","date":"2009","source":"The New England journal of medicine","url":"https://pubmed.ncbi.nlm.nih.gov/19339720","citation_count":3068,"is_preprint":false,"source_track":"gene2pubmed"},{"pmid":"19847166","id":"PMC_19847166","title":"Systematic RNA interference reveals that oncogenic KRAS-driven cancers require TBK1.","date":"2009","source":"Nature","url":"https://pubmed.ncbi.nlm.nih.gov/19847166","citation_count":2959,"is_preprint":false,"source_track":"gene2pubmed"},{"pmid":"18946061","id":"PMC_18946061","title":"K-ras mutations and benefit from cetuximab in advanced colorectal cancer.","date":"2008","source":"The New England journal of medicine","url":"https://pubmed.ncbi.nlm.nih.gov/18946061","citation_count":2921,"is_preprint":false,"source_track":"gene2pubmed"},{"pmid":"18316791","id":"PMC_18316791","title":"Wild-type KRAS is required for panitumumab efficacy in patients with metastatic colorectal cancer.","date":"2008","source":"Journal of clinical oncology : official journal of the American Society of Clinical Oncology","url":"https://pubmed.ncbi.nlm.nih.gov/18316791","citation_count":2528,"is_preprint":false,"source_track":"gene2pubmed"},{"pmid":"16189514","id":"PMC_16189514","title":"Towards a proteome-scale map of the human protein-protein interaction network.","date":"2005","source":"Nature","url":"https://pubmed.ncbi.nlm.nih.gov/16189514","citation_count":2090,"is_preprint":false,"source_track":"gene2pubmed"},{"pmid":"15741570","id":"PMC_15741570","title":"Clinical and biological features associated with epidermal growth factor receptor gene mutations in lung cancers.","date":"2005","source":"Journal of the National Cancer Institute","url":"https://pubmed.ncbi.nlm.nih.gov/15741570","citation_count":2027,"is_preprint":false,"source_track":"gene2pubmed"},{"pmid":"2453289","id":"PMC_2453289","title":"Most human carcinomas of the exocrine pancreas contain mutant c-K-ras genes.","date":"1988","source":"Cell","url":"https://pubmed.ncbi.nlm.nih.gov/2453289","citation_count":1932,"is_preprint":false,"source_track":"gene2pubmed"},{"pmid":"16618717","id":"PMC_16618717","title":"KRAS mutation status is predictive of response to cetuximab therapy in colorectal cancer.","date":"2006","source":"Cancer research","url":"https://pubmed.ncbi.nlm.nih.gov/16618717","citation_count":1813,"is_preprint":false,"source_track":"gene2pubmed"},{"pmid":"24024839","id":"PMC_24024839","title":"Panitumumab-FOLFOX4 treatment and RAS mutations in colorectal cancer.","date":"2013","source":"The New England journal of medicine","url":"https://pubmed.ncbi.nlm.nih.gov/24024839","citation_count":1802,"is_preprint":false,"source_track":"gene2pubmed"},{"pmid":"20619739","id":"PMC_20619739","title":"Effects of KRAS, BRAF, NRAS, and PIK3CA mutations on the efficacy of cetuximab plus chemotherapy in chemotherapy-refractory metastatic colorectal cancer: a retrospective consortium analysis.","date":"2010","source":"The Lancet. Oncology","url":"https://pubmed.ncbi.nlm.nih.gov/20619739","citation_count":1728,"is_preprint":false,"source_track":"gene2pubmed"},{"pmid":"23535601","id":"PMC_23535601","title":"Glutamine supports pancreatic cancer growth through a KRAS-regulated metabolic pathway.","date":"2013","source":"Nature","url":"https://pubmed.ncbi.nlm.nih.gov/23535601","citation_count":1642,"is_preprint":false,"source_track":"gene2pubmed"},{"pmid":"22589270","id":"PMC_22589270","title":"A comprehensive survey of Ras mutations in cancer.","date":"2012","source":"Cancer research","url":"https://pubmed.ncbi.nlm.nih.gov/22589270","citation_count":1612,"is_preprint":false,"source_track":"gene2pubmed"},{"pmid":"19667264","id":"PMC_19667264","title":"Clinical features and outcome of patients with non-small-cell lung cancer who harbor EML4-ALK.","date":"2009","source":"Journal of clinical oncology : official journal of the American Society of Clinical Oncology","url":"https://pubmed.ncbi.nlm.nih.gov/19667264","citation_count":1566,"is_preprint":false,"source_track":"gene2pubmed"},{"pmid":"12477932","id":"PMC_12477932","title":"Generation and initial analysis of more than 15,000 full-length human and mouse cDNA sequences.","date":"2002","source":"Proceedings of the National Academy of Sciences of the United States of America","url":"https://pubmed.ncbi.nlm.nih.gov/12477932","citation_count":1479,"is_preprint":false,"source_track":"gene2pubmed"},{"pmid":"19001320","id":"PMC_19001320","title":"Wild-type BRAF is required for response to panitumumab or cetuximab in metastatic colorectal cancer.","date":"2008","source":"Journal of clinical oncology : official journal of the American Society of Clinical Oncology","url":"https://pubmed.ncbi.nlm.nih.gov/19001320","citation_count":1310,"is_preprint":false,"source_track":"gene2pubmed"},{"pmid":"16043828","id":"PMC_16043828","title":"Mutations in the epidermal growth factor receptor and in KRAS are predictive and prognostic indicators in patients with non-small-cell lung cancer treated with chemotherapy alone and in combination with erlotinib.","date":"2005","source":"Journal of clinical oncology : official journal of the American Society of Clinical Oncology","url":"https://pubmed.ncbi.nlm.nih.gov/16043828","citation_count":1231,"is_preprint":false,"source_track":"gene2pubmed"},{"pmid":"18202412","id":"PMC_18202412","title":"KRAS mutations as an independent prognostic factor in patients with advanced colorectal cancer treated with cetuximab.","date":"2008","source":"Journal of clinical oncology : official journal of the American Society of Clinical Oncology","url":"https://pubmed.ncbi.nlm.nih.gov/18202412","citation_count":1208,"is_preprint":false,"source_track":"gene2pubmed"},{"pmid":"26627737","id":"PMC_26627737","title":"High-Resolution CRISPR Screens Reveal Fitness Genes and Genotype-Specific Cancer Liabilities.","date":"2015","source":"Cell","url":"https://pubmed.ncbi.nlm.nih.gov/26627737","citation_count":1200,"is_preprint":false,"source_track":"gene2pubmed"},{"pmid":"28514442","id":"PMC_28514442","title":"Architecture of the human interactome defines protein communities and disease networks.","date":"2017","source":"Nature","url":"https://pubmed.ncbi.nlm.nih.gov/28514442","citation_count":1085,"is_preprint":false,"source_track":"gene2pubmed"},{"pmid":"19196673","id":"PMC_19196673","title":"Chemotherapy, bevacizumab, and cetuximab in metastatic colorectal cancer.","date":"2009","source":"The New England journal of medicine","url":"https://pubmed.ncbi.nlm.nih.gov/19196673","citation_count":1065,"is_preprint":false,"source_track":"gene2pubmed"},{"pmid":"11520933","id":"PMC_11520933","title":"Nerve growth factor signaling, neuroprotection, and neural repair.","date":"2001","source":"Annual review of neuroscience","url":"https://pubmed.ncbi.nlm.nih.gov/11520933","citation_count":1029,"is_preprint":false,"source_track":"gene2pubmed"},{"pmid":"26496610","id":"PMC_26496610","title":"A human interactome in three quantitative dimensions organized by stoichiometries and abundances.","date":"2015","source":"Cell","url":"https://pubmed.ncbi.nlm.nih.gov/26496610","citation_count":1015,"is_preprint":false,"source_track":"gene2pubmed"},{"pmid":"25416956","id":"PMC_25416956","title":"A proteome-scale map of the human interactome network.","date":"2014","source":"Cell","url":"https://pubmed.ncbi.nlm.nih.gov/25416956","citation_count":977,"is_preprint":false,"source_track":"gene2pubmed"},{"pmid":"20008640","id":"PMC_20008640","title":"Prognostic role of KRAS and BRAF in stage II and III resected colon cancer: results of the translational study on the PETACC-3, EORTC 40993, SAKK 60-00 trial.","date":"2009","source":"Journal of clinical oncology : official journal of the American Society of Clinical Oncology","url":"https://pubmed.ncbi.nlm.nih.gov/20008640","citation_count":956,"is_preprint":false,"source_track":"gene2pubmed"},{"pmid":"34161704","id":"PMC_34161704","title":"Acquired Resistance to KRASG12C Inhibition in Cancer.","date":"2021","source":"The New England journal of medicine","url":"https://pubmed.ncbi.nlm.nih.gov/34161704","citation_count":949,"is_preprint":false,"source_track":"gene2pubmed"},{"pmid":"19273710","id":"PMC_19273710","title":"DPC4 gene status of the primary carcinoma correlates with patterns of failure in patients with pancreatic cancer.","date":"2009","source":"Journal of clinical oncology : official journal of the American Society of Clinical Oncology","url":"https://pubmed.ncbi.nlm.nih.gov/19273710","citation_count":888,"is_preprint":false,"source_track":"gene2pubmed"},{"pmid":"19490893","id":"PMC_19490893","title":"A genome-wide RNAi screen identifies multiple synthetic lethal interactions with the Ras oncogene.","date":"2009","source":"Cell","url":"https://pubmed.ncbi.nlm.nih.gov/19490893","citation_count":843,"is_preprint":false,"source_track":"gene2pubmed"},{"pmid":"28039262","id":"PMC_28039262","title":"Potential Predictive Value of TP53 and KRAS Mutation Status for Response to PD-1 Blockade Immunotherapy in Lung Adenocarcinoma.","date":"2016","source":"Clinical cancer research : an official journal of the American Association for Cancer Research","url":"https://pubmed.ncbi.nlm.nih.gov/28039262","citation_count":827,"is_preprint":false,"source_track":"gene2pubmed"},{"pmid":"24398677","id":"PMC_24398677","title":"Identification of double-stranded genomic DNA spanning all chromosomes with mutated KRAS and p53 DNA in the serum exosomes of patients with pancreatic cancer.","date":"2014","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/24398677","citation_count":804,"is_preprint":false,"source_track":"gene2pubmed"},{"pmid":"8493579","id":"PMC_8493579","title":"Human Sos1: a guanine nucleotide exchange factor for Ras that binds to GRB2.","date":"1993","source":"Science (New York, N.Y.)","url":"https://pubmed.ncbi.nlm.nih.gov/8493579","citation_count":772,"is_preprint":false,"source_track":"gene2pubmed"}],"cost":{"stage1":{"model":"claude-sonnet-4-6","input_tokens":43510,"output_tokens":7194,"usd":0.11922},"stage2":{"model":"claude-opus-4-6","input_tokens":10991,"output_tokens":4240,"usd":0.241432},"total_usd":0.755565,"stage1_batch_id":"msgbatch_01Mgbec4vM6c9vXht7JVJzss","stage2_batch_id":"msgbatch_01JkuuH5RB2BdJNsgnhjbC6s","note":"batch pricing = 50% of standard","round2_stage1":{"model":"claude-sonnet-4-6","input_tokens":59676,"output_tokens":8403,"usd":0.152536},"round2_rules_fired":"R3","round2_stage2":{"model":"claude-opus-4-6","input_tokens":12477,"output_tokens":3968,"usd":0.242377}},"stage1_raw":"```json\n{\n  \"discoveries\": [\n    {\n      \"year\": 2018,\n      \"finding\": \"KRAS dimerization at the α4-α5 interface mediates wild-type KRAS-dependent fitness of KRAS-mutant lung adenocarcinoma cells and underlies resistance to MEK inhibition. A dimerization-disrupting mutant (KRASD154Q) abolished these effects both in vitro and in vivo, demonstrating that dimerization is mechanistically required for oncogenic KRAS activity.\",\n      \"method\": \"Genetically inducible KRAS LOH model, KRASD154Q mutant, in vitro and in vivo functional assays\",\n      \"journal\": \"Cell\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — reciprocal genetic and functional validation with defined mutant, replicated in vivo\",\n      \"pmids\": [\"29336889\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2015,\n      \"finding\": \"GTP-bound K-Ras4B catalytic domain forms stable homodimers in solution, with two major dimer interfaces: a β-sheet interface overlapping switch I/effector binding regions (inhibitory to effectors) and a helical interface that may promote Raf activation.\",\n      \"method\": \"Analytical ultracentrifugation, molecular dynamics simulations, structural analysis\",\n      \"journal\": \"Structure\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 1/2 — biophysical reconstitution in solution with structural modeling, single lab\",\n      \"pmids\": [\"26051715\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"BI-2852 binds with nanomolar affinity to a pocket between switch I and switch II on KRAS (present in both active and inactive forms), blocking all GEF, GAP, and effector interactions with KRAS, thereby inhibiting downstream signaling and showing antiproliferative effects in KRAS-mutant cells.\",\n      \"method\": \"Structure-based drug design, X-ray crystallography, biochemical binding assays, cell-based signaling assays\",\n      \"journal\": \"Proceedings of the National Academy of Sciences of the United States of America\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 — structure-based design confirmed by crystal structure and functional assays\",\n      \"pmids\": [\"31332011\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2000,\n      \"finding\": \"Crystal structures of rat FTase ternary complexes with K-Ras4B peptide substrates reveal that the polybasic region of K-Ras4B forms a type I β-turn and binds along the rim of the hydrophobic cavity, conferring enhanced affinity for FTase; zinc is essential for productive CAAX peptide binding in the active conformation.\",\n      \"method\": \"X-ray crystallography (2 Å resolution ternary complex structures)\",\n      \"journal\": \"Structure\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 — high-resolution crystal structures with functional interpretation\",\n      \"pmids\": [\"10673434\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 1995,\n      \"finding\": \"K-Ras4B processing is highly resistant to farnesyltransferase inhibitor FTI-277 (IC50 ~10 μM) but sensitive to the geranylgeranyltransferase I inhibitor GGTI-286 (IC50 ~2 μM), demonstrating that K-Ras4B can be alternatively prenylated by geranylgeranyltransferase I and that this alternative prenylation sustains its oncogenic signaling.\",\n      \"method\": \"In vitro GGTase I assay, whole-cell prenylation assays, MAP kinase signaling assays\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 — in vitro enzyme assays with dose-response and functional downstream readout\",\n      \"pmids\": [\"7592913\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2003,\n      \"finding\": \"K-Ras4B resistance to farnesyltransferase inhibitors (FTIs) arises from two independent mechanisms: (1) its polybasic domain increases affinity for farnesyltransferase, and (2) its CAAX motif permits alternative geranylgeranylation. Either element alone confers FTI resistance on Ras prenylation and downstream signaling.\",\n      \"method\": \"Chimeric Ras protein expression, Elk-1 activation assays, anchorage-independent growth assays, microarray analysis\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — multiple orthogonal functional assays with chimeric protein dissection\",\n      \"pmids\": [\"12882980\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2009,\n      \"finding\": \"The hypervariable region (HVR) of K-Ras4B is responsible for its specific interaction with calmodulin: the HVR binds to the C-terminal domain of Ca2+-loaded calmodulin with micromolar affinity, while the GTP-loaded catalytic domain may interact with calmodulin's N-terminal domain, providing nucleotide-dependent control of the interaction.\",\n      \"method\": \"NMR spectroscopy, isothermal titration calorimetry\",\n      \"journal\": \"Biochemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 — biophysical binding measurements with domain mapping by NMR and ITC\",\n      \"pmids\": [\"19583261\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2015,\n      \"finding\": \"The C-terminal HVR of K-Ras4B directly interacts with the active site of the K-Ras4B catalytic domain; this interaction is ~100-fold tighter in the GDP-bound than GTP-bound state, interferes with Ras-Raf interaction, modulates phospholipid binding, and slightly slows nucleotide exchange, implicating HVR in autoinhibition.\",\n      \"method\": \"NMR spectroscopy, surface plasmon resonance, biochemical binding assays\",\n      \"journal\": \"Biophysical journal\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 — multiple biophysical methods demonstrating direct intramolecular interaction with functional consequences\",\n      \"pmids\": [\"26682817\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2015,\n      \"finding\": \"K-Ras4B membrane binding is driven by its farnesylated HVR: the farnesyl group preferentially inserts into disordered lipid microdomains; phosphorylation of Ser-181 prohibits spontaneous farnesyl membrane insertion; the polybasic lysine sequence modulates selective binding to anionic phospholipids.\",\n      \"method\": \"Confocal microscopy, surface plasmon resonance, molecular dynamics simulations\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1/2 — orthogonal biophysical methods with structural modeling, directly linking modifications to membrane localization\",\n      \"pmids\": [\"25713064\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2016,\n      \"finding\": \"Phosphorylation of K-Ras4B at Ser181 reduces but does not fully inhibit membrane binding and clustering; phosphorylated K-Ras4B maintains association with cytosolic shuttle PDEδ; phosphorylation does not alter localization to liquid-disordered lipid subdomains but reduces overall PM enrichment.\",\n      \"method\": \"In vitro synthesis of triply modified K-Ras4B (phospho/farnesyl/methyl), model biomembrane assays, spectroscopic and imaging techniques\",\n      \"journal\": \"ACS chemical biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 — reconstituted fully modified protein with multiple spectroscopic methods\",\n      \"pmids\": [\"28448716\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2016,\n      \"finding\": \"Ca2+/calmodulin extracts K-Ras4B from negatively charged membranes in a nucleotide-independent manner; the CaM–K-Ras4B complex shows no membrane binding; CaM and PDEδ both engage the farnesyl group but regulate K-Ras4B plasma membrane localization differently.\",\n      \"method\": \"Fluorescence spectroscopy, FRET, model biomembrane assays\",\n      \"journal\": \"Biophysical journal\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1/2 — multiple biophysical approaches with mechanistic comparison of two chaperones\",\n      \"pmids\": [\"27410739\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2017,\n      \"finding\": \"Calmodulin preferentially binds the unfolded K-Ras4B HVR (not α-helical HVR); the interaction involves all three CaM domains (both lobes and central linker); the farnesyl group of K-Ras4B docks to hydrophobic pockets in both CaM lobes, further stabilizing the complex and enabling membrane extraction of K-Ras4B to support PI3Kα activation.\",\n      \"method\": \"Molecular dynamics simulations, fluorescence binding experiments\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2/3 — MD simulations supported by fluorescence data, single lab\",\n      \"pmids\": [\"28623230\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2018,\n      \"finding\": \"A small-molecule compound (Cmpd2) inhibits K-RAS4B by a membrane-dependent mechanism: it simultaneously engages a shallow pocket on KRAS and associates with the lipid bilayer, stabilizing KRAS in an orientation where the membrane occludes its effector-binding site, reducing RAF binding and impairing RAF activation.\",\n      \"method\": \"NMR spectroscopy of membrane-associated prenylated K-RAS4B, biochemical RAF-binding assays, cell-based signaling assays\",\n      \"journal\": \"Cell chemical biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1/2 — structural NMR of membrane-embedded protein with functional validation of mechanism\",\n      \"pmids\": [\"30122370\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2016,\n      \"finding\": \"K-Ras4B impairs GAP-assisted GTP hydrolysis through oncogenic mutations at G12, G13, and Q61. Molecular dynamics simulations demonstrate that GAP not only donates its R789 arginine finger but also stabilizes the catalytically competent conformation and pre-organizes catalytic residue Q61; mutations at these positions disrupt R789/Q61 organization and impair GAP-mediated hydrolysis.\",\n      \"method\": \"Molecular dynamics simulations (6.4 μs total), structural analysis of mutant complexes with GAP\",\n      \"journal\": \"Scientific reports\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 1 computational — extensive MD without experimental validation, single study\",\n      \"pmids\": [\"26902995\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2012,\n      \"finding\": \"KRAS is enriched for rare codons relative to HRAS, which limits K-Ras protein expression. Converting rare to common codons in KRAS increases K-Ras protein levels and tumorigenicity to mirror HRas, demonstrating that synonymous codon usage differences underlie differential expression and tumorigenic activity between KRAS and HRAS isoforms.\",\n      \"method\": \"Synonymous codon-altered constructs, expression quantification, transformation assays\",\n      \"journal\": \"Current biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — isogenic constructs with orthogonal functional readouts, replicated in mouse model\",\n      \"pmids\": [\"23246410\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2014,\n      \"finding\": \"Rare codon bias in endogenous Kras limits its expression; mice with optimized Kras exon 3 (Kras^ex3op) develop fewer carcinogen-induced tumors, and the optimized allele shows tumor-suppressive activity when not mutated, while inducing growth arrest when oncogenically mutated—demonstrating that KRAS rare codon bias is integral to its role in tumorigenesis.\",\n      \"method\": \"Knock-in mouse model with synonymous codon-optimized Kras exon 3, urethane carcinogenesis, tumor burden assessment\",\n      \"journal\": \"The Journal of clinical investigation\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — in vivo knock-in mouse model with defined functional readout\",\n      \"pmids\": [\"25437878\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2014,\n      \"finding\": \"YAP1 can functionally substitute for oncogenic KRAS in maintaining survival of KRAS-dependent colon cancer cells. KRAS and YAP1 converge on the transcription factor FOS to activate a transcriptional program regulating EMT; YAP1 is required for KRAS-induced cell transformation, and increased YAP1 signaling mediates acquired resistance to KRAS suppression.\",\n      \"method\": \"ORF rescue screen (15,294 ORFs), inducible KRAS shRNA, mouse lung cancer KRAS suppression model, transcriptional analysis\",\n      \"journal\": \"Cell\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — genome-wide screen with genetic validation in vivo and mechanistic transcription factor identification\",\n      \"pmids\": [\"24954536\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2017,\n      \"finding\": \"A subset of KRAS-deficient pancreatic cancer cells survive via PI3K-dependent MAPK signaling (identified by CRISPR/Cas9-mediated KRAS deletion), demonstrating that KRAS is dispensable in some PDAC cells and that KRAS normally suppresses metastasis-related gene expression; loss of KRAS induces sensitivity to PI3K inhibitors.\",\n      \"method\": \"CRISPR/Cas9 KRAS knockout, gene expression profiling, pharmacological inhibitor assays\",\n      \"journal\": \"Nature communications\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — clean CRISPR KO with specific mechanistic readout and pharmacological validation\",\n      \"pmids\": [\"29061961\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"Upstream feedback activation of wild-type RAS (not a shift of KRASG12C to GTP-bound state) drives RAS-MAPK reactivation as a key resistance mechanism to KRASG12C inhibitors; multiple receptor tyrosine kinases can independently drive this feedback, and convergent upstream or downstream signaling blockade enhances KRASG12C inhibitor activity.\",\n      \"method\": \"Cell line signaling assays, RTK inhibitor combinations, genetic RTK knockdown, pharmacological pathway inhibition\",\n      \"journal\": \"Cell reports\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — mechanistic dissection with multiple orthogonal approaches identifying wild-type RAS as the mediator\",\n      \"pmids\": [\"35732135\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"KRAS-dependent gene transcription in KRAS-mutant cancers is driven predominantly through the ERK MAPK cascade; ERK deregulates the anaphase promoting complex/cyclosome (APC/C) and cell cycle machinery as key processes driving PDAC growth. The KRAS-dependent gene signature diverges substantially from the Hallmark KRAS signaling gene set.\",\n      \"method\": \"KRAS suppression combined with ERK inhibition, transcriptomic profiling, phosphoproteomics and total proteomics integration\",\n      \"journal\": \"Science\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — multiple orthogonal omics approaches with pharmacological and genetic loss-of-function\",\n      \"pmids\": [\"38843331\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"Global allosteric mapping of KRAS using >26,000 mutations and double-mutant cycle analysis (>22,000 free energy changes) identifies allosteric communication as particularly effective across the central β-sheet; multiple surface pockets including a distal C-terminal lobe pocket are genetically validated as allosterically active; most allosteric mutations inhibit binding to all tested effectors, but some can alter binding specificity.\",\n      \"method\": \"Deep mutational scanning, double-mutant cycle analysis, biophysical free energy inference\",\n      \"journal\": \"Nature\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 — massively parallel biophysical measurements with rigorous controls across multiple effectors\",\n      \"pmids\": [\"38109937\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2016,\n      \"finding\": \"Kras is specifically required for B cell development through the Raf-1/MEK/ERK pathway: hematopoietic-specific Kras deletion impairs pre-B cell and late B cell maturation, and B cell-specific deletion demonstrates this is cell-intrinsic. Kras deficiency selectively reduces BCR- and pre-BCR-induced Raf-1/MEK/ERK activation without affecting T cell development.\",\n      \"method\": \"Conditional Kras knockout mice, bone marrow chimeras, signaling pathway analysis by Western blot\",\n      \"journal\": \"Journal of Immunology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — clean conditional KO with specific pathway readout and cell-intrinsic validation\",\n      \"pmids\": [\"26773157\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2014,\n      \"finding\": \"Wild-type KRAS acts as a tumor suppressor in the T-cell lineage: loss of the wild-type KRAS allele is a secondary mutation found in all KRAS(G12D)-driven T-ALL cells in mice, and retention of wild-type KRAS inhibits T-ALL development by oncogenic KRAS.\",\n      \"method\": \"Mouse KRAS(G12D) model, LOH analysis, transplantation experiments\",\n      \"journal\": \"Leukemia\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — in vivo genetic model with consistent LOH finding, single lab\",\n      \"pmids\": [\"25371176\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2017,\n      \"finding\": \"USP18, a deubiquitinase, stabilizes KRAS protein and maintains its plasma membrane localization; USP18 knockdown reduces KRAS half-life (cycloheximide chase), mislocalizes KRAS from the plasma membrane, and loss of Usp18 in Kras-driven mouse lung cancer model significantly reduces tumor burden.\",\n      \"method\": \"Cycloheximide chase assay, protein stability measurements, immunofluorescence localization, Kras/Usp18-/- mouse model\",\n      \"journal\": \"Molecular cancer research\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — protein stability and localization assays in cells and in vivo model, single lab\",\n      \"pmids\": [\"28242811\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2012,\n      \"finding\": \"CD44 is required for activation of KRAS-mediated MAPK signaling in lung adenocarcinoma: CD44 deletion in KrasG12D mice attenuates tumor formation and signaling through the MAPK pathway, demonstrating CD44 acts downstream of oncogenic Kras to promote tumor cell proliferation.\",\n      \"method\": \"KrasG12D mouse model with CD44 genetic deletion, tumor burden assessment, MAPK signaling analysis\",\n      \"journal\": \"Oncogene\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — genetic epistasis in vivo with defined pathway readout, single lab\",\n      \"pmids\": [\"23208496\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2016,\n      \"finding\": \"Dual inhibition of PLK1 and ROCK is synthetically lethal in KRAS-mutant cancer cells by increasing transcription and activity of p21(WAF1/CIP1), leading to G2/M blockade specifically in KRAS-mutant cells. p21(WAF1/CIP1) overexpression alone preferentially impairs KRAS-mutant cell growth, identifying a druggable synthetic lethal interaction.\",\n      \"method\": \"Isogenic KRAS-mutant/WT cell screen, microarray, cDNA overexpression, LSL-KRAS(G12D) mouse model, patient tumor explant model\",\n      \"journal\": \"Nature communications\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — multiple validation approaches in vitro and in two in vivo models with mechanistic identification\",\n      \"pmids\": [\"27193833\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"KRAS oncogene drives ERK-mediated deregulation of the anaphase promoting complex/cyclosome (APC/C) and cell cycle machinery; loss of AMPKα in combination with KRAS(G12D) activation increases lung tumor burden in mice, placing AMPKα as a suppressor of KRAS-driven tumorigenesis, connected mechanistically through a KRAS/KIMAT1/LDHB/AMPKα axis.\",\n      \"method\": \"KrasLSLG12D/AMPKαfl/fl mouse model, in vitro growth/migration assays, lncRNA functional studies\",\n      \"journal\": \"Cell death and differentiation\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — in vivo genetic model with mechanistic pathway identification, single lab\",\n      \"pmids\": [\"34040167\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"Cancer-associated fibroblast (CAF)-derived NRG1 activates cancer cell ERBB2/ERBB3 receptor tyrosine kinases as a paracrine mechanism enabling KRAS*-independent PDAC growth upon KRAS inhibition; genetic or pharmacological inhibition of ERBB2/3 or NRG1 abolishes this bypass and synergizes with KRASG12D inhibitors.\",\n      \"method\": \"Genetic KRAS extinction, pharmacological KRAS inhibition, receptor RTK expression analysis, co-culture models, mouse and human PDAC models\",\n      \"journal\": \"Genes & development\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — genetic and pharmacological epistasis in multiple human and mouse models with defined paracrine mechanism\",\n      \"pmids\": [\"37775182\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"K-Ras4B allosterically activates PI3Kα by binding-induced conformational changes: K-Ras4B binding disrupts p110/p85 interface interactions (particularly nSH2 in p85 with C2, helical, and kinase domains of p110), exposes the kinase domain, promotes membrane association and substrate phosphorylation, and rewires allosteric signaling from the helical to the kinase domain.\",\n      \"method\": \"Accelerated molecular dynamics simulations, allosteric pathway analysis\",\n      \"journal\": \"International journal of biological macromolecules\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 4 — computational only, no experimental validation\",\n      \"pmids\": [\"31816384\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2014,\n      \"finding\": \"miR-200c directly targets KRAS mRNA: KRAS was validated as a miR-200c target by Western blot and luciferase reporter assay in breast cancer cell lines; miR-200c-mediated KRAS silencing inhibits proliferation and cell cycle progression.\",\n      \"method\": \"Luciferase reporter assay, Western blot, siRNA knockdown, cell proliferation assays\",\n      \"journal\": \"Oncotarget\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — dual validation with reporter and protein assays with siRNA dissection, single lab\",\n      \"pmids\": [\"24368337\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2018,\n      \"finding\": \"K-Ras4B lacks intrinsic dimerization capability on membranes across a wide range of surface densities and lipid compositions including cholesterol variations, as measured by fluorescence correlation spectroscopy and single-molecule tracking of fully processed native K-Ras4B in supported lipid bilayers.\",\n      \"method\": \"Fluorescence correlation spectroscopy, single-molecule tracking on supported lipid bilayers with native farnesylated/methylated K-Ras4B\",\n      \"journal\": \"Biophysical journal\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 — rigorous single-molecule biophysics with fully processed protein, multiple conditions tested\",\n      \"pmids\": [\"29320680\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"KRAS drives kinome reprogramming in PDAC: upon KRAS knockdown, 15 kinases are upregulated (including SRC, DDR1) and 13 are downregulated (cell cycle kinases including WEE1, Aurora A, PLK1). WEE1 inhibition suppresses PDAC growth; WEE1 inhibition paradoxically activates ERK, and combined WEE1+ERK inhibition achieves synergistic growth suppression and apoptosis.\",\n      \"method\": \"Multiplexed inhibitor bead/MS kinome profiling, KRAS siRNA knockdown across 6 cell lines, pharmacological inhibition\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — systems-level kinome profiling with pharmacological validation across multiple cell lines\",\n      \"pmids\": [\"34688654\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"KRAS functions as a membrane-anchored small GTPase that cycles between GDP-bound inactive and GTP-bound active states; oncogenic mutations (G12, G13, Q61) impair GAP-assisted hydrolysis by disrupting the R789/Q61 catalytic organization, locking KRAS in the active state; membrane localization requires farnesylation of the CAAX motif and the polybasic HVR (K-Ras4B), with the HVR also mediating autoinhibition in the GDP-bound state by occluding the effector-binding site; active KRAS signals predominantly through the RAF-MEK-ERK cascade (driving cell cycle machinery including APC/C deregulation) and PI3Kα (allosterically activated via disruption of the p110/p85 interface), while calmodulin selectively extracts K-Ras4B from the membrane via the farnesylated HVR to support a K-Ras4B–CaM–PI3Kα ternary complex; KRAS dimerization at the α4-α5 interface is mechanistically required for full oncogenic activity and MEK inhibitor resistance; codon usage bias in KRAS limits its protein expression relative to HRAS, modulating tumorigenic potential; and resistance to direct KRAS inhibition is driven by upstream feedback reactivation of wild-type RAS via multiple RTKs and by paracrine CAF-derived NRG1/ERBB2/3 signaling.\"\n}\n```\n\n--- ROUND 2 (R2/R3, augmented corpus) ---\n\n```json\n{\n  \"discoveries\": [\n    {\n      \"year\": 1988,\n      \"finding\": \"Oncogenic mutations at codon 12 of c-K-ras (KRAS) are present in 21 of 22 human pancreatic carcinomas, establishing KRAS mutation as a near-universal and early critical event in exocrine pancreatic oncogenesis.\",\n      \"method\": \"PCR amplification and RNase A mismatch cleavage mutation detection on primary tumor and metastasis specimens\",\n      \"journal\": \"Cell\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 — direct sequencing of primary tumors and matched metastases, replicated across >100 subsequent studies\",\n      \"pmids\": [\"2453289\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 1989,\n      \"finding\": \"KRAS2 codon 12 mutations occur early in human colon carcinoma development, preceding ploidy changes and existing in diploid cells from which aneuploid subpopulations arise; mutations can be present in histologically normal mucosa adjacent to carcinoma.\",\n      \"method\": \"Histological enrichment, cell sorting, PCR amplification, and direct DNA sequencing of colon carcinomas, adenomas, and adjacent normal tissue\",\n      \"journal\": \"Proceedings of the National Academy of Sciences of the United States of America\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — multiple orthogonal methods, staged tissue analysis establishing temporal order of mutation\",\n      \"pmids\": [\"2648401\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 1993,\n      \"finding\": \"Human SOS1 (hSos1) functions as a guanine nucleotide exchange factor (GEF) for RAS, with its CDC25-related domain specifically stimulating guanine nucleotide exchange on mammalian Ras proteins in vitro; hSos1 binds GRB2 via SH3 domain interactions, coupling receptor tyrosine kinases to RAS/KRAS signaling.\",\n      \"method\": \"In vitro GEF assay, yeast complementation, co-immunoprecipitation, overexpression in mammalian cells\",\n      \"journal\": \"Science\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 — in vitro reconstitution of GEF activity plus yeast complementation and co-IP, foundational paper\",\n      \"pmids\": [\"8493579\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 1995,\n      \"finding\": \"K-Ras4B processing is more sensitive to inhibition by geranylgeranyltransferase I (GGTase I) inhibitor GGTI-286 than to farnesyltransferase inhibitor FTI-277, demonstrating that K-Ras4B can be alternatively geranylgeranylated and that its oncogenic signaling can be disrupted by GGTase I inhibition.\",\n      \"method\": \"In vitro prenylation assays, whole-cell processing assays, MAP kinase activity assays in intact cells\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 — in vitro enzyme assays with cell-based validation, first demonstration of K-Ras4B alternative prenylation\",\n      \"pmids\": [\"7592913\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2000,\n      \"finding\": \"Crystal structures of rat farnesyltransferase (FTase) ternary complexes with farnesyl diphosphate analogs and K-Ras4B peptide substrates revealed that the K-Ras4B polybasic region forms a type I beta turn and binds along the rim of the hydrophobic cavity, conferring the highest affinity of any natural FTase substrate; zinc is essential for productive Ca1a2X peptide binding.\",\n      \"method\": \"X-ray crystallography of ternary complexes at 2 Å resolution\",\n      \"journal\": \"Structure\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 — crystal structure at 2 Å with four independent complexes, defines substrate binding mechanism\",\n      \"pmids\": [\"10673434\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2003,\n      \"finding\": \"K-Ras4B resistance to farnesyltransferase inhibitors (FTIs) arises from two independent mechanisms: (1) its polybasic domain increases affinity for FTase, and (2) its CAAX motif can be alternatively geranylgeranylated. Either the polybasic domain alone or an alternatively prenylated CAAX alone renders K-Ras4B FTI-resistant, and K-Ras4B function is independent of the identity of the prenyl group.\",\n      \"method\": \"Chimeric Ras protein constructs, Elk-1 activation assays, anchorage-independent colony formation, microarray analysis\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — multiple orthogonal methods with structure-function chimeras, mechanistically dissects two independent resistance mechanisms\",\n      \"pmids\": [\"12882980\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2009,\n      \"finding\": \"The C-terminal hypervariable region (HVR) of K-Ras4B, specifically its polybasic farnesylated tail, is responsible for isoform-specific interaction with calmodulin (CaM); the HVR binds the C-terminal domain of Ca2+-loaded CaM with micromolar affinity, while the GTP-loaded catalytic domain may additionally interact with the N-terminal CaM domain, linking nucleotide state to CaM binding.\",\n      \"method\": \"NMR spectroscopy, isothermal titration calorimetry (ITC)\",\n      \"journal\": \"Biochemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 — NMR and ITC provide quantitative binding data with domain-level resolution\",\n      \"pmids\": [\"19583261\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2009,\n      \"finding\": \"TBK1 is a synthetic lethal partner of oncogenic KRAS; TBK1 selectively activates NF-κB anti-apoptotic signals (via c-Rel and BCL-XL) in KRAS-mutant cancer cells, and its suppression induces apoptosis specifically in KRAS-dependent cells.\",\n      \"method\": \"Genome-wide RNA interference screen, mechanistic validation by epistasis and apoptosis assays in human cancer cell lines\",\n      \"journal\": \"Nature\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — genome-wide RNAi screen followed by mechanistic pathway epistasis, replicated across multiple KRAS-mutant lines\",\n      \"pmids\": [\"19847166\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2012,\n      \"finding\": \"KRAS is enriched for rare codons relative to HRAS, limiting KRAS protein expression. Converting rare to common codons increases K-Ras expression and tumorigenicity to mirror that of H-Ras, demonstrating that synonymous nucleotide differences affecting codon usage underlie differences in HRas vs KRas expression and oncogenic function.\",\n      \"method\": \"Synonymous codon-optimized transgenes expressed from identical loci, protein expression quantification, transformation assays\",\n      \"journal\": \"Current biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — isogenic codon-substituted alleles with functional readouts, identifies hardwired translational regulatory mechanism\",\n      \"pmids\": [\"23246410\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2013,\n      \"finding\": \"Oncogenic KRAS reprograms glutamine metabolism in pancreatic ductal adenocarcinoma (PDAC) via transcriptional upregulation of RREBP1 and MYC and downregulation of GLUD1: PDAC cells use a non-canonical pathway converting glutamine-derived aspartate to oxaloacetate (via GOT1), then to malate and pyruvate, maintaining NADPH/NADP+ ratio and redox balance. Knockdown of any enzyme in this pathway suppresses PDAC growth in vitro and in vivo.\",\n      \"method\": \"Isotope tracing with 13C-glutamine, siRNA knockdown, xenograft tumor models, gene expression analysis\",\n      \"journal\": \"Nature\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 — metabolic flux tracing plus genetic validation with multiple enzymes, in vivo confirmation\",\n      \"pmids\": [\"23535601\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2014,\n      \"finding\": \"YAP1 can substitute for oncogenic KRAS to rescue cell viability in KRAS-dependent cancer cells; KRAS and YAP1 converge on transcription factor FOS to activate a transcriptional program regulating the epithelial-mesenchymal transition (EMT). YAP1 is required for KRAS-induced cell transformation, and acquired resistance to Kras suppression in a murine lung cancer model involves increased YAP1 signaling.\",\n      \"method\": \"ORFeome rescue screen (15,294 ORFs) in KRAS-dependent cells with inducible KRAS shRNA, epistasis analysis, murine lung cancer model\",\n      \"journal\": \"Cell\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — genome-scale functional screen plus mechanistic in vitro and in vivo validation\",\n      \"pmids\": [\"24954536\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2014,\n      \"finding\": \"In mice, codon-optimized Kras alleles (Kras(ex3op)) producing more K-Ras protein from the endogenous locus lead to fewer carcinogen-induced tumors and induce growth arrest when oncogenically mutated, demonstrating that the rare codon bias of KRAS is a tumor-suppressive mechanism that limits oncogenic K-Ras protein levels in vivo.\",\n      \"method\": \"Knock-in mice with synonymous codon-optimized Kras exon 3, urethane carcinogenesis, tumor burden quantification\",\n      \"journal\": \"The Journal of clinical investigation\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — in vivo genetic model with clean isogenic comparison, validates translational regulation of KRAS oncogenesis\",\n      \"pmids\": [\"25437878\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2015,\n      \"finding\": \"The C-terminal HVR of K-Ras4B directly interacts with the active site/effector-binding region of the catalytic domain with ~100-fold higher affinity in the GDP-bound vs. GTP-bound state; HVR binding interferes with Ras-Raf interaction, modulates phospholipid binding, and slightly slows nucleotide exchange, establishing an autoinhibitory mechanism.\",\n      \"method\": \"NMR spectroscopy, surface plasmon resonance, isothermal titration calorimetry\",\n      \"journal\": \"Biophysical journal\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 — multiple biophysical methods (NMR, SPR, ITC) demonstrating direct intramolecular interaction with quantitative affinities\",\n      \"pmids\": [\"26682817\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2015,\n      \"finding\": \"GTP-bound K-Ras4B forms stable homodimers; two major dimer interfaces were identified: a β-sheet interface overlapping effector binding sites (potentially inhibitory) and a helical interface that may promote Raf dimerization and activation. Ras self-association can regulate effector binding and activity.\",\n      \"method\": \"Analytical ultracentrifugation, molecular dynamics simulations, small-angle X-ray scattering\",\n      \"journal\": \"Structure\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — multiple biophysical methods but single lab, without cell-based validation of the dimer interfaces\",\n      \"pmids\": [\"26051715\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2015,\n      \"finding\": \"K-Ras4B membrane binding is driven by farnesyl group insertion into disordered lipid microdomains; phosphorylation of Ser-181 prohibits spontaneous farnesyl membrane insertion; the polybasic polylysine sequence modulates specific binding to anionic phospholipids and farnesyl membrane orientation.\",\n      \"method\": \"Confocal microscopy, surface plasmon resonance, molecular dynamics simulations\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — orthogonal experimental and computational methods, single lab\",\n      \"pmids\": [\"25713064\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2016,\n      \"finding\": \"Molecular dynamics simulations reveal that oncogenic mutations G12C/G12D/G12V/G13D/Q61H differentially drive inactive-to-active conformational transitions in K-Ras4B-GTP; GAP not only donates its R789 arginine finger but stabilizes the catalytically competent conformation and pre-organizes Q61; oncogenic mutations disrupt R789/Q61 organization, impairing GAP-mediated GTP hydrolysis.\",\n      \"method\": \"6.4 μs cumulative molecular dynamics simulations of WT and mutant K-Ras4B with and without GAP\",\n      \"journal\": \"Scientific reports\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 4 for computational, but extensive simulation coverage; mechanistic model consistent with established biochemistry\",\n      \"pmids\": [\"26902995\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2016,\n      \"finding\": \"Kras is required for B cell lymphopoiesis: hematopoietic-specific deletion of Kras impairs early B cell development at the pre-B cell stage and late B cell maturation. Kras deficiency specifically impairs pre-BCR- and BCR-induced activation of the Raf-1/MEK/ERK pathway, while T cell development is unaffected, demonstrating Kras as the unique Ras family member critical for the Raf-1/MEK/ERK axis in B cells.\",\n      \"method\": \"Conditional knockout mice (hematopoietic-specific and B cell-specific Cre), bone marrow chimeras, flow cytometry, proliferation and signaling assays\",\n      \"journal\": \"Journal of Immunology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — clean conditional KO in two cell-type-specific models with defined signaling pathway readout\",\n      \"pmids\": [\"26773157\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2016,\n      \"finding\": \"Phosphorylation at Ser-181 of K-Ras4B reduces but does not fully abolish membrane binding and clustering; phosphorylated K-Ras4B maintains association with cytosolic shuttle PDEδ; phosphorylation does not alter localization to liquid-disordered lipid subdomains but facilitates dissociation from the plasma membrane.\",\n      \"method\": \"Semisynthesis of triply modified K-Ras4B (phosphate + farnesyl + methyl), supported lipid bilayer studies, fluorescence spectroscopy, cell microinjection\",\n      \"journal\": \"ACS Chemical Biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 — chemically defined post-translationally modified protein studied by multiple spectroscopic methods plus cell experiments\",\n      \"pmids\": [\"28448716\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2016,\n      \"finding\": \"Ca2+/calmodulin (CaM) extracts K-Ras4B from negatively charged membranes in a nucleotide-independent manner; the CaM/K-Ras4B complex is stable in the presence of anionic membranes and shows no membrane binding. PDEδ and CaM affect K-Ras4B membrane interaction through different mechanisms.\",\n      \"method\": \"Surface plasmon resonance, fluorescence spectroscopy (FCS, FRET), model membrane studies, FRAP\",\n      \"journal\": \"Biophysical journal\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 — multiple spectroscopic and biophysical techniques with chemically defined proteins and membranes\",\n      \"pmids\": [\"27410739\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2017,\n      \"finding\": \"Oncogenic KRAS drives a non-canonical glutamine-to-aspartate metabolic pathway through transcriptional regulation of key enzymes; the KRAS-regulated kinome in PDAC includes WEE1 among kinases downregulated upon KRAS loss, and combined WEE1 + ERK inhibition causes enhanced PDAC growth suppression and apoptosis.\",\n      \"method\": \"Multiplexed inhibitor bead/MS kinomics, siRNA knockdown, pharmacological inhibition, synergy studies in PDAC cell lines\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — system-wide kinome profiling across six cell lines with pharmacological validation\",\n      \"pmids\": [\"34688654\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2017,\n      \"finding\": \"CaM preferentially binds unfolded K-Ras4B HVR (not α-helical HVR) using all three CaM domains; interaction is stabilized by docking of farnesyl to hydrophobic pockets in both CaM lobes; CaM wraps around the polybasic anchor region of HVR, enabling membrane extraction of K-Ras4B to form a K-Ras4B–CaM–PI3Kα ternary complex that activates PI3Kα.\",\n      \"method\": \"Molecular dynamics simulations, fluorescence binding experiments\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 3 — computational modeling supported by fluorescence experiments; ternary complex model not directly confirmed by structure\",\n      \"pmids\": [\"28623230\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2017,\n      \"finding\": \"USP18 deubiquitinase stabilizes the KRAS oncoprotein; USP18 loss reduces KRAS protein half-life and mislocalizes KRAS from the plasma membrane; USP18 gain increases KRAS stability; Usp18 loss in Kras-mutant mice significantly reduces lung tumor burden.\",\n      \"method\": \"Cycloheximide chase, subcellular fractionation, conditional KO mice, immunohistochemistry\",\n      \"journal\": \"Molecular Cancer Research\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — protein stability and localization experiments with in vivo genetic confirmation\",\n      \"pmids\": [\"28242811\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2018,\n      \"finding\": \"KRAS dimerization at the α4-α5 interface mediates wild-type KRAS-dependent fitness of KRAS-mutant lung adenocarcinoma cells and underlies resistance to MEK inhibition; KRASD154Q (dimerization-deficient mutant) abrogates these effects both in vitro and in vivo; dimerization also has a critical role in the oncogenic activity of mutant KRAS.\",\n      \"method\": \"Genetically inducible KRAS LOH model, dimerization-disrupting mutant KRASD154Q, MEK inhibitor response assays, in vitro and in vivo tumor models\",\n      \"journal\": \"Cell\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — clean genetic models with structure-guided mutant, multiple in vitro and in vivo readouts, replicated across human and murine systems\",\n      \"pmids\": [\"29336889\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2018,\n      \"finding\": \"Full-length, fully processed (farnesylated + methylated) K-Ras4B lacks intrinsic dimerization capability on supported lipid bilayers across a wide range of surface densities and lipid compositions including cholesterol-containing membranes.\",\n      \"method\": \"Fluorescence correlation spectroscopy (FCS), single-molecule tracking on supported lipid bilayers with natively processed K-Ras4B\",\n      \"journal\": \"Biophysical journal\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 — single-molecule imaging with natively processed protein under multiple lipid conditions, directly contradicts other reports\",\n      \"pmids\": [\"29320680\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2018,\n      \"finding\": \"K-Ras4B mutant (G12C/G12D) HVR shows preferential interaction with phosphatidic acid (PA) over other phospholipids; in the GDP-bound state the HVR shields the effector-binding site (autoinhibition); GTP binding and oncogenic mutations release HVR, enabling calmodulin interaction.\",\n      \"method\": \"Molecular dynamics simulations, NMR, phospholipid binding assays\",\n      \"journal\": \"Current opinion in structural biology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — combines computational and NMR data; mutation-specific phospholipid binding specificity established\",\n      \"pmids\": [\"26709496\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2018,\n      \"finding\": \"A compound (Cmpd2) inhibits K-RAS4B by stabilizing membrane-dependent occlusion of the effector-binding site: it simultaneously engages a shallow pocket on KRAS and the lipid bilayer, orienting membrane-associated prenylated KRAS so the membrane sterically occludes the effector-binding site, reducing RAF binding and impairing RAF activation.\",\n      \"method\": \"NMR, lipid bilayer binding assays, cell-based RAF activation assays, structure-based mechanism elucidation\",\n      \"journal\": \"Cell Chemical Biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 — NMR structural characterization of mechanism with cell-based functional validation\",\n      \"pmids\": [\"30122370\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"BI-2852 binds with nanomolar affinity to the switch I/II pocket on KRAS (present in both active and inactive forms), blocking all GEF, GAP, and effector interactions simultaneously, demonstrating that this pocket is druggable; binding inhibits downstream signaling and has antiproliferative effects in KRAS-mutant cells.\",\n      \"method\": \"Structure-based drug design, X-ray crystallography, biochemical GEF/GAP/effector competition assays, cell-based signaling assays\",\n      \"journal\": \"Proceedings of the National Academy of Sciences of the United States of America\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 — crystal structure plus biochemical assays showing blockade of multiple interaction partners\",\n      \"pmids\": [\"31332011\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"K-Ras4B allosterically activates PI3Kα by binding-induced conformational changes that disrupt p110/p85 (nSH2) interactions, exposing the kinase domain for membrane association and substrate phosphorylation; allosteric signaling is rewired from helical to kinase domain in the K-Ras4B/PI3Kα complex.\",\n      \"method\": \"Accelerated molecular dynamics simulations, allosteric pathway analysis, community network analysis\",\n      \"journal\": \"International journal of biological macromolecules\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 4 — computational only, no in vitro or cell-based validation\",\n      \"pmids\": [\"31816384\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"A small molecule KRAS agonist (KRA-533) binds the GTP/GDP-binding pocket of KRAS and prevents GTP cleavage, accumulating active GTP-KRAS; K117A mutation in KRAS abolishes KRA-533 binding and blocks its activity; KRA-533-mediated KRAS hyperactivation promotes apoptosis and autophagic cell death preferentially in KRAS-mutant cancer cells.\",\n      \"method\": \"GDP/GTP exchange assay, site-directed mutagenesis, cell viability assays, xenograft and GEMM models\",\n      \"journal\": \"Molecular cancer\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — biochemical assay plus mutagenesis plus in vivo models from single lab\",\n      \"pmids\": [\"30971271\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"Urethane carcinogenesis specificity is determined by the sequence specificity of urethane mutagenesis coupled with transcription bias and isoform locus: the initiating Kras Q61L/R mutation was captured days after urethane exposure using error-corrected high-throughput sequencing, demonstrating that transcription rate and isoform-specific context drive RAS mutation tropism.\",\n      \"method\": \"Error-corrected high-throughput sequencing of mouse Ras genes at multiple time points post-carcinogen exposure\",\n      \"journal\": \"Nature communications\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — direct sequencing capturing the initiating mutation in vivo with mechanistic dissection of specificity determinants\",\n      \"pmids\": [\"32286309\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"KRAS-dependent transcription is driven predominantly through the ERK/MAPK cascade; KRAS-regulated ERK signaling deregulates the anaphase promoting complex/cyclosome (APC/C) and cell cycle machinery as key drivers of PDAC growth; the KRAS-dependent gene signature diverges substantially from the Hallmark KRAS signaling gene signature.\",\n      \"method\": \"Inducible KRAS knockdown, transcriptomics, phosphoproteomics, total proteomics, patient tumor data integration\",\n      \"journal\": \"Science\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — multiple omics layers integrated with genetic perturbation and patient data validation\",\n      \"pmids\": [\"38843331\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"Resistance to KRASG12C inhibitors is driven primarily by upstream feedback activation of wild-type RAS (rather than reactivation of KRASG12C to its GTP-bound state); multiple RTKs can independently drive this KRASG12C-independent RAS-MAPK reactivation; convergent upstream or downstream blockade can overcome resistance.\",\n      \"method\": \"RAS activity assays, RTK inhibitor combinations, cell signaling studies in KRASG12C-mutant cancer cells treated with G12C inhibitors\",\n      \"journal\": \"Cell reports\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — mechanistic dissection with multiple inhibitors and RAS activity measurements identifying WT RAS as the resistance driver\",\n      \"pmids\": [\"35732135\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"Cancer-associated fibroblast (CAF)-derived NRG1 activates cancer cell ERBB2/ERBB3 receptor tyrosine kinases to support KRAS*-independent pancreatic cancer growth; genetic extinction of KRAS* upregulates ERBB2/ERBB3 in cancer cells, which then utilize paracrine CAF-NRG1 as a survival factor; ERBB2/3 or NRG1 depletion abolishes KRAS* bypass.\",\n      \"method\": \"Genetic KRAS extinction, pharmacological inhibition, paracrine co-culture models, mouse and human PDAC models\",\n      \"journal\": \"Genes & development\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — genetic and pharmacological evidence in multiple model systems with defined paracrine mechanism\",\n      \"pmids\": [\"37775182\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"A comprehensive deep mutational scan quantified >26,000 mutations' effects on KRAS folding and binding to six interaction partners, mapping >22,000 causal free energy changes; allosteric propagation is particularly effective across the central β-sheet; multiple surface pockets are validated as allosterically active including a distal C-terminal lobe pocket; most allosteric mutations inhibit all effectors but some can alter binding specificity.\",\n      \"method\": \"Deep mutational scanning, double-mutant genetic interaction analysis, free energy change inference\",\n      \"journal\": \"Nature\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 — genome-scale biophysical measurements with double-mutant epistasis providing causal energy landscapes\",\n      \"pmids\": [\"38109937\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"Acquired resistance to adagrasib (KRASG12C inhibitor) involves diverse mechanisms including secondary KRAS mutations (G12D/R/V/W, G13D, Q61H, R68S, H95D/Q/R, Y96C), KRAS amplification, and bypass alterations (MET amplification, NRAS/BRAF/MAP2K1/RET mutations, oncogenic fusions involving ALK/RET/BRAF/RAF1/FGFR3, NF1/PTEN loss); deep mutational scanning systematically defined the landscape of KRAS mutations conferring inhibitor resistance.\",\n      \"method\": \"Genomic sequencing of paired pre-/post-treatment biopsies, in vitro deep mutational scanning screen\",\n      \"journal\": \"The New England journal of medicine\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — clinical genomics plus systematic in vitro resistance landscape, multiple orthogonal evidence types\",\n      \"pmids\": [\"34161704\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2017,\n      \"finding\": \"KRAS can bind numerous effector proteins (RAF, PI3K, RalGDS families, and others); combinatorial siRNA knockdown of 41 KRAS effector nodes in 92 cell lines identified two major subtypes of KRAS-mutant cancers with distinct effector dependencies, demonstrating that each cell line has a unique effector engagement pattern.\",\n      \"method\": \"Arrayed combinatorial siRNA screen of 41 effector nodes across 92 cancer cell lines, quantitative phenotype assessment\",\n      \"journal\": \"Cell reports\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — large-scale systematic knockdown across diverse cell lines with quantitative phenotypic readouts\",\n      \"pmids\": [\"29444439\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"KRAS encodes a small GTPase that cycles between GDP-bound (inactive) and GTP-bound (active) states regulated by GEFs (e.g., SOS1) and GAPs; oncogenic mutations at codons 12, 13, or 61 impair GAP-mediated GTP hydrolysis, locking KRAS in its active state to constitutively signal through multiple effector pathways (RAF-MEK-ERK, PI3K-AKT, RalGDS); membrane association via farnesylation of the C-terminal CAAX motif and the polybasic HVR is essential for oncogenic signaling; the HVR can autoinhibit the effector-binding site in the GDP-bound state; KRAS dimerization at the α4-α5 interface modulates MEK inhibitor sensitivity and oncogenic activity; calmodulin specifically extracts K-Ras4B from the membrane via HVR interaction to promote PI3Kα activation; rare codon bias in KRAS limits protein expression as a built-in tumor-suppressive mechanism; oncogenic KRAS reprograms glutamine metabolism through a non-canonical GOT1-dependent pathway to maintain redox balance; KRAS signals predominantly through ERK/MAPK to regulate APC/C and cell cycle machinery driving tumor growth; and resistance to covalent KRASG12C inhibitors occurs via secondary KRAS mutations, KRAS amplification, and bypass RTK/RAS pathway reactivation.\"\n}\n```","stage2_raw":"```json\n{\n  \"mechanistic_narrative\": \"KRAS is a membrane-anchored small GTPase that cycles between GDP-bound inactive and GTP-bound active states to relay mitogenic signals predominantly through the RAF-MEK-ERK cascade and PI3K pathway, with essential roles in processes including B cell development and cell cycle control [PMID:26773157, PMID:38843331]. Membrane localization requires farnesylation of the C-terminal CAAX motif and electrostatic interactions of the polybasic hypervariable region (HVR) with anionic phospholipids; the HVR also mediates autoinhibition in the GDP-bound state by occluding the effector-binding site, and calmodulin extracts K-Ras4B from membranes by engaging the farnesylated HVR [PMID:25713064, PMID:26682817, PMID:27410739]. Oncogenic mutations at G12, G13, and Q61 impair GAP-assisted GTP hydrolysis by disrupting the catalytic organization of R789 and Q61, locking KRAS in the active state; KRAS dimerization at the β4-β5 interface is required for full oncogenic activity, while resistance to direct KRAS inhibitors is driven by feedback reactivation of wild-type RAS through multiple receptor tyrosine kinases and by paracrine NRG1-ERBB2/3 signaling from cancer-associated fibroblasts [PMID:26902995, PMID:29336889, PMID:35732135, PMID:37775182]. Rare codon bias in KRAS limits its protein expression relative to HRAS, modulating tumorigenic potential, and wild-type KRAS can function as a tumor suppressor against oncogenic KRAS alleles [PMID:23246410, PMID:25437878, PMID:25371176].\",\n  \"teleology\": [\n    {\n      \"year\": 1995,\n      \"claim\": \"Establishing that K-Ras4B can escape farnesyltransferase inhibition through alternative geranylgeranylation explained the clinical failure of FTIs and revealed an unexpected flexibility in KRAS post-translational processing.\",\n      \"evidence\": \"In vitro prenylation assays and MAP kinase signaling readouts comparing FTI and GGTase I inhibitor sensitivity\",\n      \"pmids\": [\"7592913\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"The structural basis for GGTase I recognition of K-Ras4B CAAX was not determined\", \"In vivo relevance of alternative prenylation in tumors was not tested\"]\n    },\n    {\n      \"year\": 2000,\n      \"claim\": \"Crystal structures of FTase–K-Ras4B peptide complexes revealed how the polybasic HVR enhances farnesyltransferase affinity and how zinc coordinates productive CAAX binding, providing an atomic framework for understanding K-Ras4B prenylation.\",\n      \"evidence\": \"X-ray crystallography at 2 Å resolution of rat FTase ternary complexes with K-Ras4B peptides\",\n      \"pmids\": [\"10673434\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Full-length K-Ras4B–FTase complex was not resolved\", \"Structural basis for GGTase I cross-reactivity remained unknown\"]\n    },\n    {\n      \"year\": 2003,\n      \"claim\": \"Dissection of K-Ras4B FTI resistance into two independent elements — the polybasic domain increasing FTase affinity and the CAAX motif permitting alternative geranylgeranylation — resolved how each feature independently confers drug resistance.\",\n      \"evidence\": \"Chimeric Ras protein constructs tested by Elk-1 activation, anchorage-independent growth, and microarray analysis\",\n      \"pmids\": [\"12882980\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Therapeutic strategies to overcome both mechanisms simultaneously were not identified\"]\n    },\n    {\n      \"year\": 2009,\n      \"claim\": \"Identification of the HVR as the primary calmodulin-binding determinant on K-Ras4B, with nucleotide-dependent modulation via the catalytic domain, established the molecular basis for CaM-mediated KRAS regulation.\",\n      \"evidence\": \"NMR spectroscopy and isothermal titration calorimetry with domain-dissected K-Ras4B and Ca²⁺-loaded calmodulin\",\n      \"pmids\": [\"19583261\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Functional consequence of CaM binding for downstream KRAS signaling was not measured\", \"In vivo significance of CaM–KRAS interaction was not tested\"]\n    },\n    {\n      \"year\": 2012,\n      \"claim\": \"Discovery that rare codon bias in KRAS limits its protein expression relative to HRAS, and that equalizing codon usage equalizes tumorigenic potential, revealed a translational regulatory layer governing RAS isoform biology.\",\n      \"evidence\": \"Synonymous codon-altered KRAS constructs with expression quantification and transformation assays; validated in vivo with knock-in mouse model\",\n      \"pmids\": [\"23246410\", \"25437878\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"The ribosome-level mechanism (e.g., tRNA availability, ribosome stalling) was not directly measured\", \"Whether codon bias affects KRAS mRNA stability was not distinguished from translational effects\"]\n    },\n    {\n      \"year\": 2014,\n      \"claim\": \"Demonstration that wild-type KRAS suppresses oncogenic KRAS-driven T-ALL — with universal loss of the wild-type allele in tumors — and that YAP1 converges with KRAS on FOS to drive EMT programs and resistance, broadened the understanding of KRAS from a simple oncogene to a context-dependent tumor suppressor and revealed an alternative survival pathway.\",\n      \"evidence\": \"Mouse KRAS(G12D) T-ALL model with LOH analysis; genome-wide ORF rescue screen (15,294 ORFs) with inducible KRAS shRNA and in vivo validation\",\n      \"pmids\": [\"25371176\", \"24954536\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Mechanism by which wild-type KRAS suppresses oncogenic KRAS signaling was not elucidated\", \"Whether YAP1-FOS convergence operates in all KRAS-driven cancers was not tested\"]\n    },\n    {\n      \"year\": 2015,\n      \"claim\": \"Biophysical characterization of K-Ras4B membrane binding and intramolecular autoinhibition showed that the farnesylated HVR inserts into disordered lipid domains, Ser181 phosphorylation modulates membrane insertion, and the HVR directly occludes the effector site ~100-fold more tightly in the GDP state, establishing HVR-mediated autoinhibition as a regulatory mechanism.\",\n      \"evidence\": \"NMR, SPR, confocal microscopy, and MD simulations with prenylated K-Ras4B and model membranes\",\n      \"pmids\": [\"25713064\", \"26682817\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Autoinhibition was not demonstrated in live cells\", \"Quantitative contribution of autoinhibition to KRAS signaling dynamics in vivo was not measured\"]\n    },\n    {\n      \"year\": 2015,\n      \"claim\": \"Solution biophysics established that GTP-bound K-Ras4B forms homodimers with two distinct interfaces — a β-sheet interface overlapping the effector site and a helical interface potentially promoting RAF activation — opening the question of whether dimerization is functionally required.\",\n      \"evidence\": \"Analytical ultracentrifugation and molecular dynamics simulations of the catalytic domain\",\n      \"pmids\": [\"26051715\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Dimer interfaces were modeled but not confirmed by high-resolution structure\", \"Functional consequences of each interface were not experimentally tested\"]\n    },\n    {\n      \"year\": 2016,\n      \"claim\": \"Multiple advances clarified KRAS regulation and signaling: CaM was shown to extract K-Ras4B from membranes via the farnesyl group; oncogenic mutations were modeled to disrupt GAP-mediated R789/Q61 catalytic organization; Ser181 phosphorylation was shown to reduce but not abolish membrane binding; and KRAS was found to be specifically required for B cell development through RAF-1/MEK/ERK.\",\n      \"evidence\": \"Fluorescence/FRET membrane extraction assays; MD simulations of mutant KRAS-GAP complexes; reconstituted triply-modified K-Ras4B on model membranes; conditional Kras knockout mice with bone marrow chimeras\",\n      \"pmids\": [\"27410739\", \"26902995\", \"28448716\", \"26773157\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"GAP-hydrolysis disruption was computationally modeled but not experimentally validated for all hotspot mutations\", \"Whether CaM extraction is relevant in non-cancer physiological contexts was not tested\", \"Mechanism by which KRAS is selectively required for B cells but not T cells was not fully resolved\"]\n    },\n    {\n      \"year\": 2018,\n      \"claim\": \"Functional genetic studies established that KRAS dimerization at the β4-β5 interface is mechanistically required for oncogenic KRAS fitness and MEK inhibitor resistance, while single-molecule biophysics showed that fully processed K-Ras4B lacks intrinsic dimerization capability on membranes, suggesting scaffolding or effector-mediated dimerization in cells.\",\n      \"evidence\": \"Genetically inducible KRAS LOH model with KRASD154Q mutant in vitro and in vivo; FCS and single-molecule tracking of native farnesylated/methylated K-Ras4B on supported lipid bilayers\",\n      \"pmids\": [\"29336889\", \"29320680\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"The factor(s) that promote KRAS dimerization on native cell membranes remain unidentified\", \"Crystal structure of the β4-β5 dimer interface in the membrane context is lacking\"]\n    },\n    {\n      \"year\": 2019,\n      \"claim\": \"Structure-based inhibitor BI-2852 validated the switch I/II pocket as a druggable site on both active and inactive KRAS, blocking all GEF, GAP, and effector interactions, while a membrane-dependent compound (Cmpd2) demonstrated an orthogonal inhibition mechanism by reorienting KRAS on membranes to occlude the effector site.\",\n      \"evidence\": \"X-ray crystallography and biochemical/cell-based assays for BI-2852; NMR of membrane-associated prenylated K-RAS4B with RAF-binding assays for Cmpd2\",\n      \"pmids\": [\"31332011\", \"30122370\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"In vivo efficacy and selectivity of these compounds were not established\", \"Whether membrane-dependent inhibition generalizes across lipid environments is untested\"]\n    },\n    {\n      \"year\": 2021,\n      \"claim\": \"Systems-level kinome profiling revealed that KRAS drives coordinated upregulation of cell cycle kinases (WEE1, Aurora A, PLK1) in PDAC, and that WEE1 inhibition paradoxically reactivates ERK, necessitating combined WEE1+ERK inhibition for synergistic growth suppression.\",\n      \"evidence\": \"Multiplexed inhibitor bead/mass spectrometry kinome profiling across 6 PDAC cell lines with KRAS siRNA and pharmacological validation\",\n      \"pmids\": [\"34688654\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Whether kinome reprogramming is specific to PDAC or generalizes to other KRAS-driven cancers\", \"Direct mechanism linking KRAS to WEE1 transcriptional regulation was not identified\"]\n    },\n    {\n      \"year\": 2022,\n      \"claim\": \"The discovery that resistance to KRASG12C inhibitors is driven by feedback reactivation of wild-type RAS (not reactivation of the mutant allele) through multiple RTKs resolved a central clinical question and indicated that combination strategies must target the wild-type RAS pool.\",\n      \"evidence\": \"Cell line signaling assays, RTK inhibitor combinations, genetic RTK knockdown, pharmacological pathway inhibition\",\n      \"pmids\": [\"35732135\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Whether this resistance mechanism applies equally to non-G12C KRAS inhibitors\", \"Patient-level validation in clinical cohorts was not provided\"]\n    },\n    {\n      \"year\": 2023,\n      \"claim\": \"Two key advances: global allosteric mapping of KRAS via deep mutational scanning revealed that allosteric communication propagates across the central β-sheet and identified distal druggable surface pockets; and CAF-derived NRG1–ERBB2/3 paracrine signaling was identified as an extrinsic resistance mechanism to KRAS inhibition in PDAC.\",\n      \"evidence\": \"Deep mutational scanning with >26,000 mutations and double-mutant cycle analysis across multiple effectors; genetic KRAS extinction and pharmacological inhibition in co-culture and mouse/human PDAC models\",\n      \"pmids\": [\"38109937\", \"37775182\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Allosteric pockets have not been targeted by small molecules\", \"Whether NRG1 blockade is sufficient to prevent clinical resistance in patients is unknown\"]\n    },\n    {\n      \"year\": 2024,\n      \"claim\": \"Integrated multi-omics demonstrated that KRAS-dependent transcription in PDAC is driven predominantly through ERK-mediated deregulation of APC/C and cell cycle machinery, substantially revising the established KRAS transcriptional signature.\",\n      \"evidence\": \"KRAS suppression combined with ERK inhibition, transcriptomic profiling, phosphoproteomics and total proteomics in PDAC models\",\n      \"pmids\": [\"38843331\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Whether APC/C deregulation is directly phosphorylation-dependent or transcriptionally mediated is not fully resolved\", \"Generalizability beyond PDAC to other KRAS-driven cancers was not tested\"]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"Key unresolved questions include: what scaffolds or cofactors mediate KRAS dimerization on native cell membranes; whether allosteric surface pockets can be pharmacologically exploited; and whether combined targeting of wild-type RAS feedback and paracrine NRG1-ERBB2/3 signaling can durably overcome clinical resistance to direct KRAS inhibitors.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"Low\",\n      \"gaps\": [\"No structural model of KRAS dimer on membranes with scaffolding partners\", \"No clinical validation of allosteric pocket targeting\", \"No clinical data on triple-combination strategies addressing both intrinsic and paracrine resistance\"]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0003924\", \"supporting_discovery_ids\": [13, 2]},\n      {\"term_id\": \"GO:0098772\", \"supporting_discovery_ids\": [28, 19]},\n      {\"term_id\": \"GO:0008289\", \"supporting_discovery_ids\": [8, 9]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005886\", \"supporting_discovery_ids\": [8, 9, 10, 23, 30]},\n      {\"term_id\": \"GO:0005829\", \"supporting_discovery_ids\": [10, 9]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-162582\", \"supporting_discovery_ids\": [0, 19, 21, 28, 16, 18]},\n      {\"term_id\": \"R-HSA-1640170\", \"supporting_discovery_ids\": [19, 25, 31, 26]},\n      {\"term_id\": \"R-HSA-1643685\", \"supporting_discovery_ids\": [14, 15, 27, 18]},\n      {\"term_id\": \"R-HSA-392499\", \"supporting_discovery_ids\": [3, 4, 5]},\n      {\"term_id\": \"R-HSA-168256\", \"supporting_discovery_ids\": [21]}\n    ],\n    \"complexes\": [\n      \"KRAS-RAF complex\",\n      \"CaM-K-Ras4B complex\",\n      \"KRAS-PI3Kα complex\"\n    ],\n    \"partners\": [\n      \"RAF1\",\n      \"CALM1\",\n      \"PIK3CA\",\n      \"PIK3R1\",\n      \"PDE6D\",\n      \"YAP1\",\n      \"USP18\",\n      \"CD44\"\n    ],\n    \"other_free_text\": []\n  }\n}\n```\n\n--- ROUND 2 (R2/R3, augmented corpus) ---\n\n```json\n{\n  \"mechanistic_narrative\": \"KRAS encodes a small GTPase that cycles between GDP-bound (inactive) and GTP-bound (active) states, transducing signals from receptor tyrosine kinases through effector pathways including RAF-MEK-ERK, PI3K-AKT, and RalGDS to control cell proliferation, survival, and metabolism [PMID:8493579, PMID:29444439]. Oncogenic mutations at codons 12, 13, and 61 impair GAP-mediated GTP hydrolysis, locking KRAS in the active state and constitutively driving downstream signaling; these mutations are near-universal early events in pancreatic and colorectal carcinogenesis [PMID:2453289, PMID:2648401, PMID:26902995]. Membrane association via C-terminal farnesylation and the polybasic hypervariable region is essential for signaling, and this region also mediates nucleotide-state-dependent autoinhibition of the effector-binding site, calmodulin-dependent membrane extraction feeding PI3Kα activation, and regulation by phosphorylation at Ser-181 [PMID:26682817, PMID:19583261, PMID:27410739, PMID:28448716]. Oncogenic KRAS reprograms glutamine metabolism through a non-canonical GOT1-dependent pathway to maintain redox balance, signals predominantly through ERK to deregulate APC/C and cell-cycle machinery, and its protein output is constrained by rare codon usage that acts as a built-in tumor-suppressive mechanism [PMID:23535601, PMID:38843331, PMID:23246410, PMID:25437878].\",\n  \"teleology\": [\n    {\n      \"year\": 1988,\n      \"claim\": \"Establishing that KRAS mutations are a near-universal initiating event in pancreatic cancer answered whether a single oncogene could account for virtually all cases of this malignancy and defined KRAS as the central driver.\",\n      \"evidence\": \"PCR-based mutation detection in 22 primary pancreatic carcinomas and colon tumors with staged tissue analysis\",\n      \"pmids\": [\"2453289\", \"2648401\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Mechanism by which mutant KRAS drives tumorigenesis was undefined\", \"No effector pathway specificity was determined\", \"Functional contribution versus passenger role not formally tested\"]\n    },\n    {\n      \"year\": 1993,\n      \"claim\": \"Identification of SOS1 as the GEF coupling RTKs to RAS via GRB2 resolved how extracellular signals activate KRAS, placing it downstream of growth factor receptors.\",\n      \"evidence\": \"In vitro GEF reconstitution, yeast complementation, co-immunoprecipitation\",\n      \"pmids\": [\"8493579\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"How GAP-mediated inactivation was specifically impaired by oncogenic mutations remained structural speculation\", \"Isoform-specific GEF regulation was not addressed\"]\n    },\n    {\n      \"year\": 1995,\n      \"claim\": \"Discovery that K-Ras4B can be alternatively geranylgeranylated explained the clinical failure of farnesyltransferase inhibitors against KRAS-driven cancers, revealing redundancy in prenylation.\",\n      \"evidence\": \"In vitro prenylation assays and cell-based processing/MAPK assays with FTI and GGTase I inhibitors\",\n      \"pmids\": [\"7592913\", \"12882980\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Structural basis for K-Ras4B's dual prenylation susceptibility was not resolved\", \"In vivo pharmacological confirmation was lacking\"]\n    },\n    {\n      \"year\": 2000,\n      \"claim\": \"Crystal structures of K-Ras4B peptide–FTase complexes revealed how the polybasic HVR confers the highest FTase affinity among Ras isoforms, providing structural understanding of C-terminal processing.\",\n      \"evidence\": \"X-ray crystallography at 2 Å resolution of ternary FTase complexes\",\n      \"pmids\": [\"10673434\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Full-length K-Ras4B structure with lipid modifications was unavailable\", \"Mechanism of alternative geranylgeranylation was not structurally addressed\"]\n    },\n    {\n      \"year\": 2009,\n      \"claim\": \"NMR and calorimetry defined the calmodulin–K-Ras4B HVR interaction and a genome-wide RNAi screen identified TBK1 as a synthetic lethal partner, jointly revealing non-canonical signaling outputs and vulnerabilities of mutant KRAS.\",\n      \"evidence\": \"NMR/ITC for CaM binding; genome-wide RNAi screen with mechanistic epistasis in KRAS-mutant cell lines\",\n      \"pmids\": [\"19583261\", \"19847166\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Whether CaM-dependent extraction operates in vivo was not shown\", \"TBK1 synthetic lethality was not validated in clinical settings\"]\n    },\n    {\n      \"year\": 2012,\n      \"claim\": \"Demonstrating that KRAS rare codon bias limits protein expression and that codon optimization increases tumorigenicity uncovered a hard-wired translational tumor suppressor mechanism embedded in the coding sequence.\",\n      \"evidence\": \"Isogenic codon-optimized transgenes with transformation assays in vitro; knock-in mice with codon-optimized Kras exon 3 and carcinogenesis assays\",\n      \"pmids\": [\"23246410\", \"25437878\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"The ribosomal or tRNA mechanisms mediating rare-codon suppression were not identified\", \"Whether codon bias affects mutant vs. wild-type KRAS differentially was not resolved\"]\n    },\n    {\n      \"year\": 2013,\n      \"claim\": \"Isotope tracing revealed that oncogenic KRAS reprograms glutamine metabolism through a non-canonical GOT1-dependent pathway to maintain NADPH and redox balance, establishing metabolic rewiring as a core function of mutant KRAS in PDAC.\",\n      \"evidence\": \"13C-glutamine flux analysis, siRNA knockdown of pathway enzymes, xenograft validation\",\n      \"pmids\": [\"23535601\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Whether this pathway operates in KRAS-mutant cancers beyond PDAC was untested\", \"Direct transcriptional mechanism linking KRAS to enzyme expression was incompletely defined\"]\n    },\n    {\n      \"year\": 2015,\n      \"claim\": \"Biophysical studies established that the K-Ras4B HVR autoinhibits the effector-binding site in the GDP state with ~100-fold selectivity, and that GTP-bound K-Ras4B forms homodimers at two distinct interfaces, resolving how intramolecular and intermolecular interactions regulate effector access.\",\n      \"evidence\": \"NMR/SPR/ITC for autoinhibition; analytical ultracentrifugation, SAXS, and MD for dimerization\",\n      \"pmids\": [\"26682817\", \"26051715\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Dimerization interfaces lacked cell-based validation\", \"Whether autoinhibition is modulated by specific lipid environments was unknown\"]\n    },\n    {\n      \"year\": 2016,\n      \"claim\": \"Conditional knockout of Kras in hematopoietic cells demonstrated a non-redundant requirement for KRAS in B cell lymphopoiesis through the Raf-1/MEK/ERK axis, establishing a physiological developmental role distinct from oncogenesis.\",\n      \"evidence\": \"Hematopoietic- and B cell-specific conditional KO mice, bone marrow chimeras, signaling assays\",\n      \"pmids\": [\"26773157\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Whether KRAS has analogous non-redundant roles in other non-hematopoietic lineages was not tested\", \"Mechanism for KRAS isoform specificity in B cells was unclear\"]\n    },\n    {\n      \"year\": 2016,\n      \"claim\": \"Studies of Ser-181 phosphorylation, PDE\\u03b4 shuttling, and calmodulin-mediated membrane extraction defined three orthogonal mechanisms controlling K-Ras4B plasma membrane residence, clarifying how localization is dynamically regulated.\",\n      \"evidence\": \"Semisynthetic phosphorylated/farnesylated K-Ras4B on supported bilayers; SPR/FCS/FRET with CaM and PDE\\u03b4\",\n      \"pmids\": [\"28448716\", \"27410739\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Relative contributions of CaM, PDE\\u03b4, and phosphorylation in living cells were not quantified\", \"Whether these mechanisms are differentially engaged in mutant vs. WT KRAS was not resolved\"]\n    },\n    {\n      \"year\": 2018,\n      \"claim\": \"Genetic evidence that KRAS dimerization at the α4-α5 interface mediates fitness of KRAS-mutant cells and MEK inhibitor resistance provided a structural rationale for wild-type KRAS dependency in mutant KRAS tumors, though single-molecule studies challenged whether full-length processed K-Ras4B dimerizes on membranes.\",\n      \"evidence\": \"KRASD154Q dimerization-deficient mutant in LOH model and tumor assays; single-molecule FCS on supported lipid bilayers\",\n      \"pmids\": [\"29336889\", \"29320680\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Discrepancy between cell-based genetic dimerization data and biophysical monomeric behavior on bilayers is unresolved\", \"Whether scaffold proteins mediate dimerization in cells was not tested\"]\n    },\n    {\n      \"year\": 2019,\n      \"claim\": \"Structure-based drug design yielded compounds targeting two distinct KRAS pockets—BI-2852 at the switch I/II interface blocking all GEF/GAP/effector interactions, and Cmpd2 stabilizing membrane-occluded KRAS—proving KRAS is druggable at multiple sites.\",\n      \"evidence\": \"X-ray crystallography and NMR with biochemical competition and cell-based signaling assays\",\n      \"pmids\": [\"31332011\", \"30122370\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"In vivo efficacy and selectivity of these compounds were not demonstrated\", \"Whether these sites are accessible in all KRAS mutant contexts was untested\"]\n    },\n    {\n      \"year\": 2021,\n      \"claim\": \"Clinical genomics and deep mutational scanning systematically mapped the resistance landscape to KRASG12C inhibitors, revealing that secondary KRAS mutations, gene amplification, and RTK-driven wild-type RAS reactivation represent convergent escape mechanisms.\",\n      \"evidence\": \"Paired pre-/post-treatment biopsy sequencing; in vitro deep mutational scanning; RAS activity assays with RTK inhibitor combinations\",\n      \"pmids\": [\"34161704\", \"35732135\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Therapeutic strategies to preempt multi-mechanism resistance were not clinically validated\", \"Whether resistance mechanisms differ across tissue types was not systematically tested\"]\n    },\n    {\n      \"year\": 2021,\n      \"claim\": \"Multi-omic profiling showed that KRAS-dependent transcription in PDAC is driven predominantly through ERK/MAPK, with APC/C and cell-cycle machinery as key downstream effectors, revising the canonical KRAS transcriptional signature.\",\n      \"evidence\": \"Inducible KRAS knockdown with integrated transcriptomics, phosphoproteomics, proteomics, and patient tumor data\",\n      \"pmids\": [\"38843331\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Whether APC/C deregulation is a direct or indirect effect of ERK signaling was not distinguished\", \"Contribution of non-ERK effectors to KRAS-dependent transcription was not fully quantified\"]\n    },\n    {\n      \"year\": 2023,\n      \"claim\": \"A comprehensive deep mutational scan of >26,000 KRAS mutations mapped the allosteric network across the central β-sheet and identified multiple allosterically active surface pockets, providing a near-complete biophysical atlas of KRAS function.\",\n      \"evidence\": \"Deep mutational scanning with double-mutant epistasis and free energy inference across six binding partners\",\n      \"pmids\": [\"38109937\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Structural validation of newly identified allosteric pockets by crystallography or cryo-EM is pending\", \"Whether allosteric mutations differentially affect KRAS signaling in cellular contexts was not tested\"]\n    },\n    {\n      \"year\": 2023,\n      \"claim\": \"CAF-derived NRG1 activating ERBB2/3 was identified as a microenvironment-driven bypass mechanism enabling KRAS-independent PDAC survival, demonstrating that KRAS-targeted therapy must account for paracrine escape.\",\n      \"evidence\": \"Genetic KRAS extinction, co-culture paracrine models, pharmacological ERBB inhibition in mouse and human PDAC\",\n      \"pmids\": [\"37775182\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Whether NRG1-ERBB bypass operates in non-pancreatic KRAS-mutant cancers is unknown\", \"The full repertoire of microenvironment-mediated escape routes has not been mapped\"]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"Key unresolved questions include whether KRAS dimerization occurs on native plasma membranes under physiological conditions, the complete structural basis for isoform-specific effector engagement in vivo, and how to therapeutically target the full spectrum of resistance mechanisms arising during KRAS inhibitor treatment.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"High\",\n      \"gaps\": [\"No consensus on in vivo KRAS dimerization stoichiometry or scaffold dependency\", \"Full-length, lipid-modified KRAS structure in a membrane environment is unavailable\", \"Clinical strategies to preempt convergent resistance remain undefined\"]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0003924\", \"supporting_discovery_ids\": [15, 26, 28, 33]},\n      {\"term_id\": \"GO:0098772\", \"supporting_discovery_ids\": [30, 35]},\n      {\"term_id\": \"GO:0008289\", \"supporting_discovery_ids\": [14, 24, 25]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005886\", \"supporting_discovery_ids\": [14, 17, 18, 21, 23, 25]},\n      {\"term_id\": \"GO:0005829\", \"supporting_discovery_ids\": [17, 18]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-162582\", \"supporting_discovery_ids\": [2, 7, 10, 20, 27, 30, 35]},\n      {\"term_id\": \"R-HSA-1640170\", \"supporting_discovery_ids\": [30]},\n      {\"term_id\": \"R-HSA-1430728\", \"supporting_discovery_ids\": [9, 19]},\n      {\"term_id\": \"R-HSA-1643685\", \"supporting_discovery_ids\": [0, 1, 31, 34]}\n    ],\n    \"complexes\": [],\n    \"partners\": [\n      \"SOS1\",\n      \"BRAF\",\n      \"RAF1\",\n      \"PIK3CA\",\n      \"CALM1\",\n      \"YAP1\",\n      \"USP18\",\n      \"PDED\"\n    ],\n    \"other_free_text\": []\n  }\n}\n```"}