CoChat
Sign Up
Cited by 574Lung Cancer Treatments and MutationsHER2/EGFR in Cancer ResearchCancer Genomics and Diagnostics

Structural, Biochemical, and Clinical Characterization of Epidermal Growth Factor Receptor (EGFR) Exon 20 Insertion Mutations in Lung Cancer

Open access
Yasuda, Hiroyuki, Park, Eunyoung, Yun, Cai-Hong, Sng, Natasha J., Lucena-Araujo, Antonio R., Yeo, Wee-Lee, Huberman, Mark S., Cohen, David W. +16 more·2013·Science Translational Medicine
DOI 10.1126/scitranslmed.3007205PMC PMC3954775
Open this paper in CoChat — free account

Abstract

Crystal structure and detailed analysis of different EGFR mutants explain why some mutations in exon 20 make lung cancers resistant to EGFR inhibitors and others make them more sensitive.

Integrity signals

Registry

Crossref · 10.1126/scitranslmed.3007205

Cites

71 of 71 references verified0 citation errors · 0 not checkable

Statistics

Nothing to recompute.

Claims the paper makes

0 of 75 cited sources checked · 0 supported · 0 flagged

  • Findingown data

    Among studied exon 20 insertions, only A763_Y764insFQEA was inhibited by erlotinib below 0.1 μM, along with control delL747_P753insS and L858R.

    “only cells with EGFR-delL747_P753insS, L858R and the atypical A763_Y764insFQEA - among all EGFR exon 20 insertions - were inhibited by erlotinib concentrations below 0.1 µM (Fig. 1B, 1C and Sup. Table 2).”
    Quote in the paperSubstantial claim
  • Findingown data

    D770_N771insNPG bound gefitinib with a Ki similar to WT EGFR (25.7 nM vs 16.4 nM).

    “The D770_N771insNPG mutant bound gefitinib with a Ki similar to that of WT EGFR (Ki=25.7 nM for D770_N771insNPG and Ki=16.4 nM for WT EGFR).”
    Quote in the paperSubstantial claim
  • Findingown data

    Gefitinib and afatinib showed similar sensitivity patterns to erlotinib across the studied mutants.

    “Nearly identical results were obtained with gefitinib (Sup. Table 2), and a similar pattern of insensitivity was noted with the more potent irreversible EGFR TKI afatinib (Sup. Table 2).”
    Quote in the paperNot a substantial claim
  • Findingown data

    In cells with inhibitor-insensitive exon 20 insertions, phosphorylation of EGFR, AKT, and ERK was not fully inhibited by 1 μM erlotinib.

    “phosphorylation of EGFR, AKT and ERK were not fully inhibited by 1 µM erlotinib in cells with the inhibitor-insensitive EGFR exon 20 insertions mutations (Fig. 1D).”
    Quote in the paperSubstantial claim
  • Findingown data

    A767_V769dupASV lacked deactivation of phosphorylated EGFR, PI3K/AKT, MAPK/ERK, and upregulation of BIM with 1 μM or submicromolar erlotinib.

    “an inhibitor-insensitive EGFR exon 20 insertion (A767_V769dupASV) lacked deactivation of phosphorylated EGFR, downstream targets (PI3K/AKT and MAPK/ERK) and upregulation of the apoptotic cascade, as measured by levels of BIM”
    Quote in the paperSubstantial claim
  • May combine two claims. 33
  • Classified as probably not a substantial claim. 12

  • May combine two claims.

    Findingown data

    A763_Y764insFQEA had a Ki for gefitinib of 2.5 nM, comparable to L858R (6.4 nM).

    “The Ki value for A763_Y764insFQEA (2.5 nM) was comparable to that we measured for the L858R mutant (6.4 nM).”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    All other studied exon 20 insertions and T790M-bearing mutations had erlotinib IC50s above 2 μM; most exon 20 insertions were not inhibited by submicromolar reversible EGFR TKIs.

    “All other EGFR exon 20 insertions and T790M–bearing mutations had 50% inhibitory concentrations (IC50s) that exceeded 2 µM erlotinib (Fig. 1B, 1C and Sup. Table 2).”
    • Also stated in: § DISCUSSION · boosted
      “our data confirm that most exon 20 insertions are not inhibited by submicromolar concentrations of reversible EGFR TKIs.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    Ba/F3 cells stably expressing the studied EGFR mutations proliferate without IL3, indicating transforming ability.

    “All were able to proliferate in the absence of IL3 (Sup. Fig. 1), indicating transforming ability of these mutations.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    Both D770_N771insNPG and A763_Y764insFQEA had higher catalytic activity than WT EGFR, but less than L858R.

    “both insertion mutants exhibited higher catalytic activity than WT EGFR; although they were not as active as EGFR-L858R (Table 1).”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    A763_Y764insFQEA showed inhibition of downstream phosphorylation and upregulation of BIM similar to L858R.

    “the inhibition of these downstream phosphorylation events and upregulation of BIM were seen with EGFR-A763_Y764insFQEA in a similar fashion as with inhibitor-sensitive L858R (Fig. 1D, 1E)”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    EGFR-specific siRNAs decreased EGFR expression and suppressed growth in BID007 and H3255.

    “In both BID007 and H3255, EGFR-specific siRNAs decreased expression of EGFR and suppressed cell growth (Fig. 3B).”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    A763_Y764insFQEA had an IC50 of <0.05 μM to erlotinib or gefitinib, more than 10-fold lower than clinically achievable serum concentrations.

    “In vitro characterization of A763_Y764insFQEA demonstrated that it had an IC50 of < 0.05µM to erlotinib or gefitinib, which is more than 10-fold lower than the clinically achievable serum concentrations of these EGFR TKIs”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    Together with D770, the three inserted residues form a tight turn with a hydrogen bond between the main chain carbonyl of D770 and the amide of the inserted glycine.

    “Together with D770, the three inserted residues form a tight turn, with a hydrogen bond formed between the main chain carbonyl of D770 and the amide of the inserted glycine (Fig. 4B, Sup. Fig. 10B, 10C).”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Findingown data

    D770_N771insNPG adopts an active conformation with the C-helix in its inward position.

    “The D770_N771insNPG mutant adopts an active conformation with the C-helix in its inward position (Fig. 4A).”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    Submicromolar erlotinib inhibited EGFR phosphorylation in A763_Y764insFQEA mutant cells, deactivated PI3K/AKT and MAPK/ERK pathways, and induced apoptosis.

    “submicromolar concentrations of erlotinib inhibited phosphorylation of EGFR in A763_Y764insFQEA mutant cells, led to deactivation of the PI3K/AKT and MAPK/ERK pathways, and subsequently induced apoptosis in our models.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    Differences between D770_N771insNPG and L858R structures include a small change in relative orientation of N- and C-lobes and a fully ordered activation loop in D770_N771insNPG but not in L858R.

    “Differences, which likely arise from the divergent crystal packing in D770_N771insNPG, include a small change in the relative orientation of the N- and C-lobes, and the conformation of the activation loop, which is fully ordered in the D770_N771insNPG structure but not in the present L858R structure.”
    Quote in the paperNot a substantial claim

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Findingown data

    BID007 had slight amplification of region around chromosome 7 corresponding to EGFR.

    “The region around chromosome 7, corresponding to EGFR, demonstrated slight amplification (Sup. Fig. 9B).”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Findingown data

    A763_Y764insFQEA is activated relative to WT EGFR and much more potently inhibited by gefitinib than WT EGFR.

    “Our kinetic characterization of this mutant revealed that it is activated, relative to WT EGFR, and that it is much more potently inhibited by gefitinib than is WT EGFR.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    Homology models of A763_Y764insFQEA using active and inactive conformations show the insertion modeled in helical conformation, shifting register N-terminus and resulting in longer β3-αC loop.

    “In both models, the insertion was modeled in a helical conformation, shifting the register of the C-helix toward its N-terminus, and resulting in a longer loop at its N-terminal end (the β3-αC loop).”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    The FQEA insertion is compatible with asymmetric dimer formation, and size-exclusion chromatography revealed a monomer-dimer equilibrium in solution.

    “inspection of the model indicates that the FQEA insertion is compatible with formation of the asymmetric dimer and, consistent with this, analysis of the A763_Y764insFQEA mutant using size-exclusion chromatography revealed a monomer-dimer equilibrium in solution (Sup. Fig. 11).”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Findingown data

    E762Q_insFQEA was highly active in a ligand-independent manner, while A763_Y764insFQQA was inactive.

    “We found that E762Q_insFQEA, like A763_Y764insFQEA, was highly active in a ligand independent manner but A763_Y764insFQQA was inactive (Fig. 5B).”
    Quote in the paperSubstantial claim

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Findingown data

    BID007 had focal deletion encompassing CDKN2A.

    “BID007 had a focal area of deletion encompassing the CDKN2A tumor suppressor gene (Sup. Fig. 9C); an area commonly deleted in cancers 45.”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [45] Weir BA Woo MS Getz G Perner S Ding L Beroukhim R Lin WM Province MA Kraja A Johnson LA Shah K Sato M Thomas RK Barletta JA Borecki IB Broderick S Chang AC Chiang DY Chirieac LR Cho J Fujii Y Gazdar AF Giordano T Greulich H Hanna M Johnson BE Kris MG Lash A Lin L Lindeman N Mardis ER McPherson JD Minna JD Morgan MB Nadel M Orringer MB Osborne JR Ozenberger B Ramos AH Robinson J Roth JA Rusch V Sasaki H Shepherd F Sougnez C Spitz MR Tsao MS Twomey D Verhaak RG Weinstock GM Wheeler DA Winckler W Yoshizawa A Yu S Zakowski MF Zhang Q Beer DG Wistuba II Watson MA Garraway LA Ladanyi M Travis WD Pao

      Not checked in the free run.

  • May combine two claims.

    Findingreasoning

    D770_N771insNPG activates EGFR without markedly diminishing its affinity for ATP or increasing its affinity for EGFR TKIs.

    “Unlike EGFR-L858R and exon 19 deletion mutants, the D770_N771insNPG mutation lacks this “Achilles’ heel”, since it activates EGFR without markedly diminishing its affinity for ATP or increasing its affinity for EGFR TKIs.”
    Quote in the paperSubstantial claim

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation

    The most common EGFR mutations are exon 19 deletions around LREA (45%) and L858R (40%).

    “The most common EGFR mutations are in-frame deletions around the LREA motif (amino-acid residues 747 to 750) of exon 19 (45% of EGFR mutations) and the exon 21 L858R point mutation (40% of EGFR mutations) 5,6,8.”
    Quote in the paperNot a substantial claim0 of 3 sources checked
    • cites [5] Shigematsu H Lin L Takahashi T Nomura M Suzuki M Wistuba II Fong KM Lee H Toyooka S Shimizu N Fujisawa T Feng Z Roth JA Herz J Minna JD Gazdar AF Clinical and biological features associated with epidermal growth factor receptor gene mutations in lung cancers J Natl. Cancer Inst 2005 97 339 346 15741570

      Not checked in the free run.

    • cites [6] Sequist LV Bell DW Lynch TJ Haber DA Molecular predictors of response to epidermal growth factor receptor antagonists in non-small-cell lung cancer J Clin. Oncol 2007 25 587 595 17290067

      Not checked in the free run.

    • cites [8] Tokumo M Toyooka S Kiura K Shigematsu H Tomii K Aoe M Ichimura K Tsuda T Yano M Tsukuda K Tabata M Ueoka H Tanimoto M Date H Gazdar AF Shimizu N The relationship between epidermal growth factor receptor mutations and clinicopathologic features in non-small cell lung cancers Clin. Cancer Res 2005 11 1167 1173 15709185

      Not checked in the free run.

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation

    Exon 20 insertion mutations make up 4-10% of all EGFR mutations in NSCLC.

    “Exon 20 insertion mutations comprise 4–10% of all EGFR mutations 27,29–32.”
    Quote in the paperNot a substantial claim0 of 5 sources checked
    • cites [27] Mitsudomi T Yatabe Y Mutations of the epidermal growth factor receptor gene and related genes as determinants of epidermal growth factor receptor tyrosine kinase inhibitors sensitivity in lung cancer Cancer Sci 2007 98 1817 1824 17888036 PMC11159145

      Not checked in the free run.

    • cites [29] Choi YL Soda M Yamashita Y Ueno T Takashima J Nakajima T Yatabe Y Takeuchi K Hamada T Haruta H Ishikawa Y Kimura H Mitsudomi T Tanio Y Mano H EML4-ALK mutations in lung cancer that confer resistance to ALK inhibitors N. Engl. J Med 2010 363 1734 1739 20979473

      Not checked in the free run.

    • cites [30] Pao W Chmielecki J Rational, biologically based treatment of EGFR-mutant non-small-cell lung cancer Nat. Rev. Cancer 2010 10 760 774 20966921 PMC3072803 source

      Not checked in the free run.

    • cites [31] Oxnard GR Lo PC Nishino M Dahlberg SE Lindeman NI Butaney M Jackman DM Johnson BE Janne PA Natural history and molecular characteristics of lung cancers harboring EGFR exon 20 insertions J Thorac. Oncol 2013 8 179 184 23328547 PMC3549533

      Not checked in the free run.

    • cites [32] Arcila ME Nafa K Chaft JE Rekhtman N Lau C Reva BA Zakowski MF Kris MG Ladanyi M EGFR exon 20 insertion mutations in lung adenocarcinomas: prevalence, molecular heterogeneity, and clinicopathologic characteristics Mol. Cancer Ther 2013 12 220 229 23371856 PMC3714231

      Not checked in the free run.

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation

    Other EGFR mutations (G719A/C/S, rare exon 19 insertions, L861Q) are associated with some sensitivity to gefitinib and erlotinib.

    “Other EGFR mutations have also been associated with some sensitivity to gefitinib and erlotinib. These include exon 18 point mutations in position G719 (G719A, C or S - ∼3% of EGFR mutations), rare inframe exon 19 insertions 25, and the exon 21 L861Q mutant (∼2% of EGFR mutations) 26–28.”
    Quote in the paperNot a substantial claim0 of 4 sources checked
    • cites [25] He M Capelletti M Nafa K Yun CH Arcila ME Miller VA Ginsberg MS Zhao B Kris MG Eck MJ Janne PA Ladanyi M Oxnard GR EGFR exon 19 insertions: a new family of sensitizing EGFR mutations in lung adenocarcinoma Clin Cancer Res 2012 18 1790 1797 22190593 PMC3306520

      Not checked in the free run.

    • cites [26] Mitsudomi T Yatabe Y Epidermal growth factor receptor in relation to tumor development: EGFR gene and cancer FEBS J 2010 277 301 308 19922469

      Not checked in the free run.

    • cites [27] Mitsudomi T Yatabe Y Mutations of the epidermal growth factor receptor gene and related genes as determinants of epidermal growth factor receptor tyrosine kinase inhibitors sensitivity in lung cancer Cancer Sci 2007 98 1817 1824 17888036 PMC11159145

      Not checked in the free run.

    • cites [28] Yeh P Chen H Andrews J Naser R Pao W Horn L DNA-Mutation Inventory to Refine and Enhance Cancer Treatment (DIRECT): A Catalog of Clinically Relevant Cancer Mutations to Enable Genome-Directed Anticancer Therapy Clin Cancer Res 2013 19 1894 1901 23344264 PMC4121886

      Not checked in the free run.

  • May combine two claims.

    Interpretationreasoning

    Based on Ki/Km[ATP], A763_Y764insFQEA is predicted to be almost as sensitive to gefitinib as L858R, while D770_N771insNPG is expected to be many fold less sensitive.

    “Based on this measure, A763_Y764insFQEA is predicted to be almost as sensitive to gefitinib as the L858R mutant, whereas D770_N771insNPG is expected to be many fold less sensitive (Table 1).”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Premisecitation

    Most exon 20 insertion mutations lie near the end of the C-helix after M766, while a small subset map to the middle of the C-helix affecting E762 to Y764.

    “Most of these mutations lie near the end of the C-helix within the N-lobe of the kinase, after residue M766, but a small subset map to the middle of the C-helix (affecting amino-acids E762 to Y764) 5,33,34.”
    Quote in the paperNot a substantial claim0 of 3 sources checked
    • cites [5] Shigematsu H Lin L Takahashi T Nomura M Suzuki M Wistuba II Fong KM Lee H Toyooka S Shimizu N Fujisawa T Feng Z Roth JA Herz J Minna JD Gazdar AF Clinical and biological features associated with epidermal growth factor receptor gene mutations in lung cancers J Natl. Cancer Inst 2005 97 339 346 15741570

      Not checked in the free run.

    • cites [33] Murray S Dahabreh IJ Linardou H Manoloukos M Bafaloukos D Kosmidis P Somatic mutations of the tyrosine kinase domain of epidermal growth factor receptor and tyrosine kinase inhibitor response to TKIs in non-small cell lung cancer: an analytical database J Thorac. Oncol 2008 3 832 839 18670300

      Not checked in the free run.

    • cites [34] Linardou H Dahabreh IJ Bafaloukos D Kosmidis P Murray S Somatic EGFR mutations and efficacy of tyrosine kinase inhibitors in NSCLC Nat. Rev. Clin Oncol 2009 6 352 366 19483740

      Not checked in the free run.

  • May combine two claims.

    Premisecitation

    Most NSCLCs with EGFR exon 20 insertion mutations do not respond radiographically or clinically to gefitinib or erlotinib, with reported response rate below 5%.

    “Unlike EGFR exon 19 deletions and L858R–bearing tumors, most NSCLCs with EGFR exon 20 insertion mutations don’t respond radiographically or clinically to gefitinib or erlotinib. The reported response rate (RR) is below 5% and most patients have short intervals of disease control 35.”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites [35] Yasuda H Kobayashi S Costa DB EGFR exon 20 insertion mutations in non-small-cell lung cancer: preclinical data and clinical implications Lancet Oncol 2012 13 e23 e31 21764376 source

      Not checked in the free run.

  • May combine two claims.

    Study design and conductnone given

    The study was limited by paucity of NSCLC cell lines with exon 20 insertions, lack of in vivo models, and small number of exon 20 insertion mutated NSCLC cases treated with reversible TKIs.

    “Our study was limited by a number of factors, including a paucity of available NSCLC cell lines with EGFR exon 20 insertion mutations, a lack of in vivo models and the small number of exon 20 insertion mutated NSCLC cases that were treated with reversible EGFR TKIs in our centers.”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Interpretationown data

    Structure and kinetic data show D770_N771insNPG promotes the active conformation, reflected by enhanced catalytic activity and propensity to form asymmetric dimer.

    “our structure and kinetic data show that D770_N771insNPG promotes the active conformation, as reflected by its enhanced catalytic activity (Table 1) and a propensity to form the asymmetric dimer as seen in the crystal structure (Sup. Fig. 10A) and in solution (Sup. Fig. 11).”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Interpretationreasoning

    A shift toward the N-terminus would displace E762 by one turn but effectively replace it with a glutamic acid residue introduced by the insertion.

    “Alternatively, a shift toward the N-terminus would displace this residue by one turn, but effectively replace it with a glutamic acid residue introduced by the insertion (FQEA, Fig. 5A).”
    Quote in the paperSubstantial claim

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation

    Mutation of L747 in this cluster to proline or serine is believed to underlie the activity of EGFR-exon 19 insertions.

    “mutation of L747 in this cluster to proline or serine is believed to underlie the activity of EGFR-exon 19 insertions 25.”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [25] He M Capelletti M Nafa K Yun CH Arcila ME Miller VA Ginsberg MS Zhao B Kris MG Eck MJ Janne PA Ladanyi M Oxnard GR EGFR exon 19 insertions: a new family of sensitizing EGFR mutations in lung adenocarcinoma Clin Cancer Res 2012 18 1790 1797 22190593 PMC3306520

      Not checked in the free run.

  • May combine two claims.

    Interpretationown data

    Binding of gefitinib and erlotinib is expected to be unperturbed in D770_N771insNPG, consistent with similar Ki values for gefitinib between D770_N771insNPG and WT EGFR.

    “Likewise, we expect that binding of gefitinib and erlotinib would be unperturbed in this mutant, consistent with the closely similar Ki values for gefitinib we measure for D770_N771insNPG and WT EGFR (Table 1).”
    Quote in the paperSubstantial claim

  • Probably not a substantial claim: may not pass S1 (asserted).

    Interpretationreasoning

    The de facto I759A replacement caused by FQEA insertion may activate EGFR via destabilization of the inactive conformation.

    “Thus we hypothesize that the de facto I759A replacement caused by the FQEA insertion may activate EGFR via a similar mechanism (destabilization of the inactive conformation).”
    Quote in the paperNot a substantial claim
    • Rests on an assumption: I759, together with L747, L858, and L861, forms a cluster of interacting hydrophobic residues that helps stabilize the inactive conformation.

  • Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.

    Premisecitation

    Asymmetric dimer interaction mediates EGFR activation by ligand-induced dimerization and is typically recapitulated in crystals of active EGFR kinase.

    “This mode of interaction mediates EGFR activation by ligand-induced dimerization 48, and is typically recapitulated in crystals of the EGFR kinase in its active conformation.”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [48] Zhang X Gureasko J Shen K Cole PA Kuriyan J An allosteric mechanism for activation of the kinase domain of epidermal growth factor receptor Cell 2006 125 1137 1149 16777603

      Not checked in the free run.

  • May combine two claims.

    Interpretationreasoning

    The inserted residues are expected to form approximately one turn of helix and shift the register of adjacent residues either toward N-terminus or C-terminus.

    “the four inserted residues are expected to form approximately one turn of helix and shift the register of adjacent residues in the helix, either toward the N-terminus or toward the C-terminus (Fig. 5A).”
    Quote in the paperSubstantial claim
    • The authors state this as an assumption: The four inserted residues in A763_Y764insFQEA form approximately one turn of helix.

  • Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.

    Premisecitation

    PD168393 is a preclinical irreversible anilinoquinazoline inhibitor structurally related to afatinib and dacomitinib.

    “PD168393 is a pre-clinical irreversible anilinoquinazoline inhibitor that is structurally related to afatinib and dacomitinib 46,47.”
    Quote in the paperNot a substantial claim0 of 2 sources checked
    • cites [46] Engelman JA Zejnullahu K Gale CM Lifshits E Gonzales AJ Shimamura T Zhao F Vincent PW Naumov GN Bradner JE Althaus IW Gandhi L Shapiro GI Nelson JM Heymach JV Meyerson M Wong KK Janne PA PF00299804, an irreversible pan-ERBB inhibitor, is effective in lung cancer models with EGFR and ERBB2 mutations that are resistant to gefitinib Cancer Res 2007 67 11924 11932 18089823 source

      Not checked in the free run.

    • cites [47] Li D Ambrogio L Shimamura T Kubo S Takahashi M Chirieac LR Padera RF Shapiro GI Baum A Himmelsbach F Rettig WJ Meyerson M Solca F Greulich H Wong KK BIBW2992, an irreversible EGFR/HER2 inhibitor highly effective in preclinical lung cancer models Oncogene 2008 27 4702 4711 18408761 PMC2748240 source

      Not checked in the free run.

  • Classified as probably not a substantial claim.

    Premisecitation

    L858R and L861Q mutations within the hydrophobic cluster are well-characterized activating mutations.

    “The L858R and L861Q mutations within this cluster are well-characterized activating mutations 28”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites [28] Yeh P Chen H Andrews J Naser R Pao W Horn L DNA-Mutation Inventory to Refine and Enhance Cancer Treatment (DIRECT): A Catalog of Clinically Relevant Cancer Mutations to Enable Genome-Directed Anticancer Therapy Clin Cancer Res 2013 19 1894 1901 23344264 PMC4121886

      Not checked in the free run.

  • May combine two claims.

    Interpretationown data

    The insertion shifts the register of the C-helix in the N-terminal direction, and the glutamic acid in FQEA assumes the active site role of E762 in WT kinase.

    “Thus, we conclude that the insertion shifts the register of the helix in the N-terminal direction and that the glutamic acid residue in the FQEA insertion assumes the active site role of E762 in the WT kinase.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Premisecitation

    L858R and exon 19 deletion mutants exhibit increased Km[ATP] and higher affinity (5-10 fold) for gefitinib or erlotinib than WT EGFR.

    “L858R and exon 19 deletion mutants exhibit increased Km[ATP] and higher affinity (5–10 fold) for gefitinib or erlotinib than the WT receptor 39,50,56.”
    Quote in the paperSubstantial claim0 of 3 sources checked
    • cites [39] Carey KD Garton AJ Romero MS Kahler J Thomson S Ross S Park F Haley JD Gibson N Sliwkowski MX Kinetic analysis of epidermal growth factor receptor somatic mutant proteins shows increased sensitivity to the epidermal growth factor receptor tyrosine kinase inhibitor, erlotinib Cancer Res 2006 66 8163 8171 16912195

      Not checked in the free run.

    • cites [50] Eck MJ Yun CH Structural and mechanistic underpinnings of the differential drug sensitivity of EGFR mutations in non-small cell lung cancer Biochim. Biophys. Acta 2010 1804 559 566 20026433 PMC2859716

      Not checked in the free run.

    • cites [56] Mulloy R Ferrand A Kim Y Sordella R Bell DW Haber DA Anderson KS Settleman J Epidermal growth factor receptor mutants from human lung cancers exhibit enhanced catalytic activity and increased sensitivity to gefitinib Cancer Res 2007 67 2325 2330 17332364

      Not checked in the free run.

  • Probably not a substantial claim: may not pass S3 (load-bearing), S4 (based).

    Interpretationnone given

    Structurally, EGFR-A763_Y764insFQEA is very different from other exon 20 insertions.

    “Structurally, EGFR-A763_Y764insFQEA is very different from other exon 20 insertions.”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Interpretationcitation

    The increased Km[ATP] and higher TKI affinity of L858R and exon 19 deletion mutants, together with the oncogene-addicted state of tumors bearing these mutations, explain their sensitivity to EGFR TKIs.

    “Together with the oncogene-addicted state of tumors bearing these mutations 39,50,56, these factors explain their sensitivity to EGFR TKIs 50.”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites [50] Eck MJ Yun CH Structural and mechanistic underpinnings of the differential drug sensitivity of EGFR mutations in non-small cell lung cancer Biochim. Biophys. Acta 2010 1804 559 566 20026433 PMC2859716

      Not checked in the free run.

  • Probably not a substantial claim: may not pass S1 (asserted).

    Interpretationreasoning

    The I759A alteration may lead to catalytic activation in a manner related to L858R/L861Q.

    “we hypothesize that it leads to catalytic activation in a related manner.”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Premisecitation

    Tumors bearing L858R or exon 19 deletion mutations are oncogene-addicted.

    “Together with the oncogene-addicted state of tumors bearing these mutations 39,50,56, these factors explain their sensitivity to EGFR TKIs 50.”
    Quote in the paperSubstantial claim0 of 3 sources checked
    • cites [39] Carey KD Garton AJ Romero MS Kahler J Thomson S Ross S Park F Haley JD Gibson N Sliwkowski MX Kinetic analysis of epidermal growth factor receptor somatic mutant proteins shows increased sensitivity to the epidermal growth factor receptor tyrosine kinase inhibitor, erlotinib Cancer Res 2006 66 8163 8171 16912195

      Not checked in the free run.

    • cites [50] Eck MJ Yun CH Structural and mechanistic underpinnings of the differential drug sensitivity of EGFR mutations in non-small cell lung cancer Biochim. Biophys. Acta 2010 1804 559 566 20026433 PMC2859716

      Not checked in the free run.

    • cites [56] Mulloy R Ferrand A Kim Y Sordella R Bell DW Haber DA Anderson KS Settleman J Epidermal growth factor receptor mutants from human lung cancers exhibit enhanced catalytic activity and increased sensitivity to gefitinib Cancer Res 2007 67 2325 2330 17332364

      Not checked in the free run.

  • May combine two claims.

    Premisecitation

    T790M resistance involves alteration of the gatekeeper residue in the ATP-site, which is an important determinant of inhibitor specificity.

    “The EGFR-T790M mutation confers resistance by reversing the sensitization conferred by primary activating mutations, but it does so by altering the gatekeeper residue in the ATP-site that is known to be an important determinant of inhibitor specificity 50,57.”
    Quote in the paperSubstantial claim0 of 2 sources checked
    • cites [50] Eck MJ Yun CH Structural and mechanistic underpinnings of the differential drug sensitivity of EGFR mutations in non-small cell lung cancer Biochim. Biophys. Acta 2010 1804 559 566 20026433 PMC2859716

      Not checked in the free run.

    • cites [57] Yun CH Mengwasser KE Toms AV Woo MS Greulich H Wong KK Meyerson M Eck MJ The T790M mutation in EGFR kinase causes drug resistance by increasing the affinity for ATP Proc. Natl. Acad. Sci. U. S. A 2008 105 2070 2075 18227510 PMC2538882

      Not checked in the free run.

  • May combine two claims.

    Premisecitation

    EGFR monoclonal antibodies and EGFR WT kinase sparing covalent T790M selective TKIs are ineffective in typical EGFR exon 20 insertion mutants.

    “Indeed, it seems that EGFR monoclonal antibodies 60 and EGFR WT kinase sparing covalent T790M selective TKIs 61 are ineffective in typical EGFR exon 20 insertion mutants.”
    Quote in the paperSubstantial claim0 of 2 sources checked
    • cites [60] Cho J Chen L Sangji N Okabe T Yonesaka K Francis JM Flavin RJ Johnson W Kwon J Yu S Greulich HE Johnson BE Eck MJ Janne PA Wong KK Meyerson M Cetuximab response of lung cancer-derived EGF receptor mutants is associated with asymmetric dimerization Cancer Res 2013 PMC3903789 24063894

      Not checked in the free run.

    • cites [61] Walter AO Tjin Tham SR Haringsma HJ Ohashi K Sun J Lee K Dubrovskiy A Labenski M Zhu Z Wang Z Sheets M St Martin T Karp R van Kalken D Chaturvedi P Niu D Nacht M Petter RC Westlin W Lin K Jaw-Tsai S Raponi M Van Dyke T Etter J Weaver Z Pao W Singh J Simmons AD Harding TC Allen A Discovery of a mutant-selective covalent inhibitor of EGFR that overcomes T790M–mediated resistance in NSCLC Cancer Discov 2013 PMC4048995 24065731 source

      Not checked in the free run.

  • May combine two claims.

    Interpretationown data

    Structural analysis and supporting mutagenesis indicate that the inserted FQEA sequence shifts the register of the C-helix toward its N-terminus, altering the length of the β3-αC loop and effectively leading to an I759A replacement.

    “our structural analysis and supporting mutagenesis indicate that the inserted FQEA sequence shifts the register of the C-helix toward its N-terminus, altering the length of the β3-αC loop leading into the helix and effectively leading to an I759A replacement.”
    Quote in the paperSubstantial claim
43 of 43

  • The paper relies on this without support.

    The concentration of ATP in cells approximates 1 mM.

    “which approximates the concentration of ATP in cells”

  • The authors state this as an assumption.

    The four inserted residues in A763_Y764insFQEA form approximately one turn of helix.

    “the four inserted residues are expected to form approximately one turn of helix”

  • The paper relies on this without support.

    In the inactive state, the C-helix pivots outward and shifts by ~3.5Å toward the insertion site.

    “In the inactive state, the C-helix pivots outward, and shifts by ∼3.5Å toward the insertion site.”

  • The paper relies on this without support.

    I759, together with L747, L858, and L861, forms a cluster of interacting hydrophobic residues that helps stabilize the inactive conformation.

    “Isoleucine at position 759, together with L747, L858 and L861, is part of a cluster of interacting hydrophobic residues that helps to stabilize the inactive conformation.”

  • The paper relies on this without support.

    D770_N771insNPG is a representative EGFR TKI-insensitive exon 20 insertion mutation.

    “The structure of D770_N771insNPG, which we studied as a representative EGFR TKI-insensitive exon 20 mutation, shows how the three amino acid insertion forms a “wedge” at the end of the C-helix that may effectively lock the helix in its inward, active position.”
5 of 5

Claims other papers make about this one

Each is checked against this paper’s own text: does it say what the citing paper says it does?

Cites

Flagged first, then by citations

Cited by

Sorted by citations

Related

Authors
Yasuda, H., Park, E., Yun, C.-H., Sng, N. J., Lucena-Araujo, A. R., Yeo, W.-L., Huberman, M. S., Cohen, D. W., Nakayama, S., Ishioka, K., Yamaguchi, N., Hanna, M., Oxnard, G. R., Lathan, C. S., Moran, T., Sequist, L. V., Chaft, J. E., Riely, G. J., Arcila, M. E., Soo, R. A., Meyerson, M., Eck, M. J., Kobayashi, S. S., & Costa, D. B.
Affiliations
  • Beth Israel Deaconess Medical Center (US)
  • Harvard University (US)
  • Peking University (CN)
  • Keio University (JP)
  • Broad Institute (US)
  • Brigham and Women's Hospital (US)
  • Massachusetts General Hospital, Harvard Medical School, Boston, MA 02114, USA.
  • Memorial Sloan Kettering Cancer Center (US)
  • National University of Singapore (SG)
Corresponding author
  • Yasuda, Hiroyuki
  • Park, Eunyoung
  • Yun, Cai-Hong
  • Cohen, David W.
  • Eck, Michael J.
  • Kobayashi, Susumu S.
  • Costa, Daniel B.
ORCID
Venue
Science Translational Medicine
Publisher
American Association for the Advancement of Science (AAAS)
Published
2013-12-18
Volume
5
Issue
216
Pages
216ra177-216ra177
ISSN
  • 1946-6234 (ISSN-L)
  • 1946-6242
DOI
10.1126/scitranslmed.3007205
PMC
PMC3954775
OpenAlex
W2076883919
Cited by
571
References
71
Citations by year
2026: 24 · 2025: 41 · 2024: 46 · 2023: 60 · 2022: 52 · 2021: 64 · 2020: 65 · 2019: 43 · 2018: 40 · 2017: 43 · 2016: 34 · 2015: 41 · 2014: 17
FWCI
24.98
Type
journal article
Language
en
Open access
Locations
  • Science Translational Medicine (publishedVersion)
  • PubMed (publishedVersion)
  • PubMed Central (acceptedVersion)
  • National University of Singapore (submittedVersion)
Funders
National Cancer Institute (R01CA116020, CA090578, R00CA126026, R01CA169259, CA 116020, P50CA090578, K99CA126026, P20CA090578, P01CA154303, CA154303, CA126026, CA169259)
Topics
  • Health Sciences › Medicine › Pulmonary and Respiratory Medicine › Lung Cancer Treatments and Mutations
  • Health Sciences › Medicine › Oncology › HER2/EGFR in Cancer Research
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Cancer Research › Cancer Genomics and Diagnostics
Keywords
EGFR exon 20 insertion, non-small cell lung cancer, EGFR tyrosine kinase inhibitors, gefitinib, erlotinib, tyrosine kinase inhibitor resistance, L858R mutation, ATP binding site, kinase conformation, crystal structure, targeted therapy
MeSH terms
Erlotinib Hydrochloride, Gefitinib, Amino Acid Sequence, Antineoplastic Agents, Base Sequence, Carcinoma, Non-Small-Cell Lung, DNA, Neoplasm, Exons, Humans, Lung Neoplasms, Models, Molecular, Molecular Sequence Data, Mutation, Protein Conformation, Quinazolines, Mutagenesis, Insertional, Crystallography, X-Ray, Genes, erbB-1, Drug Resistance, Neoplasm, Structural Homology, Protein, Cell Line, Tumor, Protein Kinase Inhibitors, Translational Research, Biomedical, ErbB Receptors

About these checks

Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.

Claims read by deepseek-v4.1-flash under definition version 3 (extract-v3) on 2026-10-06.

Classified by jev-1.13-20260917 (gold-guideline-v0.2) on 2026-10-06.

Statistics and arithmetic are recomputed, not judged.

Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.

What counts as a claim ›How we measure confidence ›How we check this ›

Are you an author, or do you see a problem with this page? Request a review