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Cited by 83Protein Kinase Regulation and GTPase SignalingHippo pathway signaling and YAP/TAZMelanoma and MAPK Pathways

Focal Adhesion Kinase (FAK) Inhibition Synergizes with KRAS G12C Inhibitors in Treating Cancer through the Regulation of the FAK–YAP Signaling

Open access
Zhang, Baoyuan, Zhang, Yan, Zhang, Jiangwei, Liu, Ping, Jiao, Bo, Wang, Zaiqi, Ren, Ruibao·2021·Advanced Science
DOI 10.1002/advs.202100250PMC PMC8373085
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Abstract

KRAS mutation is one of the most prevalent genetic drivers of cancer development, yet KRAS mutations are until very recently considered undruggable. There are ongoing trials of drugs that target the KRAS G12C mutation, yet acquired drug resistance from the extended use has already become a major concern. Here, it is demonstrated that KRAS G12C inhibition induces sustained activation of focal adhesive kinase (FAK) and show that a combination therapy comprising KRAS G12C inhibition and a FAK inhibitor (IN10018) achieves synergistic anticancer effects. It can simultaneously reduce the extent of drug resistance. Diverse CDX and PDX models of KRAS G12C mutant cancer are examined and synergistic benefits from the combination therapy are consistently observed. Mechanistically, it is found that both aberrant FAK–YAP signaling and FAK‐related fibrogenesis impact on the development of KRAS G12C inhibitor resistance. This study thus illustrates the mechanism of resistance of cancer to the treatment of KRAS G12C inhibitor, as well as an innovative combination therapy to improve treatment outcomes for KRAS G12C mutant cancers.

AI snapshot

Written from the paper’s full text.

TL;DR

KRAS G12C inhibition adaptively activates FAK, and combining KRAS G12C inhibitors (AMG510 or MRTX849) with the FAK inhibitor IN10018 produces synergistic anticancer effects in KRAS G12C mutant models. The synergy is linked to repression of FAK–YAP signaling and reduced FAK-related tumor fibrosis.

Key findings

  • KRAS G12C inhibition with AMG510 or MRTX849 induced sustained activation of FAK, measured by Phospho FAK Y397, in all tested KRAS G12C mutant cell lines; no such FAK stimulation was observed in KRAS non-G12C mutant A549 and HCC827 cells.
  • AMG510 plus IN10018 was more cytotoxic than either monotherapy across KRAS G12C mutant cell lines, including SW837, Mia PaCa-2, CO-04-0070 PDC, NCI-H23, NCI-H1792, and NCI-H2122, with synergy assessed by Bliss and combination index analysis. In vivo, the combination showed synergistic tumor growth inhibition in CDX models (Mia PaCa-2, NCI-H358, NCI-H2122) and PDX models (CO-04-0315, CO-04-0070, LU-01-0030); in CO-04-0070, the combination conferred tumor regression, whereas AMG510 monotherapy showed persistent tumor growth.
  • Mechanistically, combination treatment more strongly repressed Hippo/YAP signaling than monotherapy: RNA-seq showed Hippo pathway enrichment among downregulated genes, RT-qPCR showed greater repression of 7 Hippo pathway signature genes, and Western blot/immunofluorescence showed reduced nuclear YAP and CYR61. YAP1 siRNA enhanced AMG510 killing in NCI-H358 and NCI-H1792 cells, while LATS1/LATS2 knockdown rescued the combination effect.
  • Long-term AMG510 treatment at 30 mg kg−1 increased fibrogenesis in NCI-H2122, CO-04-0070, and LU-01-0030 models by Masson and Sirius red staining; adding IN10018 markedly decreased this fibrosis.

Methods

  • In vitro assays included CellTiter-Glo viability, clonogenic survival, Western blot, RT-qPCR, RNA-seq with GO/KEGG analysis, siRNA knockdown, nuclear/cytoplasm fractionation, and immunofluorescence.
  • In vivo studies used KRAS mutant CDX and PDX models in BALB/c nude and NOD SCID mice, with oral gavage once daily; IN10018 was dosed at 25 or 50 mg kg−1, and AMG510 at 10 or 30 mg kg−1 depending on model. Tumor volume was calculated as 0.5 × long diameter × short diameter × short diameter.
  • TCGA survival analysis used cBioPortal data with PTK2 mRNA z scores < −1 or > 1; statistics included unpaired two-tailed t-tests, one-way ANOVA, log-rank tests, Bliss scores, combination index values, and in vivo synergy P values.

Limitations

  • The paper does not include a dedicated limitations section and states no specific limitations.
  • Evident from the text: all reported efficacy data are preclinical, using cancer cell lines, CDX, and PDX mouse models; no human clinical efficacy or safety outcomes are reported, and the authors state that a clinical trial is planned or underway.
  • Evident from the text: direct small-molecule targeting of YAP remains elusive, so FAK inhibition is proposed as a substitute; several in vivo experiments use small group sizes (e.g., n = 3–5), and the findings are limited to the tested KRAS G12C mutant models.

Model-written summary. Check it against the paper before relying on it.

Integrity signals

Registry

Crossref · 10.1002/advs.202100250

Cites

62 of 62 references verified0 citation errors · 0 not checkable

Statistics

Nothing to recompute.

Claims the paper makes

0 of 48 cited sources checked · 0 supported · 0 flagged

  • Findingown data

    IN10018 significantly inhibits FAK activity in examined cell lines.

    “we confirmed significant inhibition of FAK activity for the examined cell lines (Figure S3A–G, Supporting Information).”
    Quote in the paperSubstantial claim
    • Rests on an assumption: Phospho FAK Y397 is a valid marker of FAK signaling activity.
  • Findingcitation

    Within the KRAS mutant subgroup, lower RNA expression of PTK2 (FAK coding gene) correlates with better survival outcomes.

    “Within the KRAS mutant subgroup, lower RNA expression of PTK2 (FAK coding gene) correlates with better survival outcomes”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites [19] a) E. Cerami, J. Gao, U. Dogrusoz, B. E. Gross, S. O. Sumer, B. A. Aksoy, A. Jacobsen, C. J. Byrne, M. L. Heuer, E. Larsson, Y. Antipin, B. Reva, A. P. Goldberg, C. Sander, N. Schultz, Cancer Discovery 2012, 2, 401; 22588877 PMC3956037 b) C. Hutter, J. C. Zenklusen, Cell 2018, 173, 283. 29625045 source

      Not checked in the free run.

  • Findingown data

    KRAS G12C inhibitors exert effects after drug administration, indicated by a clear band shift for the KRAS protein in all KRAS G12C mutant cell lines.

    “these experiments revealed a clear band shift for the KRAS protein in all the KRAS G12C mutant cell lines, indicating that KRAS G12C inhibitors exert effects after drug administration.[ 21 ]”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [21] J. Canon, K. Rex, A. Y. Saiki, C. Mohr, K. Cooke, D. Bagal, K. Gaida, T. Holt, C. G. Knutson, N. Koppada, B. A. Lanman, J. Werner, A. S. Rapaport, T. San Miguel, R. Ortiz, T. Osgood, J. R. Sun, X. Zhu, J. D. McCarter, L. P. Volak, B. E. Houk, M. G. Fakih, B. H. O'Neil, T. J. Price, G. S. Falchook, J. Desai, J. Kuo, R. Govindan, D. S. Hong, W. Ouyang, H. Henary, T. Arvedson, V. J. Cee, J. R. Lipford, Nature 2019, 575, 217. 31666701 source

      Not checked in the free run.

  • Findingcitation

    In silico analysis showed all KRAS-mutant-dependent cell lines exhibited growth modulation related to PTK2 expression, unlike any KRAS wild-type cancer cell lines.

    “Analysis of data for shRNA modulated PTK2 knockdown showed that, unlike any of the KRAS wild‐type cancer cell lines, all of the KRAS‐mutant‐dependent cell lines exhibited growth modulation related to PTK2 expression (Figure S2B, Supporting Information).”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites [22] A. Tsherniak, F. Vazquez, P. G. Montgomery, B. A. Weir, G. Kryukov, G. S. Cowley, S. Gill, W. F. Harrington, S. Pantel, J. M. Krill‐Burger, R. M. Meyers, L. Ali, A. Goodale, Y. Lee, G. Jiang, J. Hsiao, W. F. J. Gerath, S. Howell, E. Merkel, M. Ghandi, L. A. Garraway, D. E. Root, T. R. Golub, J. S. Boehm, W. C. Hahn, Cell 2017, 170, 564. 28753430 PMC5667678 source

      Not checked in the free run.

  • Findingown data

    For the KRAS wild type subgroup, there is no difference between the PTK2 low and high expression groups.

    “while for KRAS wild type subgroup, there is no difference between the PTK2 low and high expression groups (Figure 1A,B)”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites [19] a) E. Cerami, J. Gao, U. Dogrusoz, B. E. Gross, S. O. Sumer, B. A. Aksoy, A. Jacobsen, C. J. Byrne, M. L. Heuer, E. Larsson, Y. Antipin, B. Reva, A. P. Goldberg, C. Sander, N. Schultz, Cancer Discovery 2012, 2, 401; 22588877 PMC3956037 b) C. Hutter, J. C. Zenklusen, Cell 2018, 173, 283. 29625045 source

      Not checked in the free run.

  • May combine two claims. 17
  • Classified as probably not a substantial claim. 10

  • May combine two claims.

    Findingown data

    Some KRAS G12C mutant cell lines show good sensitivity to KRAS G12C inhibition (e.g., Mia PaCa-2 and NCI-H358), while other cell lines are not obviously impacted, suggesting intrinsic resistance.

    “Some of the KRAS G12C mutant cell lines showed good sensitivity to KRAS G12C inhibition (e.g., Mia PaCa‐2 and NCI‐H358), while other cell lines were not obviously impacted by either treatment, suggesting intrinsic resistance (Figure 1C,D).”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    CO-04-0315 is relatively sensitive to 30 mg/kg AMG510 and combination of AMG510 and 25 mg/kg IN10018 exhibited better anti-tumor effects compared to either monotherapies.

    “CO‐04‐0315 is relatively sensitive to 30 mg kg‐1 AMG510 and the combination of AMG510 and 25 mg kg‐1 IN10018 exhibited better anti‐tumor effects compared to either of the monotherapies (Figure 7A,C).”
    Quote in the paperSubstantial claim
    • Rests on an assumption: The CDX and PDX models used are representative of human KRAS G12C mutant cancers for predicting clinical treatment outcomes.

  • May combine two claims.

    Findingown data

    At 48 h postadministration of AMG510-IN10018 combination, clear induction of Phospho FAK Y397 after KRAS G12C inhibition and IN10018 significantly decreased FAK activity.

    “We also monitored FAK signaling activity at the 48 h postadministration time point of the AMG510–IN10018 combination therapy in the various KRAS G12C mutant cell lines, which revealed clear induction of Phospho FAK Y397 after KRAS G12C inhibition. Further, the impact of IN10018 in significantly decreasing FAK activity in the treated cells was evident by the obviously decreased accumulation of”
    Quote in the paperSubstantial claim
    • Rests on an assumption: Phospho FAK Y397 is a valid marker of FAK signaling activity.

  • May combine two claims.

    Findingown data

    AMG510 treatment promotes nuclear YAP accumulation, whereas AMG510-IN10018 combination caused net reduction in YAP accumulation compared to AMG510 alone in NCI-H1792 cells.

    “Western blot‐based and Immunofluorescence‐based monitoring of nuclear YAP level showed that AMG510 treatment promotes nuclear accumulation, whereas the AMG510‐IN10018 combination treatment caused a net reduction in the YAP accumulation compared to AMG510 alone (Figure 5E–G).”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    AMG510 group tumors exhibited apparent inhibition of FAK activity, and IN10018 in combination efficiently reduced this activity.

    “Western blotting and pathology experiments showed that the AMG510 group tumors exhibited apparent inhibition of FAK activity, and the presence of IN10018 in the combination group efficiently reduced this activity (Figure 7G).”
    Quote in the paperSubstantial claim
    • Rests on an assumption: Phospho FAK Y397 is a valid marker of FAK signaling activity.

  • May combine two claims.

    Findingown data

    Enhanced effects from AMG510 for cancer cell killing result from either IN10018 or YAP1 siRNA.

    “We also detected that enhanced effects from AMG510 for cancer cell killing result from either IN10018 or YAP1 siRNA, further supporting that the aberrantly activated FAK–YAP signaling contributes to the compromised efficacy of KRAS G12C inhibitors.”
    Quote in the paperSubstantial claim
    • Rests on an assumption: IN10018 is a FAK inhibitor.

  • May combine two claims.

    Findingown data

    AMG510 treatment resulted in significantly more nuclear YAP accumulation compared to vehicle control in CO-04-0070 model; combination of AMG510 and IN10018 efficiently reduced nuclear YAP accumulation.

    “AMG510 treatment resulted in significantly more nuclear YAP accumulation compared to the vehicle control group in CO‐04‐0070 model and the combination of AMG510 and IN10018 efficiently reduced nuclear YAP accumulation (Figure 7H,I).”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    IN10018 treatment accumulated Phospho YAP S127 within 24 h and reduced CYR61 level in NCI-H358 cells.

    “Phospho YAP S127 which is indicative of YAP signaling inhibition,[ 30 ] was accumulated within 24 h of IN10018 treatment. Consistently, the level of CYR61, a downstream marker of YAP signaling, was apparently reduced by IN10018 treatment (Figure 4E).”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites [30] B. Zhao, X. Wei, W. Li, R. S. Udan, Q. Yang, J. Kim, J. Xie, T. Ikenoue, J. Yu, L. Li, P. Zheng, K. Ye, A. Chinnaiyan, G. Halder, Z. C. Lai, K. L. Guan, Genes Dev. 2007, 21, 2747. 17974916 PMC2045129 source

      Not checked in the free run.

  • May combine two claims.

    Findingown data

    AMG510 treatment efficiently reduced Phospho YAP S127:total YAP ratio, whereas simultaneous FAK inhibition rescued the ratio, and IN10018 reduced extent of nuclear YAP induced by AMG510.

    “we observed that the AMG510 treatment efficiently reduced this ratio, whereas simultaneous FAK inhibition rescued the ratio, and the presence of IN10018 in the treatment reduced the extent of nuclear YAP induced by AMG510.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    LATS1/LATS2 knockdown can rescue cancer cell killing effect from combination treatment of AMG510 and IN10018, suggesting Hippo kinase signaling is involved in FAK-YAP related drug response.

    “Finally, LATS1/LATS2 knockdown can rescue the cancer cell killing effect from the combination treatment of AMG510 and IN10018, suggesting Hippo kinase signaling is involved in this FAK‐YAP related drug response (Figure S6E,F, Supporting Information).”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    Excessive fibrosis was generated by AMG510 treatment, and inhibition of FAK by IN10018 sharply decreased fibrosis in CO-04-0070 model.

    “Masson and Sirius red staining indicated that excessive fibrosis was generated by AMG510 treatment, the inhibition of FAK by IN10018 sharply decreased the fibrosis (Figure 7J–L).”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Interpretationown data

    These results provide direct evidence linking KRAS mutation to altered FAK signaling and prove that FAK inhibition by IN10018 is effective to KRAS mutant cancers.

    “These results provide direct evidence linking KRAS mutation to altered FAK signaling and prove that FAK inhibition by IN10018 is effective to KRAS mutant cancers.”
    Quote in the paperSubstantial claim

  • Classified as probably not a substantial claim.

    Premisecitation

    Activation of FAK signaling has been proposed as a mechanism underlying resistance to targeted therapies.

    “activation of FAK signaling has been proposed as a mechanism underlying resistance to target therapies, specifically because of its impacts on multiple aspects of the tumor microenvironment.[ 15, 16 ]”
    Quote in the paperSubstantial claim0 of 2 sources checked
    • cites [15] C. J. Diaz Osterman, D. Ozmadenci, E. G. Kleinschmidt, K. N. Taylor, A. M. Barrie, S. Jiang, L. M. Bean, F. J. Sulzmaier, C. Jean, I. Tancioni, K. Anderson, S. Uryu, E. A. Cordasco, J. Li, X. L. Chen, G. Fu, M. Ojalill, P. Rappu, J. Heino, A. M. Mark, G. Xu, K. M. Fisch, V. N. Kolev, D. T. Weaver, J. A. Pachter, B. Gyorffy, M. T. McHale, D. C. Connolly, A. Molinolo, D. G. Stupack, D. D. Schlaepfer, eLife 2019, 8, e47327. 31478830 PMC6721800 source

      Not checked in the free run.

    • cites [16] M. C. Frame, A. Serrels, Cancer Cell 2015, 27, 429. 25873166 source

      Not checked in the free run.

  • May combine two claims.

    Premisecitation

    Acquired or adaptive resistance to KRAS G12C inhibitors is a major concern and can limit the long-term therapeutic utility of these treatments.

    “the fact that they are targeted therapies inevitably means they are vulnerable to the development of intrinsic or adaptive resistance;[ 6, 7 ] such resistance can severely limit the long‐term therapeutic utility of these treatments.”
    Quote in the paperSubstantial claim0 of 2 sources checked
    • cites [6] J. Y. Xue, Y. Zhao, J. Aronowitz, T. T. Mai, A. Vides, B. Qeriqi, D. Kim, C. Li, E. de Stanchina, L. Mazutis, D. Risso, P. Lito, Nature 2020, 577, 421. 31915379 PMC7308074 source

      Not checked in the free run.

    • cites [7] Y. Adachi, K. Ito, Y. Hayashi, R. Kimura, T. Z. Tan, R. Yamaguchi, H. Ebi, Clin. Cancer Res. 2020, 26, 5962. 32900796 source

      Not checked in the free run.

  • May combine two claims.

    Premisecitation

    FAK acts downstream of KRAS, and its inhibition is effective in suppressing the progression of KRAS mutant cancer.

    “FAK acts downstream of KRAS, and its inhibition is effective in suppressing the progression of KRAS mutant cancer.[ 12, 13, 14 ]”
    Quote in the paperSubstantial claim0 of 3 sources checked
    • cites [12] K. J. Tang, J. D. Constanzo, N. Venkateswaran, M. Melegari, M. Ilcheva, J. C. Morales, F. Skoulidis, J. V. Heymach, D. A. Boothman, P. P. Scaglioni, Clin. Cancer Res. 2016, 22, 5851. 27220963 PMC5122471 source

      Not checked in the free run.

    • cites [13] Y. Zhou, J. Dang, K. Y. Chang, E. Yau, P. Aza‐Blanc, J. Moscat, T. M. Rana, Cancer Res. 2016, 76, 5777. 27698189 PMC5155639 source

      Not checked in the free run.

    • cites [14] G. Konstantinidou, G. Ramadori, F. Torti, K. Kangasniemi, R. E. Ramirez, Y. Cai, C. Behrens, M. T. Dellinger, R. A. Brekken, I. I. Wistuba, A. Heguy, J. Teruya‐Feldstein, P. P. Scaglioni, Cancer Discovery 2013, 3, 444. 23358651 PMC3625467 source

      Not checked in the free run.

  • May combine two claims.

    Interpretationown data

    Both aberrant FAK-YAP signaling and FAK-related fibrogenesis impact the development of KRAS G12C inhibitor resistance.

    “Mechanistically, it is found that both aberrant FAK–YAP signaling and FAK‐related fibrogenesis impact on the development of KRAS G12C inhibitor resistance.”
    Quote in the paperSubstantial claim

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

    Premisecitation

    KRAS G12C inhibitors AMG510 and MRTX849 are in clinical trials and show promising antitumor activity.

    “This strategy was translated into a series of KRAS G12C inhibitors including AMG510 and MRTX849, both of which are currently in clinical trials.[ 5 ]”
    • Also stated in: § Introduction · plain
      “Although these KRAS G12C inhibitors are showing promising antitumor activity”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [5] a) B. A. Lanman, J. R. Allen, J. G. Allen, A. K. Amegadzie, K. S. Ashton, S. K. Booker, J. J. Chen, N. Chen, M. J. Frohn, G. Goodman, D. J. Kopecky, L. Liu, P. Lopez, J. D. Low, V. Ma, A. E. Minatti, T. T. Nguyen, N. Nishimura, A. J. Pickrell, A. B. Reed, Y. Shin, A. C. Siegmund, N. A. Tamayo, C. M. Tegley, M. C. Walton, H. L. Wang, R. P. Wurz, M. Xue, K. C. Yang, P. Achanta, M. D. Bartberger, J. Canon, L. S. Hollis, J. D. McCarter, C. Mohr, K. Rex, A. Y. Saiki, T. San Miguel, L. P. Volak, K. H. Wang, D. A. Whittington, S. G. Zech, J. R. Lipford, V. J. Cee, J. Med. Chem. 2020, 63, 52; 31820981

      Couldn’t be matched to a paper in Crossref, so it can’t be checked.

  • Classified as probably not a substantial claim.

    Premisecitation

    FAK positively regulates YAP activity through tyrosine phosphorylation of the YAP upstream modulator MOB1, then dissociating the MOB1/LATS complex.

    “On the other hand, FAK was reported to positively regulate YAP activity through tyrosine phosphorylation of the YAP upstream modulator MOB1, then dissociating the MOB1/LATS complex.[ 31, 32 ]”
    Quote in the paperSubstantial claim0 of 2 sources checked
    • cites [31] X. Feng, N. Arang, D. C. Rigiracciolo, J. S. Lee, H. Yeerna, Z. Wang, S. Lubrano, A. Kishore, J. A. Pachter, G. M. Konig, M. Maggiolini, E. Kostenis, D. D. Schlaepfer, P. Tamayo, Q. Chen, E. Ruppin, J. S. Gutkind, Cancer Cell 2019, 35, 457. 30773340 PMC6737937 source

      Not checked in the free run.

    • cites [32] D. Lachowski, E. Cortes, B. Robinson, A. Rice, K. Rombouts, A. E. Del Rio Hernandez, FASEB J. 2018, 32, 1099. 29070586 source

      Not checked in the free run.

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

    Premisecitation

    FAK activation is implicated in the development of resistance against various chemotherapy agents through its regulation of the tumor microenvironment.

    “Interestingly, multiple previous studies have implicated FAK activation in the development of resistance against various chemotherapy agents through its regulation of the tumor microenvironment.[ 11, 15, 44 ]”
    Quote in the paperNot a substantial claim0 of 3 sources checked
    • cites [11] F. J. Sulzmaier, C. Jean, D. D. Schlaepfer, Nat. Rev. Cancer 2014, 14, 598. 25098269 PMC4365862 source

      Not checked in the free run.

    • cites [15] C. J. Diaz Osterman, D. Ozmadenci, E. G. Kleinschmidt, K. N. Taylor, A. M. Barrie, S. Jiang, L. M. Bean, F. J. Sulzmaier, C. Jean, I. Tancioni, K. Anderson, S. Uryu, E. A. Cordasco, J. Li, X. L. Chen, G. Fu, M. Ojalill, P. Rappu, J. Heino, A. M. Mark, G. Xu, K. M. Fisch, V. N. Kolev, D. T. Weaver, J. A. Pachter, B. Gyorffy, M. T. McHale, D. C. Connolly, A. Molinolo, D. G. Stupack, D. D. Schlaepfer, eLife 2019, 8, e47327. 31478830 PMC6721800 source

      Not checked in the free run.

    • cites [44] R. A. Francois, K. Maeng, A. Nawab, F. J. Kaye, S. N. Hochwald, M. Zajac‐Kaye, J. Natl. Cancer Inst. 2015, 107, djv123. PMC4554194 25971297 source

      Not checked in the free run.

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

    Premisecitation

    YAP is required for KRAS-dependent cell transformation; extracellular supplementation of YAP activators can rescue cancer cell killing after KRAS knockdown.

    “On the one hand, it is known that YAP activity is positively correlated with the resistance to therapies targeting KRAS signaling components including RAF and MEK,[ 28, 46 ] and YAP is required for KRAS‐dependent cell transformation: extracellular supplementation of YAP activators can rescue cancer cell killing after KRAS knockdown.[ 47 ]”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [47] D. D. Shao, W. Xue, E. B. Krall, A. Bhutkar, F. Piccioni, X. Wang, A. C. Schinzel, S. Sood, J. Rosenbluh, J. W. Kim, Y. Zwang, T. M. Roberts, D. E. Root, T. Jacks, W. C. Hahn, Cell 2014, 158, 171. 24954536 PMC4110062 source

      Not checked in the free run.

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

    Interpretationreasoning

    A triplet strategy may further enhance CD8+ T cell infiltration within tumors to benefit immunotherapy-related outcomes.

    “A triplet strategy may thus further enhance CD8+ T cell infiltration within tumors to benefit immunotherapy‐related outcomes.”
    Quote in the paperNot a substantial claim

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

    Study design and conductown data

    In vivo tests were replicated at least four times.

    “The in vivo tests were replicated at least for four times.”
    Quote in the paperNot a substantial claim

  • Classified as probably not a substantial claim.

    Premisecitation

    Immunotherapy efficacy can be enhanced by KRAS G12C inhibition, owing to the resulting enhancement of tumor infiltration of CD8+ T cells, macrophages, and dendritic cells.

    “A recent report revealed that immunotherapy efficacy can be enhanced by KRAS G12C inhibition, owing specially to the resulting enhancement of tumor infiltration of CD8+ T cells, macrophages, and dendritic cells.[ 21 ]”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites [21] J. Canon, K. Rex, A. Y. Saiki, C. Mohr, K. Cooke, D. Bagal, K. Gaida, T. Holt, C. G. Knutson, N. Koppada, B. A. Lanman, J. Werner, A. S. Rapaport, T. San Miguel, R. Ortiz, T. Osgood, J. R. Sun, X. Zhu, J. D. McCarter, L. P. Volak, B. E. Houk, M. G. Fakih, B. H. O'Neil, T. J. Price, G. S. Falchook, J. Desai, J. Kuo, R. Govindan, D. S. Hong, W. Ouyang, H. Henary, T. Arvedson, V. J. Cee, J. R. Lipford, Nature 2019, 575, 217. 31666701 source

      Not checked in the free run.

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

    Study design and conductnone given

    All in vitro experiments were tested at least in triplicate.

    “All the in vitro experiments were tested at least in triplicate.”
    Quote in the paperNot a substantial claim

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

    Interpretationreasoning

    A triple combination of KRAS G12C, FAK, and PD1 inhibitors may provide further benefits to treatment outcomes against KRAS G12C mutant cancers.

    “So, we strongly suspect that a triple combination of KRAS G12C, FAK, and PD1 inhibitors may provide further benefits to treatment outcomes against KRAS G12C mutant cancers.”
    Quote in the paperNot a substantial claim
    • Rests on an assumption: IN10018 is a FAK inhibitor.
25 of 25

  • The paper relies on this without support.

    The CDX and PDX models used are representative of human KRAS G12C mutant cancers for predicting clinical treatment outcomes.

    “This study thus illustrates the mechanism of resistance of cancer to the treatment of KRAS G12C inhibitor, as well as an innovative combination therapy to improve treatment outcomes for KRAS G12C mutant cancers.”

  • The paper relies on this without support.

    Phospho FAK Y397 is a valid marker of FAK signaling activity.

    “Phospho FAK Y397 exhibited sustained stimulation in all the tested cell lines, indicating its potential function in adaptive drug resistance to KRAS G12C inhibition (Figure 1E–Q).”

  • The paper relies on this without support.

    The LU-01-0030 PDX model displays primary resistance to KRAS G12C inhibition.

    “We also employed a KRAS G12C mutant NSCLC PDX model LU‐01‐0030 which displays primary resistance to KRAS G12C inhibition for additional drug evaluation.”

  • The paper relies on this without support.

    Preclinical cancer models (cell lines, CDX, and PDX) are predictive of clinical benefit in human KRAS G12C mutant cancers.

    “Given the prevalence of the KRAS G12C mutation in diverse malignancies, it seems that this (and related) combination strategies can benefit the treatment outcomes of many cancer patients.”

  • The paper relies on this without support.

    The KRAS G12C mutation is prevalent in diverse malignancies.

    “Given the prevalence of the KRAS G12C mutation in diverse malignancies, it seems that this (and related) combination strategies can benefit the treatment outcomes of many cancer patients.”

  • The paper relies on this without support.

    IN10018 is a FAK inhibitor.

    “We focused on a clinical‐stage FAK inhibitor: IN10018, which was previously developed by Boehringer‐Ingelheim under the name BI853520.”

  • The paper relies on this without support.

    Fibrosis observed in mouse tumor models limits the anticancer effects of KRAS G12C inhibitors in human cancers.

    “The combination treatment of AMG510 and IN10018 significantly decreased the extent of fibrosis in these tumors, results both supporting that FAK‐related fibrosis may also limit the anticancer effects of KRAS G12C inhibitors and highlighting an additional mechanism through which the combination therapy we tested can help preclude development of drug resistance.”
7 of 7

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?

  • Targeting KRAS-G12C may lead to hyperactivation of focal adhesion kinase (FAK), thereby triggering FAK/SRC signaling and activating the JNK/ERK pathway.

    “In addition, targeting KRAS-G12C may lead to hyperactivation of focal adhesion kinase (FAK), thereby triggering FAK/SRC signaling and activating the JNK/ERK pathway (68).”

    From SRC kinase drives multidrug resistance induced by KRAS-G12C inhibition (2024) · cites this paper as [68]

    The Results section reports sustained stimulation of phosphorylated FAK after KRAS G12C inhibition in the tested mutant cell lines, supporting the FAK-activation part of the claim. It does not establish the proposed FAK/SRC-to-JNK/ERK sequence; the Discussion instead identifies FAK–YAP signaling as the relevant resistance mechanism. No page numbers are available in our copy. (LLM, conf=0.88)

    Experimental — accuracy not yet measured

  • Non-receptor tyrosine kinases such as FAK and SRC can promote resistance to KRAS-targeted therapy.

    “In addition to RTKs, non-receptor tyrosine kinases such as focal adhesion kinase (FAK) and SRC can promote resistance to KRAS-targeted therapy.”

    From Differential Response and Resistance to KRAS-Targeted Therapy (2025) · cites this paper as [147]

    The Discussion (page unavailable) links FAK–YAP signaling to resistance to KRAS G12C inhibitors, supporting the FAK part of the claim. The passages from our copy do not establish that SRC promotes resistance, so they do not support that part. (LLM, conf=0.88)

    Experimental — accuracy not yet measured

  • KRAS G12C inhibition can induce sustained adaptive activation of FAK, mediating therapeutic resistance via both the FAK-YAP signaling axis and FAK-mediated fibrosis.

    “KRAS G12C inhibition can induce sustained adaptive activation of FAK, mediating therapeutic resistance via both the FAK-YAP signaling axis and FAK-mediated fibrosis [147].”

    From Differential Response and Resistance to KRAS-Targeted Therapy (2025) · cites this paper as [147]

  • Targeting FAK could overcome the resistance to KRAS G12C inhibition in vitro and in vivo.

    “Targeting FAK could overcome the resistance to KRAS G12C inhibition in vitro and in vivo [147], suggesting a potential use in combing FAK and KRAS inhibition.”

    From Differential Response and Resistance to KRAS-Targeted Therapy (2025) · cites this paper as [147]

  • A synergistic effect was achieved with the combination treatment of KRAS (G12C) inhibition and a FAK inhibitor (IN10018).

    “A synergistic effect was achieved with the combination treatment of KRAS (G12C) inhibition and a FAK inhibitor (IN10018).”

    From Targeting KRAS mutant cancers: from druggable therapy to drug resistance (2022) · cites this paper as [122]

Self-check

Percentages, sums and reported differences re-computed from the paper’s own numbers.

Each contradiction between two passages listed here was confirmed by an independent check.

  • Figure 5D cell line differs between text and caption

    Minor

    The results text assigns the combination-treatment result to NCI‐H358 cells, whereas the Figure 5D caption assigns it to NCI‐H1792 cells.

  • AMG510 effect on FAK activity conflicts for CO‐04‐0070 tumors

    Minor

    The CO‐04‐0070 results say AMG510 inhibited FAK activity, but the Discussion says AMG510-induced fibrosis in that same model was accompanied by hyperactivated FAK signaling. These are opposite descriptions of a central mechanistic result.

2 of 2

Cites

Flagged first, then by citations

Research works printed but not linked: 3

  1. Molina‐Arcas M., 2019, Sci. Transl. Med.
  2. Katoh K., 2020, Cells
  3. Francois R. A., 2015, J. Natl. Cancer Inst.

Cited by

Sorted by citations

Authors
Zhang, B., Zhang, Y., Zhang, J., Liu, P., Jiao, B., Wang, Z., & Ren, R.
Affiliations
  • Shanghai Jiao Tong University (CN)
  • InxMed (Shanghai) Co., Ltd Shanghai 201202 China
Corresponding author
  • Wang, Zaiqi
  • Ren, Ruibao
ORCID
Venue
Advanced Science
Publisher
Wiley
Published
2021-06-20
Volume
8
Issue
16
Pages
e2100250-e2100250
ISSN
2198-3844 (ISSN-L)
DOI
10.1002/advs.202100250
PMC
PMC8373085
OpenAlex
W3175323447
Semantic Scholar
60ea28b981fdac8901a0efb2c27412622848c429
Cited by
83
References
62
Citations by year
2026: 10 · 2025: 14 · 2024: 33 · 2023: 10 · 2022: 12 · 2021: 4
FWCI
4.63
Type
journal article
Language
en
Open access
Licence
Locations
  • Advanced Science (publishedVersion, cc-by)
  • PubMed Central (publishedVersion, cc-by)
  • PubMed (publishedVersion)
  • DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
Funders
  • National Natural Science Foundation of China (81870112)
  • Samuel Waxman Cancer Research Foundation
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Protein Kinase Regulation and GTPase Signaling
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Cell Biology › Hippo pathway signaling and YAP/TAZ
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Melanoma and MAPK Pathways
Keywords
focal adhesion kinase, KRAS G12C, KRAS inhibitor resistance, combination therapy, fibrogenesis, drug resistance, KRAS-mutant cancer, patient-derived xenografts
MeSH terms
YAP-Signaling Proteins, Animals, Disease Models, Animal, Female, Mice, Inbred BALB C, Mice, Nude, Neoplasms, Signal Transduction, Proto-Oncogene Proteins p21(ras), Cell Line, Tumor, Mice, Focal Adhesion Kinase 1

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.

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