Research ArticleExperimental Studies
Open Access

Cetuximab Enhances the Efficacy of MRTX1133, a Novel KRASG12D Inhibitor, in Colorectal Cancer Treatment

MASAHIRO KATAOKA, MASATO KITAZAWA, SATOSHI NAKAMURA, MAKOTO KOYAMA, YUTA YAMAMOTO, SATORU MIYAZAKI, NAO HONDO, HIROKAZU TANAKA and YUJI SOEJIMA
Anticancer Research October 2023, 43 (10) 4341-4348; DOI: https://doi.org/10.21873/anticanres.16629

Abstract

Background/Aim: Kirsten Rat Sarcoma viral oncogene homolog (KRAS) has remained undruggable for decades. KRAS has predominantly been used to evaluate the applicability of anti-Epidermal Growth Factor Receptor (EGFR) antibody drugs. However, various KRAS inhibitors have recently emerged. Unfortunately, KRAS inhibitors have not been effective against colorectal cancer. Therefore, this study aimed to determine the effects of MRTX1133, a novel KRASG12D inhibitor, in combination with an anti-EGFR antibody, cetuximab, on signal transduction and cell proliferation. Materials and Methods: The KRASG12D-mutated LS513 and KRAS wild-type CACO-2 human colon cancer cell lines were utilized. The KRASG12D mutation was stably transduced into the CACO-2 cells using a retrovirus. We evaluated the effects of the drugs using the CCK-8 assay and assessed the activity of proteins related to the MAPK pathway using western blotting. Results: We demonstrated that the administration of MRTX1133, a novel KRASG12D inhibitor, to KRASG12D-mutated colorectal cancer cells led to feedback activation of the ERK pathway via EGFR activation, inducing drug resistance. Intriguingly, when MRTX1133 was used in combination with cetuximab, KRASG12D-mutant colorectal cancer growth was effectively inhibited, both in vitro and in vivo. Conclusion: The combination of MRTX1133 and cetuximab serves as a potential and promising therapeutic approach for colorectal cancer with KRASG12D mutation. KRASG12D is a frequent genetic mutation not only in colorectal cancer, but also in pancreatic and lung cancer, and the results of this study open new avenues for potential treatment of many cancer patients.

Key Words:

The Rat Sarcoma (RAS) gene family consists of the Kirsten RAS viral oncogene homolog (KRAS), Harvey RAS viral oncogene homolog (HRAS), and neuroblastoma RAS viral (v-ras) oncogene homolog (NRAS). Mutated RAS are the most common oncogenes found in 15% of all human tumors (1, 2). KRAS mutations are one of the most dominant mutations in colorectal cancer and are associated with poor prognosis and drug resistance (3). The most common KRAS mutation variants are G12D (37.5%), G13D (23.0%), G12V (21.7%), and G12C (2.8%) (4). Although KRAS mutations were discovered in the 1980s, drugs that target these mutations have not developed and direct drug targeting remains challenging due to the lack of classic drug binding sites (5).

Recently, the development of KRASG12C inhibitors have rendered KRAS mutations targetable. Sotorasib, an irreversible FDA approved KRASG12C inhibitor (6), has demonstrated high clinical efficacy against non-small cell lung cancer (NSCLC) (7). The sotorasib response rate in colorectal cancer is lower than that in NSCLC (7). This suggests that sotorasib resistance exists in colorectal cancer, which may involve multiple signaling pathways, but not in lung cancer.

Resistance signals to KRASG12C inhibitors can be overcome by the combined use of anti-EGFR antibody drug (8, 9). This indicates that EGFR activity is deeply involved in KRASG12C resistance. The combination of the KRASG12C inhibitor adagrasib and cetuximab has been effective in clinical trials (10).

MRTX1133, a novel KRASG12D inhibitor, is currently being evaluated in clinical trials. The KRASG12D mutation is the most common KRAS mutation found in colorectal cancer patients; therefore, the number of eligible patients is very large. An effective therapeutic strategy that targets KRASG12D mutations in colorectal cancer will lead to favorable outcomes for many patients. Therefore, in this study, we aimed to determine the MRTX1133-generated resistance signals, due to EGFR activation, when used to target KRASG12D mutations in colorectal cancer. Further, we aimed to determine whether KRASG12D inhibition efficacy is enhanced by the concurrent use of anti-EGFR drugs.

Materials and Methods

Cell culture. LS513, the human colon cancer cell line with a KRASG12D mutation, was purchased from American Type Culture Collection (ATCC, Manassas, VA, USA). CACO-2, a KRAS wild-type human colon cancer cell line, was purchased from RIKEN Cell Bank (Ibaraki, Japan). LS513 cells were cultured in RPMI1640 (FUJIFILM Wako Pure Chemical Corporation, Osaka, Japan) supplemented with 10% fetal bovine serum and 100 U/ml penicillin and 100 mg/ml streptomycin. CACO-2 cells were cultured in Dulbecco’s modified Eagles medium (DMEM; FUJIFILM Wako Pure Chemical Corporation) supplemented with 10% fetal bovine serum and 100 U/ml penicillin and 100 mg/ml streptomycin. Both cell lines were incubated at 37°C and 5% CO2.

Antibodies and regents. The monoclonal antibodies rabbit anti-ERK (cat. no. 4695; 1:1,000), monoclonal rabbit anti-p ERK (cat. no. 4376; 1:1,000), monoclonal rabbit anti-MEK1/2 (cat. no. 4694; 1:1,000), monoclonal rabbit anti-p MEK1/2 (cat. no. 9121; 1:1,000), monoclonal rabbit anti-AKT (cat. no. 4691; 1:1,000), monoclonal rabbit anti-p-AKT (cat. no. 4060; 1:1,000), monoclonal rabbit anti-EGFR (cat. no. 4267; 1:1,000), and monoclonal anti-p-EGFR (cat. no. 2237; 1:1,000) were purchased from Cell Signaling Technology, Inc. (Danvers, MA, USA) and used for western blotting. In addition, the monoclonal mouse anti-β actin (cat. no. sc 47787; 1:2,000) antibody was used in western blotting and was purchased from Santa Cruz Biotechnology, Inc. (Dallas, TX, USA). The secondary polyclonal antibodies goat anti mouse (cat. no. P0447; 1:5,000) IgG and goat anti rabbit (cat. no. P0448; 1:5,000) IgG conjugated with HRP were obtained from Dako; Agilent Technologies, Inc. (Santa Clara, CA, USA). MRTX1133 was purchased from Selleck Chemicals (Houston, TX, USA) and Cetuximab was purchased from Merck (Darmstadt, Germany).

Construction and retroviral transduction of the KRAS mutations. Total mRNA was extracted from CACO-2 cells using the NucleoSpin RNAplus kit (Takara Bio Inc., Shiga, Japan; #740984), according to the manufacturer’s instructions. KRAS-4B with the C-terminal FLAG was PCR amplified using CACO-2 cDNA as the template and the PrimeSTAR® Max DNA Polymerase kit (Takara Bio Inc. #R045), according to the manufacturer’s instructions. The amplified KRAS 4B was inserted into the pMXs-IRES-GFP vector by inverse PCR using the In-Fusion®HD Cloning kit (Takara Bio Inc. #639649), according to the manufacturer’s instructions. Next, this vector was used as a template to create a vector with KRASG12D mutant with a C-terminal FLAG. This was performed by inverse PCR using the In-Fusion® HD Cloning kit, according to the manufacturer’s instructions. Next, using the pMXs-IRES-GFP (KRAS wild and G12D) vector as a template, a pDON-5 Neo DNA vector (Takara Bio Inc.), with KRAS wild type and its mutations, was constructed by reverse PCR using the In-Fusion® HD Cloning Kit, according to the manufacturer’s instructions. DNA sequences of all vector constructs were verified on an ABI 3130xl Genetic Analyzer using the BigDye® Terminator v3.1 Cycle Sequencing Kit (Thermo Fisher Scientific, Inc. #4337454), according to the manufacturer’s instructions. These vectors were transfected into amphipathic packaging cells Phoenix-AMPHO (ATCC) using PEI MAX (Polyscience, Inc., Warrington, PA, USA). The transfected cells were cultured, and virus-containing supernatant was collected 24 and 48 h later. Next, CACO-2 cells were infected with retroviral particles on plates coated with RetroNectin (Takara Bio Inc. #T100). pMXs IRES GFP vector transfer efficiency was measured using a flow cytometer (BD FACSCanto II, BD, Franklin Lakes, NJ, USA) and confirmed by the GFP positivity rate, analyzed with Kaluza 2.1 software (Beckman Coulter, Inc., Brea, CA, USA). Following pDON-5Neo DNA vector transduction, transduced CACO-2 cells were selected via a 10-day culture with G418. pDON 5Neo DNA vector transduction efficiency was confirmed using western blotting. Vector creation and retroviral transduction was performed as previously described in Kitazawa et al. (11) and Koyama et al. (12).

Protein sample preparation and western blotting. After seeding and drug treatment, cells were washed with cold PBS and lysed in RIPA Lysis Buffer System (Santa Cruz Biotechnology, Inc. #sc-24948) on ice for 30 min. The lysate was separated by centrifugation at 10,000 × g for 10 min at 4°C, and the resulting supernatant was collected (total lysate). Protein (10-15 μg) was separated using NuPage 4-12% gel (Thermo Fisher Scientific, Inc., Waltham, MA, USA; #NP0322) and electrophoresed onto a PVDF membrane. Membranes were blocked with Tris-buffered saline containing 5% non-fat dry milk and 0.1% Tween-20 for 1 h at room temperature and then probed with primary antibody for 16 h at 4°C. Next, the membranes were incubated with horseradish peroxidase-conjugated secondary antibody for 1 h at room temperature. The protein was detected by chemiluminescence using Immobilon Western HRP (Amersham ECL Prime Western Blotting Detection Reagent; Cytiva #RBN2236, Tokyo, Japan). Densitometry analyses were performed using Image Lab Software version 6.0.1 (Bio-Rad Laboratories, Inc., Hercules, CA, USA).

Cell proliferation. Cell proliferation was measured using the Cell Counting Kit-8 (CCK-8) [Dojindo Molecular Technologies, Inc. (Kumamoto, Japan) #343-07623]. Cells (5.0×103/well) were seeded into a 96-well tissue culture plate and incubated at 37°C for 16 h. Subsequently, each drug (MRTX1133 100 nmol/l, cetuximab 5 μg/ml) or DMSO (control) was added to the plate and the cells were further incubated for 24 or 72 h at 37°C. Then, CCK8 reagent (10 μl/well) was added, and the plate was incubated at 37°C for 2 h. Absorbance was measured at 450 nm using a plate reader (BioTek Epoch Microplate Spectrophotometer, Agilent Technologies, Inc.), and cell viability was calculated relative to DMSO absorbance. Data are representative of three independent series of experiments.

In vivo studies. All animal experiments were performed in compliance with the Guide for the Care and Use of Laboratory Animals (8th Edition) and approved by the Institutional Animal Care and Use Committee of Shinshu University (Matsumoto, Japan, Approval No. 022040). Male BALB/c nude mice (6 to 8 weeks old; 21-26 g) were purchased from CLEA Japan, Inc. (Tokyo, Japan). Mice were housed in pathogen-free rooms on a 12-hour light/dark cycle with free access to water and food. LS531 (5×106) and CACO-2 cells (5×106) stably expressing the KRASG12D mutant in 200 μl of solution [50% Hank’s Balanced Salt Solution (Fujifilm Wako Pure Chemicals Corporation) + 50% Matrigel (Corning, Inc., Corning, NY, USA)] were injected subcutaneously into the mice flank, to create xenograft tumors. Vehicle (12.5% Cremophor, 12.5% ethanol, 75%) or MRTX1133 (0.5 mg/kg) were orally administered to mice once a day for 21 consecutive days (n=4 per group) until tumor size reached 100-200 mm3. Cetuximab (50 mg/kg) was administered intraperitoneally once a week. Tumor volume was measured twice weekly, according to the following formula: volume (mm3)=0.5 × width2 (mm) × length (mm). All mice were sacrificed by cervical dislocation under 3% sevoflurane anesthesia on day 23.

Statistical analysis. All statistical analyses were performed using EZR (Saitama Medical Center, Jichi Medical University, Japan) (13), a graphical user interface for R (The R Foundation for Statistical Computing, Vienna, Austria, version 1.51). Statistical significance was evaluated using an unpaired Student’s t-test or one-way analysis of variance followed by Bonferroni’s correction. All error bars represent the mean and its standard error. p-Values <0.05 were considered statistically significant.

Results

MRTX1133-induced RAS/MAPK pathway activation in KRAS mutant colon cancer cell lines. LS513 and CACO-2 cell lines, both with the KRASG12D mutation, were treated with MRTX1133 and cell lysates were collected at four and 24 h. Using the cell lysate and western blotting, MAPK protein expression was evaluated. At four h following MRTX1133 treatment, pERK and pAKT expression was down- and up-regulated, respectively, in both cell lines. At 24 h post MRTX1133 administration, pERK expression was upregulated in both cell lines (Figure 1A and B). These results suggest that in KRASG12D-mutated CRC cells, reactivation of the MAPK pathway plays a key role in MRTX1133 resistance.

Figure 1.
Figure 1.

The MAPK signaling was reactivated in KRASG12D-mutated CRC cells 24 h after MRTX1133 treatment. (A) LS513 cells were treated with MRTX1133 (100 nM) for 0, 4, and 24 h. Western blot analysis was performed to determine EGFR, RAS/MAPK pathway, and PI3K/AKT pathway activation. β-actin was used as a loading control. (B) KRASG12D-mutated CACO-2 cells were treated with MRTX1133 (100 nM) for 0, 4, and 24 h. Western blot analysis was performed to determine EGFR, RAS/MAPK pathway, and PI3K/AKT pathway activation. β-actin was used as a loading control.

In addition, KRASG12D-mutated LS513 and CACO-2 cells were treated with cetuximab and a combination of the two drugs (MRTX1133 and cetuximab). Again, MAPK-related proteins were evaluated. At four and 24 h post-cetuximab treatment, pERK expression, in both cell lines, was not altered. In contrast in LS513, at 24 h after combination therapy, pERK expression was completely suppressed (Figure 2A). Furthermore, in CACO-2, at 4 and 24 h after combination therapy, pERK expression was significantly down-regulated (Figure 2B). These results suggest that EFGR upstream of KRAS is involved in the KRASG12D inhibitor resistance mechanism; therefore, anti-EGFR antibodies may overcome this resistance.

Figure 2.
Figure 2.

Reactivation of the MAPK signaling was inhibited by the addition of cetuximab. (A) LS513 cells were treated with MRTX1133 (100 nM) and cetuximab (5 μg/ml) for 0, 4, and 24 h. Western blot analysis was performed to determine EGFR, RAS/MAPK pathway, and PI3K/AKT pathway activation. β-actin was used as a loading control. (B) KRASG12D-mutated CACO-2 cells were treated with MRTX1133 (100 nM) and cetuximab (5 μg/ml) for 0, 4, and 24 h. Western blot analysis was performed to determine EGFR, RAS/MAPK pathway, and PI3K/AKT pathway activation. β-actin was used as a loading control.

MRTX1133 and cetuximab combination treatment inhibits proliferation of KRASG12D-mutant colorectal cancer cell lines. Cells were treated with MRTX1133, cetuximab, and the two-drug combination. Then, cell proliferation was evaluated using the CCK-8 assay. MRTX1133 significantly inhibited LS513 proliferation and the two-drug combination (MRTX1133 and cetuximab) further inhibited proliferation (Figure 3A). In contrast, cetuximab alone did not inhibit proliferation. MRTX1133 did not inhibit KRAS wild-type CACO-2 cell proliferation but cetuximab significantly inhibited cell proliferation (Figure 3B). In MRTX1133 did not significantly inhibit KRASG12D-mutant CACO-2 cell proliferation; however, the combined treatment inhibited proliferation. In contrast, cetuximab alone did inhibit proliferation (Figure 3C).

Figure 3.
Figure 3.

Cell proliferation was most inhibited by the combination of MRTX1133 and cetuximab. (A) LS513 cells were treated with MRTX1133 (100 nM) and/or cetuximab (5 μg/ml) and proliferation was measured using the CCK-8 assay. (B) Wild-type CACO-2 cells were treated with MRTX1133 (100 nM) and/or cetuximab (5 μg/ml) and proliferation was measured using the CCK-8 assay. (C) KRASG12D-mutated CACO-2 cells were treated with MRTX1133 (100 nM) and/or cetuximab (5 μg/ml) and proliferation was measured using the CCK-8 assay. *p<0.05 and **p<0.01.

The combination of MRTX1133 and cetuximab significantly inhibited tumorigenesis. The in vivo efficacy of MRTX1133 was evaluated using mouse xenografts derived from KRASG12D-transfected LS513 and CACO-2 cells. MRTX1133 significantly inhibited tumorigenesis in both in vivo models; this effect was further enhanced when MRTX1133 was combined with cetuximab (Figure 4A and B).

Figure 4.
Figure 4.

Significant tumor shrinkage was achieved in vivo with the combination of MRTX1133 and cetuximab. (A) LS513 xenograft was treated with MRTX1133 (n=4, 1 mg/kg orally once a day), cetuximab (n=4, 50 mg/kg intraperitoneally once a week), or a combination of both drugs (n=4) for 21 days. (B) KRASG12D-mutated CACO-2 xenograft was treated with MRTX1133 (n=4, 1 mg/kg orally once a day), cetuximab (n=4, 50 mg/kg intraperitoneally once a week), or combination (n=4) for 21 days. Statistical significance was evaluated at day 21, *p<0.05 and **p<0.01.

Discussion

KRAS, a p21 protein with GTPase activity, has been reported to be present and active in 15% of all human tumors (1, 2). There are numerous types of KRAS mutations, including G12D, G13D, and G12V. The KRASG12D mutation, which is found in 10-12% of all colorectal cancers, is reported to be the most common KRAS mutation in colorectal cancer (14). The KRAS gene, a proto-oncogene, was discovered approximately 40 years ago and has been of significant clinical importance in colorectal cancer due to its association with drug resistance (15-18). However, until recently, drugs acting directly on KRAS mutations were not developed, leaving the gene largely untargeted (4). The advent of various KRAS inhibitors is now changing the treatment strategy for KRAS mutant related colorectal cancer.

Recently, Sotorasib, a KRASG12C inhibitor, was developed. It is a small molecule compound that targets the KRASG12C mutation and is currently used in clinical practice, exhibiting high therapeutic efficacy in non-small cell lung cancer. However, its efficacy in KRASG12C mutant colorectal cancer is lower than that observed in lung cancer (7, 19, 20), suggesting drug resistance. A resistance feedback signal/pathway has been reported with the use of sotorasib in KRASG12C mutant colorectal cancer; however, this resistance can be overcome by the use of anti-EGFR antibody drugs (8, 9). In addition, the use of the KRASG12C inhibitor, Adagrasib, in combination with cetuximab, showed high antitumor activity in patients with KRASG12C mutation-positive metastatic colorectal cancer who had received multiple prior therapies (10).

BRAF inhibitor monotherapy is a promising treatment of many cancers, including melanoma and NSCLC; however, BRAF inhibitor efficacy in colorectal cancer patients with BRAF V600E mutations is limited (21). However, the use of BRAF inhibitors in combination with anti-EGFR inhibitors and MEK inhibitors has been found to suppress feedback reactivation and provide high antitumor efficacy (22-24) and is now widely used in clinical practice.

Recently, MRTX1133, a highly selective inhibitor of the KRASG12D mutant, was developed. It reversibly binds to activated and inactivated KRASG12D protein to inhibit activity (25, 26). MRTX1133 could have the same resistance feedback as the KRASG12C inhibitor. Therefore, we evaluated the changes in protein expression induced by MRTX1133 treatment in KRASG12D mutant colorectal cancer. We showed that MRTX1133 administration up-regulated p-AKT and p-ERK expression at four and 24 h post-treatment, respectively. These results indicate feedback activation of the RAS/MAPK pathway-associated molecules, which transduce proliferative signals downstream of KRAS. Previous studies have shown that vertical inhibition of EGFR and SHP-2 (upstream RAS molecules), suppresses feedback reactivation and has stronger antitumor effects than monotherapy (8, 27). In this study, the combination of cetuximab, a wildly used anti-EGFR antibody in clinical practice, with MRTX1133 demonstrated a high treatment response. Since similar resistance feedback signals exist for KRASG12C and KRASG12D inhibitors, and EGFR inhibition has been shown to enhance antitumor efficacy, it is possible that a common mechanism exists for the various KRAS inhibitors.

Prognosis in colorectal cancer patients with KRAS mutations is poor due to the lack of anti-EGFR antibody efficacy (16, 17). The current study has demonstrated, both in vivo and in vitro, the effectiveness as well as resistance mechanisms of MRTX1133. MRTX1133 is a novel therapeutic that targets KRASG12D, a mutation frequently found in colorectal cancer. The combination of MRTX1133 with anti-EGFR antibodies enhances the MRTX1133 therapeutic effect and the use of anti-EGFR antibodies overcomes MRTX1133 resistance in other cancer types, such as pancreatic and lung cancer. Therefore, the combination of MRTX1133 with anti-EGFR antibodies is a potential effective therapeutic strategy for the treatment of colorectal cancer.

Conclusion

In the current study, we revealed that resistance to the novel KRASG12D inhibitor, MRTX1133, in colorectal cancer cells arises from the feedback reactivation of the MAPK pathway via EGFR. The use of anti-EGFR antibodies, in addition to MRTX1133 treatment, successfully overcomes this resistance signal, demonstrating potent anti-tumor effects both in vitro and in vivo. This suggests that MRTX1133, used in combination with anti-EGFR antibodies, could serve as an effective therapeutic strategy for colorectal cancer.

Acknowledgements

The Authors would like to thank Editage (www.editage.com) for English language editing.

Footnotes

  • Authors’ Contributions

    MKa: Data curation, Formal analysis, Investigation and Writing-original draft; MKi: Conceptualization, Methodology, Funding acquisition and Writing-review & editing; SN: Conceptualization and Methodology; MKo: Conceptualization and Methodology; YY: Conceptualization and Methodology; SM: Formal analysis and Methodology; NH: Data curation and Methodology; HT: Data curation and Investigation; YS: Supervision and Writing-review & editing. All Authors read and approved the final manuscript.

  • Conflicts of Interest

    The Authors have no competing interests to declare in relation to this study.

  • Funding

    This work was supported by the Japan Society for the Promotion of Science KAKENHI (grant no. JP22K15555, to M. Kitazawa).

  • Received July 29, 2023.
  • Revision received August 29, 2023.
  • Accepted August 30, 2023.

This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY-NC-ND) 4.0 international license (https://creativecommons.org/licenses/by-nc-nd/4.0).

References

  1. ↵
    1. Zehir A,
    2. Benayed R,
    3. Shah RH,
    4. Syed A,
    5. Middha S,
    6. Kim HR,
    7. Srinivasan P,
    8. Gao J,
    9. Chakravarty D,
    10. Devlin SM,
    11. Hellmann MD,
    12. Barron DA,
    13. Schram AM,
    14. Hameed M,
    15. Dogan S,
    16. Ross DS,
    17. Hechtman JF,
    18. DeLair DF,
    19. Yao J,
    20. Mandelker DL,
    21. Cheng DT,
    22. Chandramohan R,
    23. Mohanty AS,
    24. Ptashkin RN,
    25. Jayakumaran G,
    26. Prasad M,
    27. Syed MH,
    28. Rema AB,
    29. Liu ZY,
    30. Nafa K,
    31. Borsu L,
    32. Sadowska J,
    33. Casanova J,
    34. Bacares R,
    35. Kiecka IJ,
    36. Razumova A,
    37. Son JB,
    38. Stewart L,
    39. Baldi T,
    40. Mullaney KA,
    41. Al-Ahmadie H,
    42. Vakiani E,
    43. Abeshouse AA,
    44. Penson AV,
    45. Jonsson P,
    46. Camacho N,
    47. Chang MT,
    48. Won HH,
    49. Gross BE,
    50. Kundra R,
    51. Heins ZJ,
    52. Chen HW,
    53. Phillips S,
    54. Zhang H,
    55. Wang J,
    56. Ochoa A,
    57. Wills J,
    58. Eubank M,
    59. Thomas SB,
    60. Gardos SM,
    61. Reales DN,
    62. Galle J,
    63. Durany R,
    64. Cambria R,
    65. Abida W,
    66. Cercek A,
    67. Feldman DR,
    68. Gounder MM,
    69. Hakimi AA,
    70. Harding JJ,
    71. Iyer G,
    72. Janjigian YY,
    73. Jordan EJ,
    74. Kelly CM,
    75. Lowery MA,
    76. Morris LGT,
    77. Omuro AM,
    78. Raj N,
    79. Razavi P,
    80. Shoushtari AN,
    81. Shukla N,
    82. Soumerai TE,
    83. Varghese AM,
    84. Yaeger R,
    85. Coleman J,
    86. Bochner B,
    87. Riely GJ,
    88. Saltz LB,
    89. Scher HI,
    90. Sabbatini PJ,
    91. Robson ME,
    92. Klimstra DS,
    93. Taylor BS,
    94. Baselga J,
    95. Schultz N,
    96. Hyman DM,
    97. Arcila ME,
    98. Solit DB,
    99. Ladanyi M,
    100. Berger MF
    : Mutational landscape of metastatic cancer revealed from prospective clinical sequencing of 10,000 patients. Nat Med 23(6): 703-713, 2017. DOI: 10.1038/nm.4333
    OpenUrlCrossRefPubMed
  2. ↵
    1. Prior IA,
    2. Hood FE,
    3. Hartley JL
    : The frequency of Ras mutations in cancer. Cancer Res 80(14): 2969-2974, 2020. DOI: 10.1158/0008-5472.CAN-19-3682
    OpenUrlAbstract/FREE Full Text
  3. ↵
    1. Perkins G,
    2. Pilati C,
    3. Blons H,
    4. Laurent-Puig P
    : Beyond KRAS status and response to anti-EGFR therapy in metastatic colorectal cancer. Pharmacogenomics 15(7): 1043-1052, 2014. DOI: 10.2217/pgs.14.66
    OpenUrlCrossRefPubMed
  4. ↵
    1. Moore AR,
    2. Rosenberg SC,
    3. McCormick F,
    4. Malek S
    : RAS-targeted therapies: is the undruggable drugged? Nat Rev Drug Discov 19(8): 533-552, 2020. DOI: 10.1038/s41573-020-0068-6
    OpenUrlCrossRefPubMed
  5. ↵
    1. Chida K,
    2. Kotani D,
    3. Masuishi T,
    4. Kawakami T,
    5. Kawamoto Y,
    6. Kato K,
    7. Fushiki K,
    8. Sawada K,
    9. Kumanishi R,
    10. Shirasu H,
    11. Matsubara Y,
    12. Yuki S,
    13. Komatsu Y,
    14. Yamazaki K,
    15. Yoshino T
    : The prognostic impact of KRAS G12C mutation in patients with metastatic colorectal cancer: a multicenter retrospective observational study. Oncologist 26(10): 845-853, 2021. DOI: 10.1002/onco.13870
    OpenUrlCrossRefPubMed
  6. ↵
    1. Canon J,
    2. Rex K,
    3. Saiki AY,
    4. Mohr C,
    5. Cooke K,
    6. Bagal D,
    7. Gaida K,
    8. Holt T,
    9. Knutson CG,
    10. Koppada N,
    11. Lanman BA,
    12. Werner J,
    13. Rapaport AS,
    14. San Miguel T,
    15. Ortiz R,
    16. Osgood T,
    17. Sun J,
    18. Zhu X,
    19. Mccarter JD,
    20. Volak LP,
    21. Houk BE,
    22. Fakih MG,
    23. O’neil BH,
    24. Price TJ,
    25. Falchook GS,
    26. Desai J,
    27. Kuo J,
    28. Govindan R,
    29. Hong DS,
    30. Ouyang W,
    31. Henary H,
    32. Arvedson T,
    33. Cee VJ,
    34. Lipford JR
    : The clinical KRAS(G12C) inhibitor AMG 510 drives anti-tumour immunity. Nature 575(7781): 217-223, 2019. DOI: 10.1038/s41586-019-1694-1
    OpenUrlCrossRefPubMed
  7. ↵
    1. Skoulidis F,
    2. Li BT,
    3. Dy GK,
    4. Price TJ,
    5. Falchook GS,
    6. Wolf J,
    7. Italiano A,
    8. Schuler M,
    9. Borghaei H,
    10. Barlesi F,
    11. Kato T,
    12. Curioni-Fontecedro A,
    13. Sacher A,
    14. Spira A,
    15. Ramalingam SS,
    16. Takahashi T,
    17. Besse B,
    18. Anderson A,
    19. Ang A,
    20. Tran Q,
    21. Mather O,
    22. Henary H,
    23. Ngarmchamnanrith G,
    24. Friberg G,
    25. Velcheti V,
    26. Govindan R
    : Sotorasib for lung cancers with KRAS p.G12C mutation. N Engl J Med 384(25): 2371-2381, 2021. DOI: 10.1056/NEJMoa2103695
    OpenUrlCrossRefPubMed
  8. ↵
    1. Amodio V,
    2. Yaeger R,
    3. Arcella P,
    4. Cancelliere C,
    5. Lamba S,
    6. Lorenzato A,
    7. Arena S,
    8. Montone M,
    9. Mussolin B,
    10. Bian Y,
    11. Whaley A,
    12. Pinnelli M,
    13. Murciano-Goroff YR,
    14. Vakiani E,
    15. Valeri N,
    16. Liao WL,
    17. Bhalkikar A,
    18. Thyparambil S,
    19. Zhao HY,
    20. de Stanchina E,
    21. Marsoni S,
    22. Siena S,
    23. Bertotti A,
    24. Trusolino L,
    25. Li BT,
    26. Rosen N,
    27. Di Nicolantonio F,
    28. Bardelli A,
    29. Misale S
    : EGFR blockade reverts resistance to KRAS(G12C) inhibition in colorectal cancer. Cancer Discov 10(8): 1129-1139, 2020. DOI: 10.1158/2159-8290.CD-20-0187
    OpenUrlAbstract/FREE Full Text
  9. ↵
    1. Yaeger R,
    2. Mezzadra R,
    3. Sinopoli J,
    4. Bian Y,
    5. Marasco M,
    6. Kaplun E,
    7. Gao Y,
    8. Zhao H,
    9. Paula ADC,
    10. Zhu Y,
    11. Perez AC,
    12. Chadalavada K,
    13. Tse E,
    14. Chowdhry S,
    15. Bowker S,
    16. Chang Q,
    17. Qeriqi B,
    18. Weigelt B,
    19. Nanjangud GJ,
    20. Berger MF,
    21. Der-Torossian H,
    22. Anderes K,
    23. Socci ND,
    24. Shia J,
    25. Riely GJ,
    26. Murciano-Goroff YR,
    27. Li BT,
    28. Christensen JG,
    29. Reis-Filho JS,
    30. Solit DB,
    31. de Stanchina E,
    32. Lowe SW,
    33. Rosen N,
    34. Misale S
    : Molecular characterization of acquired resistance to KRASG12C-EGFR inhibition in colorectal cancer. Cancer Discov 13(1): 41-55, 2023. DOI: 10.1158/2159-8290.CD-22-0405
    OpenUrlCrossRefPubMed
  10. ↵
    1. Yaeger R,
    2. Weiss J,
    3. Pelster MS,
    4. Spira AI,
    5. Barve M,
    6. Ou SI,
    7. Leal TA,
    8. Bekaii-Saab TS,
    9. Paweletz CP,
    10. Heavey GA,
    11. Christensen JG,
    12. Velastegui K,
    13. Kheoh T,
    14. Der-Torossian H,
    15. Klempner SJ
    : Adagrasib with or without cetuximab in colorectal cancer with mutated KRAS G12C. N Engl J Med 388(1): 44-54, 2023. DOI: 10.1056/NEJMoa2212419
    OpenUrlCrossRefPubMed
  11. ↵
    1. Kitazawa M,
    2. Miyagawa Y,
    3. Koyama M,
    4. Nakamura S,
    5. Hondo N,
    6. Miyazaki S,
    7. Muranaka F,
    8. Tokumaru S,
    9. Yamamoto Y,
    10. Ehara T,
    11. Kuroiwa M,
    12. Tanaka H,
    13. Komatsu D,
    14. Takeoka M,
    15. Soejima Y
    : Drug sensitivity profile of minor KRAS mutations in colorectal cancer using mix culture assay: The effect of AMG-510, a novel KRAS G12C selective inhibitor, on colon cancer cells is markedly enhanced by the combined inhibition of MEK and BCL-XL. Mol Clin Oncol 15(1): 148, 2021. DOI: 10.3892/mco.2021.2310
    OpenUrlCrossRefPubMed
  12. ↵
    1. Koyama M,
    2. Kitazawa M,
    3. Nakamura S,
    4. Matsumura T,
    5. Miyazaki S,
    6. Miyagawa Y,
    7. Muranaka F,
    8. Tokumaru S,
    9. Okumura M,
    10. Yamamoto Y,
    11. Ehara T,
    12. Hondo N,
    13. Takahata S,
    14. Takeoka M,
    15. Miyagawa SI,
    16. Soejima Y
    : Low dose trametinib and Bcl xL antagonist have a specific antitumor effect in KRAS mutated colorectal cancer cells. Int J Oncol 57(5): 1179-1191, 2020. DOI: 10.3892/ijo.2020.5117
    OpenUrlCrossRefPubMed
  13. ↵
    1. Kanda Y
    : Investigation of the freely available easy-to-use software ‘EZR’ for medical statistics. Bone Marrow Transplant 48(3): 452-458, 2013. DOI: 10.1038/bmt.2012.244
    OpenUrlCrossRefPubMed
  14. ↵
    1. Li S,
    2. Balmain A,
    3. Counter CM
    : A model for RAS mutation patterns in cancers: finding the sweet spot. Nat Rev Cancer 18(12): 767-777, 2018. DOI: 10.1038/s41568-018-0076-6
    OpenUrlCrossRefPubMed
  15. ↵
    1. Schubbert S,
    2. Shannon K,
    3. Bollag G
    : Hyperactive Ras in developmental disorders and cancer. Nat Rev Cancer 7(4): 295-308, 2007. DOI: 10.1038/nrc2109
    OpenUrlCrossRefPubMed
  16. ↵
    1. Karapetis CS,
    2. Khambata-Ford S,
    3. Jonker DJ,
    4. O’Callaghan CJ,
    5. Tu D,
    6. Tebbutt NC,
    7. Simes RJ,
    8. Chalchal H,
    9. Shapiro JD,
    10. Robitaille S,
    11. Price TJ,
    12. Shepherd L,
    13. Au H,
    14. Langer C,
    15. Moore MJ,
    16. Zalcberg JR
    : K-ras mutations and benefit from cetuximab in advanced colorectal cancer. N Engl J Med 359(17): 1757-1765, 2008. DOI: 10.1056/NEJMoa0804385
    OpenUrlCrossRefPubMed
  17. ↵
    1. Amado RG,
    2. Wolf M,
    3. Peeters M,
    4. Van Cutsem E,
    5. Siena S,
    6. Freeman DJ,
    7. Juan T,
    8. Sikorski R,
    9. Suggs S,
    10. Radinsky R,
    11. Patterson SD,
    12. Chang DD
    : Wild-type KRAS is required for panitumumab efficacy in patients with metastatic colorectal cancer. J Clin Oncol 26(10): 1626-1634, 2008. DOI: 10.1200/JCO.2007.14.7116
    OpenUrlAbstract/FREE Full Text
  18. ↵
    1. Imamura Y,
    2. Lochhead P,
    3. Yamauchi M,
    4. Kuchiba A,
    5. Qian ZR,
    6. Liao X,
    7. Nishihara R,
    8. Jung S,
    9. Wu K,
    10. Nosho K,
    11. Wang YE,
    12. Peng S,
    13. Bass AJ,
    14. Haigis KM,
    15. Meyerhardt JA,
    16. Chan AT,
    17. Fuchs CS,
    18. Ogino S
    : Analyses of clinicopathological, molecular, and prognostic associations of KRAS codon 61 and codon 146 mutations in colorectal cancer: cohort study and literature review. Mol Cancer 13: 135, 2014. DOI: 10.1186/1476-4598-13-135
    OpenUrlCrossRefPubMed
  19. ↵
    1. Fakih MG,
    2. Kopetz S,
    3. Kuboki Y,
    4. Kim TW,
    5. Munster PN,
    6. Krauss JC,
    7. Falchook GS,
    8. Han S,
    9. Heinemann V,
    10. Muro K,
    11. Strickler JH,
    12. Hong DS,
    13. Denlinger CS,
    14. Girotto G,
    15. Lee M,
    16. Henary H,
    17. Tran Q,
    18. Park JK,
    19. Ngarmchamnanrith G,
    20. Prenen H,
    21. Price TJ
    : Sotorasib for previously treated colorectal cancers with KRASG12C mutation (CodeBreaK100): a prespecified analysis of a single-arm, phase 2 trial. Lancet Oncol 23(1): 115-124, 2022. DOI: 10.1016/S1470-2045(21)00605-7
    OpenUrlCrossRefPubMed
  20. ↵
    1. Hong DS,
    2. Fakih MG,
    3. Strickler JH,
    4. Desai J,
    5. Durm GA,
    6. Shapiro GI,
    7. Falchook GS,
    8. Price TJ,
    9. Sacher A,
    10. Denlinger CS,
    11. Bang YJ,
    12. Dy GK,
    13. Krauss JC,
    14. Kuboki Y,
    15. Kuo JC,
    16. Coveler AL,
    17. Park K,
    18. Kim TW,
    19. Barlesi F,
    20. Munster PN,
    21. Ramalingam SS,
    22. Burns TF,
    23. Meric-Bernstam F,
    24. Henary H,
    25. Ngang J,
    26. Ngarmchamnanrith G,
    27. Kim J,
    28. Houk BE,
    29. Canon J,
    30. Lipford JR,
    31. Friberg G,
    32. Lito P,
    33. Govindan R,
    34. Li BT
    : KRAS(G12C) inhibition with sotorasib in advanced solid tumors. N Engl J Med 383(13): 1207-1217, 2020. DOI: 10.1056/NEJMoa1917239
    OpenUrlCrossRefPubMed
  21. ↵
    1. Hyman DM,
    2. Puzanov I,
    3. Subbiah V,
    4. Faris JE,
    5. Chau I,
    6. Blay JY,
    7. Wolf J,
    8. Raje NS,
    9. Diamond EL,
    10. Hollebecque A,
    11. Gervais R,
    12. Elez-Fernandez ME,
    13. Italiano A,
    14. Hofheinz RD,
    15. Hidalgo M,
    16. Chan E,
    17. Schuler M,
    18. Lasserre SF,
    19. Makrutzki M,
    20. Sirzen F,
    21. Veronese ML,
    22. Tabernero J,
    23. Baselga J
    : Vemurafenib in Multiple Nonmelanoma Cancers with BRAF V600 Mutations. N Engl J Med 373(8): 726-736, 2015. DOI: 10.1056/NEJMoa1502309
    OpenUrlCrossRefPubMed
  22. ↵
    1. Kopetz S,
    2. Grothey A,
    3. Yaeger R,
    4. Van Cutsem E,
    5. Desai J,
    6. Yoshino T,
    7. Wasan H,
    8. Ciardiello F,
    9. Loupakis F,
    10. Hong YS,
    11. Steeghs N,
    12. Guren TK,
    13. Arkenau H,
    14. Garcia-Alfonso P,
    15. Pfeiffer P,
    16. Orlov S,
    17. Lonardi S,
    18. Elez E,
    19. Kim T,
    20. Schellens JH,
    21. Guo C,
    22. Krishnan A,
    23. Dekervel J,
    24. Morris V,
    25. Calvo Ferrandiz A,
    26. Tarpgaard L,
    27. Braun M,
    28. Gollerkeri A,
    29. Keir C,
    30. Maharry K,
    31. Pickard M,
    32. Christy-Bittel J,
    33. Anderson L,
    34. Sandor V,
    35. Tabernero J
    : Encorafenib, binimetinib, and cetuximab in BRAF V600E–mutated colorectal cancer. N Engl J Med 381(17): 1632-1643, 2019. DOI: 10.1056/NEJMoa1908075
    OpenUrlCrossRefPubMed
    1. Corcoran RB,
    2. Atreya CE,
    3. Falchook GS,
    4. Kwak EL,
    5. Ryan DP,
    6. Bendell JC,
    7. Hamid O,
    8. Messersmith WA,
    9. Daud A,
    10. Kurzrock R,
    11. Pierobon M,
    12. Sun P,
    13. Cunningham E,
    14. Little S,
    15. Orford K,
    16. Motwani M,
    17. Bai Y,
    18. Patel K,
    19. Venook AP,
    20. Kopetz S
    : Combined BRAF and MEK inhibition with dabrafenib and trametinib in BRAF V600-mutant colorectal cancer. J Clin Oncol 33(34): 4023-4031, 2015. DOI: 10.1200/JCO.2015.63.2471
    OpenUrlAbstract/FREE Full Text
  24. ↵
    1. Corcoran RB,
    2. André T,
    3. Atreya CE,
    4. Schellens JHM,
    5. Yoshino T,
    6. Bendell JC,
    7. Hollebecque A,
    8. McRee AJ,
    9. Siena S,
    10. Middleton G,
    11. Muro K,
    12. Gordon MS,
    13. Tabernero J,
    14. Yaeger R,
    15. O’Dwyer PJ,
    16. Humblet Y,
    17. De Vos F,
    18. Jung AS,
    19. Brase JC,
    20. Jaeger S,
    21. Bettinger S,
    22. Mookerjee B,
    23. Rangwala F,
    24. Van Cutsem E
    : Combined BRAF, EGFR, and MEK inhibition in patients with BRAF(V600E)-mutant colorectal cancer. Cancer Discov 8(4): 428-443, 2018. DOI: 10.1158/2159-8290.CD-17-1226
    OpenUrlAbstract/FREE Full Text
  25. ↵
    1. Wang X,
    2. Allen S,
    3. Blake JF,
    4. Bowcut V,
    5. Briere DM,
    6. Calinisan A,
    7. Dahlke JR,
    8. Fell JB,
    9. Fischer JP,
    10. Gunn RJ,
    11. Hallin J,
    12. Laguer J,
    13. Lawson JD,
    14. Medwid J,
    15. Newhouse B,
    16. Nguyen P,
    17. O’leary JM,
    18. Olson P,
    19. Pajk S,
    20. Rahbaek L,
    21. Rodriguez M,
    22. Smith CR,
    23. Tang TP,
    24. Thomas NC,
    25. Vanderpool D,
    26. Vigers GP,
    27. Christensen JG,
    28. Marx MA
    : Identification of MRTX1133, a noncovalent, potent, and selective KRASG12D inhibitor. J Med Chem 65(4): 3123-3133, 2022. DOI: 10.1021/acs.jmedchem.1c01688
    OpenUrlCrossRefPubMed
  26. ↵
    1. Hallin J,
    2. Bowcut V,
    3. Calinisan A,
    4. Briere DM,
    5. Hargis L,
    6. Engstrom LD,
    7. Laguer J,
    8. Medwid J,
    9. Vanderpool D,
    10. Lifset E,
    11. Trinh D,
    12. Hoffman N,
    13. Wang X,
    14. David Lawson J,
    15. Gunn RJ,
    16. Smith CR,
    17. Thomas NC,
    18. Martinson M,
    19. Bergstrom A,
    20. Sullivan F,
    21. Bouhana K,
    22. Winski S,
    23. He L,
    24. Fernandez-Banet J,
    25. Pavlicek A,
    26. Haling JR,
    27. Rahbaek L,
    28. Marx MA,
    29. Olson P,
    30. Christensen JG
    : Anti-tumor efficacy of a potent and selective non-covalent KRASG12D inhibitor. Nat Med 28(10): 2171-2182, 2022. DOI: 10.1038/s41591-022-02007-7
    OpenUrlCrossRefPubMed
  27. ↵
    1. Ryan MB,
    2. Fece de la Cruz F,
    3. Phat S,
    4. Myers DT,
    5. Wong E,
    6. Shahzade HA,
    7. Hong CB,
    8. Corcoran RB
    : Vertical pathway inhibition overcomes adaptive feedback resistance to KRAS(G12C) inhibition. Clin Cancer Res 26(7): 1633-1643, 2020. DOI: 10.1158/1078-0432.CCR-19-3523
    OpenUrlAbstract/FREE Full Text
View Abstract
Back to top

In this issue

Print
Download PDF
Article Alerts
Email Article
Citation Tools
Reprints and Permissions
Cetuximab Enhances the Efficacy of MRTX1133, a Novel KRASG12D Inhibitor, in Colorectal Cancer Treatment
MASAHIRO KATAOKA, MASATO KITAZAWA, SATOSHI NAKAMURA, MAKOTO KOYAMA, YUTA YAMAMOTO, SATORU MIYAZAKI, NAO HONDO, HIROKAZU TANAKA, YUJI SOEJIMA
Anticancer Research Oct 2023, 43 (10) 4341-4348; DOI: 10.21873/anticanres.16629
Twitter logo Facebook logo Mendeley logo

Jump to section

Keywords