Clinical and Functional Characterization of Atypical KRAS / NRAS Mutations in Metastatic Colorectal Cancer
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Abstract
Purpose: Mutations in KRAS/NRAS (RAS) predict lack of anti-EGFR efficacy in metastatic colorectal cancer (mCRC). However, it is unclear if all RAS mutations have similar impact, and atypical mutations beyond those in standard guidelines exist. Experimental Design: We reviewed 7 tissue and 1 cell-free DNA cohorts of 9,485 patients to characterize atypical RAS variants. Using an in vitro cell-based assay (functional annotation for cancer treatment), Ba/F3 transformation, and in vivo xenograft models of transduced isogenic clones, we assessed signaling changes across mutations. Results: KRAS exon 2, extended RAS, and atypical RAS mutations were noted in 37.8%, 9.5%, and 1.2% of patients, respectively. Among atypical variants, KRAS L19F, Q22K, and D33E occurred at prevalence ≥0.1%, whereas no NRAS codon 117/146 and only one NRAS codon 59 mutation was noted. Atypical RAS mutations had worse overall survival than RAS/BRAF wild-type mCRC (HR, 2.90; 95% confidence interval, 1.24–6.80; P = 0.014). We functionally characterized 114 variants with the FACT assay. All KRAS exon 2 and extended RAS mutations appeared activating. Of 57 atypical RAS variants characterized, 18 (31.6%) had signaling below wild-type, 23 (40.4%) had signaling between wild-type and activating control, and 16 (28.1%) were hyperactive beyond the activating control. Ba/F3 transformation (17/18 variants) and xenograft model (7/8 variants) validation was highly concordant with FACT results, and activating atypical variants were those that occurred at highest prevalence in clinical cohorts. Conclusions: We provide best available evidence to guide treatment when atypical RAS variants are identified. KRAS L19F, Q22K, D33E, and T50I are more prevalent than many guideline-included RAS variants and functionally relevant.
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Mutations at KRAS codons 12, 13, 59, 61, 117, and 146, plus atypical variants, enhance oncogenic potential.
“mutations at other sites in KRAS, including codons 12, 13, 59, 61, 117, and 146 mutations, plus some atypical variants not currently included in standard guidelines in KRAS testing, also enhance its oncogenic potential (17).”
From Expanding the Reach of Precision Oncology by Drugging All <i>KRAS</i> Mutants (2022) · cites this paper as [17]
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Cites
Comprehensive molecular characterization of human colon and rectal cancer
The Cancer Genome Atlas Network (2012) · Nature · doi:10.1038/nature11252 · Cited by 8,499 · see citing works
<i>K-ras</i> Mutations and Benefit from Cetuximab in Advanced Colorectal Cancer
Karapetis, C. S., Khambata-Ford, S., Jonker, D. J., O'Callaghan, C. J., Tu, D., Tebbutt, N. C., Simes, R. J., Chalchal, H., Shapiro, J. D., Robitaille, S., et al. (2008) · New England Journal of Medicine · doi:10.1056/nejmoa0804385 · Cited by 3,622 · see citing works
Amado, R. G., Wolf, M., Peeters, M., Van Cutsem, E., Siena, S., Freeman, D. J., Juan, T., Sikorski, R., Suggs, S., Radinsky, R., et al. (2008) · Journal of Clinical Oncology · doi:10.1200/jco.2007.14.7116 · Cited by 3,207 · see citing works
Panitumumab–FOLFOX4 Treatment and <i>RAS</i> Mutations in Colorectal Cancer
Douillard, J.-Y., Oliner, K. S., Siena, S., Tabernero, J., Burkes, R., Barugel, M., Humblet, Y., Bodoky, G., Cunningham, D., Jassem, J., et al. (2013) · New England Journal of Medicine · doi:10.1056/nejmoa1305275 · Cited by 2,263 · see citing works
AACR Project GENIE: Powering Precision Medicine through an International Consortium
The AACR Project GENIE Consortium, The AACR Project GENIE Consortium, André, F., Arnedos, M., Baras, A. S., Baselga, J., Bedard, P. L., Berger, M. F., Bierkens, M., Calvo, F., et al. (2017) · Cancer Discovery · doi:10.1158/2159-8290.cd-17-0151 · Cited by 1,964 · see citing works
Self-citationCOSMIC: somatic cancer genetics at high-resolution
Forbes, S. A., Beare, D., Boutselakis, H., Bamford, S., Bindal, N., Tate, J., Cole, C. G., Ward, S., Dawson, E., Ponting, L., et al. (2017) · Nucleic Acids Research · doi:10.1093/nar/gkw1121 · Cited by 1,922 · see citing works
Kinase-Dead BRAF and Oncogenic RAS Cooperate to Drive Tumor Progression through CRAF
Heidorn, S. J., Milagre, C. S., Whittaker, S. R., Nourry, A., Niculescu‐Duvaz, I., Dhomen, N. S., Hussain, J., Reis-Filho, J. S., Springer, C. J., Pritchard, C. A., et al. (2010) · Cell · doi:10.1016/j.cell.2009.12.040 · Cited by 1,468 · see citing works
The Frequency of Ras Mutations in Cancer
Prior, I. A., Hood, F. E., & Hartley, J. L. (2020) · Cancer Research · doi:10.1158/0008-5472.can-19-3682 · Cited by 1,130 · see citing works
Maughan, T. S., Adams, R. A., Smith, C. G., Meade, A. M., Seymour, M. T., Wilson, R. H., Idziaszczyk, S., Harris, R., Fisher, D., Kenny, S. L., et al. (2011) · The Lancet · doi:10.1016/s0140-6736(11)60613-2 · Cited by 960 · see citing works
Genomic Correlates of Immune-Cell Infiltrates in Colorectal Carcinoma
Giannakis, M., Mu, X. J., Shukla, S. A., Qian, Z. R., Cohen, O., Nishihara, R., Bahl, S., Cao, Y., Amin‐Mansour, A., Yamauchi, M., et al. (2016) · Cell Reports · doi:10.1016/j.celrep.2016.03.075 · Cited by 912 · see citing works
De Roock, W., Jonker, D. J., Di Nicolantonio, F., Sartore-Bianchi, A., Tu, D., Siena, S., Lamba, S., Arena, S., Frattini, M., Piessevaux, H., et al. (2010) · JAMA · doi:10.1001/jama.2010.1535 · Cited by 748 · see citing works
Germline KRAS mutations cause Noonan syndrome
Schubbert, S., Zenker, M., Rowe, S. L., Böll, S., Klein, C., Bollag, G., van der Burgt, I., Musante, L., Kalscheuer, V., Wehner, L.-E., et al. (2006) · Nature Genetics · doi:10.1038/ng1748 · Cited by 730 · see citing works
Palacios, R., & Steinmetz, M. (1985) · Cell · doi:10.1016/s0092-8674(85)80053-2 · Cited by 716 · see citing works
Douillard, J. Y., Siena, S., Cassidy, J., Tabernero, J., Burkes, R., Barugel, M., Humblet, Y., Bodoky, G., Cunningham, D., Jassem, J., et al. (2014) · Annals of Oncology · doi:10.1093/annonc/mdu141 · Cited by 606 · see citing works
Ras Isoforms Vary in Their Ability to Activate Raf-1 and Phosphoinositide 3-Kinase
Yan, J., Roy, S., Apolloni, A., Lane, A., & Hancock, J. F. (1998) · Journal of Biological Chemistry · doi:10.1074/jbc.273.37.24052 · Cited by 477 · see citing works
Odegaard, J. I., Vincent, J. J., Mortimer, S., Vowles, J. V., Ulrich, B. C., Banks, K. C., Fairclough, S. R., Zill, O. A., Sikora, M., Mokhtari, R. B., et al. (2018) · Clinical Cancer Research · doi:10.1158/1078-0432.ccr-17-3831 · Cited by 437 · see citing works
Meric-Bernstam, F., Brusco, L., Shaw, K., Horombe, C., Kopetz, S., Davies, M. A., Routbort, M., Piha-Paul, S. A., Janku, F., Ueno, N., et al. (2015) · Journal of Clinical Oncology · doi:10.1200/jco.2014.60.4165 · Cited by 424 · see citing works
Self-citationSepulveda, A. R., Hamilton, S. R., Allegra, C. J., Grody, W. W., Cushman‐Vokoun, A. M., Funkhouser, W. K., Kopetz, S., Lieu, C. H., Lindor, N. M., Minsky, B. D., et al. (2017) · Journal of Clinical Oncology · doi:10.1200/jco.2016.71.9807 · Cited by 365 · see citing works
Self-citationTejpar, S., Celik, I., Schlichting, M., Sartorius, U., Bokemeyer, C., & Van Cutsem, E. (2012) · Journal of Clinical Oncology · doi:10.1200/jco.2012.42.2592 · Cited by 353 · see citing works
Modest, D. P., Ricard, I., Heinemann, V., Hegewisch-Becker, S., Schmiegel, W., Porschen, R., Stintzing, S., Graeven, U., Arnold, D., von Weikersthal, L. F., et al. (2016) · Annals of Oncology · doi:10.1093/annonc/mdw261 · Cited by 351 · see citing works
Systematic Functional Annotation of Somatic Mutations in Cancer
Ng, P. K., Li, J., Jeong, K. J., Shao, S. H., Chen, H., Tsang, Y. H., Sengupta, S., Wang, Z., Bhavana, V. H., Tran, R., et al. (2018) · Cancer Cell · doi:10.1016/j.ccell.2018.01.021 · Cited by 328 · see citing works
Self-citationDynamics and Variability of ERK2 Response to EGF in Individual Living Cells
Cohen‐Saidon, C., Cohen, A. A., Sigal, A., Liron, Y., & Alon, U. S. (2009) · Molecular Cell · doi:10.1016/j.molcel.2009.11.025 · Cited by 246 · see citing works
Hong, D. S., Morris, V. K., El Osta, B., Sorokin, A. V., Janku, F., Fu, S., Overman, M. J., Piha-Paul, S., Subbiah, V., Kee, B., et al. (2016) · Cancer Discovery · doi:10.1158/2159-8290.cd-16-0050 · Cited by 219 · see citing works
Self-citationSmith, G. C., Bounds, R., Wolf, H., Steele, R. J., CAREY, F. A., & Wolf, C. R. (2010) · British Journal of Cancer · doi:10.1038/sj.bjc.6605534 · Cited by 176 · see citing works
Role of NRAS mutations as prognostic and predictive markers in metastatic colorectal cancer
Schirripa, M., Cremolini, C., Loupakis, F., Morvillo, M., Bergamo, F., Zoratto, F., Salvatore, L., Antoniotti, C., Marmorino, F., Sensi, E., et al. (2014) · International Journal of Cancer · doi:10.1002/ijc.28955 · Cited by 166 · see citing works
Peeters, M., Kafatos, G., Taylor, A., Gastanaga, V. M., Oliner, K. S., Hechmati, G., Terwey, J.-H., & van Krieken, J. H. (2015) · European Journal of Cancer · doi:10.1016/j.ejca.2015.05.017 · Cited by 163 · see citing works
Biochemical and Functional Characterization of Germ Line KRAS Mutations
Schubbert, S., Bollag, G. E., Lyubynska, N., Nguyen, H. T., Kratz, C. P., Zenker, M., Niemeyer, C. M., Molven, A., & Shannon, K. (2007) · Molecular and Cellular Biology · doi:10.1128/mcb.00965-07 · Cited by 99 · see citing works
Clinical Actionability Enhanced through Deep Targeted Sequencing of Solid Tumors
Chen, K., Meric‐Bernstam, F., Zhao, H., Zhang, Q. C., Ezzeddine, N., Tang, L.-Y., Qi, Y., Mao, Y., Chen, T., Chong, Z., et al. (2015) · Clinical Chemistry · doi:10.1373/clinchem.2014.231100 · Cited by 97 · see citing works
Self-citationWinters, I. P., Chiou, S., Paulk, N. K., McFarland, C. D., Lalgudi, P. V., Ma, R. K., Lisowski, L., Connolly, A. J., Petrov, D. A., Kay, M. A., et al. (2017) · Nature Communications · doi:10.1038/s41467-017-01519-y · Cited by 93 · see citing works
Clinical Features and Outcomes of Patients with Colorectal Cancers Harboring NRAS Mutations
Cercek, A., Braghiroli, M. I., Chou, J. F., Hechtman, J. F., Kemeny, N. E., Saltz, L. B., Capanu, M., & Yaeger, R. D. (2017) · Clinical Cancer Research · doi:10.1158/1078-0432.ccr-17-0400 · Cited by 88 · see citing works
Molecular Landscape of <i>ERBB2/ERBB3</i> Mutated Colorectal Cancer
Loree, J. M., Bailey, A. M., Johnson, A. M., Yu, Y., Wu, W., Bristow, C. A., Davis, J. S., Shaw, K. R., Broaddus, R., Banks, K. C., et al. (2018) · JNCI: Journal of the National Cancer Institute · doi:10.1093/jnci/djy067 · Cited by 68 · see citing works
Self-citationSegelov, E., Thavaneswaran, S., Waring, P. M., Desai, J., Robledo, K. P., Gebski, V., Élez, E., Nott, L. M., Karapetis, C. S., Lunke, S., et al. (2016) · Journal of Clinical Oncology · doi:10.1200/jco.2015.65.6843 · Cited by 59 · see citing works
Loree, J. M., Dowers, A., Tu, D., Jonker, D. J., Edelstein, D. L., Quinn, H., Holtrup, F., Price, T. J., Zalcberg, J. R., Moore, M. J., et al. (2020) · Clinical Cancer Research · doi:10.1158/1078-0432.ccr-20-2710 · Cited by 29 · see citing works
Self-citationKong, K., Ng, P. K., & Scott, K. L. (2017) · Oncotarget · doi:10.18632/oncotarget.17828 · Cited by 18 · see citing works
Self-citationJing, C., Li, H., Du, Y., et al. (2018) · Molecular Medicine Reports · doi:10.3892/mmr.2018.9730 · Cited by 13 · see citing works
Atypical RAS Mutations in Metastatic Colorectal Cancer
Pietrantonio, F., Yaeger, R., Schrock, A. B., Randon, G., Romero-Cordoba, S. L., Rossini, D., Fucà, G., Ross, J. S., Kotani, D., Madison, R., et al. (2019) · JCO Precision Oncology · doi:10.1200/po.19.00136 · Cited by 10 · see citing works
A Novel High-Throughput Multispectral Cell Segmentation Algorithm
Golbstein, J., Tocker, Y., Sharivkin, R., Tarcic, G., & Vidne, M. (2017) · Communications in computer and information science · doi:10.1007/978-3-319-60964-5_66 · Cited by 6 · see citing works
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Cancer Therapy Guided by Mutation Tests: Current Status and Perspectives
Aleksakhina, S. N., & Imyanitov, E. N. (2021) · International Journal of Molecular Sciences · doi:10.3390/ijms222010931 · Cited by 41 · see citing works
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Leite, L. F., Noronha, M. M., de Menezes, J. S. A., da Conceição, L. D., Almeida, L. F. C., Cappellaro, A. P., Belotto, M., Biachi de Castria, T., Peixoto, R. D., & Megid, T. B. C. (2025) · Cancers · doi:10.3390/cancers17172804 · Cited by 32 · see citing works
- Authors
- Loree, J. M., Wang, Y., Syed, M. A., Sorokin, A. V., Coker, O., Xiu, J., Weinberg, B. A., Vanderwalde, A. M., Tesfaye, A., Raymond, V. M., Miron, B., Tarcic, G., Zelichov, O., Broaddus, R. R., Ng, P. K. S., Jeong, K. J., Tsang, Y. H., Mills, G. B., Overman, M. J., Grothey, A., Marshall, J. L., & Kopetz, S.
- Affiliations
- 1BC Cancer, Vancouver, British Columbia, Canada.
- Mayo Clinic (US)
- The University of Texas MD Anderson Cancer Center (US)
- Caris Life Sciences (United States) (US)
- Georgetown University Medical Center (US)
- 6West Cancer Center, Memphis, Tennessee.
- The Barbara Ann Karmanos Cancer Institute (US)
- Guardant (United States) (US)
- 9Novellus, Jerusalem, Israel.
- University of North Carolina at Chapel Hill (US)
- Oregon Health & Science University (US)
- Corresponding author
- Kopetz, Scott
- ORCID
- Loree, Jonathan M.
- Wang, Yucai
- Sorokin, Alexey V.
- Coker, Oluwadara
- Xiu, Joanne
- Weinberg, Benjamin A.
- Vanderwalde, Ari M.
- Tesfaye, Anteneh
- Raymond, Victoria M.
- Miron, Benjamin
- Tarcic, Gabi
- Broaddus, Russell R.
- Ng, Patrick Kwok Shing
- Jeong, Kang Jin
- Tsang, Yiu Huen
- Mills, Gordon B.
- Overman, Michael J.
- Grothey, Axel
- Marshall, John L.
- Kopetz, Scott
- Venue
- Clinical Cancer Research
- Published
- 2021-06-11
- Volume
- 27
- Issue
- 16
- Pages
- 4587-4598
- ISSN
- 1078-0432 (ISSN-L)
- 1557-3265
- PMC
- PMC8364867
- OpenAlex
- W3167814974
- Cited by
- 43
- References
- 37
- Citations by year
- 2026: 7 · 2025: 12 · 2024: 11 · 2023: 8 · 2022: 3 · 2021: 2
- FWCI
- 2.19
- Type
- journal article
- Language
- en
- Open access
- bronze
- doi.org
- Locations
- Clinical Cancer Research (publishedVersion)
- PubMed (publishedVersion)
- PubMed Central (acceptedVersion)
- Funders
- CCSG (CA016672)
- NIH (R01 CA187238, R01 CA184843)
- Topics
- Health Sciences › Medicine › Oncology › Colorectal Cancer Treatments and Studies
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Protein Kinase Regulation and GTPase Signaling
- Health Sciences › Medicine › Pathology and Forensic Medicine › Genetic factors in colorectal cancer
- Keywords
- KRAS, NRAS, metastatic colorectal cancer, anti-EGFR therapy, xenograft models, overall survival
- MeSH terms
- Aged, Female, Humans, Male, Membrane Proteins, Middle Aged, Mutation, Neoplasm Metastasis, Colorectal Neoplasms, Proto-Oncogene Proteins p21(ras), GTP Phosphohydrolases
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