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Cited by 30Ubiquitin and proteasome pathwaysHER2/EGFR in Cancer ResearchProtein Degradation and Inhibitors

Epidermal growth factor receptor (EGFR) is a target of the tumor-suppressor E3 ligase FBXW7

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Boretto, Matteo, Geurts, Maarten H., Gandhi, Shashank, Ma, Ziliang, Staliarova, Nadzeya, Celotti, Martina, Lim, Sangho, He, Gui-Wei +8 more·2024·Proceedings of the National Academy of Sciences
DOI 10.1073/pnas.2309902121PMC PMC10962967
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Abstract

FBXW7 is an E3 ubiquitin ligase that targets proteins for proteasome-mediated degradation and is mutated in various cancer types. Here, we use CRISPR base editors to introduce different FBXW7 hotspot mutations in human colon organoids. Functionally, FBXW7 mutation reduces EGF dependency of organoid growth by ~10,000-fold. Combined transcriptomic and proteomic analyses revealed increased EGFR protein stability in FBXW7 mutants. Two distinct phosphodegron motifs reside in the cytoplasmic tail of EGFR. Mutations in these phosphodegron motifs occur in human cancer. CRISPR-mediated disruption of the phosphodegron motif at T693 reduced EGFR degradation and EGF growth factor dependency. FBXW7 mutant organoids showed reduced sensitivity to EGFR-MAPK inhibitors. These observations were further strengthened in CRC-derived organoid lines and validated in a cohort of patients treated with panitumumab. Our data imply that FBXW7 mutations reduce EGF dependency by disabling EGFR turnover.

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  • Due to the excellent clonal capacity of ASC organoids, these protocols enable rapid screening of various hotspot mutations within a gene in a single round of clone picking.

    “However, due to the excellent clonal capacity of ASC organoids these protocols enable the rapid screening of various hotspot mutations within a gene in a single round of clone picking.51”

    From Protocol to create isogenic disease models from adult stem cell-derived organoids using next-generation CRISPR tools (2024) · cites this paper as [51]

    The Results section, “Establishment of an Organoid Biobank of Recurrent FBXW7 Mutations” (page unavailable), reports generating several FBXW7 mutations in human colon organoids and genotyping individually picked clones. It does not establish that this was a rapid screen enabled by clonal capacity. The Methods section, “Establishment of Clonal Lines and Genotyping” (page unavailable), describes picking organoids after selection and picking growing organoids again a few days later, rather than a single round of picking. (LLM, conf=0.86)

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Authors
Boretto, M., Geurts, M. H., Gandhi, S., Ma, Z., Staliarova, N., Celotti, M., Lim, S., He, G.-W., Millen, R., Driehuis, E., Begthel, H., Smabers, L., Roodhart, J., van Es, J., Wu, W., & Clevers, H.
Affiliations
  • Royal Netherlands Academy of Arts and Sciences (NL)
  • Agency for Science, Technology and Research (SG)
  • Utrecht University (NL)
  • University Medical Center Utrecht (NL)
  • Organoid group, Oncode Institute, Hubrecht Institute, Royal Netherlands Academy of Arts and Sciences and University Medical Center, 3584 CT Utrecht, the Netherlands
Corresponding author
  • Boretto, Matteo
  • Geurts, Maarten H.
  • Gandhi, Shashank
  • Clevers, Hans
ORCID
Venue
Proceedings of the National Academy of Sciences
Publisher
National Academy of Sciences
Published
2024-03-14
Volume
121
Issue
12
Pages
e2309902121-e2309902121
ISSN
  • 0027-8424 (ISSN-L)
  • 1091-6490
DOI
10.1073/pnas.2309902121
PMC
PMC10962967
OpenAlex
W4392818837
Cited by
30
References
5
Citations by year
2026: 13 · 2025: 11 · 2024: 6
FWCI
4.08
Type
journal article
Language
en
Open access
Licence
Locations
  • Proceedings of the National Academy of Sciences (publishedVersion, cc-by-nc-nd)
  • PubMed Central (publishedVersion, cc-by-nc-nd)
  • PubMed (publishedVersion)
  • KNAW Research Portal (The Royal Netherlands Academy of Arts and Sciences) (publishedVersion, other-oa)
  • Utrecht University Repository (Utrecht University) (submittedVersion, other-oa)
Funders
European Molecular Biology Organization (ALTF 769-2019)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Ubiquitin and proteasome pathways
  • Health Sciences › Medicine › Oncology › HER2/EGFR in Cancer Research
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Protein Degradation and Inhibitors
Keywords
FBXW7, EGF receptor, E3 ubiquitin ligase, protein degradation, phosphodegron, colon organoids, colorectal cancer, panitumumab, tumor suppressor
MeSH terms
F-Box-WD Repeat-Containing Protein 7, Epidermal Growth Factor, Humans, Neoplasms, Proteomics, Ubiquitin-Protein Ligases, F-Box Proteins, ErbB Receptors

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