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

Matteo Boretto; Maarten H. Geurts; Shashank Gandhi; Ziliang Ma; Nadzeya Staliarova; Martina Celotti; Sangho Lim; Gui Wei He; Rosemary Millen; Else Driehuis; Harry Begthel; Lidwien Smabers; Jeanine Roodhart; Johan van Es; Wei Wu; Hans Clevers

doi:10.1073/pnas.2309902121

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

Matteo Boretto
, Maarten H. Geurts
, Shashank Gandhi
, Ziliang Ma
, Nadzeya Staliarova
, Martina Celotti
, Sangho Lim
, Gui Wei He
, Rosemary Millen
, Else Driehuis
, Harry Begthel
, Lidwien Smabers
, Jeanine Roodhart
, Johan van Es
, Wei Wu
, Hans Clevers

Research output: Contribution to journal › Article › Academic › peer-review

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.

Original language	English
Article number	e2309902121
Journal	Proceedings of the National Academy of Sciences of the United States of America
Volume	121
Issue number	12
DOIs	https://doi.org/10.1073/pnas.2309902121
Publication status	Published - 19 Mar 2024

Bibliographical note

Publisher Copyright:
Copyright © 2024 the Author(s). Published by PNAS.

Funding

ACKNOWLEDGMENTS. We thank Stieneke van den Brink for the production of R-spondin1-conditioned medium, essential for the organoid expansion medium. This work was supported by an award from the Cancer Research UK Grand Challenge (C6307/A29058) and the Mark Foundation for Cancer Research to the SPECIFICANCER team (H.C., M.H.G., G.-W.H., and M.C.) and by the Netherlands Organ-on-Chip Initiative, an NWO Gravitation project (024.003.001) funded by the Ministry of Education, Culture and Science of the government of the Netherlands. M.B. is a postdoctoral researcher supported by a long-term EMBO fellowship (ALTF 769-2019). We thank Stieneke van den Brink for the production of R-spondin1-conditioned medium, essential for the organoid expansion medium. This work was supported by an award from the Cancer Research UK Grand Challenge (C6307/A29058) and the Mark Foundation for Cancer Research to the SPECIFICANCER team (H.C., M.H.G., G.-W.H., and M.C.) and by the Netherlands Organ-on-Chip Initiative, an NWO Gravitation project (024.003.001) funded by the Ministry of Education, Culture and Science of the government of the Netherlands. M.B. is a postdoctoral researcher supported by a long-term EMBO fellowship (ALTF 769-2019).

Funders	Funder number
Cancer Research UK
Mark Foundation For Cancer Research
Ministerie van onderwijs, cultuur en wetenschap
Nederlandse Organisatie voor Wetenschappelijk Onderzoek	024.003.001
European Molecular Biology Organization	ALTF 769-2019
Cancer Research UK Grand Challenge	C6307/A29058

Keywords

colorectal cancer
EGFR
FBXW7
organoids

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

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10.1073/pnas.2309902121Licence: CC BY-NC-ND

boretto-et-al-2024-epidermal-growth-factor-receptor-(egfr)-is-a-target-of-the-tumor-suppressor-e3-ligase-fbxw7Final published version, 5.45 MBLicence: CC BY-NC-ND

Cite this

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. (2024). Epidermal growth factor receptor (EGFR) is a target of the tumor-suppressor E3 ligase FBXW7. Proceedings of the National Academy of Sciences of the United States of America, 121(12), Article e2309902121. https://doi.org/10.1073/pnas.2309902121

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

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 \textasciitilde{}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.",

keywords = "colorectal cancer, EGFR, FBXW7, organoids",

author = "Matteo Boretto and Geurts, \{Maarten H.\} and Shashank Gandhi and Ziliang Ma and Nadzeya Staliarova and Martina Celotti and Sangho Lim and He, \{Gui Wei\} and Rosemary Millen and Else Driehuis and Harry Begthel and Lidwien Smabers and Jeanine Roodhart and \{van Es\}, Johan and Wei Wu and Hans Clevers",

note = "Publisher Copyright: Copyright {\textcopyright} 2024 the Author(s). Published by PNAS.",

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doi = "10.1073/pnas.2309902121",

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Boretto, M, Geurts, MH, Gandhi, S, Ma, Z, Staliarova, N, Celotti, M, Lim, S, He, GW, Millen, R, Driehuis, E, Begthel, H, Smabers, L, Roodhart, J, van Es, J, Wu, W & Clevers, H 2024, 'Epidermal growth factor receptor (EGFR) is a target of the tumor-suppressor E3 ligase FBXW7', Proceedings of the National Academy of Sciences of the United States of America, vol. 121, no. 12, e2309902121. https://doi.org/10.1073/pnas.2309902121

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T1 - Epidermal growth factor receptor (EGFR) is a target of the tumor-suppressor E3 ligase FBXW7

AU - Boretto, Matteo

AU - Geurts, Maarten H.

AU - Gandhi, Shashank

AU - Ma, Ziliang

AU - Staliarova, Nadzeya

AU - Celotti, Martina

AU - Lim, Sangho

AU - He, Gui Wei

AU - Millen, Rosemary

AU - Driehuis, Else

AU - Begthel, Harry

AU - Smabers, Lidwien

AU - Roodhart, Jeanine

AU - van Es, Johan

AU - Wu, Wei

AU - Clevers, Hans

PY - 2024/3/19

Y1 - 2024/3/19

N2 - 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.

AB - 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.

KW - colorectal cancer

KW - EGFR

KW - FBXW7

KW - organoids

UR - https://www.scopus.com/pages/publications/85187847955

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DO - 10.1073/pnas.2309902121

M3 - Article

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AN - SCOPUS:85187847955

SN - 0027-8424

VL - 121

JO - Proceedings of the National Academy of Sciences of the United States of America

JF - Proceedings of the National Academy of Sciences of the United States of America

IS - 12

M1 - e2309902121

ER -

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

Abstract

Bibliographical note

Funding

Keywords

UN SDGs

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