The University of Southampton
University of Southampton Institutional Repository

MYOD-SKP2 axis boosts tumorigenesis in fusion negative rhabdomyosarcoma by preventing differentiation through p57Kip2 targeting

MYOD-SKP2 axis boosts tumorigenesis in fusion negative rhabdomyosarcoma by preventing differentiation through p57Kip2 targeting
MYOD-SKP2 axis boosts tumorigenesis in fusion negative rhabdomyosarcoma by preventing differentiation through p57Kip2 targeting

Rhabdomyosarcomas (RMS) are pediatric mesenchymal-derived malignancies encompassing PAX3/7-FOXO1 Fusion Positive (FP)-RMS, and Fusion Negative (FN)-RMS with frequent RAS pathway mutations. RMS express the master myogenic transcription factor MYOD that, whilst essential for survival, cannot support differentiation. Here we discover SKP2, an oncogenic E3-ubiquitin ligase, as a critical pro-tumorigenic driver in FN-RMS. We show that SKP2 is overexpressed in RMS through the binding of MYOD to an intronic enhancer. SKP2 in FN-RMS promotes cell cycle progression and prevents differentiation by directly targeting p27Kip1 and p57Kip2, respectively. SKP2 depletion unlocks a partly MYOD-dependent myogenic transcriptional program and strongly affects stemness and tumorigenic features and prevents in vivo tumor growth. These effects are mirrored by the investigational NEDDylation inhibitor MLN4924. Results demonstrate a crucial crosstalk between transcriptional and post-translational mechanisms through the MYOD-SKP2 axis that contributes to tumorigenesis in FN-RMS. Finally, NEDDylation inhibition is identified as a potential therapeutic vulnerability in FN-RMS.

Carcinogenesis/genetics, Cell Line, Tumor, Child, Humans, Rhabdomyosarcoma/genetics, Transcription Factors
2041-1723
Pomella, Silvia
013ada95-f338-4e94-975b-469a2a8368d2
Cassandri, Matteo
ace0bb27-109d-401f-83af-b9e7c98a5737
D'Archivio, Lucrezia
87cf125f-5a7a-482c-a32f-b7c0f1393ac2
Porrazzo, Antonella
88395ef1-38c8-4f7d-8094-a607315dd3b9
Cossetti, Cristina
3f0a83d8-fa7f-4fbd-b0d9-5526a2ba7f90
Phelps, Doris
df2911e9-f932-46b7-9ce7-00ab15119663
Perrone, Clara
d8eece9d-7f1b-40d8-8c2d-28558267faa0
Pezzella, Michele
ddb76497-bfa3-449e-8a02-f4fc8ba59916
Cardinale, Antonella
db4ee112-bca7-49e5-ad69-5a7d47d05ee3
Wachtel, Marco
1fca51f5-48dd-4ed8-9077-c559565d203d
Aloisi, Sara
ed255e69-d0fb-47bf-b098-99b40879443e
Milewski, David
46ed38a8-6fc7-4545-ab4e-54eafeb87f1b
Colletti, Marta
f90b6e09-dbf5-44e2-b77e-a7f29e0276f0
Sreenivas, Prethish
8c6274b6-9a38-48c9-bcb2-525b2517c6bb
Walters, Zoë S.
e1ccd35d-63a9-4951-a5da-59122193740d
Barillari, Giovanni
81ad76d1-141b-4d63-86b5-a39052ffdacf
Di Giannatale, Angela
fb76962e-5aa7-4b6c-95ba-1eebd724f2b2
Milano, Giuseppe Maria
ac7ffa66-fa0d-4d09-a5dd-59db36bb4900
De Stefanis, Cristiano
7e4ce844-cb4c-4667-80ae-9538fced27fc
Alaggio, Rita
b4a4bd83-4220-488b-8758-aa9b9bc6d30e
Rodriguez-Rodriguez, Sonia
79642943-1b52-4814-b2ba-6bfb7510db68
Carlesso, Nadia
3669047c-619c-4a0d-aaf4-84113a9ea1d3
Vakoc, Christopher R.
1beda014-ed82-47e2-a02b-2c1dd6f72011
Velardi, Enrico
6e67c987-7522-46aa-bd95-b6918782a171
Schafer, Beat W.
74dd0034-2f97-4fc8-ba57-0c58e2266564
Guccione, Ernesto
9c537e57-3114-40c9-850c-10d47a8f09bb
Gatz, Susanne A.
e80301a4-7fb8-4720-8f1f-7f4cc4f63cf5
Wasti, Ajla
99e0e0b9-b790-4c23-a9b1-19ed1dcdb69f
Yohe, Marielle
e4cc6c87-0c6a-49d9-9a6c-bd98df1ee1bc
Ignatius, Myron
89f035fc-c337-4f8f-a84d-2b5a5852b8cf
Quintarelli, Concetta
eb990843-93d9-4a60-bc19-f7b220d79a16
Shipley, Janet
c4bd3760-d826-42ba-8321-ce78582034ac
Miele, Lucio
8f1d5ed9-17b7-422b-8d6a-5158baaec804
Khan, Javed
32241e41-02bf-40e4-af68-e48005a8b675
Houghton, Peter J.
8f7fcbcd-164f-4b6b-8ab2-fc7a40cf9b69
Marampon, Francesco
7057c6f7-11ea-4872-a90b-5e0404a8c570
Gryder, Berkley E.
21d979c6-7aaf-4108-9959-b4d8b13c2544
De Angelis, Biagio
02f1c5ed-9e0a-41f5-924e-f8b5f534654d
Locatelli, Franco
56e5eb68-2b2c-48ca-aa67-5391dda7e36b
Rota, Rossella
ff0a8696-f3ef-4be8-86da-e1254c374236
Pomella, Silvia
013ada95-f338-4e94-975b-469a2a8368d2
Cassandri, Matteo
ace0bb27-109d-401f-83af-b9e7c98a5737
D'Archivio, Lucrezia
87cf125f-5a7a-482c-a32f-b7c0f1393ac2
Porrazzo, Antonella
88395ef1-38c8-4f7d-8094-a607315dd3b9
Cossetti, Cristina
3f0a83d8-fa7f-4fbd-b0d9-5526a2ba7f90
Phelps, Doris
df2911e9-f932-46b7-9ce7-00ab15119663
Perrone, Clara
d8eece9d-7f1b-40d8-8c2d-28558267faa0
Pezzella, Michele
ddb76497-bfa3-449e-8a02-f4fc8ba59916
Cardinale, Antonella
db4ee112-bca7-49e5-ad69-5a7d47d05ee3
Wachtel, Marco
1fca51f5-48dd-4ed8-9077-c559565d203d
Aloisi, Sara
ed255e69-d0fb-47bf-b098-99b40879443e
Milewski, David
46ed38a8-6fc7-4545-ab4e-54eafeb87f1b
Colletti, Marta
f90b6e09-dbf5-44e2-b77e-a7f29e0276f0
Sreenivas, Prethish
8c6274b6-9a38-48c9-bcb2-525b2517c6bb
Walters, Zoë S.
e1ccd35d-63a9-4951-a5da-59122193740d
Barillari, Giovanni
81ad76d1-141b-4d63-86b5-a39052ffdacf
Di Giannatale, Angela
fb76962e-5aa7-4b6c-95ba-1eebd724f2b2
Milano, Giuseppe Maria
ac7ffa66-fa0d-4d09-a5dd-59db36bb4900
De Stefanis, Cristiano
7e4ce844-cb4c-4667-80ae-9538fced27fc
Alaggio, Rita
b4a4bd83-4220-488b-8758-aa9b9bc6d30e
Rodriguez-Rodriguez, Sonia
79642943-1b52-4814-b2ba-6bfb7510db68
Carlesso, Nadia
3669047c-619c-4a0d-aaf4-84113a9ea1d3
Vakoc, Christopher R.
1beda014-ed82-47e2-a02b-2c1dd6f72011
Velardi, Enrico
6e67c987-7522-46aa-bd95-b6918782a171
Schafer, Beat W.
74dd0034-2f97-4fc8-ba57-0c58e2266564
Guccione, Ernesto
9c537e57-3114-40c9-850c-10d47a8f09bb
Gatz, Susanne A.
e80301a4-7fb8-4720-8f1f-7f4cc4f63cf5
Wasti, Ajla
99e0e0b9-b790-4c23-a9b1-19ed1dcdb69f
Yohe, Marielle
e4cc6c87-0c6a-49d9-9a6c-bd98df1ee1bc
Ignatius, Myron
89f035fc-c337-4f8f-a84d-2b5a5852b8cf
Quintarelli, Concetta
eb990843-93d9-4a60-bc19-f7b220d79a16
Shipley, Janet
c4bd3760-d826-42ba-8321-ce78582034ac
Miele, Lucio
8f1d5ed9-17b7-422b-8d6a-5158baaec804
Khan, Javed
32241e41-02bf-40e4-af68-e48005a8b675
Houghton, Peter J.
8f7fcbcd-164f-4b6b-8ab2-fc7a40cf9b69
Marampon, Francesco
7057c6f7-11ea-4872-a90b-5e0404a8c570
Gryder, Berkley E.
21d979c6-7aaf-4108-9959-b4d8b13c2544
De Angelis, Biagio
02f1c5ed-9e0a-41f5-924e-f8b5f534654d
Locatelli, Franco
56e5eb68-2b2c-48ca-aa67-5391dda7e36b
Rota, Rossella
ff0a8696-f3ef-4be8-86da-e1254c374236

Pomella, Silvia, Cassandri, Matteo, D'Archivio, Lucrezia, Porrazzo, Antonella, Cossetti, Cristina, Phelps, Doris, Perrone, Clara, Pezzella, Michele, Cardinale, Antonella, Wachtel, Marco, Aloisi, Sara, Milewski, David, Colletti, Marta, Sreenivas, Prethish, Walters, Zoë S., Barillari, Giovanni, Di Giannatale, Angela, Milano, Giuseppe Maria, De Stefanis, Cristiano, Alaggio, Rita, Rodriguez-Rodriguez, Sonia, Carlesso, Nadia, Vakoc, Christopher R., Velardi, Enrico, Schafer, Beat W., Guccione, Ernesto, Gatz, Susanne A., Wasti, Ajla, Yohe, Marielle, Ignatius, Myron, Quintarelli, Concetta, Shipley, Janet, Miele, Lucio, Khan, Javed, Houghton, Peter J., Marampon, Francesco, Gryder, Berkley E., De Angelis, Biagio, Locatelli, Franco and Rota, Rossella (2023) MYOD-SKP2 axis boosts tumorigenesis in fusion negative rhabdomyosarcoma by preventing differentiation through p57Kip2 targeting. Nature Communications, 14 (1), [8373]. (doi:10.1038/s41467-023-44130-0).

Record type: Article

Abstract

Rhabdomyosarcomas (RMS) are pediatric mesenchymal-derived malignancies encompassing PAX3/7-FOXO1 Fusion Positive (FP)-RMS, and Fusion Negative (FN)-RMS with frequent RAS pathway mutations. RMS express the master myogenic transcription factor MYOD that, whilst essential for survival, cannot support differentiation. Here we discover SKP2, an oncogenic E3-ubiquitin ligase, as a critical pro-tumorigenic driver in FN-RMS. We show that SKP2 is overexpressed in RMS through the binding of MYOD to an intronic enhancer. SKP2 in FN-RMS promotes cell cycle progression and prevents differentiation by directly targeting p27Kip1 and p57Kip2, respectively. SKP2 depletion unlocks a partly MYOD-dependent myogenic transcriptional program and strongly affects stemness and tumorigenic features and prevents in vivo tumor growth. These effects are mirrored by the investigational NEDDylation inhibitor MLN4924. Results demonstrate a crucial crosstalk between transcriptional and post-translational mechanisms through the MYOD-SKP2 axis that contributes to tumorigenesis in FN-RMS. Finally, NEDDylation inhibition is identified as a potential therapeutic vulnerability in FN-RMS.

Text
s41467-023-44130-0 - Version of Record
Available under License Creative Commons Attribution.
Download (9MB)

More information

Published date: 15 December 2023
Additional Information: Funding Information: We would like to thank Maharshi Chakraborty, Richard Sallari (Axiotl Inc, Cleveland OH, USA) and Heike Wollmann (Institute of Molecular and Cell Biology (IMCB), Agency for Science, Technology and Research (A*STAR), Singapore), for technical and bioinformatics assistance. The Myogenin (Wright WE) and MHC (Fishman DA) antibodies were obtained from the Developmental Studies Hybridoma Bank, developed under the auspices of the NICHD and maintained by The University of Iowa, Department of Biology, Iowa City, IA 52242, USA. C.P. was supported by the Italian Ministry of University and Research (MIUR) Ph.D. fellowship (Department of Science, Roma Tre University Doctorate). We are very grateful to M. Dyer and E. Stewart from the Childhood Solid Tumor Network (CSTN) at St. Jude for PDX models used in this study. M.Co. was supported by a “Fondazione Veronesi” fellowship. This project has been funded by Associazione Italiana Ricerca sul Cancro (AIRC IG #10338, #15312 and #27794), Alleanza Contro il Cancro (ACC) Italian Network-Working Group Sarcomas, and Italian Ministry of Health Fondi 5xmille (2020-2021) and Current Research to R.R.; Alleanza Contro il Cancro (ACC) Italian Network-Working Group Sarcomas and Italian Ministry of Health Ricerca Finalizzata GR-2016-02364546 to B.D.A.; Italian Ministry of Health Ricerca Finalizzata GR-2013 02359212 to C.Q.; AIRC MFGA #22889 to E.V.; AIRC IG #24696 to F.M.; AIRC 5xmille (9962) to F.L. B.E.G. is supported through the DOD’s Convergent Science Virtual Cancer Center, Reign in Sarcoma, the V Foundation and Alex’s Lemonade Stand Foundation. The funders had no role in study design, data collection and analysis, decision to publish or preparation of the manuscript. Funding Information: We would like to thank Maharshi Chakraborty, Richard Sallari (Axiotl Inc, Cleveland OH, USA) and Heike Wollmann (Institute of Molecular and Cell Biology (IMCB), Agency for Science, Technology and Research (A*STAR), Singapore), for technical and bioinformatics assistance. The Myogenin (Wright WE) and MHC (Fishman DA) antibodies were obtained from the Developmental Studies Hybridoma Bank, developed under the auspices of the NICHD and maintained by The University of Iowa, Department of Biology, Iowa City, IA 52242, USA. C.P. was supported by the Italian Ministry of University and Research (MIUR) Ph.D. fellowship (Department of Science, Roma Tre University Doctorate). We are very grateful to M. Dyer and E. Stewart from the Childhood Solid Tumor Network (CSTN) at St. Jude for PDX models used in this study. M.Co. was supported by a “Fondazione Veronesi” fellowship. This project has been funded by Associazione Italiana Ricerca sul Cancro (AIRC IG #10338, #15312 and #27794), Alleanza Contro il Cancro (ACC) Italian Network-Working Group Sarcomas, and Italian Ministry of Health Fondi 5xmille (2020-2021) and Current Research to R.R.; Alleanza Contro il Cancro (ACC) Italian Network-Working Group Sarcomas and Italian Ministry of Health Ricerca Finalizzata GR-2016-02364546 to B.D.A.; Italian Ministry of Health Ricerca Finalizzata GR-2013 02359212 to C.Q.; AIRC MFGA #22889 to E.V.; AIRC IG #24696 to F.M.; AIRC 5xmille (9962) to F.L. B.E.G. is supported through the DOD’s Convergent Science Virtual Cancer Center, Reign in Sarcoma, the V Foundation and Alex’s Lemonade Stand Foundation. The funders had no role in study design, data collection and analysis, decision to publish or preparation of the manuscript. Publisher Copyright: © 2023, The Author(s).
Keywords: Carcinogenesis/genetics, Cell Line, Tumor, Child, Humans, Rhabdomyosarcoma/genetics, Transcription Factors

Identifiers

Local EPrints ID: 485866
URI: http://eprints.soton.ac.uk/id/eprint/485866
ISSN: 2041-1723
PURE UUID: bed05144-be8e-4d2d-a614-c14aaf3dd5aa
ORCID for Zoë S. Walters: ORCID iD orcid.org/0000-0002-1835-5868

Catalogue record

Date deposited: 03 Jan 2024 17:03
Last modified: 18 Mar 2024 03:43

Export record

Altmetrics

Contributors

Author: Silvia Pomella
Author: Matteo Cassandri
Author: Lucrezia D'Archivio
Author: Antonella Porrazzo
Author: Cristina Cossetti
Author: Doris Phelps
Author: Clara Perrone
Author: Michele Pezzella
Author: Antonella Cardinale
Author: Marco Wachtel
Author: Sara Aloisi
Author: David Milewski
Author: Marta Colletti
Author: Prethish Sreenivas
Author: Zoë S. Walters ORCID iD
Author: Giovanni Barillari
Author: Angela Di Giannatale
Author: Giuseppe Maria Milano
Author: Cristiano De Stefanis
Author: Rita Alaggio
Author: Sonia Rodriguez-Rodriguez
Author: Nadia Carlesso
Author: Christopher R. Vakoc
Author: Enrico Velardi
Author: Beat W. Schafer
Author: Ernesto Guccione
Author: Susanne A. Gatz
Author: Ajla Wasti
Author: Marielle Yohe
Author: Myron Ignatius
Author: Concetta Quintarelli
Author: Janet Shipley
Author: Lucio Miele
Author: Javed Khan
Author: Peter J. Houghton
Author: Francesco Marampon
Author: Berkley E. Gryder
Author: Biagio De Angelis
Author: Franco Locatelli
Author: Rossella Rota

Download statistics

Downloads from ePrints over the past year. Other digital versions may also be available to download e.g. from the publisher's website.

View more statistics

Atom RSS 1.0 RSS 2.0

Contact ePrints Soton: eprints@soton.ac.uk

ePrints Soton supports OAI 2.0 with a base URL of http://eprints.soton.ac.uk/cgi/oai2

This repository has been built using EPrints software, developed at the University of Southampton, but available to everyone to use.

We use cookies to ensure that we give you the best experience on our website. If you continue without changing your settings, we will assume that you are happy to receive cookies on the University of Southampton website.

×