Ral GTPases promote breast cancer metastasis by controlling biogenesis and organ targeting of exosomes
- INSERM U1109, France
- Laboratoire de Spectrométrie de Masse BioOrganique, France
- Muséum National d'Histoire Naturelle de Paris, France
- Université de Bordeaux, CNRS, France
- Centre National de la Recherche Scientifique, Université de Strasbourg, France
- CNRS UPS 3156, France
- IGBMC (UMR7104)/ INSERM (U1258)/ Université de Strasbourg,, France
- CRCINA, INSERM, CNRS, Université de Nantes, Université d'Angers, France
- Icans, France
- Garvan Institute of Medical Research, Australia
- INSERM 1263, Inrae 1260, France
- Institut des Neurosciences Cellulaires et Intégratives; UPR-3212 CNRS, Strasbourg University, France
Abstract
Cancer extracellular vesicles (EVs) shuttle at distance and fertilize pre-metastatic niches facilitating subsequent seeding by tumor cells. However, the link between EV secretion mechanisms and their capacity to form pre-metastatic niches remains obscure. Using mouse models, we show that GTPases of the Ral family control, through the phospholipase D1, multi-vesicular bodies homeostasis and tune the biogenesis and secretion of pro-metastatic EVs. Importantly, EVs from RalA or RalB depleted cells have limited organotropic capacities in vivo and are less efficient in promoting metastasis. RalA and RalB reduce the EV levels of the adhesion molecule MCAM/CD146, which favors EV-mediated metastasis by allowing EVs targeting to the lungs. Finally, RalA, RalB and MCAM/CD146, are factors of poor prognosis in breast cancer patients. Altogether, our study identifies RalGTPases as central molecules linking the mechanisms of EVs secretion and cargo loading to their capacity to disseminate and induce pre-metastatic niches in a CD146 dependent manner.
Data availability
Sequencing and mass spectrometry data have been deposited to EV-Track knowledgebase
-
Comprehensive molecular portraits of human breast tumoursTCGA breast invasive carcinoma cohort (1097 patients).
Article and author information
Author details
Funding
Institut National Du Cancer (PLBIO19-291)
- Jacky G Goetz
La Ligue contre le cancer
- Jacky G Goetz
Canceropole grand est
- Jacky G Goetz
Plan Cancer
- Jacky G Goetz
Roche
- Jacky G Goetz
Agence Nationale de la Recherche (ANR-10-INBS-08-03)
- Christine Carapito
Agence Nationale de la Recherche (ANR-19-CE44-0019)
- Nicolas Vitale
Agence Nationale de la Recherche (ANR-11-INBS- 0010)
- Laetitia Fouillen
The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.
Reviewing Editor
- Erica A Golemis, Fox Chase Cancer Center, United States
Ethics
Animal experimentation: All animals were housed and han- dled according to the guidelines of INSERM and the eth- ical committee of Alsace, France (CREMEAS) (Direc- tive 2010/63/EU on the protection of animals used for scientific purposes). Animal facility agreement num- ber: C67-482-33. Experimental license for mice: Apafis 4707-20l6032416407780; experimental license for zebrafish: Apafis 16862-2018121914292754.
Human subjects: Paraffin sections of 4 µm from metastasic and non-metastasic breast tumours were obtained from CRB-Tumorothèque of the Institut de Cancérologie de l'Ouest (ICO, Saint-Herblain, France) (Heymann et al., 2020). Patients were diagnosed and treated at ICO (Integrated Center for Oncology, St Herblain) for metastatic and non-metastatic breast cancers. Samples were processed and included in the CRB-Tumorothèque ICO upon donor agreement and informed consent. Samples and related information are destroyed at the request of the donor. The CRB-Tumorothèque ICO have been declared to and authorized by the French Research Ministry (Declaration Number: DC-2018-3321). This declaration includes approval by a research ethics committee (CPP "Comité de protection des personnes"), in accordance with the French legislation of the Public Health Code.
Version history
- Received: July 28, 2020
- Accepted: January 5, 2021
- Accepted Manuscript published: January 6, 2021 (version 1)
- Version of Record published: January 22, 2021 (version 2)
Copyright
© 2021, Ghoroghi et al.
This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.
Metrics
-
- 4,349
- views
-
- 668
- downloads
-
- 73
- citations
Views, downloads and citations are aggregated across all versions of this paper published by eLife.
Download links
A two-part list of links to download the article, or parts of the article, in various formats.
Downloads (link to download the article as PDF)
Open citations (links to open the citations from this article in various online reference manager services)
Cite this article (links to download the citations from this article in formats compatible with various reference manager tools)
Ral GTPases promote breast cancer metastasis by controlling biogenesis and organ targeting of exosomes
eLife 10:e61539.
https://doi.org/10.7554/eLife.61539
Further reading
-
- Cancer Biology
- Cell Biology
Internalization from the cell membrane and endosomal trafficking of receptor tyrosine kinases (RTKs) are important regulators of signaling in normal cells that can frequently be disrupted in cancer. The adrenal tumor pheochromocytoma (PCC) can be caused by activating mutations of the rearranged during transfection (RET) receptor tyrosine kinase, or inactivation of TMEM127, a transmembrane tumor suppressor implicated in trafficking of endosomal cargos. However, the role of aberrant receptor trafficking in PCC is not well understood. Here, we show that loss of TMEM127 causes wildtype RET protein accumulation on the cell surface, where increased receptor density facilitates constitutive ligand-independent activity and downstream signaling, driving cell proliferation. Loss of TMEM127 altered normal cell membrane organization and recruitment and stabilization of membrane protein complexes, impaired assembly, and maturation of clathrin-coated pits, and reduced internalization and degradation of cell surface RET. In addition to RTKs, TMEM127 depletion also promoted surface accumulation of several other transmembrane proteins, suggesting it may cause global defects in surface protein activity and function. Together, our data identify TMEM127 as an important determinant of membrane organization including membrane protein diffusability and protein complex assembly and provide a novel paradigm for oncogenesis in PCC where altered membrane dynamics promotes cell surface accumulation and constitutive activity of growth factor receptors to drive aberrant signaling and promote transformation.
-
- Cancer Biology
- Genetics and Genomics
Enhancers are critical for regulating tissue-specific gene expression, and genetic variants within enhancer regions have been suggested to contribute to various cancer-related processes, including therapeutic resistance. However, the precise mechanisms remain elusive. Using a well-defined drug-gene pair, we identified an enhancer region for dihydropyrimidine dehydrogenase (DPD, DPYD gene) expression that is relevant to the metabolism of the anti-cancer drug 5-fluorouracil (5-FU). Using reporter systems, CRISPR genome-edited cell models, and human liver specimens, we demonstrated in vitro and vivo that genotype status for the common germline variant (rs4294451; 27% global minor allele frequency) located within this novel enhancer controls DPYD transcription and alters resistance to 5-FU. The variant genotype increases recruitment of the transcription factor CEBPB to the enhancer and alters the level of direct interactions between the enhancer and DPYD promoter. Our data provide insight into the regulatory mechanisms controlling sensitivity and resistance to 5-FU.
