Extracellular vesicle-associated tyrosine kinase-like orphan receptors ROR1 and ROR2 promote breast cancer progression.
Journal
Cell communication and signaling : CCS
ISSN: 1478-811X
Titre abrégé: Cell Commun Signal
Pays: England
ID NLM: 101170464
Informations de publication
Date de publication:
10 07 2023
10 07 2023
Historique:
received:
15
03
2023
accepted:
06
06
2023
medline:
12
7
2023
pubmed:
11
7
2023
entrez:
10
7
2023
Statut:
epublish
Résumé
Extracellular vesicles (EVs) harbor a plethora of different biomolecules, which they can transport across cells. In cancer, tumor-derived EVs thereby support the creation of a favorable tumor microenvironment. So far, EV uptake and cargo delivery into target cells have been regarded as the main mechanisms for the pro-tumoral function of EVs. To test this hypothesis, we investigated the fate of the oncogenic transmembrane Wnt tyrosine kinase-like orphan receptor 1 and 2 (ROR1, ROR2) delivered via distinct EV subpopulations to breast cancer cells and aimed to unravel their impact on tumor progression. EVs were isolated by differential ultracentrifugation from cell culture supernatant as well as plasma samples from healthy individuals (n = 27) and breast cancer patients (n = 41). EVs were thoroughly characterized by electron microscopy, nanoparticle tracking analysis, immunoblot, and flow cytometry. ROR transfer to target cells was observed using microscopy-based assays and biodistribution experiments were conducted in syngeneic mice. EV impact on cancer cell migration and invasion was tested in functional assays. We observed that the supernatant of ROR-overexpressing cells was sufficient for transferring the receptors to ROR-negative cells. Analyzing the secretome of the ROR-overexpressing cells, we detected a high enrichment of ROR1/2 on large and small EVs, but not on large oncosomes. Interestingly, the majority of ROR-positive EVs remained attached to the target cell surface after 24 h of stimulation and was quickly removed by treatment with trypsin. Nonetheless, ROR-positive EVs increased migration and invasion of breast cancer cells, even after chemically inhibiting EV uptake, in dependence of RhoA downstream signaling. In vivo, ROR-depleted EVs tended to distribute less into organs prone for the formation of breast cancer metastases. ROR-positive EVs were also significantly elevated in the plasma of breast cancer patients and allowed to separate them from healthy controls. The oncogenic Wnt receptors ROR1/2 are transferred via EVs to the surface of ROR-negative cancer cells, in which they induce an aggressive phenotype supporting tumor progression. Video Abstract.
Sections du résumé
BACKGROUND
Extracellular vesicles (EVs) harbor a plethora of different biomolecules, which they can transport across cells. In cancer, tumor-derived EVs thereby support the creation of a favorable tumor microenvironment. So far, EV uptake and cargo delivery into target cells have been regarded as the main mechanisms for the pro-tumoral function of EVs. To test this hypothesis, we investigated the fate of the oncogenic transmembrane Wnt tyrosine kinase-like orphan receptor 1 and 2 (ROR1, ROR2) delivered via distinct EV subpopulations to breast cancer cells and aimed to unravel their impact on tumor progression.
METHODS
EVs were isolated by differential ultracentrifugation from cell culture supernatant as well as plasma samples from healthy individuals (n = 27) and breast cancer patients (n = 41). EVs were thoroughly characterized by electron microscopy, nanoparticle tracking analysis, immunoblot, and flow cytometry. ROR transfer to target cells was observed using microscopy-based assays and biodistribution experiments were conducted in syngeneic mice. EV impact on cancer cell migration and invasion was tested in functional assays.
RESULTS
We observed that the supernatant of ROR-overexpressing cells was sufficient for transferring the receptors to ROR-negative cells. Analyzing the secretome of the ROR-overexpressing cells, we detected a high enrichment of ROR1/2 on large and small EVs, but not on large oncosomes. Interestingly, the majority of ROR-positive EVs remained attached to the target cell surface after 24 h of stimulation and was quickly removed by treatment with trypsin. Nonetheless, ROR-positive EVs increased migration and invasion of breast cancer cells, even after chemically inhibiting EV uptake, in dependence of RhoA downstream signaling. In vivo, ROR-depleted EVs tended to distribute less into organs prone for the formation of breast cancer metastases. ROR-positive EVs were also significantly elevated in the plasma of breast cancer patients and allowed to separate them from healthy controls.
CONCLUSIONS
The oncogenic Wnt receptors ROR1/2 are transferred via EVs to the surface of ROR-negative cancer cells, in which they induce an aggressive phenotype supporting tumor progression. Video Abstract.
Identifiants
pubmed: 37430307
doi: 10.1186/s12964-023-01186-1
pii: 10.1186/s12964-023-01186-1
pmc: PMC10331971
doi:
Substances chimiques
Protein-Tyrosine Kinases
EC 2.7.10.1
Ror1 protein, mouse
EC 2.7.10.1
Ror2 protein, mouse
EC 2.7.10.1
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Video-Audio Media
Langues
eng
Sous-ensembles de citation
IM
Pagination
171Informations de copyright
© 2023. The Author(s).
Références
Oncoimmunology. 2012 Oct 1;1(7):1074-1083
pubmed: 23170255
Carcinogenesis. 2011 Mar;32(3):434-42
pubmed: 21173432
J Clin Invest. 2016 Feb;126(2):585-98
pubmed: 26690702
J Extracell Vesicles. 2015 Apr 20;4:26316
pubmed: 25899407
Nat Methods. 2017 Feb 28;14(3):228-232
pubmed: 28245209
Semin Cancer Biol. 2022 Nov;86(Pt 1):112-126
pubmed: 35032650
Proc Natl Acad Sci U S A. 2016 Feb 23;113(8):E968-77
pubmed: 26858453
Sci Rep. 2019 Jul 19;9(1):10522
pubmed: 31324885
J Extracell Vesicles. 2012 Sep 11;1:
pubmed: 24009881
Cells. 2021 Jan 12;10(1):
pubmed: 33445713
J Mol Cell Biol. 2015 Apr;7(2):143-53
pubmed: 25503107
Med Sci (Basel). 2020 Nov 04;8(4):
pubmed: 33158117
Nat Med. 2012 Dec;18(12):1835-40
pubmed: 23142818
Curr Protoc Immunol. 2001 May;Chapter 20:Unit 20.2
pubmed: 18432775
J Extracell Vesicles. 2017 Jul 16;6(1):1340745
pubmed: 28804596
CA Cancer J Clin. 2021 May;71(3):209-249
pubmed: 33538338
Cancer Res. 2013 Jun 15;73(12):3649-60
pubmed: 23771907
Nature. 2015 Nov 19;527(7578):329-35
pubmed: 26524530
Cell Commun Signal. 2021 Apr 23;19(1):47
pubmed: 33892745
Cancer Cell. 2020 Apr 13;37(4):471-484
pubmed: 32289271
EMBO J. 2023 Apr 25;:e112614
pubmed: 37096681
Nat Commun. 2022 Aug 1;13(1):4461
pubmed: 35915084
Cancer Discov. 2013 Dec;3(12):1378-93
pubmed: 24104062
Mol Cancer. 2014 Apr 26;13:88
pubmed: 24766647
Oncotarget. 2013 Nov;4(11):2057-66
pubmed: 24185202
Mol Imaging Biol. 2020 Dec;22(6):1501-1510
pubmed: 32737655
Oncotarget. 2015 May 10;6(13):11327-41
pubmed: 25857301
Biomolecules. 2018 Sep 14;8(3):
pubmed: 30223513
J Extracell Vesicles. 2015 May 14;4:27066
pubmed: 25979354
Nat Commun. 2016 Jan 04;7:10060
pubmed: 26725982
Leukemia. 2006 May;20(5):847-56
pubmed: 16453000
Cells. 2019 Jul 25;8(8):
pubmed: 31349740
Oncogene. 2012 Nov 8;31(45):4740-9
pubmed: 22266864
Elife. 2021 Jan 06;10:
pubmed: 33404012
AIDS. 2003 Jan 3;17(1):33-42
pubmed: 12478067
Clin Exp Metastasis. 2016 Apr;33(4):309-23
pubmed: 26862065
Proc Natl Acad Sci U S A. 2006 Apr 4;103(14):5454-9
pubmed: 16569699
Nat Cell Biol. 2008 May;10(5):619-24
pubmed: 18425114
J Exp Clin Cancer Res. 2021 Dec 15;40(1):395
pubmed: 34911552
Nat Med. 2012 Jun;18(6):883-91
pubmed: 22635005
EMBO J. 2017 Oct 16;36(20):3012-3028
pubmed: 28923825
Mol Cancer. 2020 Nov 24;19(1):165
pubmed: 33234169
J Exp Med. 1996 Mar 1;183(3):1161-72
pubmed: 8642258
Nat Protoc. 2013 Nov;8(11):2281-2308
pubmed: 24157548
Cell Rep. 2022 Apr 12;39(2):110651
pubmed: 35417683
Oncogene. 2017 Oct 26;36(43):5958-5968
pubmed: 28650466
Int J Mol Sci. 2016 Feb 06;17(2):175
pubmed: 26861306
PLoS One. 2012;7(3):e31127
pubmed: 22403610
Nat Cell Biol. 2012 Oct;14(10):1036-45
pubmed: 22983114
Proc Natl Acad Sci U S A. 2019 Jan 22;116(4):1370-1377
pubmed: 30622177
Nat Cell Biol. 2015 Jun;17(6):816-26
pubmed: 25985394