DLL4/Notch3/WNT5B axis mediates bidirectional prometastatic crosstalk between melanoma and lymphatic endothelial cells

Sanni Alve; Silvia Gramolelli; Joonas Jukonen; Susanna Juteau; Anne Pink; Atte A Manninen; Satu Hänninen; Elisa Monto; Madeleine H Lackman; Olli Carpén; Pipsa Saharinen; Sinem Karaman; Kari Vaahtomeri; Päivi M Ojala

doi:10.1172/jci.insight.171821

DLL4/Notch3/WNT5B axis mediates bidirectional prometastatic crosstalk between melanoma and lymphatic endothelial cells

Sanni Alve
, Silvia Gramolelli
, Joonas Jukonen
, Susanna Juteau
, Anne Pink
, Atte A Manninen
, Satu Hänninen
, Elisa Monto
, Madeleine H Lackman
, Olli Carpén
, Pipsa Saharinen
, Sinem Karaman
, Kari Vaahtomeri
, Päivi M Ojala

Biokemi och cellbiologi

Forskningsoutput: Tidskriftsbidrag › Artikel › Vetenskaplig › Peer review

6 Citeringar (Scopus)

99 Nedladdningar (Pure)

Sammanfattning

Despite strong indications that interactions between melanoma and lymphatic vessels actively promote melanoma progression, the molecular mechanisms are not yet completely understood. To characterize molecular factors of this crosstalk, we established human primary lymphatic endothelial cell (LEC) cocultures with human melanoma cell lines. Here, we show that coculture with melanoma cells induced transcriptomic changes in LECs and led to multiple changes in their function. WNT5B, a paracrine signaling molecule upregulated in melanoma cells upon LEC interaction, was found to contribute to the functional changes in LECs. Moreover, WNT5B transcription was regulated by Notch3 in melanoma cells following the coculture with LECs, and Notch3 and WNT5B were coexpressed in melanoma patient primary tumor and metastasis samples. Moreover, melanoma cells derived from LEC coculture escaped efficiently from the primary site to the proximal tumor-draining lymph nodes, which was impaired upon WNT5B depletion. This supported the role of WNT5B in promoting the metastatic potential of melanoma cells through its effects on LECs. Finally, DLL4, a Notch ligand expressed in LECs, was identified as an upstream inducer of the Notch3/WNT5B axis in melanoma. This study elucidated WNT5B as a key molecular factor mediating bidirectional crosstalk between melanoma cells and lymphatic endothelium and promoting melanoma metastasis.

Originalspråk	Engelska
Artikelnummer	e171821
Antal sidor	21
Tidskrift	JCI Insight
Volym	9
Nummer	1
DOI	https://doi.org/10.1172/jci.insight.171821
Status	Publicerad - 9 jan. 2024
MoE-publikationstyp	A1 Tidskriftsartikel-refererad

Finansiering

We thank Nadezhda Zinovkina, Julia Härme, and Noora Andersson for excellent technical assistance. The FIMM Institute for Molecular Medicine, Finland, is acknowledged for performing scRNA-seq. The Biomedicum Imaging Unit and Genome Biology Unit, University of Helsinki, provided expert imaging services. We are grateful to the HiLife Flow Cytometry Unit, University of Helsinki, for flow cytometry services. We thank the HiLIFE Laboratory Animal Centre Core Facility, University of Helsinki, Finland, for support in mouse work. We are grateful to Timo Otonkoski (Research Programs Unit, University of Helsinki, Helsinki, Finland) for providing mice for our studies. We are grateful to Jenni Lahtela and FIMM, University of Helsinki, Finland, for the scRNA-seq services. We thank Cecilia Sahlgren (Åbo Akademi, Turku, Finland) for advice and reagents on experiments using the Notch ligands. This study was supported by the Finnish Cancer Foundation (to PMO) and Sigrid Juselius Foundation (to PMO). SA was supported by the Doctoral Programme in Bio-medicine, University of Helsinki and fellowships from the Instrumentarium Science Foundation, Finnish Cultural Foundation, Magnus Ehrnrooth Foundation, K. Albin Johansson Foundation, and Cancer Foundation Finland. JJ was supported by fellowships from the Finnish Cultural foundation, Emil Aaltonen Foundation, and Magnus Ehrnrooth Foundation. PS was supported by the European Research Council (ERC) under the EU Horizon 2020 research and innovation programme (grant agreement 773076), Sigrid Jusélius Foundation, Cancer Foundation Finland, and Academy of Finland (grant 346134). MHL was supported by Nylands Nation. SK was supported by the Academy of Finland (grants 330053 and 336126). KV was supported by the Academy of Finland (grant 315710), Sigrid Juselius foundation, and Wihuri Research Institute. These additional funders had no role in the study design, data collection, interpretation, or decision to submit the work for publication. Open access was funded by the Helsinki University Library. We thank Nadezhda Zinovkina, Julia Härme, and Noora Andersson for excellent technical assistance. The FIMM Institute for Molecular Medicine, Finland, is acknowledged for performing scRNA-seq. The Biomedi-cum Imaging Unit and Genome Biology Unit, University of Helsinki, provided expert imaging services. We are grateful to the HiLife Flow Cytometry Unit, University of Helsinki, for flow cytometry services. We thank the HiLIFE Laboratory Animal Centre Core Facility, University of Helsinki, Finland, for support in mouse work. We are grateful to Timo Otonkoski (Research Programs Unit, University of Helsinki, Helsinki, Finland) for providing mice for our studies. We are grateful to Jenni Lahtela and FIMM, University of Helsinki, Finland, for the scRNA-seq services. We thank Cecilia Sahlgren (Åbo Akademi, Turku, Finland) for advice and reagents on experiments using the Notch ligands. This study was supported by the Finnish Cancer Foundation (to PMO) and Sigrid Juselius Foundation (to PMO). SA was supported by the Doctoral Programme in Biomedicine, University of Helsinki and fellowships from the Instrumentarium Science Foundation, Finnish Cultural Foundation, Magnus Ehrnrooth Foundation, K. Albin Johansson Foundation, and Cancer Foundation Finland. JJ was supported by fellowships from the Finnish Cultural foundation, Emil Aaltonen Foundation, and Magnus Ehrnrooth Foundation. PS was supported by the European Research Council (ERC) under the EU Horizon 2020 research and innovation programme (grant agreement 773076), Sigrid Jusélius Foundation, Cancer Foundation Finland, and Academy of Finland (grant 346134). MHL was supported by Nylands Nation. SK was supported by the Academy of Finland (grants 330053 and 336126). KV was supported by the Academy of Finland (grant 315710), Sigrid Juselius foundation, and Wihuri Research Institute. These additional funders had no role in the study design, data collection, interpretation, or decision to submit the work for publication. Open access was funded by the Helsinki University Library.

Nyckelord

Cancer
Vascular Biology
Melanoma
Lymph
Cell Biology

Åtkomst av dokument

10.1172/jci.insight.171821

171821.2-20231212042920-covered-e0fd13ba177f913fd3156f593ead4cfdSlutlig publicerad version, 9,89 MBLicens: CC BY

https://urn.fi/URN:NBN:fi-fe2024040113815

Citera det här

Alve, S., Gramolelli, S., Jukonen, J., Juteau, S., Pink, A., Manninen, A. A., Hänninen, S., Monto, E., Lackman, M. H., Carpén, O., Saharinen, P., Karaman, S., Vaahtomeri, K., & Ojala, P. M. (2024). DLL4/Notch3/WNT5B axis mediates bidirectional prometastatic crosstalk between melanoma and lymphatic endothelial cells. JCI Insight, 9(1), Artikel e171821. https://doi.org/10.1172/jci.insight.171821

@article{8389b03966f24898a58c113591dd816b,

title = "DLL4/Notch3/WNT5B axis mediates bidirectional prometastatic crosstalk between melanoma and lymphatic endothelial cells",

abstract = "Despite strong indications that interactions between melanoma and lymphatic vessels actively promote melanoma progression, the molecular mechanisms are not yet completely understood. To characterize molecular factors of this crosstalk, we established human primary lymphatic endothelial cell (LEC) cocultures with human melanoma cell lines. Here, we show that coculture with melanoma cells induced transcriptomic changes in LECs and led to multiple changes in their function. WNT5B, a paracrine signaling molecule upregulated in melanoma cells upon LEC interaction, was found to contribute to the functional changes in LECs. Moreover, WNT5B transcription was regulated by Notch3 in melanoma cells following the coculture with LECs, and Notch3 and WNT5B were coexpressed in melanoma patient primary tumor and metastasis samples. Moreover, melanoma cells derived from LEC coculture escaped efficiently from the primary site to the proximal tumor-draining lymph nodes, which was impaired upon WNT5B depletion. This supported the role of WNT5B in promoting the metastatic potential of melanoma cells through its effects on LECs. Finally, DLL4, a Notch ligand expressed in LECs, was identified as an upstream inducer of the Notch3/WNT5B axis in melanoma. This study elucidated WNT5B as a key molecular factor mediating bidirectional crosstalk between melanoma cells and lymphatic endothelium and promoting melanoma metastasis.",

keywords = "Humans, Adaptor Proteins, Signal Transducing/metabolism, Calcium-Binding Proteins/metabolism, Endothelial Cells/metabolism, Lymphatic Metastasis/pathology, Lymphatic Vessels/pathology, Melanoma/pathology, Signal Transduction, Wnt Proteins/metabolism, Cancer, Vascular Biology, Melanoma, Lymph, Cell Biology",

author = "Sanni Alve and Silvia Gramolelli and Joonas Jukonen and Susanna Juteau and Anne Pink and Manninen, \{Atte A\} and Satu H{\"a}nninen and Elisa Monto and Lackman, \{Madeleine H\} and Olli Carp{\'e}n and Pipsa Saharinen and Sinem Karaman and Kari Vaahtomeri and Ojala, \{P{\"a}ivi M\}",

year = "2024",

month = jan,

day = "9",

doi = "10.1172/jci.insight.171821",

language = "English",

volume = "9",

journal = "JCI Insight",

issn = "2379-3708",

publisher = "American Society for Clinical Investigation",

number = "1",

}

Alve, S, Gramolelli, S, Jukonen, J, Juteau, S, Pink, A, Manninen, AA, Hänninen, S, Monto, E, Lackman, MH, Carpén, O, Saharinen, P, Karaman, S, Vaahtomeri, K & Ojala, PM 2024, 'DLL4/Notch3/WNT5B axis mediates bidirectional prometastatic crosstalk between melanoma and lymphatic endothelial cells', JCI Insight, vol. 9, nr. 1, e171821. https://doi.org/10.1172/jci.insight.171821

TY - JOUR

T1 - DLL4/Notch3/WNT5B axis mediates bidirectional prometastatic crosstalk between melanoma and lymphatic endothelial cells

AU - Alve, Sanni

AU - Gramolelli, Silvia

AU - Jukonen, Joonas

AU - Juteau, Susanna

AU - Pink, Anne

AU - Manninen, Atte A

AU - Hänninen, Satu

AU - Monto, Elisa

AU - Lackman, Madeleine H

AU - Carpén, Olli

AU - Saharinen, Pipsa

AU - Karaman, Sinem

AU - Vaahtomeri, Kari

AU - Ojala, Päivi M

PY - 2024/1/9

Y1 - 2024/1/9

N2 - Despite strong indications that interactions between melanoma and lymphatic vessels actively promote melanoma progression, the molecular mechanisms are not yet completely understood. To characterize molecular factors of this crosstalk, we established human primary lymphatic endothelial cell (LEC) cocultures with human melanoma cell lines. Here, we show that coculture with melanoma cells induced transcriptomic changes in LECs and led to multiple changes in their function. WNT5B, a paracrine signaling molecule upregulated in melanoma cells upon LEC interaction, was found to contribute to the functional changes in LECs. Moreover, WNT5B transcription was regulated by Notch3 in melanoma cells following the coculture with LECs, and Notch3 and WNT5B were coexpressed in melanoma patient primary tumor and metastasis samples. Moreover, melanoma cells derived from LEC coculture escaped efficiently from the primary site to the proximal tumor-draining lymph nodes, which was impaired upon WNT5B depletion. This supported the role of WNT5B in promoting the metastatic potential of melanoma cells through its effects on LECs. Finally, DLL4, a Notch ligand expressed in LECs, was identified as an upstream inducer of the Notch3/WNT5B axis in melanoma. This study elucidated WNT5B as a key molecular factor mediating bidirectional crosstalk between melanoma cells and lymphatic endothelium and promoting melanoma metastasis.

AB - Despite strong indications that interactions between melanoma and lymphatic vessels actively promote melanoma progression, the molecular mechanisms are not yet completely understood. To characterize molecular factors of this crosstalk, we established human primary lymphatic endothelial cell (LEC) cocultures with human melanoma cell lines. Here, we show that coculture with melanoma cells induced transcriptomic changes in LECs and led to multiple changes in their function. WNT5B, a paracrine signaling molecule upregulated in melanoma cells upon LEC interaction, was found to contribute to the functional changes in LECs. Moreover, WNT5B transcription was regulated by Notch3 in melanoma cells following the coculture with LECs, and Notch3 and WNT5B were coexpressed in melanoma patient primary tumor and metastasis samples. Moreover, melanoma cells derived from LEC coculture escaped efficiently from the primary site to the proximal tumor-draining lymph nodes, which was impaired upon WNT5B depletion. This supported the role of WNT5B in promoting the metastatic potential of melanoma cells through its effects on LECs. Finally, DLL4, a Notch ligand expressed in LECs, was identified as an upstream inducer of the Notch3/WNT5B axis in melanoma. This study elucidated WNT5B as a key molecular factor mediating bidirectional crosstalk between melanoma cells and lymphatic endothelium and promoting melanoma metastasis.

KW - Humans

KW - Adaptor Proteins, Signal Transducing/metabolism

KW - Calcium-Binding Proteins/metabolism

KW - Endothelial Cells/metabolism

KW - Lymphatic Metastasis/pathology

KW - Lymphatic Vessels/pathology

KW - Melanoma/pathology

KW - Signal Transduction

KW - Wnt Proteins/metabolism

KW - Cancer

KW - Vascular Biology

KW - Melanoma

KW - Lymph

KW - Cell Biology

U2 - 10.1172/jci.insight.171821

DO - 10.1172/jci.insight.171821

M3 - Article

C2 - 37971882

SN - 2379-3708

VL - 9

JO - JCI Insight

JF - JCI Insight

IS - 1

M1 - e171821

ER -

DLL4/Notch3/WNT5B axis mediates bidirectional prometastatic crosstalk between melanoma and lymphatic endothelial cells

Sammanfattning

Finansiering

Nyckelord

Åtkomst av dokument

Fingeravtryck

Citera det här