Identification of novel, immune-mediating extracellular vesicles in human lymphatic effluent draining primary cutaneous melanoma

Rachel L.G. Maus, James W. Jakub, Tina J. Hieken, Wendy K. Nevala, Trace A. Christensen, Shari L. Sutor, Thomas J. Flotte, Svetomir N. Markovic

Research output: Contribution to journalArticle

Abstract

Epithelial tumors including melanoma often first metastasize to regional, sentinel lymph nodes (SLN). Thus, the presence of SLN metastases is a critical prognostic factor of survival. Prior to metastasis, accumulating evidence suggests the SLN is immunologically compromised; however, the process by which pre-metastatic niche formation occurs remains unknown. In this prospective study, freshly dissected, afferent lymphatic fluid was obtained during SLN biopsy in three patients with primary cutaneous melanoma. Lymphatic extracellular vesicles (L-EV) were visualized by transmission electron microscopy and proteomic cargo profiled by mass spectrometry. Flow cytometry assessed L-EV effects on autologous dendritic cell maturation in vitro. Immunogold electron microscopy and immunohistochemistry visualized expression of EV cargo within the primary tumor and SLN. Lymphatic extracellular vesicles from each afferent lymphatic channel demonstrated inhibition of autologous dendritic cell maturation. Proteomic profiling identified 81 peptides shared among the L-EV preparations including a signature of 18 immune-modulating proteins including previously established inhibitor of dendritic cell maturation, S100A9. Immunohistochemistry and immunogold electron microscopy confirmed S100A9 tracking along the lymphatic path, from keratinocytes in the primary tumor to sub-capsular macrophages in the SLN. Our findings suggest L-EV cargo may serve as early mediators of tumor-induced immune subversion in regional lymph nodes, by preceding malignant cells and trafficking within the lymphatic vasculature to harbor the first pre-metastatic niche.

Original languageEnglish (US)
Article numbere1667742
JournalOncoImmunology
Volume8
Issue number12
DOIs
StatePublished - Dec 2 2019

Fingerprint

Melanoma
Skin
Dendritic Cells
Proteomics
Neoplasms
Electron Microscopy
Immunohistochemistry
Neoplasm Metastasis
Sentinel Lymph Node Biopsy
Transmission Electron Microscopy
Keratinocytes
Mass Spectrometry
Flow Cytometry
Lymph Nodes
Macrophages
Extracellular Vesicles
Sentinel Lymph Node
Prospective Studies
Peptides
Survival

Keywords

  • dendritic cells
  • extracellular vesicles
  • lymphatic
  • pre-metastatic niche
  • sentinel lymph node

ASJC Scopus subject areas

  • Immunology and Allergy
  • Immunology
  • Oncology

Cite this

Identification of novel, immune-mediating extracellular vesicles in human lymphatic effluent draining primary cutaneous melanoma. / Maus, Rachel L.G.; Jakub, James W.; Hieken, Tina J.; Nevala, Wendy K.; Christensen, Trace A.; Sutor, Shari L.; Flotte, Thomas J.; Markovic, Svetomir N.

In: OncoImmunology, Vol. 8, No. 12, e1667742, 02.12.2019.

Research output: Contribution to journalArticle

@article{e86ada45354244a0b2f3f72a931524b0,
title = "Identification of novel, immune-mediating extracellular vesicles in human lymphatic effluent draining primary cutaneous melanoma",
abstract = "Epithelial tumors including melanoma often first metastasize to regional, sentinel lymph nodes (SLN). Thus, the presence of SLN metastases is a critical prognostic factor of survival. Prior to metastasis, accumulating evidence suggests the SLN is immunologically compromised; however, the process by which pre-metastatic niche formation occurs remains unknown. In this prospective study, freshly dissected, afferent lymphatic fluid was obtained during SLN biopsy in three patients with primary cutaneous melanoma. Lymphatic extracellular vesicles (L-EV) were visualized by transmission electron microscopy and proteomic cargo profiled by mass spectrometry. Flow cytometry assessed L-EV effects on autologous dendritic cell maturation in vitro. Immunogold electron microscopy and immunohistochemistry visualized expression of EV cargo within the primary tumor and SLN. Lymphatic extracellular vesicles from each afferent lymphatic channel demonstrated inhibition of autologous dendritic cell maturation. Proteomic profiling identified 81 peptides shared among the L-EV preparations including a signature of 18 immune-modulating proteins including previously established inhibitor of dendritic cell maturation, S100A9. Immunohistochemistry and immunogold electron microscopy confirmed S100A9 tracking along the lymphatic path, from keratinocytes in the primary tumor to sub-capsular macrophages in the SLN. Our findings suggest L-EV cargo may serve as early mediators of tumor-induced immune subversion in regional lymph nodes, by preceding malignant cells and trafficking within the lymphatic vasculature to harbor the first pre-metastatic niche.",
keywords = "dendritic cells, extracellular vesicles, lymphatic, pre-metastatic niche, sentinel lymph node",
author = "Maus, {Rachel L.G.} and Jakub, {James W.} and Hieken, {Tina J.} and Nevala, {Wendy K.} and Christensen, {Trace A.} and Sutor, {Shari L.} and Flotte, {Thomas J.} and Markovic, {Svetomir N.}",
year = "2019",
month = "12",
day = "2",
doi = "10.1080/2162402X.2019.1667742",
language = "English (US)",
volume = "8",
journal = "OncoImmunology",
issn = "2162-4011",
publisher = "Landes Bioscience",
number = "12",

}

TY - JOUR

T1 - Identification of novel, immune-mediating extracellular vesicles in human lymphatic effluent draining primary cutaneous melanoma

AU - Maus, Rachel L.G.

AU - Jakub, James W.

AU - Hieken, Tina J.

AU - Nevala, Wendy K.

AU - Christensen, Trace A.

AU - Sutor, Shari L.

AU - Flotte, Thomas J.

AU - Markovic, Svetomir N.

PY - 2019/12/2

Y1 - 2019/12/2

N2 - Epithelial tumors including melanoma often first metastasize to regional, sentinel lymph nodes (SLN). Thus, the presence of SLN metastases is a critical prognostic factor of survival. Prior to metastasis, accumulating evidence suggests the SLN is immunologically compromised; however, the process by which pre-metastatic niche formation occurs remains unknown. In this prospective study, freshly dissected, afferent lymphatic fluid was obtained during SLN biopsy in three patients with primary cutaneous melanoma. Lymphatic extracellular vesicles (L-EV) were visualized by transmission electron microscopy and proteomic cargo profiled by mass spectrometry. Flow cytometry assessed L-EV effects on autologous dendritic cell maturation in vitro. Immunogold electron microscopy and immunohistochemistry visualized expression of EV cargo within the primary tumor and SLN. Lymphatic extracellular vesicles from each afferent lymphatic channel demonstrated inhibition of autologous dendritic cell maturation. Proteomic profiling identified 81 peptides shared among the L-EV preparations including a signature of 18 immune-modulating proteins including previously established inhibitor of dendritic cell maturation, S100A9. Immunohistochemistry and immunogold electron microscopy confirmed S100A9 tracking along the lymphatic path, from keratinocytes in the primary tumor to sub-capsular macrophages in the SLN. Our findings suggest L-EV cargo may serve as early mediators of tumor-induced immune subversion in regional lymph nodes, by preceding malignant cells and trafficking within the lymphatic vasculature to harbor the first pre-metastatic niche.

AB - Epithelial tumors including melanoma often first metastasize to regional, sentinel lymph nodes (SLN). Thus, the presence of SLN metastases is a critical prognostic factor of survival. Prior to metastasis, accumulating evidence suggests the SLN is immunologically compromised; however, the process by which pre-metastatic niche formation occurs remains unknown. In this prospective study, freshly dissected, afferent lymphatic fluid was obtained during SLN biopsy in three patients with primary cutaneous melanoma. Lymphatic extracellular vesicles (L-EV) were visualized by transmission electron microscopy and proteomic cargo profiled by mass spectrometry. Flow cytometry assessed L-EV effects on autologous dendritic cell maturation in vitro. Immunogold electron microscopy and immunohistochemistry visualized expression of EV cargo within the primary tumor and SLN. Lymphatic extracellular vesicles from each afferent lymphatic channel demonstrated inhibition of autologous dendritic cell maturation. Proteomic profiling identified 81 peptides shared among the L-EV preparations including a signature of 18 immune-modulating proteins including previously established inhibitor of dendritic cell maturation, S100A9. Immunohistochemistry and immunogold electron microscopy confirmed S100A9 tracking along the lymphatic path, from keratinocytes in the primary tumor to sub-capsular macrophages in the SLN. Our findings suggest L-EV cargo may serve as early mediators of tumor-induced immune subversion in regional lymph nodes, by preceding malignant cells and trafficking within the lymphatic vasculature to harbor the first pre-metastatic niche.

KW - dendritic cells

KW - extracellular vesicles

KW - lymphatic

KW - pre-metastatic niche

KW - sentinel lymph node

UR - http://www.scopus.com/inward/record.url?scp=85073994904&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85073994904&partnerID=8YFLogxK

U2 - 10.1080/2162402X.2019.1667742

DO - 10.1080/2162402X.2019.1667742

M3 - Article

AN - SCOPUS:85073994904

VL - 8

JO - OncoImmunology

JF - OncoImmunology

SN - 2162-4011

IS - 12

M1 - e1667742

ER -