Anti-human CD73 monoclonal antibody inhibits metastasis formation in human breast cancer by inducing clustering and internalization of CD73 expressed on the surface of cancer cells

Mikkel G Terp, Kristina A Olesen, Eva Arnspang Christensen, Rikke R Lund, B Christoffer Lagerholm, Henrik J Ditzel, Rikke Leth-Larsen

Research output: Contribution to journalJournal articleResearchpeer-review

Abstract

Recent studies have shown that Abs that target the cell-surface enzyme CD73 (ecto-5'-nucleotidase) reduce growth of primary tumors and metastasis in syngenic mice by inhibiting the catalytic activity of CD73, and thus increasing the activity of cytotoxic T lymphocytes. In this article, we report another anticancer mechanism of anti-CD73 Abs and show that an anti-CD73 mAb (AD2) inhibits metastasis formation by a mechanism independent of CD73 catalytic activity and inhibition of primary tumor growth. This mechanism involves clustering and internalization of CD73, but does not require cross-linking of CD73, because both whole IgG anti-CD73 AD2 mAb and Fab' fragments thereof exhibited this effect. Ex vivo treatment of different breast cancer cell lines with anti-CD73 AD2 mAb before i.v. injection into mice inhibited extravasation/colonization of circulating tumor cells and significantly reduced metastasis development. This effect was also observed when the cancer cell-surface expression of CD73 was significantly reduced by small interfering RNA knockdown. The antimetastatic activity is epitope specific, as another Ab that efficiently binds CD73-expressing live cancer cells did not lead to CD73 internalization and metastasis inhibition. Furthermore, anti-CD73 AD2 mAb inhibited development of metastasis in a spontaneous animal model of human metastatic breast cancer. Our study shows that some anti-CD73 mAbs cause cell-surface clustering of CD73 followed by internalization, thus inhibiting the ability of circulating tumor cells to extravasate and colonize, leading to inhibition of metastasis. Ab-based CD73 cancer therapy should include a combination of Abs that target the catalytic activity of CD73, as well as those with the characteristics described in this article.
Original languageEnglish
JournalJournal of Immunology
Volume191
Issue number8
Pages (from-to)4165-4173
ISSN0022-1767
DOIs
Publication statusPublished - 2013

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Cluster Analysis
Circulating Neoplastic Cells
Neoplasms
5'-Nucleotidase
Growth
Small Interfering RNA
Cell Line
Enzymes

Keywords

  • 5'-Nucleotidase
  • Animals
  • Antibodies, Monoclonal
  • Biological Transport
  • Breast Neoplasms
  • Cell Line, Tumor
  • Cell Movement
  • Female
  • Humans
  • Immunoglobulin Fab Fragments
  • Mice
  • Neoplasm Metastasis
  • Neoplasm Transplantation
  • Neoplastic Cells, Circulating
  • RNA Interference
  • RNA, Small Interfering
  • Xenograft Model Antitumor Assays

Cite this

@article{95f74dec2dad4a858d33e8c87fc781dd,
title = "Anti-human CD73 monoclonal antibody inhibits metastasis formation in human breast cancer by inducing clustering and internalization of CD73 expressed on the surface of cancer cells",
abstract = "Recent studies have shown that Abs that target the cell-surface enzyme CD73 (ecto-5'-nucleotidase) reduce growth of primary tumors and metastasis in syngenic mice by inhibiting the catalytic activity of CD73, and thus increasing the activity of cytotoxic T lymphocytes. In this article, we report another anticancer mechanism of anti-CD73 Abs and show that an anti-CD73 mAb (AD2) inhibits metastasis formation by a mechanism independent of CD73 catalytic activity and inhibition of primary tumor growth. This mechanism involves clustering and internalization of CD73, but does not require cross-linking of CD73, because both whole IgG anti-CD73 AD2 mAb and Fab' fragments thereof exhibited this effect. Ex vivo treatment of different breast cancer cell lines with anti-CD73 AD2 mAb before i.v. injection into mice inhibited extravasation/colonization of circulating tumor cells and significantly reduced metastasis development. This effect was also observed when the cancer cell-surface expression of CD73 was significantly reduced by small interfering RNA knockdown. The antimetastatic activity is epitope specific, as another Ab that efficiently binds CD73-expressing live cancer cells did not lead to CD73 internalization and metastasis inhibition. Furthermore, anti-CD73 AD2 mAb inhibited development of metastasis in a spontaneous animal model of human metastatic breast cancer. Our study shows that some anti-CD73 mAbs cause cell-surface clustering of CD73 followed by internalization, thus inhibiting the ability of circulating tumor cells to extravasate and colonize, leading to inhibition of metastasis. Ab-based CD73 cancer therapy should include a combination of Abs that target the catalytic activity of CD73, as well as those with the characteristics described in this article.",
keywords = "5'-Nucleotidase, Animals, Antibodies, Monoclonal, Biological Transport, Breast Neoplasms, Cell Line, Tumor, Cell Movement, Female, Humans, Immunoglobulin Fab Fragments, Mice, Neoplasm Metastasis, Neoplasm Transplantation, Neoplastic Cells, Circulating, RNA Interference, RNA, Small Interfering, Xenograft Model Antitumor Assays",
author = "Terp, {Mikkel G} and Olesen, {Kristina A} and Christensen, {Eva Arnspang} and Lund, {Rikke R} and Lagerholm, {B Christoffer} and Ditzel, {Henrik J} and Rikke Leth-Larsen",
year = "2013",
doi = "10.4049/jimmunol.1301274",
language = "English",
volume = "191",
pages = "4165--4173",
journal = "Journal of Immunology",
issn = "0022-1767",
publisher = "American Association of Immunologists",
number = "8",

}

Anti-human CD73 monoclonal antibody inhibits metastasis formation in human breast cancer by inducing clustering and internalization of CD73 expressed on the surface of cancer cells. / Terp, Mikkel G; Olesen, Kristina A; Christensen, Eva Arnspang; Lund, Rikke R; Lagerholm, B Christoffer; Ditzel, Henrik J; Leth-Larsen, Rikke.

In: Journal of Immunology, Vol. 191, No. 8, 2013, p. 4165-4173.

Research output: Contribution to journalJournal articleResearchpeer-review

TY - JOUR

T1 - Anti-human CD73 monoclonal antibody inhibits metastasis formation in human breast cancer by inducing clustering and internalization of CD73 expressed on the surface of cancer cells

AU - Terp, Mikkel G

AU - Olesen, Kristina A

AU - Christensen, Eva Arnspang

AU - Lund, Rikke R

AU - Lagerholm, B Christoffer

AU - Ditzel, Henrik J

AU - Leth-Larsen, Rikke

PY - 2013

Y1 - 2013

N2 - Recent studies have shown that Abs that target the cell-surface enzyme CD73 (ecto-5'-nucleotidase) reduce growth of primary tumors and metastasis in syngenic mice by inhibiting the catalytic activity of CD73, and thus increasing the activity of cytotoxic T lymphocytes. In this article, we report another anticancer mechanism of anti-CD73 Abs and show that an anti-CD73 mAb (AD2) inhibits metastasis formation by a mechanism independent of CD73 catalytic activity and inhibition of primary tumor growth. This mechanism involves clustering and internalization of CD73, but does not require cross-linking of CD73, because both whole IgG anti-CD73 AD2 mAb and Fab' fragments thereof exhibited this effect. Ex vivo treatment of different breast cancer cell lines with anti-CD73 AD2 mAb before i.v. injection into mice inhibited extravasation/colonization of circulating tumor cells and significantly reduced metastasis development. This effect was also observed when the cancer cell-surface expression of CD73 was significantly reduced by small interfering RNA knockdown. The antimetastatic activity is epitope specific, as another Ab that efficiently binds CD73-expressing live cancer cells did not lead to CD73 internalization and metastasis inhibition. Furthermore, anti-CD73 AD2 mAb inhibited development of metastasis in a spontaneous animal model of human metastatic breast cancer. Our study shows that some anti-CD73 mAbs cause cell-surface clustering of CD73 followed by internalization, thus inhibiting the ability of circulating tumor cells to extravasate and colonize, leading to inhibition of metastasis. Ab-based CD73 cancer therapy should include a combination of Abs that target the catalytic activity of CD73, as well as those with the characteristics described in this article.

AB - Recent studies have shown that Abs that target the cell-surface enzyme CD73 (ecto-5'-nucleotidase) reduce growth of primary tumors and metastasis in syngenic mice by inhibiting the catalytic activity of CD73, and thus increasing the activity of cytotoxic T lymphocytes. In this article, we report another anticancer mechanism of anti-CD73 Abs and show that an anti-CD73 mAb (AD2) inhibits metastasis formation by a mechanism independent of CD73 catalytic activity and inhibition of primary tumor growth. This mechanism involves clustering and internalization of CD73, but does not require cross-linking of CD73, because both whole IgG anti-CD73 AD2 mAb and Fab' fragments thereof exhibited this effect. Ex vivo treatment of different breast cancer cell lines with anti-CD73 AD2 mAb before i.v. injection into mice inhibited extravasation/colonization of circulating tumor cells and significantly reduced metastasis development. This effect was also observed when the cancer cell-surface expression of CD73 was significantly reduced by small interfering RNA knockdown. The antimetastatic activity is epitope specific, as another Ab that efficiently binds CD73-expressing live cancer cells did not lead to CD73 internalization and metastasis inhibition. Furthermore, anti-CD73 AD2 mAb inhibited development of metastasis in a spontaneous animal model of human metastatic breast cancer. Our study shows that some anti-CD73 mAbs cause cell-surface clustering of CD73 followed by internalization, thus inhibiting the ability of circulating tumor cells to extravasate and colonize, leading to inhibition of metastasis. Ab-based CD73 cancer therapy should include a combination of Abs that target the catalytic activity of CD73, as well as those with the characteristics described in this article.

KW - 5'-Nucleotidase

KW - Animals

KW - Antibodies, Monoclonal

KW - Biological Transport

KW - Breast Neoplasms

KW - Cell Line, Tumor

KW - Cell Movement

KW - Female

KW - Humans

KW - Immunoglobulin Fab Fragments

KW - Mice

KW - Neoplasm Metastasis

KW - Neoplasm Transplantation

KW - Neoplastic Cells, Circulating

KW - RNA Interference

KW - RNA, Small Interfering

KW - Xenograft Model Antitumor Assays

U2 - 10.4049/jimmunol.1301274

DO - 10.4049/jimmunol.1301274

M3 - Journal article

C2 - 24043904

VL - 191

SP - 4165

EP - 4173

JO - Journal of Immunology

JF - Journal of Immunology

SN - 0022-1767

IS - 8

ER -