Digital PCR-Based T-cell Quantification-Assisted Deconvolution of the Microenvironment Reveals that Activated Macrophages Drive Tumor Inflammation in Uveal Melanoma

Mark J de Lange, Rogier J Nell, Rajshri N Lalai, Mieke Versluis, Ekaterina S Jordanova, Gre P M Luyten, Martine J Jager, Sjoerd H van der Burg, Willem H Zoutman, Thorbald van Hall, Pieter A van der Velden

Research output: Contribution to journalArticleAcademicpeer-review

Abstract

Uveal melanoma progression can be predicted by gene expression profiles enabling a clear subdivision between tumors with a good (class I) and a poor (class II) prognosis. Poor prognosis uveal melanoma can be subdivided by expression of immune-related genes; however, it is unclear whether this subclassification is justified; therefore, T cells in uveal melanoma specimens were quantified using a digital PCR approach. Absolute T-cell quantification revealed that T-cell influx is present in all uveal melanomas associated with a poor prognosis. However, this infiltrate is only accompanied by differential immune-related gene expression profiles in uveal melanoma with the highest T-cell infiltrate. Molecular deconvolution of the immune profile revealed that a large proportion of the T-cell-related gene expression signature does not originate from lymphocytes but is derived from other immune cells, especially macrophages. Expression of the lymphocyte-homing chemokine CXCL10 by activated macrophages correlated with T-cell infiltration and thereby explains the correlation of T-cell numbers and macrophages. This was validated by in situ analysis of CXCL10 in uveal melanoma tissue with high T-cell counts. Surprisingly, CXCL10 or any of the other genes in the activated macrophage-cluster was correlated with reduced survival due to uveal melanoma metastasis. This effect was independent of the T-cell infiltrate, which reveals a role for activated macrophages in metastasis formation independent of their role in tumor inflammation.Implications: The current report uses an innovative digital PCR method to study the immune environment and demonstrates that absolute T-cell quantification and expression profiles can dissect disparate immune components. Mol Cancer Res; 1-10. ©2018 AACR.

Original languageEnglish
Pages (from-to)1902-1911
Number of pages10
JournalMolecular Cancer Research
Volume16
Issue number12
Early online date9 Aug 2018
DOIs
Publication statusPublished - Dec 2018

Cite this

de Lange, Mark J ; Nell, Rogier J ; Lalai, Rajshri N ; Versluis, Mieke ; Jordanova, Ekaterina S ; Luyten, Gre P M ; Jager, Martine J ; van der Burg, Sjoerd H ; Zoutman, Willem H ; van Hall, Thorbald ; van der Velden, Pieter A. / Digital PCR-Based T-cell Quantification-Assisted Deconvolution of the Microenvironment Reveals that Activated Macrophages Drive Tumor Inflammation in Uveal Melanoma. In: Molecular Cancer Research. 2018 ; Vol. 16, No. 12. pp. 1902-1911.
@article{44cc87e539af46deb66eab6a14786b7c,
title = "Digital PCR-Based T-cell Quantification-Assisted Deconvolution of the Microenvironment Reveals that Activated Macrophages Drive Tumor Inflammation in Uveal Melanoma",
abstract = "Uveal melanoma progression can be predicted by gene expression profiles enabling a clear subdivision between tumors with a good (class I) and a poor (class II) prognosis. Poor prognosis uveal melanoma can be subdivided by expression of immune-related genes; however, it is unclear whether this subclassification is justified; therefore, T cells in uveal melanoma specimens were quantified using a digital PCR approach. Absolute T-cell quantification revealed that T-cell influx is present in all uveal melanomas associated with a poor prognosis. However, this infiltrate is only accompanied by differential immune-related gene expression profiles in uveal melanoma with the highest T-cell infiltrate. Molecular deconvolution of the immune profile revealed that a large proportion of the T-cell-related gene expression signature does not originate from lymphocytes but is derived from other immune cells, especially macrophages. Expression of the lymphocyte-homing chemokine CXCL10 by activated macrophages correlated with T-cell infiltration and thereby explains the correlation of T-cell numbers and macrophages. This was validated by in situ analysis of CXCL10 in uveal melanoma tissue with high T-cell counts. Surprisingly, CXCL10 or any of the other genes in the activated macrophage-cluster was correlated with reduced survival due to uveal melanoma metastasis. This effect was independent of the T-cell infiltrate, which reveals a role for activated macrophages in metastasis formation independent of their role in tumor inflammation.Implications: The current report uses an innovative digital PCR method to study the immune environment and demonstrates that absolute T-cell quantification and expression profiles can dissect disparate immune components. Mol Cancer Res; 1-10. {\circledC}2018 AACR.",
author = "{de Lange}, {Mark J} and Nell, {Rogier J} and Lalai, {Rajshri N} and Mieke Versluis and Jordanova, {Ekaterina S} and Luyten, {Gre P M} and Jager, {Martine J} and {van der Burg}, {Sjoerd H} and Zoutman, {Willem H} and {van Hall}, Thorbald and {van der Velden}, {Pieter A}",
note = "{\circledC}2018 American Association for Cancer Research.",
year = "2018",
month = "12",
doi = "10.1158/1541-7786.MCR-18-0114",
language = "English",
volume = "16",
pages = "1902--1911",
journal = "Molecular Cancer Research",
issn = "1541-7786",
publisher = "American Association for Cancer Research Inc.",
number = "12",

}

de Lange, MJ, Nell, RJ, Lalai, RN, Versluis, M, Jordanova, ES, Luyten, GPM, Jager, MJ, van der Burg, SH, Zoutman, WH, van Hall, T & van der Velden, PA 2018, 'Digital PCR-Based T-cell Quantification-Assisted Deconvolution of the Microenvironment Reveals that Activated Macrophages Drive Tumor Inflammation in Uveal Melanoma' Molecular Cancer Research, vol. 16, no. 12, pp. 1902-1911. https://doi.org/10.1158/1541-7786.MCR-18-0114

Digital PCR-Based T-cell Quantification-Assisted Deconvolution of the Microenvironment Reveals that Activated Macrophages Drive Tumor Inflammation in Uveal Melanoma. / de Lange, Mark J; Nell, Rogier J; Lalai, Rajshri N; Versluis, Mieke; Jordanova, Ekaterina S; Luyten, Gre P M; Jager, Martine J; van der Burg, Sjoerd H; Zoutman, Willem H; van Hall, Thorbald; van der Velden, Pieter A.

In: Molecular Cancer Research, Vol. 16, No. 12, 12.2018, p. 1902-1911.

Research output: Contribution to journalArticleAcademicpeer-review

TY - JOUR

T1 - Digital PCR-Based T-cell Quantification-Assisted Deconvolution of the Microenvironment Reveals that Activated Macrophages Drive Tumor Inflammation in Uveal Melanoma

AU - de Lange, Mark J

AU - Nell, Rogier J

AU - Lalai, Rajshri N

AU - Versluis, Mieke

AU - Jordanova, Ekaterina S

AU - Luyten, Gre P M

AU - Jager, Martine J

AU - van der Burg, Sjoerd H

AU - Zoutman, Willem H

AU - van Hall, Thorbald

AU - van der Velden, Pieter A

N1 - ©2018 American Association for Cancer Research.

PY - 2018/12

Y1 - 2018/12

N2 - Uveal melanoma progression can be predicted by gene expression profiles enabling a clear subdivision between tumors with a good (class I) and a poor (class II) prognosis. Poor prognosis uveal melanoma can be subdivided by expression of immune-related genes; however, it is unclear whether this subclassification is justified; therefore, T cells in uveal melanoma specimens were quantified using a digital PCR approach. Absolute T-cell quantification revealed that T-cell influx is present in all uveal melanomas associated with a poor prognosis. However, this infiltrate is only accompanied by differential immune-related gene expression profiles in uveal melanoma with the highest T-cell infiltrate. Molecular deconvolution of the immune profile revealed that a large proportion of the T-cell-related gene expression signature does not originate from lymphocytes but is derived from other immune cells, especially macrophages. Expression of the lymphocyte-homing chemokine CXCL10 by activated macrophages correlated with T-cell infiltration and thereby explains the correlation of T-cell numbers and macrophages. This was validated by in situ analysis of CXCL10 in uveal melanoma tissue with high T-cell counts. Surprisingly, CXCL10 or any of the other genes in the activated macrophage-cluster was correlated with reduced survival due to uveal melanoma metastasis. This effect was independent of the T-cell infiltrate, which reveals a role for activated macrophages in metastasis formation independent of their role in tumor inflammation.Implications: The current report uses an innovative digital PCR method to study the immune environment and demonstrates that absolute T-cell quantification and expression profiles can dissect disparate immune components. Mol Cancer Res; 1-10. ©2018 AACR.

AB - Uveal melanoma progression can be predicted by gene expression profiles enabling a clear subdivision between tumors with a good (class I) and a poor (class II) prognosis. Poor prognosis uveal melanoma can be subdivided by expression of immune-related genes; however, it is unclear whether this subclassification is justified; therefore, T cells in uveal melanoma specimens were quantified using a digital PCR approach. Absolute T-cell quantification revealed that T-cell influx is present in all uveal melanomas associated with a poor prognosis. However, this infiltrate is only accompanied by differential immune-related gene expression profiles in uveal melanoma with the highest T-cell infiltrate. Molecular deconvolution of the immune profile revealed that a large proportion of the T-cell-related gene expression signature does not originate from lymphocytes but is derived from other immune cells, especially macrophages. Expression of the lymphocyte-homing chemokine CXCL10 by activated macrophages correlated with T-cell infiltration and thereby explains the correlation of T-cell numbers and macrophages. This was validated by in situ analysis of CXCL10 in uveal melanoma tissue with high T-cell counts. Surprisingly, CXCL10 or any of the other genes in the activated macrophage-cluster was correlated with reduced survival due to uveal melanoma metastasis. This effect was independent of the T-cell infiltrate, which reveals a role for activated macrophages in metastasis formation independent of their role in tumor inflammation.Implications: The current report uses an innovative digital PCR method to study the immune environment and demonstrates that absolute T-cell quantification and expression profiles can dissect disparate immune components. Mol Cancer Res; 1-10. ©2018 AACR.

U2 - 10.1158/1541-7786.MCR-18-0114

DO - 10.1158/1541-7786.MCR-18-0114

M3 - Article

VL - 16

SP - 1902

EP - 1911

JO - Molecular Cancer Research

JF - Molecular Cancer Research

SN - 1541-7786

IS - 12

ER -