Differential expression of immune-regulatory genes associated with PD-L1 display in melanoma: Implications for PD-1 pathway blockade

Janis M Taube, Geoffrey D. Young, Tracee L. McMiller, Shuming Chen, January T. Salas, Theresa S. Pritchard, Haiying Xu, Alan Keith Meeker, Jinshui Fan, Chris Cheadle, Alan E. Berger, Andrew Mark Pardoll, Suzanne Topalian

Research output: Contribution to journalArticle

Abstract

Purpose: Blocking the immunosuppressive PD-1/PD-L1 pathway has antitumor activity in multiple cancer types, and PD-L1 expression on tumor cells and infiltrating myeloid cells correlates with the likelihood of response. We previously found that IFNG (interferon-gamma) was overexpressed by tumor-infiltrating lymphocytes in PD-L1+ versus PD-L1(-) melanomas, creating adaptive immune resistance by promoting PD-L1 display. This study was undertaken to identify additional factors in the PD-L1+ melanoma microenvironment coordinately contributing to immunosuppression. Experimental Design: Archived, formalin-fixed paraffin-embedded melanoma specimens were assessed for PD-L1 protein expression at the tumor cell surface with IHC. Whole-genome expression analysis, quantitative (q)RT-PCR, IHC, and functional in vitro validation studies were used to assess factors differentially expressed in PD-L1+ versus PD-L1(-) melanomas. Results: Functional annotation clustering based on whole-genome expression profiling revealed pathways upregulated in PD-L1+ melanomas, involving immune cell activation, inflammation, and antigen processing and presentation. Analysis by qRT-PCR demonstrated overexpression of functionally related genes in PD-L1+ melanomas, involved in CD8+ T-cell activation (CD8A, IFNG, PRF1, and CCL5), antigen presentation (CD163, TLR3, CXCL1, and LYZ), and immunosuppression [PDCD1 (PD-1), CD274 (PD-L1), and LAG3, IL10]. Functional studies demonstrated that some factors, including IL10 and IL32-gamma, induced PD-L1 expression on monocytes but not tumor cells. Conclusions: These studies elucidate the complexity of immune checkpoint regulation in the tumor microenvironment, identifying multiple factors likely contributing to coordinated immunosuppression. These factors may provide tumor escape mechanisms from anti-PD-1/PD-L1 therapy, and should be considered for cotargeting in combinatorial immunomodulation treatment strategies.

Original languageEnglish (US)
Pages (from-to)3969-3976
Number of pages8
JournalClinical Cancer Research
Volume21
Issue number17
DOIs
StatePublished - Sep 1 2015

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Regulator Genes
Melanoma
Antigen Presentation
Immunosuppression
Interleukin-10
Interferon-gamma
Neoplasms
Genome
Tumor Escape
Tumor-Infiltrating Lymphocytes
Polymerase Chain Reaction
Tumor Microenvironment
Immunomodulation
Validation Studies
Myeloid Cells
Immunosuppressive Agents
Paraffin
Formaldehyde
Cluster Analysis
Monocytes

ASJC Scopus subject areas

  • Cancer Research
  • Oncology

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Differential expression of immune-regulatory genes associated with PD-L1 display in melanoma : Implications for PD-1 pathway blockade. / Taube, Janis M; Young, Geoffrey D.; McMiller, Tracee L.; Chen, Shuming; Salas, January T.; Pritchard, Theresa S.; Xu, Haiying; Meeker, Alan Keith; Fan, Jinshui; Cheadle, Chris; Berger, Alan E.; Pardoll, Andrew Mark; Topalian, Suzanne.

In: Clinical Cancer Research, Vol. 21, No. 17, 01.09.2015, p. 3969-3976.

Research output: Contribution to journalArticle

Taube, Janis M ; Young, Geoffrey D. ; McMiller, Tracee L. ; Chen, Shuming ; Salas, January T. ; Pritchard, Theresa S. ; Xu, Haiying ; Meeker, Alan Keith ; Fan, Jinshui ; Cheadle, Chris ; Berger, Alan E. ; Pardoll, Andrew Mark ; Topalian, Suzanne. / Differential expression of immune-regulatory genes associated with PD-L1 display in melanoma : Implications for PD-1 pathway blockade. In: Clinical Cancer Research. 2015 ; Vol. 21, No. 17. pp. 3969-3976.
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abstract = "Purpose: Blocking the immunosuppressive PD-1/PD-L1 pathway has antitumor activity in multiple cancer types, and PD-L1 expression on tumor cells and infiltrating myeloid cells correlates with the likelihood of response. We previously found that IFNG (interferon-gamma) was overexpressed by tumor-infiltrating lymphocytes in PD-L1+ versus PD-L1(-) melanomas, creating adaptive immune resistance by promoting PD-L1 display. This study was undertaken to identify additional factors in the PD-L1+ melanoma microenvironment coordinately contributing to immunosuppression. Experimental Design: Archived, formalin-fixed paraffin-embedded melanoma specimens were assessed for PD-L1 protein expression at the tumor cell surface with IHC. Whole-genome expression analysis, quantitative (q)RT-PCR, IHC, and functional in vitro validation studies were used to assess factors differentially expressed in PD-L1+ versus PD-L1(-) melanomas. Results: Functional annotation clustering based on whole-genome expression profiling revealed pathways upregulated in PD-L1+ melanomas, involving immune cell activation, inflammation, and antigen processing and presentation. Analysis by qRT-PCR demonstrated overexpression of functionally related genes in PD-L1+ melanomas, involved in CD8+ T-cell activation (CD8A, IFNG, PRF1, and CCL5), antigen presentation (CD163, TLR3, CXCL1, and LYZ), and immunosuppression [PDCD1 (PD-1), CD274 (PD-L1), and LAG3, IL10]. Functional studies demonstrated that some factors, including IL10 and IL32-gamma, induced PD-L1 expression on monocytes but not tumor cells. Conclusions: These studies elucidate the complexity of immune checkpoint regulation in the tumor microenvironment, identifying multiple factors likely contributing to coordinated immunosuppression. These factors may provide tumor escape mechanisms from anti-PD-1/PD-L1 therapy, and should be considered for cotargeting in combinatorial immunomodulation treatment strategies.",
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T1 - Differential expression of immune-regulatory genes associated with PD-L1 display in melanoma

T2 - Implications for PD-1 pathway blockade

AU - Taube, Janis M

AU - Young, Geoffrey D.

AU - McMiller, Tracee L.

AU - Chen, Shuming

AU - Salas, January T.

AU - Pritchard, Theresa S.

AU - Xu, Haiying

AU - Meeker, Alan Keith

AU - Fan, Jinshui

AU - Cheadle, Chris

AU - Berger, Alan E.

AU - Pardoll, Andrew Mark

AU - Topalian, Suzanne

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