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Infiltration of CD163-positive macrophages in glioma tissues after treatment with anti-PD-L1 antibody and role of PI3Kγ inhibitor as a combination therapy with anti-PD-L1 antibody in in vivo model using temozolomide-resistant murine glioma-initiating cells

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Abstract

Although chemoimmunotherapy often lengthens glioblastoma (GBM) survival, early relapses remain problematic as immunosuppressive M2 macrophages (Mϕ) that function via inhibitory cytokine and PD-L1 production cause immunotherapy resistance. Here, we detail anti-PD-L1 antibody effects on the tumor microenvironment, including Mϕ infiltration, using a temozolomide (TMZ)-treated glioma model. In addition, we tested combinations of anti-PD-L1 antibody and the M2Mϕ inhibitor IPI-549 on tumor growth. We simulated late TMZ treatment or relapse stage, persistent GBM cells by generating TMZ-resistant TS (TMZRTS) cells. M2Mϕ-associated cytokine production and PD-L1 expression in these cells were investigated. TMZRTS cells were then subcutaneously implanted into C57BL/6 mice to determine the effectiveness of an anti-PD-L1 antibody and/or IPI-549 treatment on infiltration of CD163-positive Mϕ, usually considered as an M2Mϕ marker into tumor tissues. CD163 expression in samples from human GBM patients were also evaluated. CD163-positive Mϕ heavily infiltrated TMZRS tumor tissues after in vivo anti-PD-L1 antibody treatment. Tumor growth was strongly inhibited by anti-PD-L1 antibody and IPI-549 combination therapy. Anti-PD-L1 antibody treatment significantly reduced infiltration of CD163-positive Mϕ into tumors, while combined PD-L1 antibody and IPI-549 therapy remarkably inhibited tumor growth. These therapies may be useful for recurrent or chronic GBM after TMZ treatment, but clinical safety and effectiveness studies are needed.

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Abbreviations

FRT:

Fractionated radiotherapy

GBM:

Glioblastoma multiforme

IHC:

Immunohistochemistry

MGMT:

O6-Methylguanine-DNA methyltransferase

PD-1:

Programmed cell death 1

PD-L1:

PD-1 ligand

TIL:

Tumor-infiltrating lymphocytes

Mϕ:

Macrophage

TMZ:

Temozolomide

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Acknowledgements

The authors thank Dr. Alexander Zaboronok, Department of Neurosurgery, for critical revision. The authors would also like to thank Dr. Bryan J. Mathis of the Medical English Communications Center of the Faculty of Medicine, University of Tsukuba, for language revision.

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Author notes

  1. Tsubasa Miyazaki and Eiichi Ishikawa are co-first authors.

Authors and Affiliations

  1. Department of Neurosurgery, Faculty of Medicine, University of Tsukuba, 1-1-1 Tennodai, Tsukuba, Ibaraki, 305-8575, Japan

    Tsubasa Miyazaki, Eiichi Ishikawa, Masahide Matsuda, Narushi Sugii, Hedihiro Kohzuki, Hiroyoshi Akutsu, Shingo Takano & Akira Matsumura

  2. Cell-Medicine, Inc, Sengen 2-1-6, Tsukuba Science City, Ibaraki, 305-0047, Japan

    Tsubasa Miyazaki

  3. Department of Pathology, Faculty of Medicine, University of Tsukuba, 1-1-1 Tennoudai, Tsukuba, Ibaraki, 305-8575, Japan

    Noriaki Sakamoto

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Correspondence to Eiichi Ishikawa.

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Conflict of interest

This study was supported by a Grant-in-Aid for Scientific Research (Research No. 18K08962) from the Japanese Ministry of Education, Culture, Sports, Science and Technology (MEXT) and the project for promotion of practical applications of advanced medical technologies in Tsukuba University Hospital.

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10014_2020_357_MOESM1_ESM.tif

Supplemental Figure 1 The mRNA expression levels of DNA mismatch repair genes (MLH1, PMS2, MSH2, MSH3, MSH6) in TS and TMZRTS cells. (n = 3, mean ± SD) (TIF 48 kb)

10014_2020_357_MOESM2_ESM.tif

Supplemental Figure 2 CD163 mRNA expression in GBM patients and outcomes A. Datasets of CD163 mRNA expression in GBM and normal brain tissues using GlioVis. B. Survival outcome of GBM patients with high and low CD163 mRNA expression (TIF 112 kb)

10014_2020_357_MOESM3_ESM.tif

Supplemental Table S-Table 1 GBM patient characteristics, treatment histories and pathological status determined by immunohistochemistry (IHC) (TIF 85 kb)

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Miyazaki, T., Ishikawa, E., Matsuda, M. et al. Infiltration of CD163-positive macrophages in glioma tissues after treatment with anti-PD-L1 antibody and role of PI3Kγ inhibitor as a combination therapy with anti-PD-L1 antibody in in vivo model using temozolomide-resistant murine glioma-initiating cells. Brain Tumor Pathol 37, 41–49 (2020). https://doi.org/10.1007/s10014-020-00357-z

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