Dual targeting macrophages and microglia is a therapeutic vulnerability in models of PTEN-deficient glioblastoma
Yang Liu, … , Amy B. Heimberger, Peiwen Chen
Yang Liu, … , Amy B. Heimberger, Peiwen Chen
Published October 1, 2024
Citation Information: J Clin Invest. 2024;134(22):e178628. https://doi.org/10.1172/JCI178628.
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Research Article Oncology

Dual targeting macrophages and microglia is a therapeutic vulnerability in models of PTEN-deficient glioblastoma

Abstract

Tumor-associated macrophages and microglia (TAMs) are critical for tumor progression and therapy resistance in glioblastoma (GBM), a type of incurable brain cancer. We previously identified lysyl oxidase (LOX) and olfactomedin like-3 (OLFML3) as essential macrophage and microglia chemokines, respectively, in GBM. Here, single-cell transcriptomics and multiplex sequential immunofluorescence followed by functional studies demonstrate that macrophages negatively correlate with microglia in the GBM tumor microenvironment. LOX inhibition in PTEN-deficient GBM cells upregulates OLFML3 expression via the NF-κB-PATZ1 signaling pathway, inducing a compensatory increase of microglia infiltration. Dual targeting macrophages and microglia via inhibition of LOX and the CLOCK-OLFML3 axis generates potent antitumor effects and offers a complete tumor regression in more than 60% of animals when combined with anti-PD1 therapy in PTEN-deficient GBM mouse models. Thus, our findings provide a translational triple therapeutic strategy for this lethal disease.

Authors

Yang Liu, Junyan Wu, Hinda Najem, Yiyun Lin, Lizhi Pang, Fatima Khan, Fei Zhou, Heba Ali, Amy B. Heimberger, Peiwen Chen

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Figure 1

LOX inhibition improves the efficacy of anti-PD1 therapy.

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LOX inhibition improves the efficacy of anti-PD1 therapy. (A) High-resol...
(A) High-resolution uniform manifold approximation and projection (UMAP) dimensional reduction of different subtypes, including mesenchymal-like (MES-like), neural-progenitor-like (NPC-like), astrocyte-like (AC-like) and oligodendrocyte-progenitor-like (OPC-like), of tumor cells from tumors from patients with GBM based on the scRNA-seq dataset (GSE182109). (B) Pattern representing single-cell gene expression of LOX in distinct subtypes of tumor cells based on above scRNA-seq dataset. (C) Percentage MES-like GBM cells out of total GBM cells, and normalized LOX gene expression in different subtypes of malignant cells in tumors from patients with GBM based on above scRNA-seq dataset. (D) GSEA analysis for various types of immune cells in LOX-high (n = 123) and LOX-low (n = 122) patient tumors from the TCGA GBM database. (E and F) IF (E) and quantification (F) of relative CD8+CD69+ T cells in tumors from CT2A tumor-bearing mice treated with or without LOX inhibitor BAPN (2 g/L in drinking water) on day 4. Scale bar: 50 μm. n = 3 independent samples. Student’s t test. (G and H) Immunoblots for PD-L1 and LOX in lysates of U87 (G) and PTEN-KO SF763 (H) cells expressing shRNA control (shC) and LOX shRNAs (shLOX). (I) Immunoblots for PD-L1 in lysates of U87 and PTEN-KO SF763 cells treated with BAPN at indicated concentrations. (J) Immunoblots for PD-L1 in lysates of CT2A cells and 005 GSCs treated with BAPN at indicated concentration. (K and L) Survival curves of C57BL/6J mice implanted with CT2A cells (2 × 104 cells/mouse, K) or 005 GSCs (2 × 105 cells/mouse, L). Mice were treated with BAPN (2 g/L in drinking water) on day 4, and then received the treatment with IgG or anti-PD1 (10 mg/kg body weight, i.p.) on days 11, 14, and 17. n = 5–7 mice per group. Log-rank test. *P < 0.05; **P < 0.01; ***P < 0.001.