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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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 5

LOX regulates OLFML3 expression through regulating the NF-κB-PATZ1 signaling axis.

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LOX regulates OLFML3 expression through regulating the NF-κB-PATZ1 signa...
(A) GSEA analysis on RNA-seq data of U87 cells with LOX shRNA knockdown (shLOX) versus shRNA control (shC) shows top 10 enriched oncogenic signaling pathways. (B) Immunoblots for P-P65, P65, and LOX in lysates of U87 and PTEN-KO SF763 cells expressing shC and shLOX. (C) Relative mRNA expression of OLFML3 in PTEN-KO SF763 cells expressing shC and shLOX treated with or without P65 inhibitor (P65i) SC75741 (5 μM). n = 3 independent samples. Student’s t test. (D) Identification of 22 overlapping transcription factors (TFs) in TCGA GBM tumors (LOX-low versus -high) and U87 cells (shLOX versus shC). (E) Relative mRNA expression of 10 TFs in PTEN-KO SF763 cells expressing shC and shLOX. n = 3 independent samples. 1-way ANOVA test. (F) Relative mRNA expression of the 10 TFs in U87 cells treated with or without LOX inhibitor BAPN (200 μM). n = 3 independent samples. Student’s t test. (G) Immunoblots for PATZ1 in lysates of PTEN-KO SF763 cells expressing shC and shLOX. (H) Relative mRNA expression of PATZ1 in PTEN-KO SF763 cells expressing shC and shLOX and treated with or without P65i SC75741 (5 μM). n = 3 independent samples. Student’s t test. (I) Schematic of designing ChIP-qPCR primers based on 3 potential binding sites. (J) Quantification of PATZ1 ChIP-qPCR in the OLFML3 promoter of PTEN-KO SF763 cells. IgG was used as the control. n = 3 independent samples. Student’s t test. (K) Immunoblots for OLFML3 in lysates of PTEN-KO SF763 cells with or without PATZ1 overexpression (OE) and treated with or without P65 activator (+). (L) Immunoblots for OLFML3 in lysates of PTEN-WT SF763 cells expressing shC and shPATZ1 treated with or without P65i SC75741 (5 μM). *P < 0.05; **P < 0.01; ***P < 0.001; n.s., not significant (P > 0.05).