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. 2022 Nov:155:113843.
doi: 10.1016/j.biopha.2022.113843. Epub 2022 Oct 8.

Histone deacetylase inhibitors enhance oncolytic herpes simplex virus therapy for malignant meningioma

Yoichiro Kawamura  1 , Lingyang Hua  2 , Alessandra Gurtner  1 , Ego Wong  1 , Juri Kiyokawa  1 , Nadia Shah  1 , Joshua Gorham  3 , Hiroko Wakimoto  3 , Samuel D Rabkin  1 , Robert L Martuza  1 , Hiroaki Wakimoto  4
Affiliations

Histone deacetylase inhibitors enhance oncolytic herpes simplex virus therapy for malignant meningioma

Yoichiro Kawamura et al. Biomed Pharmacother. 2022 Nov.

Abstract

Approximately 20% of meningiomas are not benign (higher grade) and tend to relapse after surgery and radiation therapy. Malignant (anaplastic) meningioma (MM) is a minor subset of high-grade meningioma that is lethal with no effective treatment options currently. Oncolytic herpes simplex virus (oHSV) is a powerful anti-cancer modality that induces both direct cell death and anti-tumor immunity, and has shown activity in preclinical models of MM. However, clinically meaningful efficacy will likely entail rational mechanistic combination approaches. We here show that epigenome modulator histone deacetylase inhibitors (HDACi) increase anti-cancer effects of oHSV in human MM models, IOMM-Lee (NF2 wild-type) and CH157 (NF2 mutant). Minimally toxic, sub-micromolar concentrations of pan-HDACi, Trichostatin A and Panobinostat, substantively increased the infectability and spread of oHSV G47Δ within MM cells in vitro, resulting in enhanced oHSV-mediated killing of target cells when infected at low multiplicity of infection (MOI). Transcriptomics analysis identified selective alteration of mRNA processing and splicing modules that might underlie the potent anti-MM effects of combining HDACi and oHSV. In vivo, HDACi treatment increased intratumoral oHSV replication and boosted the capacity of oHSV to control the growth of human MM xenografts. Thus, our work supports further translational development of the combination approach employing HDACi and oHSV for the treatment of MM.

Keywords: Histone deacetylase inhibitor; Malignant meningioma; Meningioma; Oncolytic herpes simplex virus.

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Conflict of interest statement

Conflict of interest statement SDR and RLM are co-inventors on patents relating to oncolytic herpes simplex viruses, owned and managed by Georgetown University and Massachusetts General Hospital, which have received royalties from Amgen and ActiVec Inc. SDR acted as a consultant and received honoraria from Replimune, Cellinta, and Greenfire Bio, and honoraria and equity from EG 427. RLM. is on the S.A.B. and receives payment from Virogin Biotech Ltd.

Figures

Fig. 1.
Fig. 1.
Low dose HDAC inhibitors enhance G47Δ-mediated killing of malignant meningioma cells in vitro. A and B, MTS cell viability assay showing dose-dependent responses to oHSV G47Δ in IOMM-Lee (A) and CH157 (B) cells. Assay was done 3 days post-infection. C and D, MTS cell viability assay showing dose-dependent responses to HDAC inhibitors Trichostatin A (TSA) and Panobinostat (Pano) in IOMM-Lee (C) and CH157 (D) cells. Assay was done 3 days after exposure to each drug. E-H, MTS cell viability assay showing the ability of a minimally toxic dose of HDAC inhibitors to sensitize malignant meningioma cells (IOMM-Lee, E and G; CH157, F and H) to G47Δ-mediated killing. Cells were pre-exposed to the indicated HDAC inhibitor for 5 h when cells were infected with G47Δ. MTS assay was done 3 days later. Mean and standard deviation (bars) of cell viability relative to untreated control cells are presented. *, p< 0.05; **, p< 0.01; and ***, p< 0.001 compared with no HDAC inhibitor in each group (Student t-test).
Fig. 2.
Fig. 2.
Pan-HDAC inhibitors enhance entry, replication, and spread of G47Δ in malignant meningioma cells in vitro. A-D, Virus infectability assay. A and B, IOMM-Lee cells. C and D, CH157 cells. A and C, Representative microscopic pictures showing x-gal positive cells in dark (black). Scale bars: 100 μm. B and D, Quantification of triplicate images. E and F, Virus yield assay. IOMM-Lee cells (E) and CH157 cells (F) were exposed to HDAC inhibitors at indicated concentration (nM), followed 5 h later by infection with G47Δ at MOI= 0.05 (IOMM-Lee) or 0.2 (CH157). Cells and culture media were collected at 24 h (IOMM-Lee) or 40 h (CH157) post-infection for determining virus yield. Dotted line: virus input. G and H, Virus spread assay. IOMM-Lee (G) or CH157 cells (H) were infected with G47Δ-mCherry at MOI= 0.1. Microscopic images for mCherry fluorescence were captured at 28 h (IOMM-Lee) and 45 h (CH157) post infection. Scale bars: 100 μm. *, p< 0.05; **, p< 0.01; ***, p< 0.001 compared with no HDAC inhibitor in each group (Student t-test).
Fig. 3.
Fig. 3.
RNA sequencing analysis of the impacts of HDAC inhibitors on G47Δ-induced transcriptomics. A, Principal component analysis. IOMM-Lee cells (IO, IOMM), CH157 cells (CH). Numbers that follow cell type denote: mock-treated control (C), 1 and 2; G47Δ alone, 3 and 4; Panobinostat (Pano), 5 and 6; Trichostatin A (TSA), 7 and 8; Pano+G47Δ, 9 and 10; and TSA+G47Δ, 11 and 12. G47Δ, G; HDAC inhibitors (Pano and TSA), Hi. Symbol coloring on right is based on grouping in which Pano and TSA groups were combined as Hi. B and C, Volcano plots, comparing HDAC inhibitors+G47Δ (Hi_G) vs. G47Δ alone (G) in IOMM-Lee cells (B) and CH157 cells (C). D and E, Gene Ontology (GO) enrichment analysis, comparing HDAC inhibitors+G47Δ vs. G47Δ alone in. D, IOMM-Lee cells (D) and CH157 cells (E). The most significant 30 GO terms are displayed. padj, p adjusted.
Fig. 4.
Fig. 4.
Impact of HDACis on the expression of genes involved in interferon-mediated innate anti-viral responses. Analysis of the RNA sequencing data presented in Fig. 3. A, IRF3, B, IRF7, C, STAT1, D, IFIT1, E, IFIT3, F, EIF2AK2 (PKR), G, OAS1 and H, PML.
Fig. 5.
Fig. 5.
Pan HDAC inhibitor enhances replication and therapeutic effects of oHSV in malignant meningioma. A, Tumor growth curves of IOMM-Lee flank xenografts treated with vehicle or Panobinostat (Pano). B, Schema showing the experimental design; Pano, 30 mg/kg on Days 0 and 1, and G47ΔUs11fluc injected intra-tumorally at 3 × 105 pfu on Day 0, 5 h after the first Pano dosing. BLI, Bioluminescence. Created by BioRender.com. C, BLI on Day 3. The group that received Pano alone was not subjected to BLI. D, Changes in total flux signal from Day 1 to Day 3. E, Tumor growth curves of the 3 groups: Pano, G47ΔUs11fluc (oHSV) and Pano+G47ΔUs11fluc. Tumor volume relative to Day 0 is presented to assist comparison between groups. F, Tumor volume ratio (Day 12/Day 0), comparing the 3 groups. Each dot represents a tumor. G, Spearman correlation coefficients to show relationship between Day 3 BLI total flux (TF)(x-axis) and tumor volume ratio (Day 12/Day 0)(y-axis) from the G47ΔUs11fluc (oHSV) and Pano+oHSV groups together. Each dot represents a tumor.
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