The Department of Neurosurgery, University of Texas Health Science Center at Houston, Houston, Texas.
Rice University, Houston, Texas.
Clin Cancer Res. 2020 May 15;26(10):2381-2392. doi: 10.1158/1078-0432.CCR-19-3420. Epub 2020 Mar 5.
To examine the effect of oncolytic herpes simplex virus (oHSV) on NOTCH signaling in central nervous system tumors.
Bioluminescence imaging, reverse phase protein array proteomics, fluorescence microscopy, reporter assays, and molecular biology approaches were used to evaluate NOTCH signaling. Orthotopic glioma-mouse models were utilized to evaluate effects .
We have identified that herpes simplex virus-1 (HSV-1; oncolytic and wild-type)-infected glioma cells induce NOTCH signaling, from inside of infected cells into adjacent tumor cells (inside out signaling). This was canonical NOTCH signaling, which resulted in activation of RBPJ-dependent transcriptional activity that could be rescued with dnMAML. High-throughput screening of HSV-1-encoded cDNA and miRNA libraries further uncovered that HSV-1 miR-H16 induced NOTCH signaling. We further identified that factor inhibiting HIF-1 (FIH-1) is a direct target of miR-H16, and that FIH-1 downregulation by virus encoded miR-H16 induces NOTCH activity. FIH-1 binding to Mib1 has been reported, but this is the first report that shows FIH-1 sequester Mib1 to suppress NOTCH activation. We observed that FIH-1 degradation induced NOTCH ligand ubiquitination and NOTCH activity. REMBRANDT and The Cancer Genome Atlas data analysis also uncovered a significant negative regulation between FIH-1 and NOTCH. Furthermore, combination of oHSV with NOTCH-blocking gamma secretase inhibitor (GSI) had a therapeutic advantage in two different intracranial glioma models treated with oncolytic HSV, without affecting safety profile of the virus .
To our knowledge this is the first report to identify impact of HSV-1 on NOTCH signaling and highlights the significance of combining oHSV and GSI for glioblastoma therapy.
研究溶瘤单纯疱疹病毒（oHSV）对中枢神经系统肿瘤中 NOTCH 信号通路的影响。
采用生物发光成像、反相蛋白阵列蛋白质组学、荧光显微镜、报告基因检测和分子生物学方法来评估 NOTCH 信号通路。利用原位胶质瘤-小鼠模型来评估作用。
我们发现单纯疱疹病毒 1（HSV-1；溶瘤和野生型）感染的神经胶质瘤细胞会诱导 NOTCH 信号从受感染的细胞内部传递到相邻的肿瘤细胞（内向信号）。这是典型的 NOTCH 信号，导致依赖 RBPJ 的转录活性激活，可通过 dnMAML 挽救。对 HSV-1 编码 cDNA 和 miRNA 文库的高通量筛选进一步揭示了 HSV-1miR-H16 诱导 NOTCH 信号。我们进一步确定了缺氧诱导因子 1（HIF-1）抑制剂（FIH-1）是 miR-H16 的直接靶标，病毒编码的 miR-H16 下调 FIH-1 可诱导 NOTCH 活性。已经报道了 FIH-1 与 Mib1 的结合，但这是首次报道 FIH-1 将 Mib1 隔离以抑制 NOTCH 激活。我们观察到 FIH-1 降解诱导 NOTCH 配体泛素化和 NOTCH 活性。REMBRANDT 和癌症基因组图谱数据分析还揭示了 FIH-1 和 NOTCH 之间存在显著的负调控关系。此外，在两种不同的颅内神经胶质瘤模型中，溶瘤单纯疱疹病毒联合 NOTCH 阻断γ分泌酶抑制剂（GSI）治疗具有治疗优势，而不会影响病毒的安全性。
据我们所知，这是首次报道 HSV-1 对 NOTCH 信号通路的影响，并强调了联合使用 oHSV 和 GSI 治疗胶质母细胞瘤的重要性。
