PO.TB10.09 · 肿瘤生物学
抗病毒治疗逆转 CMV 诱导的致癌信号并增强 NK 细胞对胶质母细胞瘤的细胞毒性，伴随 PD-L1 上调和免疫调节
Antiviral therapy reverses CMV-induced oncogenic signaling and enhances NK cell cytotoxicity in glioblastoma with associated PD-L1 upregulation and iImmune modulation
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中文摘要
背景：巨细胞病毒（CMV）已被认为通过促进干性、血管生成和免疫逃逸参与胶质母细胞瘤（GBM）的发病机制。在 GBM 中靶向 CMV 的临床试验已显示出初步的前景，然而其分子机制仍不明确。我们假设 CMV 增强 GBM 的致癌信号和免疫抵抗，而抗病毒治疗可能逆转这些效应并提高对免疫介导细胞毒性的易感性。
材料与方法：将人 GBM 细胞系（U251、U87、U138、T98G、LN229）和正常星形胶质细胞用 mCherry-人 CMV TB40 株（MOI 0.5）感染。进行了包括球体和集落形成在内的表型实验。使用 60 重细胞因子谱和 Western 印迹评估细胞因子谱和信号通路活化（IL-6/STAT3、Akt、SOX2、Survivin、p-RB）。使用 CMFDA 和乙锭同型二聚体染料，与 NK-92mi 细胞进行共培养实验以评估细胞毒性。
结果：CMV 感染上调 IL-6/STAT3、SOX2、Survivin、p-AKT 和 p-RB，促进 GBM 细胞增殖、干性和凋亡抵抗。感染的细胞表现出支持肿瘤的细胞因子谱，且在更昔洛韦处理后 VEGF 和 Angiopoietin-2 水平降低。CMV 感染的星形胶质细胞也表现出致癌信号和集落形成增加，提示其在早期转化中发挥作用。更昔洛韦部分逆转了这些效应，使细胞因子表达向免疫许可状态转变。虽然单独的 NK 细胞未能裂解 GBM 细胞，但经更昔洛韦处理的 CMV 感染细胞表现出显著增强的 NK 介导的细胞毒性。CMV 感染后 PD-L1 表达增加，且在治疗后仍保持升高，提示免疫调节和检查点阻断的潜力。
结论：CMV 感染通过活化 IL-6/STAT3、p-AKT、SOX2 和血管生成通路，将胶质母细胞瘤细胞和正常星形胶质细胞重编程为更具侵袭性、免疫逃逸的表型。更昔洛韦治疗部分逆转了这些效应，降低 VEGF 和 Angiopoietin-2 水平，改变细胞因子谱，并且——关键的是——增强了 NK 细胞介导的细胞毒性。感染后持续的 PD-L1 上调提示一种新的免疫检查点易感性，为抗病毒与免疫治疗的协同方法打开了大门。这些发现不仅将 CMV 确定为肿瘤进展的驱动因素，也将其确定为 GBM 中一个可干预的治疗靶点。
查看英文原文 English abstract
Background: Cytomegalovirus (CMV) has been implicated in glioblastoma (GBM) pathogenesis by promoting stemness, angiogenesis&immune evasion. Clinical trials targeting CMV in GBM have shown preliminary promise, However, the molecular mechanisms remain unclear. We hypothesized that CMV enhances oncogenic signaling and immune resistance in GBM, and that antiviral therapy may reverse these effects and increase susceptibility to immune-mediated cytotoxicity
Materials and Methods: Human GBM cell lines (U251, U87, U138, T98G, LN229) & normal astrocytes were infected with a mCherry-human CMV TB40 strain (MOI 0.5). Phenotypic assays including spheroid & colony formation were performed. A 60-plex cytokine panel and Western blotting were used to evaluate cytokine profiles & signaling pathway activation (IL-6/STAT3, Akt, SOX2, Survivin, p-RB). Co-culture experiments with NK-92mi cells were conducted using CMFDA and Eth-homodimer dyes to assess cytotoxicity.
Results: CMV infection upregulated IL-6/STAT3, SOX2, Survivin, p-AKT, and p-RB, promoting GBM cell proliferation, stemness, and resistance to apoptosis. Infected cells exhibited a tumor-supportive cytokine profile and, after ganciclovir treatment, reduced VEGF and Angiopoietin-2 levels. CMV-infected astrocytes also showed increased oncogenic signaling and colony formation, suggesting a role in early transformation. Ganciclovir partially reversed these effects, shifting cytokine expression toward an immune-permissive state. While NK cells alone failed to lyse GBM cells, ganciclovir-treated, CMV-infected cells showed significantly enhanced NK-mediated cytotoxicity. PD-L1 expression increased after CMV infection and remained elevated post-treatment, indicating immune modulation and potential for checkpoint blockade.
Conclusions: CMV infection reprograms both glioblastoma cells and normal astrocytes toward a more aggressive, immune-evasive phenotype through activation of IL-6/STAT3, p-AKT, SOX2, and angiogenic pathways. Ganciclovir treatment partially reverses these effects, reducing VEGF and Angiopoietin-2 levels, shifting cytokine profiles, and-critically-enhancing NK cell-mediated cytotoxicity. Persistent PD-L1 upregulation following infection suggests a novel immune checkpoint vulnerability, opening the door for synergistic antiviral and immunotherapy approaches. These findings identify CMV not only as a driver of tumor progression but as a tractable therapeutic target in GBM.
利益披露 Disclosure
V. Tajiknia, None.