Department of Radiation Oncology, Medical Center, Faculty of Medicine, University of Freiburg, German Cancer Consortium (DKTK), partner site DKTK-Freiburg, Robert-Koch-Str. 3, 79106, Freiburg, Germany.
Department of Neurosurgery, Medical Center, Faculty of Medicine, University of Freiburg, Freiburg, Germany.
J Neurooncol. 2024 Sep;169(2):329-340. doi: 10.1007/s11060-024-04740-0. Epub 2024 Jun 20.
Mesenchymal stromal cells (MSCs) within the glioblastoma microenvironment have been shown to promote tumor progression. Tumor Treating Fields (TTFields) are alternating electric fields with low intensity and intermediate frequency that exhibit anti-tumorigenic effects. While the effects of TTFields on glioblastoma cells have been studied previously, nothing is known about the influence of TTFields on MSCs.
Single-cell RNA sequencing and immunofluorescence staining were employed to identify glioblastoma-associated MSCs in patient samples. Proliferation and clonogenic survival of human bone marrow-derived MSCs were assessed after TTFields in vitro. MSC' characteristic surface marker expression was determined using flow cytometry, while multi-lineage differentiation potential was examined with immunohistochemistry. Apoptosis was quantified based on caspase-3 and annexin-V/7-AAD levels in flow cytometry, and senescence was assessed with ß-galactosidase staining. MSCs' migratory potential was evaluated with Boyden chamber assays.
Single-cell RNA sequencing and immunofluorescence showed the presence of glioblastoma-associated MSCs in patient samples. TTFields significantly reduced proliferation and clonogenic survival of human bone marrow-derived MSCs by up to 60% and 90%, respectively. While the characteristic surface marker expression and differentiation capacity were intact after TTFields, treatment resulted in increased apoptosis and senescence. Furthermore, TTFields significantly reduced MSCs' migratory capacity.
We could demonstrate the presence of tumor-associated MSCs in glioblastoma patients, providing a rationale to study the impact of TTFields on MSCs. TTFields considerably increase apoptosis and senescence in MSCs, resulting in impaired survival and migration. The results provide a basis for further analyses on the role of MSCs in glioblastoma patients receiving TTFields.
已证实神经胶质瘤微环境中的间充质基质细胞（MSCs）可促进肿瘤进展。肿瘤治疗电场（TTFields）是一种低强度、中频的交替电场，具有抗肿瘤作用。虽然先前已经研究了 TTFields 对神经胶质瘤细胞的影响，但对于 TTFields 对 MSCs 的影响却一无所知。
采用单细胞 RNA 测序和免疫荧光染色技术，鉴定患者样本中与神经胶质瘤相关的 MSCs。体外 TTFields 作用后，检测人骨髓来源 MSCs 的增殖和克隆存活能力。采用流式细胞术检测 MSC 特征性表面标志物表达，用免疫组织化学法检测多系分化潜能。采用流式细胞术根据 caspase-3 和 annexin-V/7-AAD 水平定量检测细胞凋亡，通过β-半乳糖苷酶染色评估衰老。采用 Boyden 室测定法评估 MSCs 的迁移能力。
单细胞 RNA 测序和免疫荧光显示患者样本中存在与神经胶质瘤相关的 MSCs。TTFields 可使人类骨髓来源的 MSCs 的增殖和克隆存活分别减少高达 60%和 90%。尽管 TTFields 后 MSC 特征性表面标志物表达和分化能力保持完整，但治疗导致细胞凋亡和衰老增加。此外，TTFields 还显著降低了 MSCs 的迁移能力。
我们能够在神经胶质瘤患者中证实肿瘤相关 MSCs 的存在，为研究 TTFields 对 MSCs 的影响提供了依据。TTFields 可显著增加 MSCs 中的细胞凋亡和衰老，导致细胞存活和迁移受损。这些结果为进一步分析接受 TTFields 治疗的神经胶质瘤患者中 MSCs 的作用提供了基础。
