PO.ET06.04 · 实验与分子治疗
肿瘤电场治疗（TTFields）可能削弱胶质母细胞瘤网络连接性，对恶性程度具有影响
Tumor Treating Fields (TTFields) may weaken glioblastoma network connectivity, with implications for malignancy
作者与单位 Authors & Affiliations
摘要 Abstract
中文摘要
背景：胶质母细胞瘤的恶性程度在很大程度上由肿瘤微管（TMs）的形成所驱动，后者促进细胞间连接和钙（Ca²⁺）波传播。肿瘤电场治疗（TTFields）影响这些TM介导过程的机制尚未完全阐明。本研究旨在阐明TTFields对肿瘤细胞网络结构和功能组织的影响，并评估信号通路（包括NF-κB）潜在的频率依赖性调节。
方法：正在使用一套全面的生物模型系统，包括2D胶质母细胞瘤细胞单层、3D脑类器官，以及具有慢性颅窗用于纵向成像的体内清醒头部固定小鼠模型。共聚焦和多光子显微镜的活细胞成像可实时观察肿瘤的形态学和功能动态。使用基于Cellpose的分割和自定义Python分析流程对Ca²⁺信号进行定量。目前采用免疫组织化学和空间转录组学（Visium HD）来解析分子机制；计划采用基于COMET的免疫荧光和RNAscope FISH以实现空间分辨的多组学。
结果：TTFields诱导了胶质母细胞瘤网络结构和功能的显著破坏。具体而言，治疗导致全局GCaMP8s介导的Ca²⁺活性降低>50%，起搏样细胞群减少，以及在2D和3D模型中S24胶质母细胞瘤细胞内同步化和Ca²⁺共激活的显著降低。
结论与展望：TTFields破坏了胶质母细胞瘤网络的完整性和Ca²⁺信号，可能降低肿瘤侵袭性。来自3D脑肿瘤类器官和体内模型的初步数据支持我们此前关于TTFields诱导活性变化的体外结果。在患者来源类器官中的平行研究通过空间转录组学分析探索频率依赖性信号效应——包括NF-κB和MAPK通路。总之，这些工作旨在进一步阐明TTFields作用的机制基础及其对胶质母细胞瘤可塑性和网络组织的影响。
查看英文原文 English abstract
Background: Glioblastoma malignancy is strongly driven by the formation of tumor microtubes (TMs), which promote intercellular connectivity and calcium (Ca²⁺) wave propagation. The mechanisms by which Tumor Treating Fields (TTFields) influence these TM-mediated processes remain incompletely understood.
This study aims to elucidate the effects of TTFields on the structural and functional organization of tumor cell networks and to assess potential frequency-dependent modulation of signaling pathways, including NF-κB.
Methods: A comprehensive set of biological model systems is being utilized, including 2D glioblastoma cell monolayers, 3D brain organoids, and in vivo , awake, head-fixed mouse models with chronic cranial windows for longitudinal imaging. Live-cell imaging with confocal and multiphoton microscopy enables real-time observation of morphological and functional tumor dynamics. Quantification of Ca²⁺ signaling is being performed using Cellpose-based segmentation and custom Python analysis pipelines. Immunohistochemistry and spatial transcriptomics (Visium HD) are currently employed to dissect molecular mechanisms; COMET-based immunofluorescence and RNAscope FISH are planned to enable spatially resolved multi-omics.
Results: TTFields induced a marked disruption of glioblastoma network architecture and function. Specifically, treatment resulted in >50% reduction in global GCaMP8s-mediated Ca²⁺ activity, a decrease in pacemaker-like cell populations, and significant reductions in synchronization and Ca²⁺ co-activity within S24 glioblastoma cells in both 2D and 3D models.
Conclusion & Outlook: TTFields disrupt glioblastoma network integrity and Ca²⁺ signaling, potentially reducing tumor aggressiveness. Preliminary data from 3D brain tumor organoids and in vivo models support our previous in vitro results regarding TTFields-induced activity changes. Parallel studies in patient-derived organoids explore frequency-dependent signaling effects - including NF-κB and MAPK pathways - via spatial transcriptomic profiling. Together, these efforts aim to further elucidate the mechanistic underpinnings of TTFields action and their impact on glioblastoma plasticity and network organization.
利益披露 Disclosure
J. Beichert,
Novocure Other, received financial compensation (speaker honorarium) from the company organizing the conference for delivering a presentation.
J. A. Kretz, None..
J. Kim, None..
X. Ma, None..
D. D. Azorín, None..
A. E. Moor, None.
H. Liu,
aiPTO CoFounder.
M. Ratliff, None.