多形性膠質母細胞瘤（Glioblastoma multiforme, GBM）是一種常見且高度惡性的腦瘤，被世界衛生組織（WHO）歸類為第四級，其特徵為快速生長與高度侵襲性。儘管接受了手術切除、化學治療等方式積極治療，GBM仍常復發，大多數患者在診斷後的12至18個月內死亡。有趣的是，GBM經常侵入如胼胝體（corpus callosum）與內囊（internal capsule）等富含髓鞘軸突與寡突膠質細胞（oligodendrocytes, OLs）的白質區域，但其偏好侵入白質的機制至今尚不清楚。由於腫瘤進展與微環境密切相關，包括發炎反應與缺氧狀態，OLs可能參與GBM向白質擴散的過程。在本研究中，我們透過活體電穿孔（in vivo electroporation, IVE）剔除三個腫瘤抑制基因Nf1、Tp53與Pten，建立一個侵入白質的GBM小鼠模型。我們證實IVE所產生的GBM細胞會沿著胼胝體遷移生長，並觀察到OLs出現在腦瘤組織中。另一方面，直接將小鼠膠瘤細胞株CT2A注射至大腦時，我們也發現OLs譜系細胞會被招募至腫瘤周邊區域，顯示GBM與OLs之間具有密切的關聯性。為了探討OLs分泌物是否具有促進腫瘤生長的功能，我們收集了初級OLs培養液的條件培養液（conditioned media, CM）。結果顯示，OL-CM促進CT2A細胞增生的效果優於來自星狀膠質細胞或神經元的CM。利用質譜分析與已知GBM單細胞基因表現資料，我們鑑定出在OL-CM中富集的兩種蛋白質：Tenascin-R（Tnr）與Thymosin beta-4（Tmsb4x）。當我們在OLs中剔除Tnr時，IVE-GBM在胼胝體的擴散更加劇烈，而剔除Tmsb4x則抑制了腫瘤的生長。然而，直接剔除CT2A細胞的Tnr或Tmsb4x基因，會使其增生能力均低於對照組，顯示OLs分泌物在GBM侵入白質的過程中具有重要角色。總結以上數據，我們的研究指出OLs分泌的Tnr可能對GBM在白質中的擴散具有抑制作用，而Tmsb4x則可能促進膠瘤的進展。本研究顯示OL的確參與調節腫瘤惡化進程，惟不同釋放因子有不同效果，故還需研究其他分子對GBM病理的影響。

Glioblastoma multiforme (GBM) is a common and highly malignant brain tumor classified as grade IV by the World Health Organization (WHO), characterized by rapid growth and aggressive invasiveness. Despite surgical resection, chemotherapy, and other treatments, GBM frequently recurs, and most patients die within 12 to 18 months of diagnosis. Interestingly, GBM often infiltrates white matter tracts such as the corpus callosum and internal capsule, which are rich in myelinated axons and oligodendrocytes (OLs). However, the mechanisms underlying this preferential invasion into white matter regions remain unclear. Since tumor progression is closely associated with the microenvironment, including inflammation and hypoxia, OLs may contribute to GBM infiltration into white matter. In this study, we established a white matter-invading GBM model by knocking out three tumor suppressor genes, Nf1, Tp53, and Pten, using in vivo electroporation (IVE). We confirmed that IVE-GBM cells migrated along the mouse corpus callosum and observed the presence of OLs within glioma tissues. In contrast, when we directly injected the mouse glioma cell line CT-2A into the brain, OL lineage cells were recruited to the tumor mass, suggesting a close relationship between glioma and OLs. To examine the pro-tumoral OL secretome, we collected conditioned media (CM) from primary OL cultures. OL-CM enhanced CT-2A cell growth more effectively than CM from astrocytes or neurons. Mass spectrometry analysis identified two proteins enriched in OL-CM: Tenascin-R (TNR) and Thymosin beta-4 (TMSB4X). Knockout of Tnr in OLs led to more aggressive IVE-GBM expansion across the corpus callosum, while Tmsb4x knockout suppressed tumor growth. When Tnr was deleted in oligodendrocytes (OLs), IVE-GBM spread more aggressively in the corpus callosum, whereas deletion of Tmsb4x inhibited tumor growth. When Tnr was deleted in oligodendrocytes (OLs), IVE-GBM spread more aggressively in the corpus callosum, whereas deletion of Tmsb4x inhibited tumor growth. However, direct deletion of the Tnr or Tmsb4x gene in CT2A cells reduced their proliferative capacity compared to the control group, suggesting that OL-derived secreted factors play a crucial role in GBM invasion into white matter. In summary, our findings suggest that Tnr secreted by oligodendrocytes may inhibit the spread of GBM within the white matter, whereas Tmsb4x may facilitate glioma progression. This study demonstrates that oligodendrocytes are indeed involved in regulating tumor progression; however, different secreted factors exert distinct effects, highlighting the need to investigate the roles of other molecules in GBM pathology.

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