Heterogeneous Glioma Cell Invasion Under Interstitial Flow Depending on Their Differentiation Status
Heterogeneous Glioma Cell Invasion Under Interstitial Flow Depending on Their Differentiation Status
复制标题
间质流下的异质胶质瘤细胞侵袭取决于其分化状态
DOI：
10.1089/ten.tea.2020.0280
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发表时间：
2021
影响因子：
4.1
通讯作者：
Sudo Ryo
中科院分区：
文献类型：
--
作者：
Namba Naoko;Chonan Yuta;Nunokawa Takehito;Sampetrean Oltea;Saya Hideyuki;Sudo Ryo
胶质母细胞瘤（GBM）是最常见和致命的恶性脑肿瘤。更深入地了解异质性GBM细胞群体的侵袭机制对于开发治疗策略至关重要。GBM细胞侵袭的关键调节因子是间质流。然而，间质流对由胶质瘤起始细胞（GIC）和相对分化的子代细胞组成的异质GBM细胞群的侵袭的影响仍不清楚。在本研究中，我们研究了GICs如何侵入三维（3D）水凝胶，以响应其分化状态的间质流。微流体培养系统用于向从细胞聚集体迁移到3D水凝胶中的细胞施加间质流。相差显微镜显示，在分化的细胞条件下的GIC的侵入和突起的形成显着增强了向前的间质流，其方向是相同的细胞侵入，而在干细胞条件下没有增强的间质流。流动诱导的入侵的机制进一步研究通过集中在分化的细胞条件。免疫荧光图像显示，细胞-细胞外基质粘附相关分子，如整合素β1，粘着斑激酶和磷酸化Src的表达在前向间质流条件下上调。然后，我们证实，细胞侵袭和突起形成显着抑制PP 2，Src抑制剂。最后，我们观察到在流动诱导的细胞侵袭之前，在侵袭前沿是nestin阳性的未成熟GIC，然后是微管蛋白β3阳性的分化细胞。我们的研究结果提供了新的治疗策略的发展，以抑制流量诱导的胶质瘤invasion.Impact statementA异质性胶质母细胞瘤细胞的侵袭机制的理解是至关重要的发展治疗策略。我们观察到，胶质瘤起始细胞（GICs）的侵袭和突起形成显着增强的分化细胞条件下的前向间质流。整合素β1、粘着斑激酶和磷酸化Src的表达上调，并且流动诱导的侵袭被Src抑制剂显著抑制。流动诱导的异质性细胞侵袭之前，在侵袭前沿的nestin阳性GIC和随后的微管蛋白β3阳性分化细胞。我们的研究结果提供了新的治疗策略的发展，以抑制流动诱导的胶质瘤侵袭的见解。
Glioblastoma (GBM) is the most common and lethal type of malignant brain tumor. A deeper mechanistic understanding of the invasion of heterogeneous GBM cell populations is crucial to develop therapeutic strategies. A key regulator of GBM cell invasion is interstitial flow. However, the effect of an interstitial flow on the invasion of heterogeneous GBM cell populations composed of glioma initiating cells (GICs) and relatively differentiated progeny cells remains unclear. In the present study, we investigated how GICs invade three-dimensional (3D) hydrogels in response to an interstitial flow with respect to their differentiation status. Microfluidic culture systems were used to apply an interstitial flow to the cells migrating from the cell aggregates into the 3D hydrogel. Phase-contrast microscopy revealed that the invasion and protrusion formation of the GICs in differentiated cell conditions were significantly enhanced by a forward interstitial flow, whose direction was the same as that of the cell invasion, whereas those in stem cell conditions were not enhanced by the interstitial flow. The mechanism of flow-induced invasion was further investigated by focusing on differentiated cell conditions. Immunofluorescence images revealed that the expression of cell–extracellular matrix adhesion-associated molecules, such as integrin β1, focal adhesion kinase, and phosphorylated Src, was upregulated in forward interstitial flow conditions. We then confirmed that cell invasion and protrusion formation were significantly inhibited by PP2, a Src inhibitor. Finally, we observed that the flow-induced cell invasion was preceded by nestin-positive immature GICs at the invasion front and followed by tubulin β3-positive differentiated cells. Our findings provide insights into the development of novel therapeutic strategies to inhibit flow-induced glioma invasion.Impact statementA mechanistic understanding of heterogeneous glioblastoma cell invasion is crucial for developing therapeutic strategies. We observed that the invasion and protrusion formation of glioma initiating cells (GICs) were significantly enhanced by forward interstitial flow in differentiated cell conditions. The expression of integrin β1, focal adhesion kinase, and phosphorylated Src was upregulated, and the flow-induced invasion was significantly inhibited by a Src inhibitor. The flow-induced heterogeneous cell invasion was preceded by nestin-positive GICs at the invasion front and followed by tubulin β3-positive differentiated cells. Our findings provide insights into the development of novel therapeutic strategies to inhibit flow-induced glioma invasion.