Role of NKCC1 on Brain Tumor Stem Cell Migration After EGF and Slit-2 Stimulation

描述（由申请人提供）：多形性胶质母细胞瘤（GBM）是最常见和最具破坏性的颅内恶性肿瘤，占所有原发性脑肿瘤的20%，中位生存率仅为14个月。癌细胞通常远离原发性肿瘤扩散，并且单个胶质瘤细胞从大体肿瘤迁移到周围的实质中，使得完全手术切除几乎不可能。恶性胶质瘤的这种迁移能力代表了对任何潜在治疗的最大挑战，尽管手术、化疗和放疗取得了进展，并且剩余侵袭性细胞的生长导致99%的复发率。究竟是什么调节了脑肿瘤细胞的迁移能力还没有完全了解，需要进行研究。该提案的主要目标是了解已知的促迁移信号如表皮生长因子（EGF）和Slit蛋白与细胞体积调节之间的联系。EGF和Slit蛋白可能通过Akt途径调节离子共转运蛋白NKCC1的活化，从而在调节GBM源性干细胞的侵袭和迁移能力中发挥重要作用。我们建议使用各种细胞迁移试验和测量细胞内阴离子浓度来研究侵入性模式和细胞体积变化，从而导致迁移细胞的领先过程的延长。从这项工作中获得的结果将有助于我们了解参与激活级联机制负责脑肿瘤细胞迁移的下游信号通路。此外，这些知识无疑将导致更好的治疗替代目前的次优治疗这一毁灭性疾病。 公共卫生相关性：多形性胶质母细胞瘤（GBM）是最常见和最具破坏性的原发性恶性肿瘤。我们的项目旨在研究GBM衍生的脑肿瘤干细胞（BTSC）的迁移。BTSC被认为负责维持肿瘤的体积并诱导手术切除后的复发，然而调节其迁移的分子机制尚不清楚。在这项研究中，我们建议了解促迁移信号在脑肿瘤侵袭中的作用，以增加可用的靶点，以防止脑肿瘤扩散。

DESCRIPTION (provided by applicant): Glioblastoma multiforme (GBM), the most common and devastating intracranial malignant tumor accounts for 20% of all primary brain tumors and has a median survival rate of only 14 months. Cancer cells often disseminate far from primary tumors and individual glioma cells migrate from the gross tumor into the surrounding parenchyma, making complete surgical resection nearly impossible. This migratory capacity of malignant gliomas represents the greatest challenge to any potential therapy in spite of advances in surgery, chemotherapy and radiotherapy and growth of the remaining invasive cells leads to a recurrence incidence of 99%. What exactly regulates the migratory capacity of brain tumor cells is not fully understood and need to be studied. The main goal of this proposal is to understand the link between known pro- migratory signals such as epidermal growth factor (EGF) and Slit proteins with cell volume regulation. EGF and Slit proteins may play an important role in the modulation of invasive and migratory ability of GBM derived stem cells through Akt pathway that in turn regulates the activation of ion cotransport NKCC1. We propose to study invasive patterns and cell volume changes resulting in the extension of a leading process of a migrating cell, using various cell migration assays and measuring intracellular anion concentration. The results obtained from this work will help us understand the downstream signaling pathways involved in the activation of cascade mechanism responsible for brain tumor cell migration. Further, such knowledge will undoubtedly result in better therapeutic alternatives to current sub-optimal treatments for this devastating disease. PUBLIC HEALTH RELEVANCE: Glioblastoma multiforme (GBM) is the most common and devastating primary malignant tumor. Our project aims to study the migration of GBM-derived Brain Tumor Stem Cells (BTSCs). BTSCs are thought to be responsible for maintaining the bulk of the tumor and to induce recurrence after surgical resection, nevertheless the molecular mechanisms that regulate their migration are not known. In this study, we propose to understand the role of pro-migratory signals in brain tumor invasion in order to increase the available targets to prevent brain tumor dispersal.