Identifying the Mechanisms Governing Mena-Induced Tumor Cell Dissemination

确定 Mena 诱导的肿瘤细胞传播的控制机制

• 批准号: 8366273
• 负责人: Minna Roh
• 金额: $ 5.22万
• 依托单位: ALBERT EINSTEIN COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-09-01 至 2014-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8366273
• 关键词: Accounting; Actins; Affect; Alternative Splicing; Apoptotic; Behavior; Biological Assay; Blocking Antibodies; Blood Vessels; Cancer Etiology; Cells; Cessation of life; Chemotactic Factors; Chemotaxis; Coupled; Data; Development; Early identification; Endothelial Cells; Environment; Epidermal Growth Factor; Event; Extracellular Matrix; Family; Gene Expression Profile; Goals; Hand; Heregulin; Immunohistochemistry; In Vitro; Knowledge; Laser Scanning Confocal Microscopy; Lead; Length; Life; Light; Malignant Epithelial Cell; Malignant Neoplasms; Mammary Neoplasms; Microscopy; Migration Assay; Mind; Molecular; Movement; Needles; Neoplasm Metastasis; Patients; Phenotype; Play; Process; Protein Isoforms; Proteins; Role; Signal Pathway; Signal Transduction; Source; Specificity; System; Testing; United States; Work; base; cell motility; cell type; cellular imaging; design; high risk; in vivo; insight; macrophage; member; migration; neoplastic cell; novel marker; research study; response; tumor; vasodilator-stimulated phosphoprotein

DESCRIPTION (provided by applicant): More than 1 in 3 people will develop cancer in their lifetime, and approximately 1,500 people die from cancer each day in the United States. Metastases, which require invasion and intravasation, are the major causes of cancer-related deaths. Key cytoskeletal, proliferative and apoptotic proteins have been shown to dynamically alter their gene expression patterns during cell invasion and intravasation, suggesting their involvement in regulating some or all of the processes involved in these cell migration phenotypes. These proteins have been termed collectively as the "Invasion Signature". One of the components of the "Invasion Signature" is Mena, a member of the Enabled (Ena)/vasodilator-stimulated phosphoprotein (VASP) family. Ena/VASP proteins are highly conserved regulators of actin dynamics, known to play critical roles in cell migration. Ena/VASP proteins contain several conserved domains that are thought to elicit specificity of Mena function through alternative splicing. Expression of a Mena isoform, MenaINV, sensitizes carcinoma cells to epidermal growth factor (EGF)- induced carcinoma cell invasion and metastasis. In response to lower levels of EGF, expression of MenaINV leads to enhanced metastasis in vivo. Furthermore, expression of MenaINV results in directional movement of carcinoma cells toward blood vessels, increased invadopodium stability, and a 200-fold increase in transendothelial migration (TEM). Expression of another Mena isoform, Mena11a, also results in increased tumor cell migration. Therefore, Mena plays critical roles in invasion and intravasation. Although many roles for Mena in tumor cell metastasis have been discovered, the mechanisms by which Mena-expressing carcinoma cells use to carry out these functions are still unknown. Thus, we will take advantage of a simplified in vitro system for specific control of parameters, as well as an in vivo system developed in our lab to take into account the tumor microenvironment. With these approaches in hand, specifically, I will test the requirement of invadopodia during Mena-induced chemotaxis, invasion and intravasation, and the upstream signaling events that regulate these processes. In particular, I propose the following aims: 1) Determining whether Mena isoform-expressing cells can autonomously chemosense a chemoattractant, and what step(s) of invadopodium formation is critical for chemosensing; 2) Investigating the relationship between Mena-induced chemotaxis and invadopodium formation in vivo; and 3) Identifying the mechanism underlying the 200 fold increase in TEM activity in MenaINV-expressing cells. Ultimately, these experiments will provide insight into the mechanisms governing the observed Mena-induced metastasis in vivo, which may be critical to the development of new anti-metastatic therapies.

描述（由申请人提供）：超过三分之一的人将在他们的一生中发展癌症，每天约有1,500人死于癌症。需要入侵和静脉内的转移是与癌症相关死亡的主要原因。已经证明，关键的细胞骨架，增殖和凋亡蛋白可以在细胞浸润和插入过程中动态改变其基因表达模式，这表明它们参与调节这些细胞迁移表型中涉及的某些或所有过程。这些蛋白质已被称为“入侵签名”。 “入侵签名”的组成部分之一是MENA，MENA是启用（ENA）/血管扩张剂刺激的磷蛋白（VASP）家族的成员。 ENA/VASP蛋白是肌动蛋白动力学的高度保守调节剂，已知在细胞迁移中起关键作用。 ENA/VASP蛋白包含几个保守的域，这些域被认为是通过替代剪接引起MENA功能的特异性。 MENAINV的MENA同工型的表达使癌细胞对表皮生长因子（EGF） - 诱导的癌细胞侵袭和转移。响应于较低的EGF水平，MENAINV的表达导致体内转移增强。此外，MENAINV的表达会导致癌细胞向血管的方向运动，稳定性增加，而跨内皮迁移（TEM）增加了200倍。 MENA11A的另一种MENA同工型的表达也导致肿瘤细胞迁移增加。因此，北非在入侵和侵入中扮演着关键的作用。尽管已经发现了MENA在肿瘤细胞转移中的许多作用，但表达MENA的癌细胞用于执行这些功能的机制仍然未知。因此，我们将利用一个简化的体外系统，用于特定控制参数，以及在实验室中开发的体内系统，以考虑肿瘤微环境。特别是这些方法，我将在MENA诱导的趋化性，侵袭和入侵以及调节这些过程的上游信号事件中测试Invadopodia的需求。特别是，我提出以下目的：1）确定ernaa同工型的细胞是否可以自主化化学素化趋化剂，以及什么步骤（S）的Invadopodium形成的哪一步对于化学效应至关重要； 2）研究MENA诱导的趋化性与体内的Invadopodium形成之间的关系； 3）识别表达MENAINV细胞中TEM活性200倍的基础机制。最终，这些实验将提供有关管理观察到的MENA诱导转移体内的机制，这对于新的抗转移疗法的发展至关重要。