Wnt-Dependent Neurite Outgrowth in Ewing Tumor Cells

尤因肿瘤细胞中 Wnt 依赖性神经突生长

• 批准号: 7966250
• 负责人: Jeffrey Rubin
• 金额: $ 28.44万
• 依托单位: DIVISION OF BASIC SCIENCES - NCI
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/7966250
• 关键词: Accounting; Cell model; Cells; Centrosome; Cilia; Disease; Embryo; Embryonic Development; Ewings sarcoma; JUN gene; Knowledge; Mediating; Neoplasm Metastasis; Neural Tube Defects; Neurites; Phosphorylation; Phosphorylation Site; Phosphotransferases; Physiological; Polycystic Kidney Diseases; Process; Regulation; Role; Signal Transduction; Site; Situs Inversus; Small Interfering RNA; Testing; Work; base; beta-arrestin; casein kinase I; cell motility; inhibitor/antagonist; insight; neoplastic cell; receptor; research study; response

We established that cells from Ewing tumors form neurites in response to Wnt-3a and have begun to define the mechanisms that account for this effect. Frizzled3 (Fzd3) was identified as the primary Wnt receptor that mediates the process, which also requires Dishevelled-2 (Dvl-2), Dishevelled-3 (Dvl-3), and amino-terminal c-Jun kinase (JNK). We showed that Dickkopf-1 also promotes neurite outgrowth in these cells, apparently by facilitating Fzd3/JNK activation by endogenous Wnts. Neurite outgrowth induced by Wnt-3a was associated with Dvl-2/3 phosphorylation; both neurite formation and Dvl phosphorylation were blocked by the casein kinase I delta/epsilon (CKId/e) inhibitor, IC261. Knockdown of CKId with small interfering RNA suppressed Wnt-3a-dependent neuritogenesis, whereas knockdown of CKIe stimulated neurite formation in the absence of exogenous Wnt-3a. The contrasting effects of CKId and CKIe on neurite outgrowth might be due to the preferential localization of CKId at the centrosome, which is thought to have a critical role in neurite formation. Knockdown of the atypical PKCiota also blocked Wnt-3a-dependent neurite outgrowth. Preliminary experiments suggest that PKCiota may be regulated by CKId and/or Dvl. This work is significant not only because it provides insights about mechanisms involved in the formation of neurites. Many of the factors that participate in neurite outgrowth also have important roles in the formation of cellular extensions critical for cell migration. The information obtained in the Ewing tumor cell model is likely to enhance our understanding of cell movement in physiological contexts and metastasis. Moreover, we have determined that CKId and Dvl also are required for the formation of primary cilia. Defective primary cilia are responsible for several disorders including neural tube defects, polycystic kidney disease and situs inversus. Aberrant Wnt signaling also can elicit these abnormalities. Thus, our studies of CKId and Dvl should extend our knowledge about the ways in which Wnt signaling controls embryonic development.

我们确定尤文瘤细胞对Wnt-3a的反应是形成神经突， 已经开始定义解释这种效应的机制。Frizzled 3（Fzd 3）是 被鉴定为介导该过程的主要Wnt受体，这也需要 Dishevelled-2（Dvl-2）、Dishevelled-3（Dvl-3）和氨基末端c-Jun激酶（JNK）。我们展示 Dickkopf-1也促进这些细胞中的神经突生长，显然是通过促进 内源性Wnt激活Fzd 3/JNK。Wnt-3a诱导的神经突起生长与 Dvl-2/3磷酸化;轴突形成和Dvl磷酸化都被阻断， 酪蛋白激酶I δ/ε（CKId/e）抑制剂，IC 261。CKId的敲低， 干扰RNA抑制Wnt-3a依赖的轴突发生，而CKIe的敲低则抑制Wnt-3a依赖的轴突发生。 在不存在外源性Wnt-3a的情况下刺激神经突形成。对比效果 CKId和CKIe对神经突生长的影响可能是由于CKId的优先定位在 中心体，被认为在神经突形成中起关键作用。敲低 非典型PKC 1 ota也阻断Wnt-3a依赖性神经突生长。初步实验 提示PKC 10 α可能受CKId和/或Dvl调节。这项工作不仅意义重大， 因为它提供了关于神经突形成机制的见解。许多 参与神经突生长的因子也在神经突的形成中起重要作用。 对细胞迁移至关重要。在尤因获得的信息 肿瘤细胞模型可能会增强我们对生理学中细胞运动的理解。 背景和转移。此外，我们已经确定CKId和Dvl也是用于 初级纤毛的形成。有缺陷的初级纤毛是负责几个疾病 包括神经管缺陷、多囊肾和内脏逆位。异常Wnt 信号传导也可以引起这些异常。因此，我们对CKId和Dvl的研究应该扩展到 我们对Wnt信号控制胚胎发育的方式的了解。