CONTROL OF SCHWANN CELL DIFFERENTIATION

描述(摘自申请人的摘要)： 这项提议的目的是了解雪旺细胞是如何 在开发过程中整合不同的环境信号，以及 周围神经的康复、疾病或创伤。长期的 目标是确定雪旺细胞如何对来自 细胞外基质(ECM)、轴突和邻近的雪旺细胞 协调生长和分化为髓鞘形成 细胞。为了实现这一目标，费尔南德斯-瓦莱博士和她的同事们 雪旺细胞黏附于细胞外基质激活的细胞外基质依赖性信号转导研究 层粘连蛋白，雪旺细胞基底膜中的一种细胞外基质蛋白。在.期间 最后一次授权期，粘着斑激酶，巴西林，fyn激酶，和 Merlin(由神经纤维瘤病2型基因编码的肿瘤抑制因子 产品)被发现与β-1整合素依赖有关 雪旺细胞体外髓鞘形成过程中的信号转导。这个机制是通过 神经系统尚不清楚梅林是哪种药物抑制肿瘤形成。 结果表明，Merlin参与了β-1整合素依赖的信号转导 雪旺细胞分化过程中的信号传导途径。假设将是 测试了Merlin在信号通路之间的“对话”；一 由胶质生长因子与HER2/HER3受体结合激活 另一种是通过层粘连蛋白与雪旺上的β-1整合素结合而激活的。 细胞表面。在本申请中，建议：1)MAP Beta-1 通过检测Rho GTP酶及其调节因子，整合素上游事件； 2)通过检测丝裂原激活来定位β-1整合素下游事件 蛋白激酶和β-1整合素内吞作用；3)寻找“串扰” 通过传导HER2/HER3受体和帕西林、梅林、 和FAK相互免疫共沉淀；4)进行功能性 显性负性和反义寡核苷酸分析 战略。 雪旺细胞的分离培养及体外髓鞘形成模型 将使用由雪旺细胞与神经元共同培养组成的细胞。这些 是有用的范例，允许蛋白质和蛋白质的功能分析 所有主要生化、分子和细胞学的应用 接近了。这一信息可能与治疗方法的开发有关 A)对周围神经的损伤，b)将雪旺细胞用作 中枢神经再生和再髓鞘形成的促进剂 创伤或疾病后的系统，以及3)治疗或预防 神经胶质瘤或其他神经系统肿瘤零星发生在 人群或由神经纤维瘤病-2型突变引起 吉恩。

DESCRIPTION (from the applicant's abstract): The objective of this proposal is to understand how Schwann cells integrate diverse environmental signals during development of, and recovery from, disease or trauma to peripheral nerve. The long-term goals are to determine how Schwann cells respond to signals from extracellular matrix (ECM), axons, and neighboring Schwann cells in order to coordinate growth and differentiation into myelin-forming cells. Toward this goal, the Dr. Fernandez-Valle and her colleagues have studied ECM dependent signaling activated by Schwann cell adhesion to laminin, an ECM protein in the Schwann cell basal lamina. During the last grant period, focal adhesion kinase, paxillin, fyn kinase, and merlin (a tumor suppressor encoded by the neurofibromatosis type-2 gene product) were found to be involved in beta-1 integrin-dependent signaling during Schwann cell myelination in vitro. The mechanism by which merlin suppresses tumor formation the nervous system is unknown. The results implicate merlin in a beta-1 integrin dependent signaling pathway during Schwann cell differentiation. The hypothesis will be tested that merlin mediates a "dialogue" between signaling pathways; one activated by glial growth factor binding to HER2/HER3 receptors and another activated by laminin binding to beta-1 integrins on the Schwann cell surface. In this application, it is proposed to: 1) map beta-1 integrin upstream events by examining rho GTPases and their regulators; 2) map beta-1 integrin downstream events by examining mitogen-activated protein kinase and beta-1 integrin endocytosis; 3) look for "cross-talk" between pathways by conducting HER2/HER3 receptor and paxillin, merlin, and FAK reciprocal co-immunoprecipitations; and 4) carry out functional analyses using dominant negative and antisense oligonucleotide strategies. Isolated Schwann cell cultures and the in vitro myelination model consisting of Schwann cells co-cultured with neurons will be used. These are useful paradigms, allowing for functional analyses of proteins and application of all major biochemical, molecular, and cytological approaches. This information may relate to development of therapies for a) injuries to peripheral nerve, b) efforts to use Schwann cells as promoters of regeneration and remyelination in the central nervous system following trauma or disease, and 3) treatment or prevention of glial or other nervous system tumors occurring sporadically in the population or resulting from mutations in the neurofibromatosis type-2 gene.