Regulation of an Angiogenic Phenotype by Jagged 1

描述(申请人提供)：Noch受体及其配体为 进化上保守的调节细胞命运的跨膜多肽 并与生理和病理过程相关联 例如血管生成和肿瘤发生。为了更好地理解 这些过程中的锯齿状1/Notch信号转导，NIH3T3细胞的表型 稳定转染非跨膜型分泌型人Jagge1 检测血管生成过程中上调的(SJ1)基因。SJ1 转染者表现出转化的表型，包括细胞的变化 形态，在克隆密度下增加存活率，与锚定无关的生长 在软琼脂中，FGF1的存在和迁移行为的改变。SJ1 细胞也能在绒毛尿囊膜中引起血管生成反应。 SJ1转基因细胞的细胞膜分析及裸鼠移植 小鼠产生了高度血管生成的肿块。SJ1的诱导机制 这些表型变化是未知的，并通过两个特定的目标来解决 1)以确定NIH 3T3 sJ1表型是否源于 特异性Notch受体信号通路的激活或抑制2) 确定足以诱导sJ1表型的sJ1的最小结构域 在NIH3T3细胞中。这些目标将通过表征NIH3T3细胞来实现 利用一种新的生物学方法稳定地转染J1和Notch缺失突变体 基于实验室的检测？S对sJ1转染体的鉴定。 阐明sj1调节细胞表型的机制可能有助于 在用于治疗病理性疾病的治疗剂的开发中 癌症等病症。

DESCRIPTION (provided by applicant): Notch receptors and their ligands are evolutionarily conserved transmembrane polypeptides that regulate cell fate determination and are associated with physiological and pathological processes such as angiogenesis and tumorigenesis. In order to understand the role of Jagged 1/Notch signaling in these processes, the phenotype of NIH 3T3 cells stably transfected with a non-transmembrane and secreted form of human Jagged 1 (sJ1) found to be upregulated during angiogenesis was examined. sJ1 transfectants exhibit a transformed phenotype that includes changes in cell morphology, increased survival at clonal density, anchorage-independent growth in soft agar in the presence of FGF1 and alterations in migratory behavior. sJ1 cells are also able to elicit an angiogenic response in the chorioallantoic membrane assay (CAM) and implantation of sJ1 transfected cells into athymic mice yielded highly angiogenic masses. The mechanisms by which sJ1 induces these phenotypic changes are unknown and are addressed by the two specific aims of this proposal 1) to determine if the NIH 3T3 sJ1 phenotype results from activation or inhibition of specific Notch receptor signaling pathways and 2) to determine the minimal domain of sJ1 sufficient to induce the sJ1 phenotype in NIH 3T3 cells. These objectives will be met by characterizing NIH 3T3 cells stably transfected with J 1 and Notch deletion mutants using a novel biological assay based on the laboratory?s characterization of sJ1 transfectants. Elucidation of the mechanisms by which sJ1 regulates cellular phenotype may aid in the development of therapeutic agents used for the treatment of pathological conditions such as cancer.