CARDIOTOXICITY OF EOSINOPHIL GRANULE CATIONIC PROTEINS

嗜酸性肌内膜纤维化是一种常见的， 特发性嗜酸性粒细胞增多综合征和 某些肿瘤和热带疾病继发的嗜酸性粒细胞。 最近的研究表明，心血管损伤是由 在嗜酸性粒细胞的特定颗粒中发现的阳离子蛋白; 这些蛋白质诱导心肌和 内部损坏情况尚不清楚。 根据教育和研究的目标， 医生科学家奖，该计划分为两个阶段。 在第一阶段，主要研究者的基本科学技能 将在细胞和分子生物学领域进行更新。 cDNA 嗜酸性粒细胞阳离子蛋白，嗜酸性粒细胞颗粒 主要碱性蛋白和嗜酸性粒细胞衍生的神经毒素将是 分离并测序。 一旦完成，第二阶段 将引入更复杂的重组DNA技术。 的cDNA 克隆将被引入原核或真核系统 允许重组蛋白的表达; 重组蛋白将被纯化并评估其 结合和/或损伤心肌和内皮细胞的能力 在文化中。 体外诱变将用于引入定义的 cDNA序列的连续缺失和点突变， 和突变重组嗜酸性粒细胞蛋白无法进行 将鉴定结合和/或细胞毒性。 这些实验 目的是鉴定功能上重要的氨基酸序列， 为了阐明嗜酸性粒细胞颗粒蛋白 与它们的心血管靶点相互作用。 从教育 从这个角度来看，计划中的方案将提供技能和 主要研究者从事研究所需的经验 作为独立的临床科学家。

Eosinophilic endomyocardial fibrosis is a common and often lethal complication of both the idiopathic hypereosinophilic syndrome and eosinophilla secondary to certain neoplastic and tropical diseases. Recent work suggests that the cardiovascular damage is mediated by cationic proteins found in the eosinophil's specific granules; the mechanisms by which these proteins induce myocardial and endocardial damage are not known. In accordance with the educational and research goals of the Physician Scientist Award, the plan is divided into two phases. n phase I, the basic science skills of the principal investigator will be updated in the areas of cell and molecular biology. cDNA clones for the eosinophil cationic protein, the eosinophil granule major basic protein and the eosinophil-derived neurotoxin will be isolated and sequenced. Once this has been accomplished, phase II will introduce more complex recombinant DNA techniques. The cDNA clones will be introduced into prokaryotic or eukaryotic systems permitting the expression of the recombinant proteins; the recombinant proteins will be purified and evaluated for their ability to bind to and /or damage myocardial and endothelial cells in culture. In vitro mutagenesis will be used to introduce defined sequential deletions and point mutations into the cDNA sequences, and mutant recombinant eosinophil proteins unable to carry out binding and/or cytotoxicity will be identified. These experiments aim to identify functionally significant amino acid sequences and to clarify the mechanisms by which the eosinophil granule proteins interact with their cardiovascular targets. From an educational perspective, the planned program will provide the skills and experience necessary for the principal investigator to pursue a career as an independent clinician-scientist.