CARDIOTOXICITY OF EOSINOPHIL GRANULE CATIONIC PROTEINS

嗜酸性心内膜心肌纤维化是一种常见且常常致命的疾病。 特发性高嗜酸性粒细胞综合征和 继发于某些肿瘤和热带疾病的嗜酸性粒细胞增多症。 最近的研究表明，心血管损害是由 在嗜酸性粒细胞的特殊颗粒中发现的阳离子蛋白； 这些蛋白诱导心肌梗死的机制 心内膜损伤尚不清楚。 根据该委员会的教育和研究目标 医师科学家奖，该计划分为两个阶段。 第一阶段，首席调查员的基本科学技能 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--嗜酸性粒细胞的克隆 主要的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。一旦完成此操作，第二阶段 将引入更复杂的重组DNA技术。CDNA3 克隆将被引入原核或真核系统 允许表达重组蛋白； 重组蛋白将被纯化并对其进行评估 结合和/或损伤心肌和内皮细胞的能力 在文化上。体外诱变将被用来引入明确的 序列缺失和点突变进入cDNAs序列， 和突变型重组嗜酸性粒细胞蛋白无法进行 将确定结合和/或细胞毒性。这些实验 目的鉴定具有重要功能的氨基酸序列和 阐明嗜酸性粒细胞颗粒蛋白的作用机制 与他们的心血管目标相互作用。来自教育性的 从长远来看，计划中的计划将提供技能和 首席调查员进行调查所需的经验 作为一名独立的临床医生兼科学家的职业。

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.