Rheological and Adherence Properties of Sickle Cells

描述（由申请人提供）：本申请的研究目的是对循环镰状红细胞的流变和粘附特性的异质性进行详细的分子和机制理解，并定义这些细胞变化的体内病理后遗症。为了实现我们的目标，我们提出了以下一系列研究：1)确定循环镰状红细胞不同亚群的异质流变特性的结构基础，并研究这些亚群的体内流动行为和存活。2)确定K-CI共转运蛋白和Gardos通道对神学上受损的致密镰状细胞产生的贡献。3)评估特异性黏附受体、配体和血浆蛋白在体外介导镰状红细胞与内皮细胞黏附中的作用。4)通过使用镰状小鼠细胞进行黏附研究，验证各种黏附配体和受体对镰状细胞黏附的贡献，其中编码特定黏附配体或受体的基因被灭活。对于拟议的研究，我们将使用多种技术在生物物理学，分子生物学，细胞生物学，小鼠遗传学和循环生理学。我们开发的镰状小鼠完全表达人类镰状血红蛋白，并表现出几种人类疾病的临床特征，这是提出的实验策略的关键组成部分。我们应用新颖的实验策略和探索创新的生物学概念，为阐明疼痛性血管闭塞和慢性器官损伤的原因提供了希望。我们预计，我们的发现将导致确定有用的治疗策略，有效的临床管理这种衰弱的人类疾病。

DESCRIPTION (provided by applicant): The research objective of this application is to develop a detailed molecular and mechanistic understanding of the heterogeneity in the rheological and adherence properties of circulating sickle red cells and define the in vivo pathologic sequelae of these cellular changes. To achieve our objective we propose the following series of studies: 1) Determine the structural basis for the heterogeneous rheological properties of distinct subpopulations of circulating sickle red cells and study the in vivo flow behavior and survival of these subpopulations. 2) Determine the contributions of the K-CI cotransporter and the Gardos channel to the generation of theologically compromised dense sickle cells. 3) Evaluate the contributions of specific adhesive receptors, ligands, and plasma proteins in mediating adhesion of sickle red cells to endothelial cells in vitro. 4) Validate the contribution of various adhesive ligands and receptors to sickle cell adhesion by performing adhesion studies using cells from sickle mice in which genes encoding specific adhesive ligands or receptors are inactivated. For the proposed studies, we will use multiple techniques in biophysics, molecular biology, cell biology, mouse genetics and circulatory physiology. The sickle mice that we developed expressing exclusively human sickle hemoglobin and exhibiting several clinical features of human disease are a key component of the proposed experimental strategy. Our application of novel experimental strategies and exploration of innovative biological concepts offers promise for elucidating the causes of painful vaso-occlusion and chronic organ damage. We anticipate that our findings will lead to the identification of useful therapeutic strategies for the effective clinical management of this debilitating human disease.