Rheological and Adherence Properties of Sickle Cells

描述(申请人提供)：本申请的研究目标是对循环中镰状红细胞的流变性和粘附性的异质性进行详细的分子和机理了解，并确定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)确定不同亚群的循环镰状红细胞的异质性流变性的结构基础，并研究这些亚群的体内流动行为和存活率。2)确定K-CI共转运体和Gardos通道在神学上受损的致密镰状细胞的产生中的作用。3)评价特异性黏附受体、配体和血浆蛋白在介导镰状红细胞与血管内皮细胞黏附中的作用。4)通过使用编码特定黏附配体或受体的基因失活的镰刀鼠细胞进行黏附研究，验证各种黏附配体和受体对镰刀细胞黏附的贡献。对于拟议的研究，我们将使用生物物理学、分子生物学、细胞生物学、小鼠遗传学和循环生理学的多种技术。我们开发的仅表达人类镰状血红蛋白并表现出人类疾病的几个临床特征的镰状小鼠是拟议的实验策略的关键组成部分。我们新的实验策略的应用和对创新生物学概念的探索为阐明痛苦的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效的治疗策略，以有效地临床管理这种令人衰弱的人类疾病。

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.