Molenaar Tom J M, Twisk Jaap, de Haas Sonja A M, Peterse Niels, Vogelaar Bram J C P, van Leeuwen Steven H, Michon Ingrid N, van Berkel Theo J C, Kuiper Johan, Biessen Erik A L
Division of Biopharmaceutics, Leiden/Amsterdam Center for Drug Research, Leiden University, P.O. Box 9502, 2300 RA Leiden, The Netherlands.
Biochem Pharmacol. 2003 Sep 1;66(5):859-66. doi: 10.1016/s0006-2952(03)00387-3.
P-selectin is of critical importance in early atherogenesis by initiating leukocyte rolling at the site of endothelial injury. In order to validate P-selectin as a candidate target for the development of anti-atherogenic strategies, we wanted to obtain quantitative information on P-selectin expression, and identify novel peptide-based lead structures that interact with P-selectin. P-selectin mRNA expression in the aortic arch and in other tissues of apoE-deficient (apoE-/-) mice was determined by real-time PCR technology. P-selectin mRNA expression of apoE-/- mice increased steadily with age to levels 14-fold higher than that of control animals. The onset and level of P-selectin expression correlated well with the extent of lesion development, and was more specific for atherosclerotic tissue as compared with other adhesion molecules. Phage display technology was used to obtain novel P-selectin antagonists. Phage display selections resulted in the isolation of a highly P-selectin-specific phage clone. Synthetic peptide-equivalents of this clone displaced the binding of the parent phage and antagonized the binding of a sialyl Lewis(x) analogue to P-selectin. In conclusion, P-selectin expression correlates with early and advanced atherosclerotic lesion development. P-selectin ligands, like the lead structure we have developed here, can therefore be considered as promising tools to identify, target or antagonize P-selectin function within the chronically inflamed arterial wall.
P选择素在早期动脉粥样硬化形成过程中起着至关重要的作用，它可在内皮损伤部位启动白细胞滚动。为了验证P选择素作为抗动脉粥样硬化策略开发的候选靶点，我们希望获得有关P选择素表达的定量信息，并鉴定与P选择素相互作用的新型基于肽的先导结构。通过实时PCR技术测定载脂蛋白E缺陷（apoE-/-）小鼠主动脉弓及其他组织中P选择素mRNA的表达。apoE-/-小鼠的P选择素mRNA表达随年龄稳步增加，达到比对照动物高14倍的水平。P选择素表达的起始和水平与病变发展程度密切相关，与其他黏附分子相比，对动脉粥样硬化组织更具特异性。采用噬菌体展示技术获得新型P选择素拮抗剂。噬菌体展示筛选分离出一个高度P选择素特异性的噬菌体克隆。该克隆的合成肽类似物取代了亲本噬菌体的结合，并拮抗了唾液酸化路易斯（x）类似物与P选择素的结合。总之，P选择素表达与早期和晚期动脉粥样硬化病变发展相关。因此，像我们在此开发的先导结构这样的P选择素配体可被视为在慢性炎症动脉壁内鉴定、靶向或拮抗P选择素功能的有前景的工具。
