Computationally Designed Heparan Sulfates and Pathophysiology

天然生物聚合物肝素和硫酸乙酰肝素在许多生物学过程中起着关键作用。 包括凝血、生长和形态学、血管生成、免疫应答、炎症 和病原体入侵。事实上，肝素及其衍生物，低分子量肝素和磺达肝素， 在临床上用作许多血栓性疾病的抗凝剂。使用的基本依据 肝素在这些疾病中的作用是其与抗凝血酶（一种血浆）的高亲和力和高特异性相互作用。 糖蛋白和凝固酶抑制剂，尤其是凝血酶、因子Xa和因子IXa。尽管 由于肝素的长期使用，它仍然存在许多问题。更好的基于肝素的 抗凝治疗是迫切需要的，特别是在它的异质性也可以给 引起与污染相关的问题。此外，尽管肝素和硫酸乙酰肝素在 虽然在其他生理和病理过程中起重要作用，但尚未设计出临床药物。 这种状态的主要原因是H/HS的惊人结构多样性，这导致a） 制备具有确定结构组成的HS制剂的困难和B）研究 大量HS结构与多种蛋白质的相互作用。在这方面需要取得重大进展 解码H/HS结构-功能关系的两个区域，以便能够设计激动剂和/或 用于调节生物过程的拮抗剂。 PEG的这个项目11解决了研究所有可能的相互作用的基本困难。 HS序列与任何蛋白质（即，上述区域B））通过实验室开发的独特技术 组合虚拟文库筛选（CVLS）技术。结合项目一、三 第四，我们建议：1）破译HS结构-功能关系的基本方面， 凝血和炎症系统，以及2）为了在体内动物模型中测试这种增强的理解， 尤其是对于凝血系统。因此，我们建议：1）研究具体和非具体的重要性， 使用计算方法研究硫酸乙酰肝素与蛋白质的特异性相互作用， 2）开发计算设计的HS结构， 肝素辅因子II的活化剂;和3）研究设计的H/HS与凝血蛋白的相互作用 在分子水平上开发为新的临床有用的抗凝剂。 相关性（参见说明）： 血栓性疾病影响美国1/3的个体。拟议的研究涉及计算 硫酸乙酰肝素作为血栓性疾病调节剂的设计、生物化学评价和动物试验。

Natural biopolymers heparin and heparan sulfate play critical roles in a large number of biological processes including coagulation, growth and morphology, angiogenesis, immune response, inflammation, and pathogen invasion. In fact, heparin and its derivatives, low molecular weight heparins and fondaparinux, are clinically used as anticoagulants in a number of thrombotic disorders. The fundamental basis for the use of heparin in these disorders is its high affinity and high specificity interaction with antlthrombin, a plasma glycoprotein and inhibitor of coagulation enzymes, especially thrombin, factor Xa and factor IXa. Despite the longstanding use of heparin, it continues to suffer from a number of problems. Better heparin-based anticoagulation therapy is critically needed, especially at a time when its heterogeneous nature can also give rise to problems associated with contaminations. Additionally, although heparin and heparan sulfate play important roles in other physiological and pathological processes, no clinical agent has as yet been devised. The major reason for this state is the phenomenal structural diversity of H/HS, which results in a) the difficulty of preparing HS preparations with defined structural composition and b) the difficulty of studying the interaction of a large number of HS structures with multiple proteins. Major advances are necessary in these two areas to decode H/HS structure - function relationships so as to enable the design of agonists and/or antagonists for modulation of biological processes. This Project 11 of the PEG addresses the fundamental difficulty of studying the interactions of all possible HS sequences with any protein (i.e., area b) above) through a unique technology developed in the laboratory of the PL called combinatorial virtual library screening (CVLS) technology. In combination with Projects I, III and IV, we propose 1) to decipher fundamental aspects of HS structure - function relationships in the coagulation and inflammation system, and 2) to test this enhanced understanding in in vivo animal models, especially for the coagulation system. Thus, we propose to 1) study the importance of specific and non- specific interactions of heparan sulfate with proteins using computational approaches and identify promising structures for in vitro and in vivo investigation; 2) develop computationally designed HS structures as specific activators of heparin cofactor II; and 3) investigate the interaction of designed H/HS with coagulation proteins at a molecular level for development as new clinically useful anticoagulants. RELEVANCE (See instructions): Thrombotic disorders affect 1 in 3 individuals in the US. The proposed research involves the computational design, biochemical evaluation and animal testing of heparan sulfates as modulators of thrombotic disorders.