RNA Aptamer-directed Anticoagulant Therapy

描述（由申请方提供）：历史上，普通肝素已用于治疗和预防血栓栓塞性疾病。然而，由于与肝素相关的并发症（如出血），低分子量肝素（LMWH）已被开发出来，并正在取代普通肝素。LMWH已被证明是有效的抗血栓药物;然而，LMWH的作用不易逆转，而普通肝素可被硫酸鱼精蛋白逆转。该提案的总体目标是开发一种抗凝血酶（ATIII）特异性RNA适体，该适体将联合收割机的安全性和有效性与普通肝素的可逆性结合起来。为了实现这一目标，将开发用于快速逆转适体的抗凝特性的该适体的寡核苷酸解毒剂。 通过开发针对凝血因子Xa的RNA适体和RNA适体解毒剂来证明这一概念。与该小组合作，将开发一种解毒剂控制的ATIII特异性RNA适体。该适体将通过结合ATIII的D-螺旋来模拟LMWH的作用，并增强因子Xa的ATIII抑制。这种适体/解毒剂对将具有优于LMWH的优点，因为它不仅能够类似于LMWH控制血栓形成，而且解毒剂的可用性将允许更好的治疗调节和干预。本提案中提出的实验将使用体外和体内方法来表征这个抗凝血酶特异性RNA适体家族。

DESCRIPTION (provided by applicant): Historically, unfractionated heparin has been used in the treatment and prevention of thromboembolic disorders. However, due to complications associated with heparin (such as bleeding), low molecular weight heparins (LMWHs) have been developed and are being used in place of unfractionated heparin. LMWHs have proven effective as antithrombotic agents; however, the effects of LMWHs cannot be readily reversed while unfractionated heparin can be reversed by protamine sulfate. The overall goal of this proposal is to develop an antithrombin (ATIII)-specific RNA aptamer that will combine the safely and efficacy of LMWH with the reversibility of unfractionated heparin. To accomplish this goal, an oligonucleotide antidote for this aptamer that will be used to rapidly reverse the anticoagulant properties of the aptamer will be developed. Proof of this concept is established by the development of an RNA aptamer and RNA aptamer antidote directed against coagulation factor Xa. In collaboration with this group, an antidote-controlled ATIII-specific RNA aptamer will be developed. This aptamer will mimic the action of LMWH by binding to the D-helix of ATIII and enhance ATIII inhibition of factor Xa. This aptamer/antidote pair will have advantages over LMWHs because it will not only be able to control thrombosis similar to LMWHs but the availability of an antidote will allow for better therapeutic regulation and intervention. The experiments proposed in this proposal will characterize this family of antithrombin-specific RNA aptamers using both in vitro and in vivo approaches.