Rho GTPase inhibitors for refrigerated platelet storage

用于冷藏血小板储存的 Rho GTPase 抑制剂

• 批准号: 8712846
• 负责人: Jose A Cancelas
• 金额: $ 22.47万
• 依托单位: P2D, INC.
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-08-01 至 2016-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8712846
• 关键词: Actins; Actomyosin; Autologous; Bacteria; Biological Assay; Blood; Blood Banks; Blood Circulation; Blood Platelets; Boxing; Cells; Cellular biology; Chemicals; Cytoskeleton; DNA Sequence Rearrangement; Data; Development; Dreams; Drug Targeting; Electron Microscopy; Enzymes; Erythrocytes; Event; Excision; F-Actin; Family; Family member; GTP Binding; Glycoprotein Ib; Glycoproteins; Goals; Guanine; Guanosine Triphosphate; Guanosine Triphosphate Phosphohydrolases; Health; Hepatocyte; Hour; Human; In Vitro; Integrins; Intellectual Property; Intervention; Label; Lead; Lesion; Licensing; Lipids; Longevity; Malignant Neoplasms; Mediating; Membrane; Membrane Glycoproteins; Membrane Lipids; Membrane Microdomains; Methods; Mus; Myosin ATPase; Nucleotides; Patients; Phagocytosis; Platelet Count measurement; Platelet Glycoproteins; Platelet Inhibitors; Platelet Transfusion; Preparation; Prevention; Process; Property Rights; Publications; Publishing; Receptor Activation; Recovery; Refrigeration; Signal Transduction; Spectrin; Staining method; Stains; Technology; Time; Transfusion; Trauma; Tubulin; Work; authority; base; chemical addition; cold temperature; commercialization; congenic; drug discovery; improved; in vivo; inhibitor/antagonist; innovation; macrophage; member; new technology; novel; platelet preservation; polymerization; prevent; protein protein interaction; receptor; rho; rho GTP-Binding Proteins; small molecule

ABSTRACT The use of platelet transfusions has increased dramatically since 1980s, but a safe, long-term platelet storage method remains missing. Current practice has platelets stored at 20 to 24¿C after preparation, which has a limited lifetime up to 5 days primarily due to concerns about bacterial contamination. Refrigerated storage reduces platelet life-span because it causes glycoprotein-Ib (GPIb) receptors to cluster on specific microdomains of the platelet membrane. Recognition of specific glycated/syalylated residues on clustered glycoproteins by macrophage b2 integrins and hepatocyte Ashwell-Morell receptors results in platelet phagocytosis by the host and removal from circulation. Thus, prevention of glycoprotein clustering represents a useful target for chemical intervention. Platelet glycoproteins are intimately associated with intracellular cytoskeleton. Their clustering depends on the formation of lipid raft in the platelet membrane which in turn depends on the dynamics of the highly regulated processes of actomyosin assembly/disassembly. Rho family GTPases, including RhoA, Rac1 and Cdc42, are a class of GTP-binding enzymes that are central regulators of F-actin polymerization/depolymarization, and have been shown to control lipid raft formation and composition. Therefore, changes in Rho GTPase activities may influence platelet membrane lipid raft assembly and glycoprotein composition. Based on preliminary and published data, we hypothesize that reversible targeting of Rho family GTPases by small molecule inhibitors can prevent cytoskeleton-dependent refrigeration storage lesions in platelets and result in increased platelet survival. In this project, we will analyze the cytoskeletal functions and in vivo viability of murine and human platelets after refrigeration in the presence or absence of various combinations of specific inhibitors for Cdc42, Rac1 and RhoA GTPases. Combination of cell biology, drug discovery and platelet analysis expertise as demonstrated by multiple co-publications of the co-PIs resulting in an innovative, outside-the-box method for intervention in platelet storage technology. Chemical drug targeting of Rho GTPase activities through interference of protein-protein interaction is a revolutionary approach allowing for reversible inhibition of cytoskeletal rearrangements for which our group has pioneered and own the intellectual property licensed to P2D Inc.

摘要 自20世纪80年代以来，血小板输注的使用急剧增加，但安全，长期的血小板输注 存储方法仍然缺失。目前的做法是在制备后将血小板储存在20至24 ℃， 主要由于担心细菌污染，其使用寿命最多为5天。 冷藏会缩短血小板寿命，因为它会导致糖蛋白Ib（GPIb）受体 聚集在血小板膜的特定微区上。特异性糖化/糖基化的识别 巨噬细胞b2整联蛋白和肝细胞Ashwell-Morell的成簇糖蛋白上的残基 受体导致宿主吞噬血小板并从循环中除去。因此，预防 糖蛋白成簇代表了化学干预的有用目标。血小板糖蛋白是 与细胞内细胞骨架密切相关。它们的聚集依赖于脂质的形成 血小板膜中的筏，这反过来又取决于高度调节的过程的动力学 肌动球蛋白组装/拆卸的过程。Rho家族GTP酶，包括RhoA、Rac 1和Cdc 42，是一种具有生物学活性的蛋白质。 一类GTP结合酶，是F-肌动蛋白聚合/解聚的中心调节剂， 并且已经显示控制脂筏的形成和组成。因此，Rho的变化 GT3活性可能影响血小板膜脂筏组装和糖蛋白组成。 基于初步和已发表的数据，我们假设Rho家族的可逆靶向作用 小分子抑制剂对GTP酶抑制作用的研究 并导致血小板存活增加。在这个项目中，我们将分析细胞骨架， 在存在或不存在血小板的条件下冷藏后， 不存在Cdc 42、Rac 1和RhoA GTP酶的特异性抑制剂的各种组合。 细胞生物学、药物研发和血小板分析专业知识的结合， 共同PI的共同出版物，从而产生了一种创新的、开箱即用的干预方法， 血小板储存技术通过干扰Rho GT3活性的化学药物靶向 蛋白质-蛋白质相互作用是一种革命性的方法， 我们集团率先进行的重新安排，并拥有许可给P2 D的知识产权 Inc.

项目成果

Antioxidant prevents clearance of hemostatically competent platelets after long-term cold storage.

• DOI: 10.1111/trf.16200
• 发表时间: 2021-03
• 期刊: Transfusion
• 影响因子: 2.9
• 作者: Hegde S;Wellendorf AM;Zheng Y;Cancelas JA
• 通讯作者: Cancelas JA