Hemostasis and Thrombosis: Chemistry, Biology and Physiology

止血和血栓形成：化学、生物学和生理学

• 批准号: 10439604
• 负责人: Sriram Krishnaswamy
• 金额: $ 163.71万
• 依托单位: CHILDREN'S HOSP OF PHILADELPHIA
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-09-01 至 2024-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10439604
• 关键词: Address; Area; Biochemical; Biochemistry; Biological; Biological Process; Biology; Blood Vessels; Blood coagulation; Chemistry; Coagulation Process; Complex; Deuterium; Disease; Ensure; Enzymes; Factor V; Factor Va; Foundations; Goals; Hemophilia A; Hemorrhage; Hemostatic Agents; Hemostatic function; Hydrogen; Individual; Kinetics; Knowledge; Lead; Life; Membrane; Membrane Proteins; Molecular; Molecular Biology; Multienzyme Complexes; Myocardial Infarction; Participant; Peptide Hydrolases; Physiology; Play; Proteins; RNA; Reaction; Reagent; Recombinants; Records; Regulation of Proteolysis; Research; Research Personnel; Roentgen Rays; Role; Series; Serine Protease; Specificity; Stroke; Structure; Substrate Interaction; Substrate Specificity; Testing; Therapeutic; Thrombosis; Thrombus; Trypsin; Variant; Western World; X-Ray Crystallography; aptamer; base; biophysical analysis; biophysical techniques; cancer procoagulant; cofactor; enzyme substrate; human disease; human morbidity; human mortality; insight; interdisciplinary approach; interest; mouse model; novel strategies; novel therapeutics; programs; protein expression; prothrombinase complex; response; structural biology

Overall Program-Abstract Blood coagulation derives from a series of specific proteolytic activation reactions that are catalyzed with narrow and defined specificity by trypsin-like serine proteinases. In several instances, these proteinases function in membrane assembled enzyme complexes. Distinctive protein substrate specificities and the modulation of enzymic function by interactions with membranes and cofactors are hallmarks of the proteolytic reactions of blood coagulation. There are major gaps in the current understanding of the molecular bases for these unique features that underlie the function of the hemostatic reactions. This program proposes an integrated approach focused on the modulation of enzymic function and specificity that uniquely arises from macromolecular interactions that underlie the action of the hemostatic enzymes. Project 1 (Krishnaswamy) uses the prothrombinase complex as a paradigm to investigate structural and functional mechanisms deriving from the membrane-dependent interactions between cofactor, proteinase and substrate in enzyme function. Project 2 (Camire) will investigate molecular mechanisms at play in the conversion of factor V to the cofactor, factor Va and the surprising new biological insights that these mechanisms reveal. Project 3 (Sullenger) employs RNA aptamers as unique probes for the macromolecular interactions essential for coagulation enzyme function and to show the way forward for novel approaches to either interfere with or enhance these interactions for therapeutic gain. Project 4 (Arruda) investigates the biochemistry of intrinsic Xase with a focus on characterizing proteinase and cofactor variants with superior function and potential for use as the next generation of therapeutics in the treatment of hemophilia. The objectives of the four projects will be supported by an administrative core (Core A), a core that provides support for molecular biology, protein expression and structural biology (Core B) and a core that supports the program with mouse models of hemostasis and thrombosis (Core C). Overall, this project applies the expertise of the individual investigators towards addressing major unanswered questions in hemostasis and thrombosis extending from biochemical insights to biological function and physiology. The proposed approaches will provide new insights into the chemistry and biology of the blood coagulation reactions with implications for an understanding of normal hemostasis and thrombosis.

总体程序提取 血液凝血来自一系列特定的蛋白水解激活反应，这些反应与 通过胰蛋白酶样丝氨酸蛋白酶狭窄而定义的特异性。在某些情况下，这些蛋白酶 膜组装酶复合物中的功能。独特的蛋白质底物特异性和 通过与膜和辅因子相互作用的酶功能调节是蛋白水解的标志 血液凝血的反应。当前对分子碱基的理解存在主要差距 这些独特的特征是止血反应功能的基础。该程序提出了一个 集成方法着重于酶功能和特异性的调节 止血酶作用的基础的大分子相互作用。项目1（克里希纳斯瓦米） 使用凝血酶酶复合物作为范式来研究得出的结构和功能机制 从酶功能中的辅因子，蛋白酶和底物之间的膜依赖性相互作用。 项目2（Camire）将研究因子V向辅因子转化时的分子机制， 因子VA和这些机制所揭示的令人惊讶的新生物学见解。项目3（Sullenger） 利用RNA适体作为对凝结必不可少的大分子相互作用的独特探针 酶功能并显示出新颖方法干扰或增强这些方法的前进方向 治疗增益的相互作用。项目4（Arruda）研究了内在Xase的生物化学，重点 在表征具有优质功能且潜在使用的蛋白酶和辅因子变体作为下一个 治疗血友病的治疗剂。这四个项目的目标将得到支持 通过行政核心（核心A），该核心为分子生物学，蛋白质表达和 结构生物学（核心B）和核心止血模型和核心的核心 血栓形成（核心C）。总体而言，该项目将个人调查人员的专业知识应用于 解决止血和血栓形成中的主要未解决问题，从生化见解延伸到 生物功能和生理。拟议的方法将为化学和 血液凝血反应的生物学，对正常止血和 血栓形成。

项目成果

Gene therapy matures to medicines.

基因疗法成熟为药物。

• DOI: 10.1093/hmg/ddz182
• 发表时间: 2019
• 期刊: Human molecular genetics
• 影响因子: 3.5
• 作者: Davidson,BeverlyL;Arruda,ValderR
• 通讯作者: Arruda,ValderR

Blood coagulation factor X: molecular biology, inherited disease, and engineered therapeutics.

• DOI: 10.1007/s11239-021-02456-w
• 发表时间: 2021-08
• 期刊: Journal of thrombosis and thrombolysis
• 影响因子: 4
• 作者: Camire RM

Therapeutic Aptamers: Evolving to Find their Clinical Niche.

• DOI: 10.2174/0929867326666191001125101
• 发表时间: 2020
• 期刊: Current medicinal chemistry
• 影响因子: 4.1
• 作者: Nimjee SM;Sullenger BA
• 通讯作者: Sullenger BA

Regulation of factor V and factor V-short by TFPIα: Relationship between B-domain proteolysis and binding.

• DOI: 10.1074/jbc.ra120.016341
• 发表时间: 2021-01
• 期刊: The Journal of biological chemistry
• 作者: Petrillo T;Ayombil F;Van't Veer C;Camire RM
• 通讯作者: Camire RM

Aptamer-based factor IXa inhibition preserves hemostasis and prevents thrombosis in a piglet model of ECMO.

• DOI: 10.1016/j.omtn.2021.12.011
• 发表时间: 2022-03-08
• 期刊: Molecular therapy. Nucleic acids
• 作者: Reed CR;Bonadonna D;Otto JC;McDaniel CG;Chabata CV;Kuchibhatla M;Frederiksen J;Layzer JM;Arepally GM;Sullenger BA;Tracy ET
• 通讯作者: Tracy ET