权益分类	功能权益	普通用户	{{item.name}}会员
{{category.name}}	{{benefitItem.name}}

Age-Related Decrease in Mitofusin 1 Results in Platelet Dysfunction and Thrombosis

与年龄相关的线粒体融合蛋白 1 减少导致血小板功能障碍和血栓形成

基本信息

批准号：
10054773
负责人：
Andrea Catherine Braganza
金额：
$ 16.27万
依托单位：
UNIVERSITY OF PITTSBURGH AT PITTSBURGH
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-09-10 至 2022-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10054773
关键词：
Adenoviruses Adult Advisory Committees Age Aging Attenuated Automobile Driving Award Bioenergetics Blood Platelets Blood Vessels Bortezomib CD34 gene Calcium Career Mobility Cell Culture System Chimeric Proteins Clinical Complement Data Deep Vein Thrombosis Development Plans Drug usage Exhibits Functional disorder Generations Half-Life Hemostatic Agents Homeostasis Human In Vitro Incidence Individual Knock-out Knockout Mice Laboratories Lasers Lead Longevity Measurement Measures Megakaryocytes Membrane Potentials Mentors Mitochondria Modeling Molecular Morbidity - disease rate Morphology Mus Myocardial Infarction Oxidants Pathway interactions Pharmaceutical Preparations Phase Phenotype Platelet Activation Platelet aggregation Postdoctoral Fellow Predisposition Process Production Proteasome Inhibitor Proteins Reactive Oxygen Species Research Personnel Risk Risk Factors Scheme Stroke Subfamily lentivirinae System Testing Therapeutic Thrombopoiesis Thrombosis Thrombus Training Ubiquitin Universities Wild Type Mouse age related aged aging population attenuation base career career development drug testing experience healthspan healthy aging human subject improved in vivo knock-down mitochondrial dysfunction molecular modeling mortality mouse model multicatalytic endopeptidase complex new therapeutic target novel overexpression platelet function prevent protein degradation rate of change stem cells therapeutic target ubiquitin-protein ligase vascular injury venous thromboembolism

项目摘要

PROJECT SUMMARY/ABSTRACT Aging is one of the greatest risk factors for thrombotic diseases such as deep vein thrombosis, myocardial infarction and stroke. Platelets are central to thrombus formation through their activation and aggregation. Prior studies show age-dependent increases in platelet activation, but the molecular mechanisms remain unknown. Though platelets are anucleate they contain functional mitochondria and an active ubiquitin-proteosome system (UPS). The UPS removes old and damaged proteins to maintain cellular homeostasis. Our preliminary data suggests that platelets isolated from healthy aged (>75 years) human subjects show increased UPS activity and decreased expression of the key mitochondrial fusion protein mitofusin 1 (MFN1) compared to platelets isolated from healthy young subjects (18-35 years). Platelets from aged individuals also exhibited increased mitochondrial oxidant production and platelet activation. Based on these observations, we will test the hypothesis that age-dependent changes in the UPS lead to increased mitochondrial dysfunction resulting in mitochondrial reactive oxygen species (mtROS) production to stimulate platelet activation and subsequent thrombosis. In Aim 1, we will characterize the UPS in isolated platelets from young and aged healthy human subjects. Concomitant MFN1 levels and platelet activation measurements will be made to determine how these systems are altered by the age-related changes in the UPS. To establish a murine model for molecular mechanism, we will perform identical measurements in young (8-10 week) and aged (>1.5 year) wildtype (WT) mice. These studies will be complemented by an in vitro cell culture system of human derived CD34+ progenitor cells which can be differentiated into platelets and used for mechanistic studies that utilize lentivirus to specifically target the UPS components. In Aim 2, we generated platelet-specific MFN1 knockout (KO) mice to determine whether decreased MFN1 in platelets results in increased mtROS and susceptibility to thrombosis. We will measure changes in platelet morphology, mtROS production, and mitochondrial bioenergetics in young and aged platelet-specific MFN1 KO mice and compare it to young and aged WT mice. Adenovirus will be used to overexpress MFN1 in old WT and platelet-specific MFN1 KO mice to determine if it rescues the aged phenotype. Additionally, young and aged WT and platelet-specific MFN1 (KO) mice will be subject to laser-induced vascular injury to determine if platelet activation and thrombosis susceptibility increases when MFN1 is absent. Lentivirus knockdown and overexpression of MFN1 in our CD34+ cell culture system will complement these mechanistic studies. Finally, Aim 3 will determine whether the clinically used drugs MitoQ (mtROS scavenger) or Bortezomib (proteasome inhibitor) can attenuate age-dependent platelet activation and thrombosis in WT and platelet-specific MFN1 KO murine models. Successful completion of this project will unveil a novel mechanism by which platelet activation and thrombosis occur during healthy aging and new strategies to repurpose clinically available drugs to mitigate age-related thrombosis.

项目总结/文摘