Ubiquitination and Endothelial Fibrosis in Early Pulmonary Arterial Hypertension

早期肺动脉高压中的泛素化和内皮纤维化

基本信息

批准号：
10207768
负责人：
Bradley Wertheim
金额：
$ 16.89万
依托单位：
BRIGHAM AND WOMEN'S HOSPITAL
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：
2020-07-01 至 2025-06-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10207768
关键词：
Advisory Committees Anabolism Area Arterial Disorder Arteries Atomic Force Microscopy Award Binding Biochemistry Biological Biological Assay Blood Vessels Cells Cessation of life Clinical Clinical Investigator Award Collagen Computer Models Coupling Data Deposition Development Disease Doctor of Medicine Doctor of Philosophy Early treatment Endothelium Failure Fibrillar Collagen Fibrosis Functional disorder Funding Gel Gene Expression Profile Genetic Transcription Heart failure Hospitalization Hospitals Human Hydrogen Peroxide Hypoxia Immunofluorescence Immunologic Immunoprecipitation Impairment In Vitro Inflammatory Investigation K-Series Research Career Programs Lung Mass Spectrum Analysis Mediating Mediator of activation protein Medical Medicine Mentors Mentorship Modeling Modification Molecular Monocrotaline Outcome Oxidation-Reduction Pathogenesis Pathogenicity Pathway Analysis Pathway interactions Phenotype Phosphotransferases Physicians Physiologic intraventricular pressure Premature Mortality Principal Investigator Progressive Disease Proteasome Inhibition Protein Tyrosine Kinase Proteins Proteomics Proto-Oncogenes Publications Pulmonary Hypertension Pulmonary Vascular Resistance Pulmonary artery structure Regulation Research Research Training Resistance Right Ventricular Dysfunction Right Ventricular Function Right ventricular structure Role SU 5416 Scientist Shortness of Breath Signal Transduction Site-Directed Mutagenesis Small Interfering RNA Smooth Muscle Myocytes Substrate Interaction Sulfhydryl Compounds Testing Time Training Training Programs Treatment Efficacy Ubiquitin Ubiquitination Vascular Diseases Vascular remodeling Ventricular Woman advanced disease arterial stiffness arteriole career development didactic education effective therapy endothelial dysfunction functional disability improved in silico in vivo lung hypoxia medical schools mortality multicatalytic endopeptidase complex novel novel strategies overexpression oxidant stress oxidation predictive modeling pressure prevent programs protein-tyrosine kinase c-src pulmonary arterial hypertension pulmonary artery endothelial cell scaffold src-Family Kinases symposium targeted treatment transcriptomics ubiquitin ligase ubiquitin-protein ligase

项目摘要

PROJECT SUMMARY/ABSTRACT This K08 Mentored Clinical Scientist Research Career Development Award describes a five-year research and training program to establish the principal investigator (PI) as an independent, R01-funded physician-scientist in the field of pulmonary vascular medicine. Co-mentorship by Bradley Maron, M.D., an expert in cysteinyl thiol biochemistry, vascular fibrosis, and PAH pathobiology and pathophysiology, and Joseph Loscalzo, M.D., Ph.D., an expert in redox biochemistry, network medicine, and proteomics, will oversee the PI’s research and professional development activities at Brigham and Women’s Hospital and Harvard Medical School. Scientific publication, guidance from an advisory committee, didactic education, and presentation at scientific conferences will support the scientific aims of the project and provide career development in three key training areas: the pathobiology of PAH inception, kinase ubiquitination, and experimental PAH models. The PI is guaranteed >80% protected academic time to dedicate to the proposed K08 program. PAH is characterized by endothelial dysfunction and oxidant stress that promotes fibrotic remodeling of the pulmonary arterioles, leading to right heart failure, and death. Mortality and hospitalization increase incrementally beginning at a mean pulmonary artery pressure (mPAP) below values that were previously thought to be pathogenic. Therefore, treatment of early-stage PAH may improve clinical outcome, but effective therapies do not exist because the pathobiology of endothelial dysfunction and vascular remodeling in early PAH is not known. We provide novel data that early-stage PAH is characterized by pulmonary arteriolar fibrosis, impaired right ventricle-pulmonary artery (RV-PA) coupling, and increased endothelial C-terminal src kinase (Csk) expression occurring prior to the development of elevated mPAP in vivo. Computational modeling predicts that the Csk binding-partner and E3 ubiquitin ligase Casitas b-lineage proto-oncogene (Cbl) is susceptible to oxidative modification at cysteinyl thiol 396 (Cys396). We hypothesize that oxidation of endothelial Cbl-Cys396 prevents Csk ubiquitination to increase Csk-dependent fibrosis and impair RV-PA coupling in early-stage PAH. To test this hypothesis, we propose the following specific aims: 1) Determine the impact of Cbl-Cys396 oxidation on Csk ubiquitination and expression in cultured human PAECs, 2) Assess the role of increased endothelial Csk on fibrillar collagen expression in cultured human PAECs, and 3) Test the effect of Csk inhibition on RV-PA coupling in early- and advanced-stage PAH in vivo. We anticipate that findings from this K08 will establish Csk- targeted therapy as a novel strategy by which to prevent or reverse endothelial fibrosis and vascular remodeling prior to the development of elevated mPAP and right heart failure in PAH. Furthermore, this research program will establish kinase-ubiquitin ligase dysregulation in early-stage PAH as a plane of separation from the PI’s current co-mentors.

项目概要/摘要 K08 指导临床科学家研究职业发展奖描述了五年研究和培训计划，将主要研究者 (PI) 确立为独立的、由 R01 资助的肺血管医学领域的医师科学家布拉德利·马龙 (Bradley Maron) 医学博士共同指导。半胱氨酰硫醇生物化学、血管纤维化、PAH 病理生物学和病理生理学专家，Joseph Loscalzo 博士是氧化还原生物化学、网络医学和蛋白质组学方面的专家，他将监督 PI 的工作布莱根妇女医院和哈佛医学院的研究和专业发展活动学校。科学出版物、咨询委员会的指导、教学教育和演讲。科学会议将支持该项目的科学目标，并在三个关键领域提供职业发展培训领域：PAH 起始的病理学、激酶泛素化和实验性 PAH 模型。保证 >80% 的受保护学术时间用于拟议的 K08 课程。 PAH 的特点是内皮功能障碍和氧化应激，促进血管纤维化重塑肺小动脉，导致右心衰竭，死亡率和住院率增加。从平均肺动脉压 (mPAP) 开始逐渐低于之前认为的值因此，早期PAH的治疗可能会改善临床结果，但有效的治疗方法并不重要。不存在，因为早期 PAH 的内皮功能障碍和血管重塑的病理学并不存在我们提供的新数据表明，早期 PAH 的特点是肺小动脉纤维化、受损。右心室-肺动脉 (RV-PA) 耦合，以及内皮 C 端 src 激酶 (Csk) 增加计算模型预测体内 mPAP 升高之前的表达。 Csk 结合伴侣和 E3 泛素连接酶 Casitas b 谱系原癌基因 (Cbl) 易受我们捕获了内皮 Cbl-Cys396 的氧化修饰。阻止 Csk 泛素化，增加 Csk 依赖性纤维化并损害早期 PAH 中的 RV-PA 偶联。为了检验这一假设，我们提出以下具体目标：1) 确定 Cbl-Cys396 氧化的影响培养的人 PAEC 中 Csk 泛素化和表达的影响，2) 评估内皮 Csk 增加的作用对培养的人 PAEC 中纤维状胶原蛋白表达的影响，以及 3) 测试 Csk 抑制对 RV-PA 的影响我们预计 K08 的发现将建立 Csk- 体内早期和晚期 PAH 的耦合。靶向治疗作为预防或逆转内皮纤维化和血管重塑的新策略在 PAH 患者出现 mPAP 升高和右心衰竭之前。将在早期 PAH 中建立激酶泛素连接酶失调作为与 PI 分离的平面现任共同导师。