The Therapeutic Role for LOLX2 in Pulmonary Hypertension

LOLX2 在肺动脉高压中的治疗作用

基本信息

批准号：
10326830
负责人：
Jochen Steppan
金额：
$ 17.17万
依托单位：
JOHNS HOPKINS UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-01-01 至 2023-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10326830
关键词：
Address Adhesions Animal Model Animals Arteries Attenuated Award Biological Biological Assay Biology Blood Vessels CRISPR/Cas technology Cardiac Catheterization Procedures Cell Adhesion Cell model Cell physiology Cells Cessation of life Chronic Collagen Complex Coupling Cytometry Data Deposition Development Disease Echocardiography Elastin Elements Extracellular Matrix Foundations Generations Genes Goals Heart failure Human Hypertrophy Hypoxia Immunohistochemistry Impairment Interruption Knock-out LOXL2 gene Lung MADH2 gene MATRICS Consensus Cognitive Battery Magnetism Measurement Mechanics Medial Mentors Modeling Molecular Monocrotaline Motion Myography Pathway interactions Patients Pharmacology Phosphorylation Phosphotransferases Property Protein Isoforms Protein-Lysine 6-Oxidase Pulmonary Circulation Pulmonary Hypertension Pulmonary artery structure Rattus Research Research Personnel Resolution Right Ventricular Dysfunction Right Ventricular Function Right ventricular strain Rodent Model Role SU 5416 Signal Pathway Signal Transduction Smooth Muscle Myocytes Stress Structure Testing Therapeutic Tissues Torsion Tracer Training Transforming Growth Factor alpha Vascular remodeling Ventricular Western Blotting amine oxidase arterial stiffness career cell behavior cell motility coronary fibrosis crosslink effective therapy electric impedance experimental study extracellular immunocytochemistry improved improved outcome in vivo Model inhibitor interest knock-down mortality novel novel strategies novel therapeutic intervention novel therapeutics pressure prevent pulmonary arterial hypertension pulmonary arterial stiffening receptor right ventricular failure skills small hairpin RNA small molecule inhibitor symptom treatment therapeutic target therapy outcome

项目摘要

PROJECT SUMMARY In patients with pulmonary hypertension (PH) and especially with pulmonary arterial hypertension (PAH), pulmonary artery (PA) stiffness increases right ventricular strain and ultimately leads to death from heart failure. However, despite pulmonary vasculature stiffening with increased impedance being a paramount mechano-biological regulator of PAH, no therapies currently target this mechanism. Therapy for PAH continues to focus on symptomatic treatment, vasodilatory therapy, or treatment of the underlying disease (if known). None of these addresses the fundamental changes in vascular remodeling. My long-term career goal is to become an expert in understanding the mechanisms underlying PAH, thus facilitating the identification of more effective therapies and improving outcomes. This goal directly builds on my prior research and extends my interests from the systemic to the pulmonary circulation. Our preliminary data has clearly demonstrated that Lysyl oxidase like-2 (LOXL2) is an attractive gene in pulmonary arterial stiffness and cardiac fibrosis. LOXL2, is an amine oxidase that catalyzes collagen crosslinking and contributes to matrix remodeling and stiffening. Our core hypothesis is that increased LOXL2 expression, secretion, and function contributes to the development of PA stiffening. Our objective here is to study LOXL2 biology, mechanism, and pathobiology in a hierarchical manner from cells to rodent models. We will determine how LOXL2 influences cell behavior, tissue mechanics, function, and matric generation during hypoxia. We will utilize lentiviral shRNA and CRISPR/Cas9 in rat PA smooth muscle cells. We will use magnetic torsion cytometry, cell substrate impedance sensing, and tensile testing; test collagen assembly, vasoreactivity, and compliance; and evaluate adhesion, motility, and proliferation. Furthermore, we will study the relationships between LOXL2 and the TGF-β1 pathway, as well as its role in rat models of PH and PAH, using multiple models to induce pulmonary vascular stiffening in the presence an absence of a LOXL2 inhibitor. Measurements will include PA stiffness using pressure myography and stress strain relationships in isolated vessels, right ventricular function using high-resolution echocardiography, and right ventricular function using pressure-volume loops. We will establish the therapeutic potential of LOXL2 inhibition in PAH and test its effects in models of PH and PAH. This could entail not only a new drug within an existing paradigm but a whole new approach to the treatment of these patients - altering the structural composition of the extracellular matrix. With the support of my mentors, my goal during the K08 award period is to acquire the expertise and training in a nurturing academic setting that will enable me to attain the proficiency necessary to conduct these experiments, and the independence to continue beyond the award period.

项目概要肺动脉高压（PH）患者，尤其是肺动脉高压患者（PAH）、肺动脉（PA）僵硬会增加右心室应变并最终导致死亡然而，尽管肺血管硬化和阻抗增加是一个原因。 PAH 的重要机械生物调节剂，目前尚无针对该机制的治疗方法。对于PAH，继续关注对症治疗、血管舒张治疗或治疗潜在疾病（如果已知）。这些都不能解决血管的根本变化。重塑。我的长期职业目标是成为了解 PAH 潜在机制的专家，从而促进更有效的治疗方法的确定并直接改善这一目标。以我之前的研究为基础，并将我的兴趣从全身循环扩展到肺循环。我们的初步数据清楚地表明赖氨酰氧化酶样 2 (LOXL2) 是一个有吸引力的基因 LOXL2 是一种催化胶原蛋白的胺氧化酶。我们的核心假设是交联并有助于基质重塑和硬化。 LOXL2 的表达、分泌和功能有助于 PA 硬化的发生。这里的目标是以分层方式研究 LOXL2 生物学、机制和病理学我们将确定 LOXL2 如何影响细胞行为、组织力学、我们将在大鼠中利用慢病毒 shRNA 和 CRISPR/Cas9。 PA 平滑肌细胞。我们将使用磁扭转细胞术、细胞基质阻抗传感和拉伸测试；测试胶原蛋白组装、血管反应性和顺应性并评估粘附性、运动性；此外，我们将研究 LOXL2 和 TGF-β1 通路之间的关系，及其在 PH 和 PAH 大鼠模型中的作用，使用多种模型诱导肺血管在不存在 LOXL2 抑制剂的情况下的硬度测量将包括使用 PA 硬度。压力肌动描记法和孤立血管的应力应变关系，使用右心室功能我们将使用压力-容量环进行高分辨率超声心动图和右心室功能检查。确定 LOXL2 在 PAH 中的治疗抑制潜力，并测试其在 PH 和 PH 模型中的效果这不仅需要在现有范式中开发一种新药，而且需要一种全新的治疗方法。这些患者的治疗 - 改变细胞外基质的结构组成。在导师的支持下，我在 K08 奖励期间的目标是获得专业知识和在培育性学术环境中接受培训，使我能够获得必要的熟练程度进行这些实验，并在获奖期限后继续独立。