Viral Gene Expression to Therapeutic Levels in Models of Heart Failure

心力衰竭模型中病毒基因表达达到治疗水平

基本信息

批准号：
7841837
负责人：
JOSEPH E RABINOWITZ
金额：
$ 38.63万
依托单位：
THOMAS JEFFERSON UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2008
资助国家：
美国
起止时间：
2008-08-01 至 2013-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7841837
关键词：
Absenteeism at work Accounting Actins Adenovirus Vector American Heart Association Amino Acid Substitution Amino Acids Animal Disease Models Animal Model Atrial Natriuretic Factor Automobile Driving Biological Assay Budgets C-terminal Ca(2+)-Transporting ATPase Capsid Cardiac Cardiac Myocytes Cardiomyopathies Cardiovascular Diseases Cessation of life Clinical Coronary Coronary artery Coupled Cytomegalovirus Data Discipline Disease Dose EFRAC Elongation Factor Enhancers Failure Fetal Reduction Future GTP-Binding Proteins Gene Delivery Gene Expression Gene Targeting Generations Genes Genetic Goals Green Fluorescent Proteins Heart Heart Diseases Heart failure Heating Hospitalization Human Hypertrophy Immunohistochemistry In Vitro Infarction Injection of therapeutic agent Introns Kinetics Knock-out Ligation Light Link Liver Luciferases MM form creatine kinase Measures Mediating Methods Modeling Molecular Mus Mutation Myocardium Myoglobin Nature Neonatal Organ Pattern Performance Phenotype Phosphotransferases Property Pump Rattus Recombinant adeno-associated virus (rAAV)Regulation Regulator Genes Reporter Genes Route S100A1 protein SERCA2a Safety Sarcoplasmic Reticulum Serotyping Skeletal Muscle Specificity System Tail Testing Therapeutic Thoracic cavity structure Time Tissues Transgenes Transgenic Animals Transgenic Organisms Translating Treatment Efficacy Tropism United States United States National Institutes of Health Veins Ventricular Viral Viral Genes Viral Genome Virion adeno-associated viral vector base cost design dosage ds-DNA gene transfer vector heart function improved in vivo interest mortality particle promoter rat atrial natriuretic peptide recombinant virus response targeted delivery therapeutic gene therapeutic transgene transduction efficiency transgene expression vector vector genome

项目摘要

DESCRIPTION (provided by applicant): Cardiovascular disease accounts for nearly 40% of all deaths that occur in the United States each year, and the cost to the US economy, $394 billion (American Heart Association; 2005), is almost 14 times greater than the NIH total annual budget (FY 2007 $28.4 billion). With respect to that scope, small improvements in therapeutics may reflect enormous benefits. The overall goal of this application is to control recombinant Adeno-associated virus (rAAV) therapeutic transgene expression in the heart after systemic delivery. To examine expression in failing hearts we will employ a standard model of heart disease. The LIM knockouts will be used as our model of hypertrophy. Two methods will be employed to restrict rAAV mediated transgene expression to the heart: 1) Comparisons between the best parental and transcapsidation (combination of capsid subunits from different AAV serotypes into one virion) serotypes, will permit examination of cardiac tropism of the recombinant virus after systemic injection; and 2) Heart specific enhancer/promoter configurations that confer gene expression induced by heart failure will be used to further aid in the specificity of rAAV delivery to the target tissue. Both AAV serotypes 6 and 9 demonstrate significant transgene expression in the heart after systemic tail vein and direct intra-coronary delivery. Quantification of luciferase expression and viral genome copy (vg) number in the heart after delivery of equivalent numbers of viral particles demonstrates that AAV9 delivery results in two-fold greater vg after tail vein injection than AAV6, while AAV6 has greater vg after intra-coronary delivery. AAV6 has a more restricted natural tropism for cardiac tissue, with up to 15% of total light units in the thoracic cavity compared with less than 2% for AAV9. Understanding the amino acids that govern AAV6s' natural tropism for the heart is one objective of this application. A second objective is to combine properties through viral transcapsidation to synergize advantageous phenotypes into one virion. Taking advantage of the cardiac tropism of AAV6 and robust expression from AAV9 as well as the best transcapsidation mixtures we will assess the therapeutic function of 2ARKct and S100A1 transgenes, after systemic injection, in a dose dependent manner. Finally, with the goal of further limiting cardiac gene expression, we have designed enhancer/promoter configurations to bias transgene expression to cardiac tissue. NARRATIVE In the United States heart failure is a major cause of mortality. In addition, the cost of hospitalization and missed work is a drain on our economy. As the molecular changes that underpin the disease are elucidated genetic treatments are beginning to show benefit in animal models. The expression of several genes in animal models of Heart Failure improves the hearts ability to pump without ventricular dilation. However, transgenic and knockout models cannot be easily adapted to human heart failure, therefore, mechanism for gene delivery to the heart become more relevant to these future therapeutic efforts. An end point for this application is to determine the therapeutic dose of a therapeutic gene delivered by an experimental gene transfer vector.

描述（由申请人提供）：心血管疾病占美国每年发生的所有死亡人数的近40％，美国经济的成本为3940亿美元（美国心脏协会； 2005年），是NIH年度总预算的近14倍（2007亿美元284亿美元）。关于该范围，治疗剂的较小改善可能反映出巨大的好处。该应用的总体目标是控制全身传递后心脏中与心脏中与腺相关的病毒（RAAV）治疗转基因表达。为了检查心脏失败的表达，我们将采用心脏病的标准模型。 LIM敲除将用作我们的肥大模型。将采用两种方法将RAAV介导的转基因表达限制为心脏：1）最佳父母和超肽之间的比较（将不同AAV血清型的衣壳血清型与一种病毒粒子结合在一起），将允许检查系统注射后重组病毒的心脏性心动作用； 2）赋予由心力衰竭诱导的基因表达的心脏特异性增强子/启动子构型将用于进一步帮助RAAV递送到目标组织的特异性。 AAV血清型6和9均表现出全身性尾静脉和冠状内递送后心脏中明显的转基因表达。在递送等效的病毒颗粒后，对心脏中心脏中荧光素酶表达和病毒基因组拷贝（VG）的数量进行定量表明，尾静脉注射后AAV9的递送比AAV6导致VG高两个倍，而AAV6在术中递送后具有更大的VG。 AAV6对心脏组织具有更大的自然巨型疗法，胸腔腔中的总光单元占15％，而AAV9的自然组织占心脏组织的15％。了解该应用是该应用的一个目标的一个目标，了解AAV6S自然的自然对心脏的氨基酸是一个目标。第二个目标是通过病毒超肽结合特性，以协同有利的表型协同为一个病毒体。利用AAV6的心脏向量和来自AAV9的鲁棒表达以及最佳的超杀解混合物，我们将以剂量依赖性的方式评估2ARKCT和S100A1转基因的治疗功能。最后，为了进一步限制心脏基因表达，我们设计了增强子/启动子构型以偏向转基因表达对心脏组织。美国心力衰竭的叙事是死亡率的主要原因。此外，住院和失业的成本是我们经济的消耗。随着疾病为基础的分子变化阐明遗传处理，开始在动物模型中显示出益处。在心力衰竭动物模型中，几种基因的表达提高了心脏在无心室扩张而泵送的能力。但是，转基因和基因敲除模型不能轻易适应人类心力衰竭，因此，将基因传递到心脏的机制与这些未来的治疗努力更加相关。该应用的终点是确定实验基因转移载体传递的治疗基因的治疗剂量。