MMP-targeted viral gene delivery vectors for treatment of infarcted heart

用于治疗梗塞心脏的 MMP 靶向病毒基因递送载体

基本信息

批准号：
9105416
负责人：
Junghae Suh
金额：
$ 18.76万
依托单位：
RICE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2015
资助国家：
美国
起止时间：
2015-07-15 至 2018-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9105416
关键词：
Adenoviruses Adverse effects Affect Anterior Arteries Atherosclerosis Behavior Binding Capsid Cardiac Cardiovascular Diseases Cell Surface Receptors Cells Congestive Heart Failure Contrast Media Data Dependovirus Detection Development Device or Instrument Development Devices Digestion Disease Dose Engineering Exhibits Exposure to Functional disorder Gelatinase A Gelatinase B Gene Delivery Gene Expression Generations Genes Goals Health Heart Heart Diseases High temperature of physical object Image Imaging Device Immune response In Vitro Knowledge Lead Left Ligation Liver Matrix Metalloproteinases Modeling Molecular Models Mus Mutagenesis Myocardial Infarction Organ PET/CT scan Pathogenesis Patient Care Peptide Hydrolases Performance Positioning Attribute Property RNA Interference Research Personnel Route Sensitivity and Specificity Serotyping Site Skeletal Muscle Solid Specificity Testing Therapeutic Tissues Transcriptional Regulation Transgenes Trypsin Variant Viral Vector Virus adeno-associated viral vector base cellular transduction design extracellular gene therapy heart cell in vivo in vivo Model in vivo imaging innovation molecular imaging molecular modeling mouse model overexpression pre-clinical programs promoter receptor receptor binding skills targeted delivery therapeutic gene transduction efficiency vector viral gene delivery

项目摘要

DESCRIPTION: Adeno-associated viruses (AAV) have demonstrated great promise for gene therapy of a variety of heart diseases. Several AAV variants, such as AAV serotype 9 (AAV9), have been identified with high delivery efficiencies in cardiac cells. Unfortunately, the vectors still suffer from significant delivery to non-target sites, such as liver and skeletal muscles. Consequently, AAV vectors may need to be delivered via invasive administration routes to physically localize gene delivery to the heart. Most current strategies to achieve targeted delivery focus on enabling vectors to bind cell surface receptors that are overexpressed on target diseased cells. Unfortunately, identification of single receptors that lead to high delivery specificity has been difficult to achieve. As an alternate strategy, we hypothesize AAV vectors can be designed to target extracellular proteases - a hallmark of several cardiovascular diseases, including myocardial infarction. The protease-activated viruses (PAVs) will be unable to transduce cardiac cells until they detect pathological concentrations of specific extracellular proteases. To enhance the gene delivery specificity, we will combine transcriptional control with protease activation. In particular, the PAVs will carry a transgene driven by a heart- specific promoter. If only transcriptional control is used, the gene expression will occur in all heart tissues. If only protease-activation is used, then gene expression may occur in off-target sites also exhibiting elevated protease levels. By combining transcriptional control with protease-activation, we aim to limit gene expression to heart tissues that are also diseased. We have promising proof-of-concept data both in vitro and in vivo showing we have successfully created the first generation of PAVs. At low levels of proteases, the vectors are unable to bind their cell surface receptor and display negligible cellular transduction. Upon exposure to extracellular proteases, either a single target protease or a combination of two different proteases, the PAVs "switch on" their cell receptor binding behavior and dramatically increase their transduction efficiency. In aim 1, we will generate a toolkit of single- or dual-input AAV9-based PAVs that are responsive to extracellular proteases known to be elevated in myocardial infarction. In aim 2, we will conduct molecular imaging with protease-activatable contrast agents to detect protease activity in the infarcted heart. The imaging studies will help guide and validate the design and performance of protease-targeted AAV9 vectors. If successful, the studies proposed here will lay the preclinical groundwork for the development of vectors that can deliver genetic therapies specifically to diseased heart tissues.

描述：腺相关病毒 (AAV) 在多种心脏疾病的基因治疗中表现出巨大的前景，例如 AAV 血清型 9 (AAV9)，已被鉴定为在心脏递送细胞中具有高效率。载体仍然受到显着的递送到非目标部位的影响，例如经测试的肝脏和骨骼肌，AAV载体可能需要通过侵入性给药途径将基因递送到心脏。当前实现靶向递送的策略集中于使载体结合在目标患病细胞上过度表达的细胞表面受体，不幸的是，识别出导致高递送的单一受体。作为一种替代策略，我们勇敢地设计了AAV载体来靶向细胞外蛋白酶——这是包括心肌梗塞在内的多种心血管疾病的标志，蛋白酶激活病毒（PAV）将无法转导心肌细胞。直到它们检测到特定细胞外蛋白酶的病理浓度。为了增强基因传递的特异性，我们将转录控制与蛋白酶激活结合起来。特别是，PAV 将携带转基因驱动。如果仅使用转录控制，则基因表达将发生在所有心脏组织中。如果仅使用蛋白酶激活，则基因表达可能发生在也表现出升高的蛋白酶水平的脱靶位点。通过蛋白酶激活进行转录控制，我们的目标是限制同样患病的心脏组织的基因表达，我们在体外和体内的概念验证数据表明我们已经成功地创造了第一代低水平的 PAV。的蛋白酶，载体无法结合其细胞在暴露于细胞外蛋白酶（单一目标蛋白酶或两种不同蛋白酶的组合）时，PAV“开启”其细胞受体结合行为并显着提高其转导效率。将生成一个基于 AAV9 的单输入或双输入 PAV 工具包，该工具包对已知在心肌梗塞中升高的细胞外蛋白酶有反应。我们将使用蛋白酶激活造影剂进行分子成像，以检测梗塞心脏中的蛋白酶活性。成像研究将有助于指导和验证蛋白酶靶向 AAV9 载体的设计和性能。如果成功，这里提出的研究将为临床前奠定基础。为开发可以专门针对患病心脏组织进行基因治疗的载体奠定了基础。