VP22 AND TAT mediated gene therapy for the CNS

VP22 和 TAT 介导的中枢神经系统基因治疗

基本信息

批准号：
7922857
负责人：
RAJENDRA KUMAR-SINGH
金额：
$ 15.93万
依托单位：
TUFTS UNIVERSITY BOSTON
依托单位国家：
美国
项目类别：
财政年份：
2009
资助国家：
美国
起止时间：
2009-09-30 至 2010-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7922857
关键词：
Address Adenovirus Infections Adenovirus Vector Adenoviruses Amino Acids Biochemical Biological Models Blindness Cell Culture Techniques Cells Central Nervous System Diseases Central Nervous System Part Chimeric Proteins Cyclic GMP Data Development Disease Eye Gene Delivery Gene Transfer Generations Genetic Green Fluorescent Proteins Hereditary Disease Histologic Infection Length Macular degeneration Mediating Mus Mutate Mutation Nerve Degeneration Neuraxis Neurons Peptides Phenotype Photoreceptors Physiological Property Proteins Relative (related person)Rest Retina Retinal Retinal Degeneration Retinitis Pigmentosa Serotyping Simplexvirus Structure of retinal pigment epithelium Technology Testing Therapeutic Tropism United States Virus Work base cell type gene therapy gutted adenoviral vector helper-dependent adenoviral vector in vivo inherited retinal degeneration mouse model mutant novel particle phosphoric diester hydrolase photoreceptor degeneration research study success therapeutic protein trafficking vector

项目摘要

DESCRIPTION (provided by applicant): Adenovirus (Ad) mediated gene transfer is a promising technology for the treatment of many genetic disorders of the Central Nervous System (CNS). Many of the drawbacks that exist with first-generation Ad vectors have been resolved through the use of 'gutted' or 'helper-dependent' vectors. The genetic basis of neurodegeneration is currently best understood in the retina. In addition, relative to the rest of the CNS, the physical accessibility of the eye makes the retina an excellent model system for studying gene therapy for the CNS. Upon ocular delivery, Ad vectors primarily infect the retinal pigment epithelium (RPE) and the Mueller cells of the retina. However, the diseases that most frequently cause blindness are associated with the expression of mutant proteins in the photoreceptor neurons. In our preliminary studies we have found that adenovirus-delivered green fluorescent protein fused to the full length Herpes Simplex Virus (HSV) tegument protein VP22 can translocate from infected cells to uninfected cells in culture or from the RPE to photoreceptors in vivo. This has led to the hypothesis we wish to test in this study: Can HSV VP22 be used to deliver therapeutic proteins to photoreceptor neurons via the RPE? This will be answered using a mouse model of inherited retinal degeneration (rd). Specifically, in this study we propose to 1) Construct an adenovirus vector expressing the protein transduction domains (PTD) of HSV VP22 fused to GFP and compare their ability to traffic GFP in cell culture and in murine retina. 2) Construct an adenovirus vector expressing a fusion between beta PDE and the PTD of HSV VP22. Assess the ability of this virus to express a functional PDE. 3) Administer a gutted adenovirus vector expressing either a VP22-beta PDE fusion to the retina of rd mice (which have a naturally occurring mutation in beta PDE) and assess the effects upon photoreceptor degeneration.

描述（由申请人提供）：腺病毒（AD）介导的基因转移是治疗中枢神经系统（CNS）许多遗传疾病的有前途的技术。第一代AD矢量存在的许多缺点通过使用“沟渠”或“辅助依赖性”向量解决了。目前，在视网膜中最好理解神经退行性的遗传基础。此外，相对于CNS的其余部分，眼睛的物理可及性使视网膜成为研究中枢神经系统基因治疗的出色模型系统。眼部传递后，AD载体主要感染视网膜色素上皮（RPE）和视网膜的Mueller细胞。但是，最常见的失明的疾病与光感受器神经元中突变蛋白的表达有关。在我们的初步研究中，我们发现腺病毒传递的绿色荧光蛋白融合到全长疱疹单纯疱疹病毒（HSV）TEGUMENT蛋白VP22可以从感染细胞转移到培养物中的未感染细胞，或者从RPE到光感受器在体内。这导致了我们希望在这项研究中检验的假设：可以使用HSV VP22通过RPE将治疗蛋白传递给感受器神经元？这将使用遗传性视网膜变性（RD）的鼠标模型来回答。具体而言，在这项研究中，我们提出了1）构建腺病毒载体，该腺病毒载体表达HSV VP22的蛋白质转导域（PTD）融合到GFP中，并比较其在细胞培养和鼠视网膜中交通GFP的能力。 2）构建一个表达βPDE和HSV VP22 PTD之间融合的腺病毒载体。评估该病毒表达功能性PDE的能力。 3）施用肠道腺病毒载体，表达对RD小鼠的视网膜（在βPDE中发生自然发生的突变）的VP22-beta PDE融合，并评估对感光受体退化的影响。