DNA Nanoparticle Gene Therapy in Brain

DNA纳米颗粒脑基因治疗

基本信息

批准号：
8179538
负责人：
David M. Yurek
金额：
$ 33.17万
依托单位：
UNIVERSITY OF KENTUCKY
依托单位国家：
美国
项目类别：
财政年份：
2011
资助国家：
美国
起止时间：
2011-07-01 至 2016-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8179538
关键词：
Affect Age Animal Model Animals Astrocytes Biodistribution Brain Brain region Caliber Cells Corpus striatum structure DNA DNA delivery Data GDNF gene Gene Transfer Genes Glial Fibrillary Acidic Protein Human Incidence Infusion procedures Injection of therapeutic agent Knowledge Laboratories Lead Lesion Methods Monitor Nerve Degeneration Neuraxis Neurodegenerative Disorders Nucleic Acids Parkinson Disease Patients Phase I Clinical Trials Plasmids Population Production Proteins Publications Rattus Replacement Therapy Reporter Genes Rodent Series Site Structure Techniques Technology Testing Therapeutic Time Transfection Transgenes Translating Tropism Up-Regulation Viral Vector Virus Work aged aging brain brain cell brain tissue design dopaminergic neuron experience gene therapy glial cell line derived neurotrophic factor, rat glial cell-line derived neurotrophic factor immunogenicity improved juvenile animal nanometer nanoparticle nervous system disorder neurotoxic neurotrophic factor nigrostriatal pathway non-viral gene therapy novel particle plasmid DNA prevent promoter research study response therapeutic gene transgene expression translational study vector young adult

项目摘要

DESCRIPTION (provided by applicant): The proposed studies will determine the feasibility of compacting plasmid DNA into "nanoparticles" and using these nanoparticles to deliver their payload into cells of the central nervous system (CNS) as a non-viral, gene therapy technique. DNA compacting techniques will be used to form nanoparticles containing condensed DNA plasmids with diameters in the range of 8-12 nanometers. In a series of recent publications coming out of our laboratories, we have shown that synthetic nanoparticles containing DNA plasmids can be used to transfect brain cells and establish both short- and long-term transgene activities following a single injection of DNA nanoparticles (DNP) directly into brain tissue. These encouraging results have lead us to propose a series of studies to further characterize and test the potential of using synthetic vectors to deliver therapeutic genes to the brain as a possible treatment for neurodegenerative disorders. My laboratory has considerable experience testing neurotrophic factor therapy as well as cellular replacement therapies in animal models of Parkinson's disease (PD), and we propose to examine the feasibility of delivering a gene encoding for the neurotrophic factor glial cell line-derived neurotrophic factor (GDNF) to brain cells as a means to protect the brain against neuronal degeneration that occurs in an animal model of PD. The first set of experiments will expand upon our current knowledge of DNP technology. In Specific Aim 1, we will attempt to further optimize plasmids design and mode of intracerebral delivery of compacted DNA nanoparticles (DNPs), and then assess the immunogenicity of this treatment. In the second specific aim, we will determine if DNP transfection of the lesion brain is greater than in the intact brain, and whether or not the aged brain is more susceptible to DNP transfection than younger brain; this studies will determine if an up-regulation of astrocytes at the site of neurodegeneration actually benefits transfection efficiency of DNPs because our preliminary studies indicate DNPs have a tropism for astrocytes. Finally, our third specific aim will determine whether or not intracerebral infusion of modified hGDNF DNPs prevent neurodegeneration of dopaminergic neurons following a neurotoxic lesion of the nigrostriatal pathway. Successful results in these studies could then be applied to animal models of neurodegenerative disorders and possibly lead to translational studies for the treatment of neurological disorders, such as Parkinson's disease. PUBLIC HEALTH RELEVANCE: The proposed studies will use a novel nanoparticle technology that allows nucleic acids (DNA) to be compacted near their theoretical limit; this technology almost duplicates the compaction efficiency of viruses. We present preliminary data showing proof-of-concept that these nanoparticles can be used as a non-viral gene therapy for transfecting cells in the brain. Successful results from our animal studies could then be translated to human studies using these targeted nanoparticles as a form of non-viral, gene therapy to treat various neurological disorders.

描述（由申请人提供）：拟议的研究将确定将质粒DNA压入“纳米颗粒”的可行性，并使用这些纳米颗粒将其有效载荷作为非病毒，基因治疗技术传递到中枢神经系统（CNS）的细胞中。 DNA压实技术将用于形成含有直径在8-12纳米范围内的凝结DNA质粒的纳米颗粒。在我们实验室的一系列最近出版物中，我们表明，含有DNA质粒的合成纳米颗粒可用于转染脑细胞，并在单个注射DNA纳米颗粒（DNP）直接注射DNA纳米颗粒（DNP）后同时建立短期和长期的转基因活性。这些令人鼓舞的结果使我们提出了一系列研究，以进一步表征和测试使用合成矢量将治疗基因传递给大脑的潜力，以作为神经退行性疾病的可能治疗。我的实验室在帕金森氏病动物模型（PD）中具有测试神经营养因素治疗以及细胞替代疗法的丰富经验，我们建议研究提供编码基因编码基因的可行性PD。第一组实验将扩展我们当前对DNP技术的知识。在特定的目标1中，我们将尝试进一步优化紧凑型DNA纳米颗粒（DNP）的质粒设计和脑内递送模式，然后评估该处理的免疫原性。在第二个特定目的中，我们将确定病变大脑的DNP转染是否大于完整的大脑，以及老年大脑是否比年轻大脑更容易受到DNP转染的影响；这项研究将确定神经变性部位的星形胶质细胞的上调是否会受益于DNP的转染效率，因为我们的初步研究表明DNP对星形胶质细胞具有向流。最后，我们的第三个特定目标将确定脑毒性途径神经毒性病变后，脑内输注是否会阻止多巴胺能神经元的神经退行性。然后，这些研究的成功结果可以应用于神经退行性疾病的动物模型，并可能导致转化研究以治疗神经系统疾病，例如帕金森氏病。公共卫生相关性：拟议的研究将使用一种新型的纳米颗粒技术，该技术允许将核酸（DNA）压实在其理论极限附近；这项技术几乎复制了病毒的压实效率。我们提供了初步数据，表明概念证明这些纳米颗粒可以用作用于大脑中转染细胞的非病毒基因治疗。然后，我们的动物研究的成功结果可以使用这些靶向纳米颗粒作为一种非病毒，基因疗法的一种形式来转化为人类研究，以治疗各种神经系统疾病。