DNA Nanoparticle Gene Therapy in Brain

DNA纳米颗粒脑基因治疗

基本信息

批准号：
8471215
负责人：
David M. Yurek
金额：
$ 30.99万
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/8471215
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.

描述（由申请人提供）：拟议的研究将确定将质粒 DNA 压缩成“纳米颗粒”并使用这些纳米颗粒将其有效负载递送到中枢神经系统（CNS）细胞中作为非病毒基因治疗技术的可行性。 DNA压缩技术将用于形成包含直径在8-12纳米范围内的浓缩DNA质粒的纳米粒子。在我们实验室最近发表的一系列出版物中，我们表明，含有 DNA 质粒的合成纳米颗粒可用于转染脑细胞，并在直接单次注射 DNA 纳米颗粒 (DNP) 后建立短期和长期转基因活性进入脑组织。这些令人鼓舞的结果促使我们提出一系列研究，以进一步表征和测试使用合成载体将治疗基因传递到大脑作为神经退行性疾病的可能治疗方法的潜力。我的实验室在帕金森病 (PD) 动物模型中测试神经营养因子疗法以及细胞替代疗法方面拥有丰富的经验，我们建议研究提供编码神经营养因子胶质细胞系衍生神经营养因子 (GDNF) 的基因的可行性）将脑细胞作为保护大脑免受帕金森病动物模型中发生的神经元变性的一种手段。第一组实验将扩展我们目前对 DNP 技术的了解。在具体目标 1 中，我们将尝试进一步优化质粒设计和压缩 DNA 纳米颗粒 (DNP) 的脑内递送模式，然后评估这种治疗的免疫原性。在第二个具体目标中，我们将确定病变大脑中的 DNP 转染是否大于完整大脑中的 DNP 转染，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星形胶质细胞的上调是否实际上有利于 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性。最后，我们的第三个具体目标是确定脑内输注修饰的 hGDNF DNP 是否可以预防黑质纹状体通路神经毒性损伤后多巴胺能神经元的神经变性。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，并可能导致治疗帕金森病等神经系统疾病的转化研究。