Non-Viral Gene Therapy for Retinal Degeneration

视网膜变性的非病毒基因疗法

基本信息

批准号：
8536453
负责人：
RAJENDRA KUMAR-SINGH
金额：
$ 18.15万
依托单位：
TUFTS UNIVERSITY BOSTON
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：
2011-09-01 至 2015-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8536453
关键词：
Acute Address Adenoviruses Adult American Animal Model Binding Blindness Cell Culture Techniques Cell Nucleus Cell surface Cells Chemicals Chemistry Clinical Clinical Trials Cloning Complex Cytosol DNA DNA Integration DNA delivery Development Disadvantaged Disease Electroretinography Elements Endosomes Eye Part Gene Expression Gene Transfer Genes Genetic Genetic Heterogeneity Glial Fibrillary Acidic Protein Goals Histology Human Immune Immune response In Situ Nick-End Labeling Individual Insertional Mutagenesis Integrase Investigational Drugs Laboratories LacZ Genes Light Longevity Luciferases Lysosomes Malignant Neoplasms Matrix Attachment Regions Measures Mediating Mitosis Mitotic Modeling Modification Monophenol Monooxygenase Muller&apos s cell Mus Neonatal Non-Viral Vector Nuclear Nucleic Acids Nucleosomes Outcome Study Patients Peptides Phase I Clinical Trials Photoreceptors Process Production Proteins Public Opinion Poll Publishing Quantum Dots Recombinant Proteins Recombinants Research Retina Retinal Retinal Degeneration Retinal Diseases Retinitis Pigmentosa Serious Adverse Event Source Staining method Stains Structure of retinal pigment epithelium Surface System Technology Testing Therapeutic Time Tissues Toxic effect Toxicology Transgenes Transgenic Animals Translating Viral Vector Virus Vision research Work body system clinical application conditioned fear design fluorophore gene delivery system gene therapy gene therapy clinical trial gene transfer vector glial cell-line derived neurotrophic factor glutamylalanine immunogenicity improved in vivo knockout animal large scale production leucyl-alanine nanoparticle nanoscale neurotrophic factor non-viral gene delivery non-viral gene therapy novel nucleolin photoreceptor degeneration plasmid DNA pre-clinical recombinant viral vector recombinant virus retinal apoptosis scaffold small molecule subretinal injection trafficking transgene expression uptake vector

项目摘要

Project Description Retinal degeneration is one of the most genetically heterogeneous groups of disorders known, involving over 184 loci. Several ocular gene therapy clinical trials have remarkably demonstrated that gene therapy is a valid approach to treat retinal diseases. Each of these clinical trials and almost every preclinical gene therapy study thus far have utilized viruses as the gene transfer vector. Viruses have significant advantages as gene transfer vectors- primarily their ability to efficiently deliver genes to post-mitotic retinal cells in vivo. However, viruses also have some disadvantages, including induction of host immune responses, a limited transgene capacity, insertional mutagenesis and difficulty in production. Despite these disadvantages, viruses are the current vector of choice in almost all ocular gene therapy studies because of a lack of alternatives. If the above disadvantages could be resolved by the development of non-viral gene transfer vectors that could deliver genes to post-mitotic tissues such as adult retina, it would have substantial impact on the field of preclinical and clinical ocular gene therapy. Unfortunately, non-viral vectors only work efficiently in cell culture or in neonatal retina where mitosis is ongoing. Hence, unlike viruses, non-viral vectors generally fail to rescue animal models of retinal degeneration unless applied in neonatal murine retina - results from which cannot be directly translated to post-mitotic human retina. Recently, we developed a 3.5 Kd peptide (POD) that can form nanoparticles resembling viruses in size (136nm) when complexed with DNA and enable transgene expression in post-mitotic retina. Although gene transfer with POD nanoparticles was not as efficient as with viruses, it was sufficient to enable a short-term delay in retinal degeneration in vivo. This is only one of two studies thus far demonstrating a delay in retinal degeneration in an adult mouse using a non-viral vector. The major limitation of our study was that of short-term transgene expression from POD nanoparticles. The primary objective of this study is to prolong transgene expression from POD nanoparticles by use of nuclear DNA integration or DNA retention elements. The second objective of this study is to improve the efficiency of gene transfer of POD such that it could be more potent and the third objective is to validate the improvements in POD in two relevant animal models of retinal degeneration. The high level of genetic heterogeneity observed in retinal degeneration hampers the timely availability of therapies for patients as each gene and virus combination needs to be developed through a lengthy process. Such approaches are not economically feasible for the >184 loci. Hence, we propose to use POD nanoparticles not to deliver individual genes but instead, genes encoding neurotrophic factors such as to develop a non-viral, non gene-specific approach to treat retinal degeneration. Upon completion of these studies we will have a novel non-viral vector ready for use in clinical trials pending toxicology studies. If successful, these studies would be a paradigm shift in ocular gene therapy.

项目描述视网膜变性是已知的最遗传异质性疾病组之一，涉及 184基因座。几项眼基因疗法临床试验已明显证明基因治疗是有效的治疗视网膜疾病的方法。这些临床试验中的每一个以及几乎所有临床前基因治疗研究到目前为止，已经将病毒用作基因转移载体。病毒作为基因转移具有显着优势载体 - 主要是它们在体内有效输送基因的能力。但是，病毒也有一些缺点，包括诱导宿主免疫反应，有限的转基因能力，插入诱变和生产困难。尽管这些缺点，病毒是当前的由于缺乏替代方案，几乎所有眼科基因疗法研究中的首选载体。如果以上可以通过开发非病毒基因转移向量来解决缺点基因对后有线组织，例如成年视网膜，将对临床前的领域产生重大影响和临床眼基因治疗。不幸的是，非病毒载体仅在细胞培养或在有丝分裂正在进行的新生儿视网膜。因此，与病毒不同，非病毒载体通常无法营救视网膜变性的动物模型除非应用于新生儿鼠视网膜的结果 - 无法从中获得直接转化为有丝分裂后的人视网膜。最近，我们开发了一个3.5 kD的肽（POD），可以形成当与DNA复合并启用转基因表达时，纳米颗粒类似于大小（136nm）的病毒（136nm）在有丝分裂后视网膜中。尽管用POD纳米颗粒转移基因不如病毒高效，但它是足以使体内视网膜变性的短期延迟。这只是到目前为止的两项研究之一使用非病毒载体证明成年小鼠的视网膜变性延迟。主要限制我们的研究是来自POD纳米颗粒的短期转基因表达。主要目标这项研究是通过使用核DNA整合或 DNA保留元件。这项研究的第二个目标是提高基因转移的效率 POD使其可能更有效，第三个目标是验证POD的改进视网膜变性的相关动物模型。在视网膜中观察到的高水平的遗传异质性退化阻碍了患者的及时可用性，因为每个基因和病毒组合需要通过漫长的过程来开发。这种方法在经济上对 > 184个基因座。因此，我们建议使用POD纳米颗粒不传递单个基因，而是基因编码神经营养因素，例如开发非病毒，非基因特异性方法来治疗视网膜退化。完成这些研究后，我们将有一个新型的非病毒载体准备在临床上使用试验即将进行毒理学研究。如果成功，这些研究将是眼部基因治疗的范式转移。