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：
8160322
负责人：
RAJENDRA KUMAR-SINGH
金额：
$ 41.25万
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/8160322
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

DESCRIPTION (provided by applicant): 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. PUBLIC HEALTH RELEVANCE: According to public opinion polls, blindness is the second most feared condition amongst Americans after cancer. Gene therapy for blindness requires the application of viruses that can cause serious adverse events in patients. The objective of this proposal is to develop a non-viral approach to gene therapy. Upon completion, this study will be ready to advance a non-viral gene therapy approach to clinical trials, pending standard toxicology studies as required by the FDA.

描述（由申请人提供）：视网膜变性是已知的最遗传异质性疾病组之一，涉及184个基因座。几项眼基因治疗临床试验表明，基因治疗是治疗视网膜疾病的有效方法。到目前为止，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都将病毒用作基因转移载体。病毒作为基因转移载体具有显着的优势 - 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。尽管存在这些缺点，但由于缺乏替代方法，病毒是目前选择的载体。如果可以通过非病毒基因转移载体的发展来解决上述缺点，这些载体可以将基因传递到成人视网膜等有丝分裂后组织，则将对临床前和临床眼基因治疗领域产生重大影响。不幸的是，非病毒载体仅在正在进行有丝分裂的细胞培养或新生儿视网膜中有效地工作。因此，与病毒不同，非病毒载体通常无法挽救视网膜变性的动物模型，除非在新生儿鼠视网膜上应用 - 无法将其直接转化为有丝分裂后人类视网膜。最近，我们开发了一个3.5 kD的肽（POD），该肽可以形成类似于大小（136nm）的病毒（136nm）的纳米颗粒，并在有丝分裂后视网膜中启用转基因表达。尽管用POD纳米颗粒转移基因不如病毒高效，但足以使体内视网膜变性短期延迟。这只是迄今为止的两项研究之一，表明使用非病毒载体的成年小鼠视网膜变性延迟。我们研究的主要局限性是来自POD纳米颗粒的短期转基因表达。这项研究的主要目的是通过使用核DNA整合或DNA保留元件来延长POD纳米颗粒的转基因表达。这项研究的第二个目标是提高POD基因转移的效率，使其可能更有效，第三个目标是验证两个相关视网膜变性动物模型中POD的改善。视网膜变性中观察到的高遗传异质性阻碍了患者的及时可用性，因为每个基因和病毒组合都需要通过冗长的过程来开发。对于184个基因座，这种方法在经济上是不可行的。因此，我们建议使用POD纳米颗粒不传递单个基因，而是使用编码神经营养因子的基因，例如开发非病毒，非基因特异性方法来治疗视网膜变性。完成这些研究后，我们将有一个新型的非病毒载体，准备在临床试验中使用，待毒理学研究。如果成功，这些研究将是眼部基因治疗的范式转移。公共卫生相关性：根据公众舆论民意测验，失明是癌症之后第二最令人担忧的状况。失明的基因疗法需要应用可能在患者中引起严重不良事件的病毒。该提案的目的是开发一种非病毒治疗方法。完成后，这项研究将准备好推进非病毒基因治疗方法进行临床试验，并按照FDA的要求进行标准毒理学研究。