Hybrid synthetic targeted lenti-VLP nanoparticle for gene delivery

用于基因递送的混合合成靶向慢病毒颗粒纳米颗粒

基本信息

批准号：
7611050
负责人：
MARTIN C WOODLE
金额：
$ 29.96万
依托单位：
APARNA BIOSCIENCES CORPORATION
依托单位国家：
美国
项目类别：
财政年份：
2008
资助国家：
美国
起止时间：
2008-09-30 至 2010-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7611050
关键词：
Acute Address Affect Animal Model Antibodies Binding Biological Biomedical Research Cell Survival Cell membrane Cell physiology Cells Charge Condition Coupling Cultured Cells Dependence Development Disease Dose Elements Endosomes Exhibits Eye diseases Facility Construction Funding Category Foundations GTP-Binding Proteins Gene Delivery Gene Expression Genes Genetic Engineering Hand Hybrids Hydrogen Bonding In Vitro Interphase Cell Length Lentivirus Vector Ligands Localized Malignant Neoplasms Mediating Membrane Fusion Methods MicroRNAs Modeling Morphology Mus N-terminal Nanotechnology Non-Viral Vector Nucleic Acids Numbers Particle Size Peptides Phase Phase I Clinical Trials Polymers Production Proteins Public Health RGD (sequence)Range Reporter Reporter Genes Research Research Design Retroviral Vector Series Serum Source Structure Structure-Activity Relationship Surface Therapeutic Tissues Tumor-Associated Vasculature Vesicular stomatitis Indiana virus Viral Virus-like particle angiogenesis base cell type cyclo(S,S)KYGCRGDWPC density directed evolution env Gene Products gene delivery system gene function gene therapy in vivo interest nanoparticle neovascularization non-viral gene delivery particle polycation polypeptide protein expression protein function receptor size small hairpin RNA surface coating tumor vector

项目摘要

DESCRIPTION (provided by applicant): A need for better gene delivery has blocked the potential advancement in biomedical research and most critically development of better therapeutics that operate at the gene function level. This problem is most acute for non-dividing cells, which in fact are the predominant cell type. Thus the considerable interest in gene therapy has narrowed considerably to efforts largely developing ex-vivo approaches and use of lentiviral vectors with several important unique capabilities that include delivery to non-dividing cells. A major limitation that applies to all the lentiviral vectors developed is a lack of applicability for parenteral administration, largely due to their dependence on a finicky envelope protein that provides the critical functions of cell binding and then cell membrane fusion and intracellular delivery. Despite many efforts, including high throughput genetic engineering (e.g. "directed evolution"), the natural envelope G protein from VSV used to pseudotype retroviral vectors for many years remains one of the most effective envelope for lentiviral vectors. This problem has posed an essentially insurmountable barrier to the development of versatile targeted lentiviral vectors. Interestingly, our development of targeted nanoparticles for non-viral gene delivery has demonstrated versatile ligand-mediated tissue targeting using ligand-polymer conjugation but activity on non-dividing cells remains a major hurdle. Thus we are developing a hybrid combining our successful synthetic tissue selective targeting with lentiviral particles adept for non-dividing cells. We are developing a ligand-targeted polymer coating to replace the lentiviral envelope protein function missing in lenti-viral like particles (lenti-VLP) produced without envelope protein. Since this protein is one of the major sources of lentiviral vector instability, greatly enhanced stability is expected to be an added benefit. Our preliminary results indicate that simple homopolymer cationic polypeptides can replace lentiviral vector envelope protein function but the potency needs to be enhanced substantially. To determine feasibility of a hybrid synthetic ligand- medated targeted lenti-VLP in this Phase I research plan, we first will construct and screen a number of cationic polypeptides with defined structures to identify an SAR for binding lenti-VLP and replacing envelope protein function but without causing instability of the lenti-VLP. We then will identify methods for coupling targeting ligands to selected polypeptides without adversely affecting the cell delivery function and optimize cell selective ligand-mediated gene deliver in cell culture and for neovasculature tissue in an animal model. Public Health Relevance: A need for better gene delivery is a major barrier to development of critically needed therapeutic treatment options that operate at the gene function level. Many efforts are focused on ex-vivo uses of lentiviral vectors with many unique advantages but a major limitation is an inability for parenteral administration, due to dependence on a finicky envelope protein that provides cell binding and intracellular delivery. Conversely, our targeted nanoparticles for nucleic acids have demonstrated ligand- mediated tissue targeting but efficiency, especially for non-dividing cells remains a major hurdle. The planned study is to determine feasibility of a hybrid synthetic ligand-targeted lenti-virus like particle lacking envelope protein and optimization of cell selective ligand-mediated gene delivery for neovasculature tissue in an animal model.

描述（由申请人提供）：需要更好的基因递送的需求阻碍了生物医学研究的潜在进步，以及在基因功能水平上运作的更好疗法的最关键发展。对于非分散细胞而言，此问题最为急性，实际上，这是主要的细胞类型。因此，对基因疗法的极大兴趣已大大缩小了在很大程度上开发前体内方法和使用具有几种重要独特能力的慢病毒载体的努力，其中包括向非分散细胞递送。适用于所有慢病毒载体的主要局限性缺乏适用于肠胃外给药的能力，这在很大程度上是由于它们依赖于挑剔的包膜蛋白，该蛋白提供了细胞结合的关键功能，然后是细胞膜融合和细胞内递送的关键功能。尽管许多努力，包括高吞吐量基因工程（例如“定向进化”），但多年来用于逆转录病毒载体的VSV的天然包膜G蛋白仍然是慢病毒载体最有效的包膜之一。这个问题为开发多功能针对性的慢病毒载体的发展带来了根本无法克服的障碍。有趣的是，我们开发用于非病毒基因递送的靶向纳米颗粒已经证明了使用配体 - 聚合物偶联的多功能配体介导的组织靶向，但对非分散细胞的活性仍然是一个主要障碍。因此，我们正在开发一种混合动力，将成功的合成组织选择性靶向与耐植物颗粒相结合，熟练的细胞。我们正在开发一个靶向配体的聚合物涂层，以替换宽叶病毒中缺少的慢病毒包膜蛋白功能，例如颗粒（lenti-vlp），而没有包膜蛋白。由于该蛋白是慢病毒载体不稳定性的主要来源之一，因此预计稳定性将是一个额外的好处。我们的初步结果表明，简单的均聚物阳离子多肽可以替代慢病毒载体包膜蛋白功能，但需要大大增强效力。为了确定在本I阶段研究计划中混合合成配体的靶向Lenti-VLP的可行性，我们首先将构建和筛选具有具有定义结构的许多阳离子多肽，以识别用于结合的Lenti-vlp的SAR，并替换了包膜蛋白质功能，但没有引起Lenti-i-vlp的不稳定性。然后，我们将识别将靶向配体偶联到选定多肽的方法，而不会对细胞递送功能产生不利影响，并优化细胞选择性配体介导的基因在细胞培养和动物模型中的新生血管形组织中递送。公共卫生相关性：需要更好的基因递送是开发在基因功能水平上运行的急需治疗方法的主要障碍。许多努力都集中在具有许多独特优势的慢病毒载体的外部使用，但主要的限制是肠胃外给药，这是由于依赖于提供细胞结合和细胞内递送的挑剔的包膜蛋白。相反，我们用于核酸的靶向纳米颗粒已显示出配体介导的组织靶向但效率，尤其是对于非分散细胞的效率仍然是一个主要障碍。计划的研究是为了确定涉及含蛋白质的粒子蛋白质的杂种合成配体靶向的小病毒的可行性，并优化了动物模型中神经组织的细胞选择性配体介导的基因递送。