WELL-DEFINED MULTIFUNCTIONAL POLYMERIC NANOCARRIERS FOR EFFECTIVE GENE DELIVERY

明确的多功能聚合物纳米载体可实现有效的基因传递

• 批准号: 8257580
• 负责人: You Han Bae
• 金额: $ 27.51万
• 依托单位: UNIVERSITY OF UTAH
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-05-01 至 2014-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8257580
• 关键词: Biological; Blood; Buffers; Cell Cycle; Cell Nucleus; Cell model; Cells; Characteristics; Charge; Chemistry; Clinical Trials; Complex; Custom; Cytosol; DNA; Development; Dissociation; Drug Formulations; Effectiveness; Electrostatics; Endocytosis; Endosomes; Environment; Evaluation; Family; Future; Gene Delivery; Gene Expression; Genes; Goals; Growth; Guidelines; Hepatocyte; In Vitro; Injection of therapeutic agent; Interphase Cell; Investigation; Left; Ligands; Link; Liver; Liver diseases; Lysosomes; Membrane; Methods; Modeling; Mutation; Nature; Non-Viral Vector; Nuclear; Nuclear Envelope; Nuclear Import; Nuclear Pore; Organ; Output; Performance; Pharmacologic Substance; Phase; Polymers; Preparation; Process; Production; Proliferating; Protons; Relative (related person); Research; Role; Serum; Serum Proteins; Site; Solubility; Stream; Sulfonamides; Surface; System; Testing; Time; Toxic effect; Transfection; Viral; Viral Vector; Work; aqueous; base; cell growth; controlled release; cytotoxicity; design; extracellular; flexibility; functional group; gene therapy; gene therapy clinical trial; in vivo; nanocarrier; nanoscale; nanosized; novel; novel strategies; nucleocytoplasmic transport; polyanion; polycation; public health relevance; receptor; tool; trafficking; translational study; tumor; vector; viral gene delivery

DESCRIPTION (provided by applicant): Despite several advantages of non-viral vectors over viral vectors in gene therapy, few clinical trials have investigated non-viral systems. A major reason for the suboptimal performance of non-viral gene approaches is their low transfection efficiency especially in vivo. Extensive investigations into the major limiting factors during the cellular trafficking processes have provided spectra of tools anticipated to circumvent the extracellular and intracellular obstacles in successful use of non-viral systems. Polymeric vectors demonstrate great flexibility in their design and functionalization. Biodegradable or non-degradable polycations are often used as DNA condensing vectors and modifying a few functional groups helps targeting and trafficking into the nucleus. However, current approaches are not truly well-defined because the modified polycations are electrostatically and randomly complexed with anionic genes. A poorly defined vector preparation with a few functionalities and inadequate exposure of the functional groups at the right places are primarily responsible for low efficiency. A novel approach proposed in this application is to accommodate all functionalities (as many as needed) for gene trafficking from an injection site to the nucleus, which include shielding the positively charged surface in the blood stream, cell targeting ligands, endosomolytic agents, controlled release of DNA from polyplex, nuclear pore dilation and nuclear import. All these functionalities will be incorporated into nanoscaled assemblies constructed by a layer-by-layer method, resulting in triple layered gene vectors; the core, mantle, and the shell layers. More importantly the nanoconstruct is expected to expose the functional groups at the place required in the intracellular compartments in a well defined manner and each layer will be peeled-off after serving its due role, leaving minimal components for nuclear import of the incorporated gene. This unique assembly approach assures high potential for pharmaceutical gene formulations for future translational studies. The long term goal will be accomplished in four specific aims: first three aims for core layer, mantle layer and shell layer respectively. Each layer will be customized for target cells; proliferating and non-proliferating cells, endosomal pH, and ligand receptor. The fourth specific aim will test and prove the proposed concept in vivo. PUBLIC HEALTH RELEVANCE: Despite several advantages of non-viral vectors over viral owns in gene therapy, particularly in reproducible production of pharmaceutical formulations, most clinical trials for gene therapy employed viral systems. This application is for development of polymeric gene carriers based on a new design concept, which is anticipated to present a high potential for a translational study, for acceptable transfection efficiency in vivo and have minimal toxicity. The novel and unique design principle will accommodate all functionalities required for effective gene transfection and these functionalities will be exposed in the appropriate intracellular compartments where needed the most in a well-defined fashion. The nano-sized gene carrier will be constructed from pharmaceutically-friendly polymers.

描述（由申请人提供）：尽管非病毒载体比基因治疗中的病毒载体有几个优势，但很少有临床试验研究了非病毒系统。非病毒基因方法次优性能的主要原因是其低转染效率，尤其是体内。对细胞运输过程中主要限制因素的广泛研究提供了预期的工具光谱，以绕过成功使用非病毒系统的细胞外和细胞内障碍。聚合物向量在其设计和功能化方面表现出极大的灵活性。可生物降解或不可降解的多阳离子通常用作DNA冷凝矢量，并修改一些官能团有助于靶向和运输到核中。但是，当前的方法并不是真正定义的，因为修饰的多阳离子与阴离子基因静电和随机复合。较差的矢量制备具有少数功能，并且在正确位置的官能团的暴露不足主要负责低效率。本应用中提出的一种新颖的方法是适应从注射部位到细胞核的基因运输的所有功能（尽可能多），其中包括屏蔽血流中带正电荷的表面，细胞靶向配体，内溶血剂，控制释放，控制释放来自流行文素的DNA，核孔的扩张和核进口。所有这些功能将被纳入由逐层方法构建的纳米级组件中，从而产生三层基因载体。核心，地幔和外壳层。更重要的是，预计纳米构造将以明确的方式将官能团在细胞内隔室中所需的位置暴露，并且在发挥其适当作用后，每一层都将剥落，从而使掺杂基因的核进口最小成分。这种独特的组装方法可确保用于未来翻译研究的药物基因制剂的高潜力。长期目标将以四个特定的目的来实现：核心层，地幔层和壳层的前三个目标。每层将针对目标细胞定制；增殖和非增殖细胞，内体pH和配体受体。第四个特定目标将在体内测试并证明拟议的概念。 公共卫生相关性：尽管非病毒载体在基因疗法中拥有几个优势，尤其是在可再现的药物配方中，大多数用于基因治疗的临床试验用于基因疗法。该应用是基于新的设计概念来开发聚合基因载体的，该概念预计将对体内可接受的转染效率具有很高的转化研究潜力，并且具有最小的毒性。新颖而独特的设计原理将适应有效基因转染所需的所有功能，这些功能将在需要的情况下以适当的细胞内隔室暴露在适当的细胞内隔室中。纳米大小的基因载体将由药物友好的聚合物构建。

项目成果

Reconstitutable charged polymeric (PLGA)(2)-b-PEI micelles for gene therapeutics delivery.

• DOI: 10.1016/j.biomaterials.2011.01.077
• 发表时间: 2011-05
• 期刊: BIOMATERIALS
• 影响因子: 14
• 作者: Mishra, Deepa;Kang, Han Chang;Bae, You Han
• 通讯作者: Bae, You Han

Bioreducible polymers as a determining factor for polyplex decomplexation rate and transfection.

• DOI: 10.1021/bm301794d
• 发表时间: 2013-02-11
• 期刊: Biomacromolecules
• 影响因子: 6.2
• 作者: Hwang HS;Kang HC;Bae YH
• 通讯作者: Bae YH

Role of polymeric endosomolytic agents in gene transfection: a comparative study of poly(L-lysine) grafted with monomeric L-histidine analogue and poly(L-histidine).

• DOI: 10.1021/bm500843r
• 发表时间: 2014-10-13
• 期刊: BIOMACROMOLECULES
• 影响因子: 6.2
• 作者: Hwang, Hee Sook;Hu, Jun;Na, Kun;Bae, You Han
• 通讯作者: Bae, You Han

Trafficking microenvironmental pHs of polycationic gene vectors in drug-sensitive and multidrug-resistant MCF7 breast cancer cells.

• DOI: 10.1016/j.biomaterials.2010.01.001
• 发表时间: 2010-04
• 期刊: Biomaterials
• 影响因子: 14
• 作者: Kang HC;Samsonova O;Bae YH
• 通讯作者: Bae YH

DNA Polyplexes as Combinatory Drug Carriers of Doxorubicin and Cisplatin: An in Vitro Study.

• DOI: 10.1021/mp500873k
• 发表时间: 2015-08-03
• 期刊: Molecular pharmaceutics
• 影响因子: 4.9
• 作者: Kang HC;Cho H;Bae YH
• 通讯作者: Bae YH