Multifunctional Nanoparticles for Intracellular Delivery

用于细胞内递送的多功能纳米颗粒

• 批准号: 6678925
• 负责人: STEFAN FRANZEN
• 金额: $ 22.58万
• 依托单位: NORTH CAROLINA STATE UNIVERSITY RALEIGH
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-06-01 至 2007-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6678925
• 关键词: atomic force microscopy; biotechnology; cell line; cell nucleus; chemical aggregate; chemical conjugate; cytotoxicity; drug delivery systems; endocytosis; intermolecular interaction; intracellular transport; light scattering; molecular dynamics; nanotechnology; neoplasm /cancer chemotherapy; neoplastic cell; oligonucleotides; particle; peptide library; protein quantitation /detection; protein sequence; surface property; synthetic peptide; technology /technique development; transfection /expression vector; transmission electron microscopy

This proposal describes the synthesis of a general class of nanoparticle delivery vectors based upon hybrid biomolecule-gold nanoparticle complexes. The vectors are designed to transport therapeutic oligonucleotides across cell membranes to target cancer cell nuclei. Therapeutic oligonucleotides are synthetic single stranded nucleic acid molecules that are resistant to digestion by nucleases. A multifunctional approach has been developed which combines cell-specific recognition ligands, endosomal escape peptides, nuclear localization signals and controlled release of therapeutic oligonucleotides to cells. The ability to target biomolecule-nanoparticle complexes to specific cell types, through a relatively simple attachment of cell-specific ligands or peptides, provides the potential for diagnosis and treatment of cancer. A quantitative test for nuclear delivery of an oligonucleotide drug to the nuclei of HeLa cells is proposed as an assay for the efficiency of delivery of a therapeutic agent. The long term goal of this research is to develop a non-viral vector capable of delivering therapeutic oligonucleotides to cancer cells in vivo. Achieving this long term goal requires the following steps. 1. Formulate stable nanoparticle-bioconjugates that are resistant to aggregation and chemical exchange in biological fluids and cells. 2. Develop microscopy techniques to monitor nanoparticle trajectories and quantitate localization in cells. 3. identify the best combination of peptides for performing the functions of endocytosis, endosomal escape and nuclear uptake. 4. Characterize fundamental interactions of protein-peptide conjugates with nanoparticles including their susceptibility to exchange or replacement. 5. Develop strategies for covalent attachment of proteins and oligonucleotides to nanoparticles. 6. Determine of the efficiency of regulated in vitro protein expression using a well-characterized model system.

该提案描述了基于杂种生物分子金纳米颗粒复合物的一般纳米颗粒递送载体的合成。载体旨在将治疗性寡核苷酸转运到细胞膜上以靶向癌细胞核。治疗性寡核苷酸是合成的单链核酸分子，可抗核酸酶消化。已经开发了一种多功能方法，该方法结合了细胞特异性识别配体，内体逃生肽，核定位信号以及将治疗性寡核苷酸的受控释放到细胞中。通过相对简单的 细胞特异性配体或肽的附着，为诊断和治疗癌症提供了潜力。 提出了将寡核苷酸药物核输送到HeLa细胞核的定量测试，以作为治疗剂递送效率的测定法。这项研究的长期目标是开发一个能够将治疗性寡核苷酸输送到体内癌细胞的非病毒载体。 实现这个长期目标需要以下步骤。 1。制定稳定的纳米颗粒 - 双缀合物，对生物液和细胞中的聚集和化学交换具有抗性。 2。开发显微镜技术来监测纳米颗粒轨迹并定量细胞中的定位。 3。确定肽的最佳组合，用于执行内吞作用，内体逃生和核摄取的功能。 4。表征蛋白质肽结合物与纳米颗粒的基本相互作用，包括其易感性交换或替代。 5。制定蛋白质和寡核苷酸与纳米颗粒的共价附着的策略。 6。使用特征良好的模型系统确定调节体外蛋白表达的效率。