Functionalized Lipid Carriers for Nucleic-Acid and Drug Therapeutics

用于核酸和药物治疗的功能化脂质载体

• 批准号: 10020420
• 负责人: CYRUS R SAFINYA
• 金额: $ 29.56万
• 依托单位: UNIVERSITY OF CALIFORNIA SANTA BARBARA
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-09-20 至 2022-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10020420
• 关键词: Achievement; Acids; Adhesions; Albumin-Stabilized Nanoparticle Paclitaxel; Biological; Biological Assay; Biophysics; Cancer cell line; Capsid; Cations; Cell Survival; Cells; Charge; Chemicals; Chemistry; Chemotherapy-Oncologic Procedure; Clinical Trials; Code; Communities; Complex; Cone; Custom; Cyclic Peptides; Cytosol; DNA; Data; Dependence; Development; Distal; Drug Delivery Systems; Electron Microscopy; Electrostatics; Encapsulated; Endosomes; Engineering; Excision; Exons; Fluorescence Microscopy; Gene Delivery; Gene Silencing; Genes; Goals; Homing; Hospitals; Human; Hydrophobicity; Hypersensitivity; In Vitro; Introns; Knowledge; Laboratories; Ligands; Lipid Bilayers; Lipids; Liposomes; Malignant Neoplasms; Mediating; Membrane; Methods; Micelles; Modernization; Non-Viral Vector; Nucleic Acids; Organelles; Organic Chemistry; Paclitaxel; Penetration; Peptides; Pharmaceutical Preparations; Phase; Probability; Property; Proteins; RNA; Reaction; Research; Research Activity; Roentgen Rays; Safety; Science; Shapes; Small Interfering RNA; Solid; Solubility; Structure; Synchrotrons; Tail; Techniques; Testing; Therapeutic; Transfection; Tumor Tissue; Untranslated RNA; Vesicle; Viral Vector; Virus; X ray diffraction analysis; base; biophysical analysis; biophysical properties; biophysical techniques; cancer cell; chemical property; cryogenics; cytotoxic; cytotoxicity; design; disorder control; efficacy testing; ethylene glycol; gene therapy; human model; immunogenic; immunogenicity; in vivo; liposome vector; malignant stomach neoplasm; mouse model; nanoparticle; nanoscale; novel; nucleic acid delivery; nucleic acid-based therapeutics; physical property; tool; tumor; uptake; vector

Project Summary/Abstract The current level of research activity involving gene therapy with either synthetic vectors (carriers) or engineered viruses is unprecedented. Liposomes are the most widely studied nonviral carriers worldwide for nucleic acid (NA) and drug delivery applications. Cationic liposomes (CLs) are relatively safe nonviral vectors used in ongoing clinical trials. CLs may either be complexed via electrostatic interactions with therapeutic NAs (anionic DNA or short interfering RNA) for gene delivery and silencing, or used as vectors of potent cytotoxic hydrophobic drugs, encapsulated within their lipid bilayer, in cancer therapeutics. Among the biggest advantages of nonviral vectors (over viral vectors which are currently more efficient in in vivo settings) are their safety, their low immunogenicity and their ability to transfer entire genes (containing coding and noncoding sequences) and regulatory sequences into cells (currently not feasible with engineered viruses because of capsid size limitations). The development of nonviral lipid-based vectors with efficacy competitive with viral vectors in vivo will require a mechanistic understanding of how synthetic vectors may be functionalized to overcome the major intracellular hurdle of endosomal escape. Successful endosomal escape is required for release of therapeutic nucleic acid within the cell cytosol and therefore maximum efficacy. The first aim of this research application is to employ modern biophysical and synthetic approaches to the rational design of functionalized CL–NA nanoparticles (NPs) with synergistic, complementary dual-function PEG-lipid and fusogenic components for optimized endosomal escape. Modern methods of organic and solid phase chemistry will be employed to synthesize dual-function PEG-lipids with cell targeting and endosome escaping properties. The second aim of this research application is to optimize efficacy of a new class of CL-based carriers of the hydrophobic drug paclitaxel (PTXL) for cancer therapeutics. This will be achieved by developing a mechanistic understanding of the relation between physical and chemical properties of the carrier (i.e. size of the functionalized CL carrier, membrane spontaneous curvature, and lipid tail structure) and functional efficacy (i.e. PTXL membrane solubility, cell uptake of vector and PTXL delivery leading to cytotoxicity against human cancer cells). The structures of CL-based vectors of NAs and hydrophobic drugs will be characterized using cryogenic electron microscopy and synchrotron x-ray diffraction techniques. The interactions between CL vectors and cell organelles will be directly visualized with spinning disk confocal fluorescence microscopy. Their structures will be correlated to their biological activity in human cancer cells. The broad, long-term objective of our research is to develop a fundamental science base through mechanistic studies that will lead to the design and synthesis of nonviral vectors of nucleic acids and hydrophobic drugs for gene and cancer therapeutics.

项目概要/摘要 目前涉及使用合成载体（载体）或工程病毒进行基因治疗的研究活动水平 是前所未有的。脂质体是全球研究最广泛的核酸（NA）和药物非病毒载体 交付应用程序。阳离子脂质体（CL）是正在进行的临床试验中使用的相对安全的非病毒载体。化学发光 可以通过静电相互作用与治疗性 NA（阴离子 DNA 或短干扰 RNA）复合， 基因传递和沉默，或用作强效细胞毒性疏水性药物的载体，封装在其脂质内 双层，用于癌症治疗。非病毒载体的最大优点之一（相对于目前流行的病毒载体） 在体内环境中更有效）是它们的安全性、低免疫原性和转移整个基因的能力 （包含编码和非编码序列）和调节序列进入细胞（目前对于工程改造不可行） 病毒，因为衣壳大小限制）。具有功效竞争性的非病毒脂质载体的开发 在体内使用病毒载体需要对合成载体如何功能化有机械性的了解 克服内体逃逸的主要细胞内障碍。释放需要成功的内体逃逸 治疗性核酸位于细胞胞浆内，因此具有最大功效。本研究应用的第一个目标 是采用现代生物物理和合成方法来合理设计功能化的CL-NA纳米粒子 (NP) 具有协同、互补的双功能 PEG-脂质和融合成分，可优化内体 逃脱。现代有机和固相化学方法将用于合成双功能PEG-脂质 具有细胞靶向和内体逃逸特性。该研究应用的第二个目标是优化功效 用于癌症治疗的疏水性药物紫杉醇 (PTXL) 的一类新型基于 CL 的载体的研究。这将是 通过发展对物理和化学性质之间关系的机械理解来实现 载体（即功能化 CL 载体的大小、膜自发曲率和脂质尾结构）和功能 功效（即 PTXL 膜溶解度、载体的细胞摄取和 PTXL 递送导致对人类的细胞毒性） 癌细胞）。基于 CL 的 NA 和疏水性药物载体的结构将使用低温进行表征 电子显微镜和同步加速器X射线衍射技术。 CL载体与细胞之间的相互作用 细胞器将通过转盘共聚焦荧光显微镜直接可视化。他们的结构将是 与其在人类癌细胞中的生物活性相关。我们研究的广泛、长期目标是开发一种 通过机制研究建立基础科学基础，这将导致非病毒载体的设计和合成 用于基因和癌症治疗的核酸和疏水性药物。