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

基本信息

批准号：
10242074
负责人：
CYRUS R SAFINYA
金额：
$ 29.53万
依托单位：
UNIVERSITY OF CALIFORNIA SANTA BARBARA
依托单位国家：
美国
项目类别：
财政年份：
2018
资助国家：
美国
起止时间：
2018-09-20 至 2024-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10242074
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

项目摘要

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.

项目摘要/摘要目前涉及合成载体（载体）或工程病毒的基因治疗的研究水平是前所未有的。脂质体是全球研究最广泛的核酸（NA）和药物的非病毒载体交付申请。阳离子脂质体（CLS）是正在进行的临床试验中使用的相对安全的非病毒载体。 CLS 可以通过静电相互作用与治疗性NAS（阴离子DNA或短干扰RNA）进行复合基因输送和沉默，或用作封装在其脂质中的有效细胞毒性疏水药物的载体双层，癌症治疗。非病毒载体的最大优势之一（与目前的病毒媒介相比在体内设置中更有效）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）和调节序列到细胞中（目前不可行由于衣壳尺寸的限制）。具有有效竞争力的非病毒基基载体的开发使用病毒载体在体内将需要机械理解合成矢量如何功能化到克服内体逃生的主要细胞内障碍。成功释放需要成功的内体逃生细胞细胞质内的治疗核酸，因此最大效率。该研究申请的第一个目的是利用现代的生物物理和合成方法来官能化CL – NA纳米颗粒的理性设计（NPS）具有协同，完整的双功能PEG脂质和融合成分，以优化内体逃脱。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脱特性。该研究应用的第二个目的是优化效率用于疏水药物紫杉醇（PTXL）的新型基于CL的基于CL的载体用于癌症治疗。这将是通过对物理和化学特性之间的关系的机械理解来实现载体（即功能化CL载体，膜赞助商和脂质尾巴结构的大小）和功能性功效（即PTXL膜溶解度，载体的细胞摄取和PTXL递送，导致对人的细胞毒性癌细胞）。 NAS和疏水性药物基于Cl的载体的结构将使用低温来表征电子显微镜和同步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旋转盘共聚焦荧光显微镜直接可视化。他们的结构将是与它们在人类癌细胞中的生物学活性相关。我们研究的广泛，长期目标是开发基本科学基础通过机械研究，将导致设计和合成非病毒载体用于基因和癌症治疗的核酸和疏水药物。