Engineering Effective Polymers to Enable Gene Based Therapies

工程有效的聚合物以实现基于基因的治疗

• 批准号: RGPIN-2019-04244
• 负责人: Uludag, Hasan
• 金额: $ 4.01万
• 依托单位: University of Alberta
• 依托单位国家: 加拿大
• 项目类别: Discovery Grants Program - Individual
• 财政年份: 2021
• 资助国家: 加拿大
• 起止时间: 2021-01-01 至 2022-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=737509
• 关键词: Engineering; Effective; Polymers; Enable; Gene

Molecular therapy utilizing nucleic acids offers an exciting approach over conventional drugs. Diseased organs can be better restored to normal state with the use of gene-based medicines derived from nucleic acids. The success, however, is absolutely dependent on the use of nucleic acid carriers, since nucleic acids alone cannot be internalized by cells due to their large size and anionic nature. Cationic polymers are safe carriers and they can readily neutralize nucleic acids and condense them into nanoparticles suitable for uptake. However, they suffer from low transfection efficiencies, short duration of activity and high toxicities. Overall goal of my research to create `engineered' polymers that can self-assemble with nucleic acids to create nanoparticles ideal for cellular delivery. We have been engineering a class of gene carriers from lipid-substituted cationic polymers. Such polymers were found to create stable, self-assembled nanoparticles, enhancing delivery of the cargo into cells by as much as 100-fold. We identified several promising lipid substitutions and charted their beneficial in the delivery of particular types of nucleic acids in particular types of cells. No universal carrier design emerged in our past work, but we identified effective carriers for different types of cells. Our overall hypothesis is: amphiphilic polymers can be engineered from cationic polymers and lipids for controlled delivery and release of nucleic acids for specific cells, ultimately leading to improved functional outcomes in a safe manner. Three aims will be pursued for next generation carriers: Aim-1. To create `tertiary' nanoparticles for superior delivery of nucleic acids. We recently discovered that adding a tertiary polyanion into nanoparticles improved the functional delivery. We will design superior tertiary nanoparticles by using in-house prepared polyanionic molecules with tailored features. Aim-2. To create multiple lipid-bearing polymers for delivery of nucleic acids to broader population of cells. We identified key lipid substituents effective for nucleic acid delivery in different patient cells. We propose to create `hybrid' lipopolymers, where different lipid substituents will be methodically varied on the polymer backbone, to create `universal' carriers capable of acting on wider source of cells. Aim-3. To determine functional performance of polymers for nucleic acid delivery. Using several in vitro models, we will investigate the functional performance of improved delivery systems from Aim-1 and -2. Systematic comparisons between current and new systems, and commercial reagents will be undertaken. My goal is to establish a foundation for polymer-guided delivery of genetic agents and develop functional materials for effective and safe delivery of nucleic acids. We envision the carriers to serve as R&D reagents in biomedical research enterprise as well as functional carriers to implement safe molecular therapies for human diseases.

利用核酸的分子疗法提供了一种优于传统药物的令人兴奋的方法。使用基于核酸的基因药物可以更好地恢复受损器官的正常状态。然而，成功绝对依赖于核酸载体的使用，因为核酸由于其大尺寸和阴离子性质而不能单独被细胞内化。阳离子聚合物是安全的载体，并且它们可以容易地中和核酸并将它们凝聚成适合于摄取的纳米颗粒。然而，它们的缺点是转染效率低、活性持续时间短和毒性高。我的研究的总体目标是创建“工程”聚合物，可以与核酸自组装，以创建理想的细胞输送纳米粒子。 我们已经从脂质取代的阳离子聚合物工程一类基因载体。发现这种聚合物可以产生稳定的自组装纳米颗粒，将货物输送到细胞中的能力提高了100倍。我们确定了几种有希望的脂质替代品，并绘制了它们在特定类型细胞中递送特定类型核酸的有益效果。在我们过去的工作中没有出现通用的载体设计，但我们确定了不同类型细胞的有效载体。我们的总体假设是：两亲性聚合物可以由阳离子聚合物和脂质工程化，用于特定细胞的核酸的受控递送和释放，最终以安全的方式导致改善的功能结果。下一代航母将追求三个目标：Aim-1。为了产生用于核酸的上级递送的“三级”纳米颗粒。我们最近发现，将叔聚阴离子添加到纳米颗粒中改善了功能性递送。我们将设计上级三级纳米粒子通过使用内部制备的聚阴离子分子与定制功能。 目标二本发明的目的是产生用于将核酸递送至更广泛的细胞群体的多个载脂聚合物。我们鉴定了在不同患者细胞中对核酸递送有效的关键脂质取代物。我们建议创建“混合”脂质聚合物，其中不同的脂质取代基将有条不紊地变化的聚合物骨架，以创建“通用”载体能够作用于更广泛的细胞来源。目标三确定用于核酸递送的聚合物的功能性能。使用几种体外模型，我们将研究来自Aim-1和Aim-2的改进的递送系统的功能性能。将对现有系统和新系统以及商业试剂进行系统比较。我的目标是建立一个基础，为聚合物引导的遗传物质的交付和开发功能材料的有效和安全的核酸交付。我们设想这种载体可以作为生物医学研究企业的研发试剂，也可以作为功能性载体，对人类疾病进行安全的分子治疗。