EAPSI: Development of Amphiphilic, pH Responsive Cationic Copolypeptoids for Nonviral Cell Transfection in Serum

EAPSI：开发用于血清中非病毒细胞转染的两亲性、pH 响应型阳离子共聚肽

• 批准号: 1712805
• 负责人: Jessica Simpson
• 金额: $ 0.54万
• 依托单位: Simpson Jessica
• 依托单位国家: 美国
• 项目类别: Fellowship Award
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-06-01 至 2018-05-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1712805&HistoricalAwards=false
• 关键词: EAPSI; Development; Amphiphilic; pH; Responsive

Carried out by the process of cell transfection (CT), the practice of gene therapy is a promising approach in addressing rare diseases and genetic disorders (e.g. hemophilia, SCID, and Kippel Trenauny). Among non-viral gene carriers used for CT polymers have received significant attention primarily because their chemical versatility, large-scale production, and cytocompatibility make them potential candidates for biomedical applications. However, drawbacks such as low transfection efficiency and size related cytotoxicity limiting full usage of these systems. In this research, responsive, cationic polypeptoid copolymers will be developed. Under the guidance of Professor Lichen Yin, an expert in cell transfection at Soochow University, synthesized copolymers will be used to investigate the particle stability and behavior of nucleic acid condensed polyplexes during in vitro cell transfection. This research will provide a strategy to stabilize genetic material during cell transfection. This approach will also permit the effective release genetic material intracellularly via endosomal escape.Poly(N-substituted glycines) are a special class of peptidomimetic polymers bearing substituents on the nitrogen present in the backbone. Commonly referred to as polypeptoids, this group of polymers are known for their minimal cytotoxicity, enhanced proteolytic stability and structural tunability, making them potential candidates for a variety of biomedical and biotechnological applications including smart-coatings, antimicrobial agents, and drug delivery. Currently, the cell transfection of polypeptoid/DNA complexes in serum-containing media is problematic due to the instability of the complexed particles in serum. The manifestation of charge shielding, in which interaction with anionic serum components (lipids, albumin, salts, etc.) reduces polyplex surface charge, is mainly responsible for this outcome. To address this challenge, block polypeptoid copolymers containing hydrophilic and hydrophobic segments will be developed in this project. In this research synthesized copolymers will be tested for their ability to effectively protect and transport therapeutic gene p53 during in vitro cell transfection in nuclease. Under extracellular conditions where charge shielding is permissible (pH= 5.0-5.5), it is anticipated that copolymers will respond to this biological stimuli by adopting a micellar structure with a charged, hydrophobic inner core and charge neutral outer shell. Dr. Yin's group proficiency in the field of cell transfection will contribute significantly towards efforts to understand and elucidate the behavior of these polymeric systems in biological environments. This award, under the East Asia and Pacific Summer Institutes program, supports summer research by a U.S. graduate student and is jointly funded by NSF and the Chinese Ministry of Science and Technology.

基因治疗通过细胞转染（CT）的过程进行，是治疗罕见疾病和遗传疾病（如血友病、SCID和Kippel Trenauny）的一种很有前途的方法。在用于CT的非病毒基因载体中，聚合物受到了极大的关注，主要是因为它们的化学通用性、大规模生产和细胞相容性使它们成为生物医学应用的潜在候选者。然而，低转染效率和大小相关的细胞毒性等缺点限制了这些系统的充分利用。在这项研究中，反应性阳离子多肽共聚物将被开发。在苏州大学细胞转染专家尹立辰教授的指导下，合成的共聚物将用于研究核酸凝聚多聚体在体外细胞转染过程中的颗粒稳定性和行为。本研究将提供一种在细胞转染过程中稳定遗传物质的策略。这种方法也允许遗传物质通过胞内体逃逸有效释放。聚（n -取代甘氨酸）是一类特殊的类肽聚合物，其主链上的氮具有取代基。通常被称为多肽，这组聚合物以其最小的细胞毒性，增强的蛋白质水解稳定性和结构可调性而闻名，使其成为各种生物医学和生物技术应用的潜在候选者，包括智能涂层，抗菌剂和药物输送。目前，多肽/DNA复合物在含血清培养基中的细胞转染存在问题，因为复合物颗粒在血清中的不稳定性。电荷屏蔽的表现，即与阴离子血清成分（脂质、白蛋白、盐等）的相互作用减少了复合表面电荷，是导致这一结果的主要原因。为了应对这一挑战，本项目将开发含有亲疏水段的嵌段多肽共聚物。在本研究中，将测试合成的共聚物在核酸酶体外细胞转染过程中有效保护和转运治疗基因p53的能力。在允许电荷屏蔽的胞外条件下（pH= 5.0-5.5），预计共聚物将通过采用带电荷的疏水内核和带电荷的中性外壳的胶束结构来响应这种生物刺激。尹博士在细胞转染领域的团队精通将对理解和阐明这些聚合物系统在生物环境中的行为做出重大贡献。该奖项由美国国家科学基金会和中国科技部共同资助，隶属于东亚和太平洋暑期研究所项目，支持美国研究生的暑期研究。