Collaborative Research: Development of an exosome based lipoproteoplex (E-LPP) for siRNA delivery

合作研究：开发用于 siRNA 递送的基于外泌体的脂蛋白复合物 (E-LPP)

• 批准号: 2203680
• 负责人: Jin Montclare
• 金额: $ 25万
• 依托单位: New York University
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-06-01 至 2026-05-31
• 项目状态: 未结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=2203680&HistoricalAwards=false
• 关键词: Collaborative; Research; Development; exosome; based

NON-TECHNICAL SUMMARY The development of safe and effective means to deliver nucleic acids has become increasingly important, especially in light of the COVID pandemic. mRNA vaccines that can provide our immune system the tools to fight COVID-19 are crucial in helping us fight this devastative virus. This project supports fundamental research focused on creating better delivery agents that can better protect and carry the important nucleic acids like mRNA so it can do its job. We explore the use of exosomes as potential delivery vehicles. Exosomes are essentially little compartments that are derived from our own cells. Because of this, they have key components that allow them to enter other cells and deliver cargo. While exosomes have this unique advantage, loading them with the target therapeutic is challenging. To address this, the research team will develop small, charged proteins that can gently hold onto the negatively charged nucleic acids, which then helps improve the loading of exosomes. This project will not only train the next generation graduate student and researcher in these new approaches, but also help inspire high school students as they will be able to carry out meaningful research experience both virtually and in person. Both the PI and co-PI have a strong record of training women and underrepresented minorities and plan to continue to recruit and train them as part of this work with the goal of developing and diversifying the future scientific workforce.TECHNICAL SUMMARY Nucleic acids possess critical instructions for controlling biological processes with tangible implications in fundamental cell and organismal studies, biotechnology and medicine especially in light of delivery of vaccines for Covid-19. There is a need to develop a modular platform for the widespread delivery of nucleic acids, and exosomes have recently been identified as promising delivery agents. The mechanisms of assembly between membrane-derived lipids and recombinant proteins is poorly understood but provides an opportunity to optimize and explore the fundamental material properties of exosome-based delivery vehicles. The research team proposes to develop an engineered protein-exosome system or lipoproteoplex capable of: 1) encapsulating siRNA at a high efficiency into the exosome; 2) increasing siRNA release once inside the cell through increased endosomal escape; 3) performing efficient siRNA delivery with a favorable safety profile. For education and outreach, the team aims to focus on verified virtual platforms to provide students safe, meaningful research experiences. Courses will focus on hybrid in-person and virtual lessons with an emphasis on group-based projects to facilitate active learning. For outreach, Dr. Montclare will participate in the Applied Research Innovations in Science and Engineering (ARISE) program and engage local NYC 10th and 11th grade students in computational design of supercharged proteins. Dr. Rabbani will implement as part of the Wyss Sumer Research Fellowship, a biochemistry training that involves exosome preparation, characterization and use for transfection as well as recording protocol video and journal club discussion online. This will democratize research experience for students during the pandemic and beyond.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

非技术总结，递送核酸的安全有效手段的发展变得越来越重要，尤其是鉴于相关大流行。 可以为我们的免疫系统提供对抗COVID-19的工具的mRNA疫苗对于帮助我们抗击这种破坏性病毒至关重要。该项目支持基础研究，重点是创建更好的递送剂，可以更好地保护和携带重要的核酸，例如mRNA，以便它可以完成工作。我们探索使用外泌体作为潜在送货车的使用。 外泌体本质上是从我们自己的细胞中得出的几乎没有隔室。因此，它们具有关键组件，使它们可以进入其他单元格并提供货物。 尽管外泌体具有这种独特的优势，但将它们加载到目标治疗方面是具有挑战性的。为了解决这个问题，研究团队将开发出小的，带电的蛋白质，这些蛋白质可以轻轻地固定在带负电荷的核酸上，从而有助于改善外泌体的负载。 该项目不仅将在这些新方法中培训下一代研究生和研究人员，而且还可以帮助启发高中生，因为他们能够实际和亲自进行有意义的研究经验。 PI和Co-Pi都有良好的培训妇女和人为少数群体不足的记录，并计划继续招募和培训她们作为这项工作的一部分，其目的是发展和多样化未来的科学劳动力。技术摘要核酸具有控制生物学过程的关键指导，在生物学细胞和生物学研究中，对生物学细胞和生物学研究进行了有效的影响，并尤其是在生物学研究中，尤其是在生物学上进行生物学研究。需要开发一个模块化平台，以广泛递送核酸，而外泌体最近被确定为有前途的递送剂。据了解，膜衍生的脂质与重组蛋白之间的组装机制知之甚少，但提供了一个机会，可以优化和探索基于外泌体的递送工具的基本材料特性。研究小组建议开发一种能够：1）以高效率封装siR​​NA的工程蛋白质 - 异位系统； 2）通过增加的内体逃生，增加siRNA释放一次； 3）以有利的安全性进行有效的siRNA传递。对于教育和宣传，该团队旨在专注于经过验证的虚拟平台，以为学生提供安全，有意义的研究经验。 课程将重点放在混合机构和虚拟课程上，重点是基于小组的项目，以促进积极学习。对于外展活动，蒙特卡莱博士将参加科学与工程（ARISE）计划的应用研究创新，并与纽约市当地的10年级和11年级学生一起参与增压蛋白的计算设计。 Rabbani博士将作为Wyss Sumer Research奖学金的一部分实施，这是一种生物化学培训，涉及外部准备，表征和转染以及在线录制协议视频和期刊俱乐部讨论。这将使学生在大流行期间的研究经验民主化。该奖项反映了NSF的法定使命，并被认为是值得通过基金会的知识分子优点和更广泛影响的评论标准来评估值得支持的。