EAGER-Harnessing natural killer cell-derived extracellular vesicles as anti-viral nanomaterials

摘要：自然杀伤细胞衍生的细胞外囊泡（NKE）是一种天然纳米颗粒，具有生物相容性、体内稳定性和抗病毒成分。因此，研究NKE的材料特性对于设计合成的仿生纳米颗粒至关重要，这些纳米颗粒可以扩大用于抗病毒应用，包括COVID-19。为了满足这些需求，生物材料计划的这个EAGER奖项的总体目标是研究纳米材料的特性、膜成分和NKE的载货量。利用这些新知识，PI和她的实验室将测试这些材料特性如何影响膜融合，以及细胞内化和抗SARS-CoV-2感染的抗病毒功能。由于NK细胞活性已被发现在不同的患者群体中存在差异，因此PI还将阐明细胞外囊泡的材料特性如何随生物因素而变化，并可用于获得对药物输送系统的相容性、稳定性和整体性能至关重要的新知识。这些研究工作将与K-12、本科生、研究生和公众的教育和推广活动相结合。大学和研究生阶段的活动主要是通过深入的研究经验来招募、指导和培训女性、URM和LGBTQ学生，而K-12阶段的活动则是基于课堂讲座和实践演示。此外，PI将通过与加州科学中心的既定合作以及K-12学生的社交媒体平台与公众互动。其目标是激励、吸引和培养来自不同背景的下一代科学家和工程师，以开发有利于全球公共卫生的创新解决方案。技术摘要：自然杀伤细胞（NK）是抵御病毒感染的第一道防线，有助于清除不健康的宿主细胞。尽管NK细胞已被提出作为一种基于细胞的治疗方法来治疗由病毒感染（如COVID-19）引起的疾病，但细胞治疗存在细胞在体内存活有限、保质期短和成本高等挑战。相反，赋予NK细胞自然杀伤特性的细胞毒性细胞外囊泡（EVs）有可能被扩大为长期的、现成的药物递送载体，并可用于设计稳定的、用于抗病毒应用的仿生药物递送载体。为此，PI和她的实验室将首先1)表征材料特性，以及来自几种人类来源的NK ev （NKE）的脂质、蛋白质、RNA膜和货物组成。然后，PI及其实验室将2)评估感染SARS-CoV-2的细胞中NKE细胞的内化和抗病毒活性，并评估NKE成分和材料特性的差异如何影响其功能。研究计划的成功有望提高NKE的基础知识，并阐明其与抗病毒功能相关的材料特性。因此，在当前的大流行（例如COVID-19）期间，研究计划的结果可能对评估NKE作为抗病毒治疗的新来源具有重要意义，但对其他病毒感染的免疫治疗也有潜在的益处。研究计划与教育和推广计划相结合，其中包括：1)通过深入的研究经验，招聘、培训和指导来自不同背景的本科生和研究生，包括女性、URM和LGBTQ群体；2)关于生物纳米材料的课堂讲座和实践活动，包括最近COVID-19疫苗中的纳米颗粒载体（洛杉矶联合学区32街学校）；3)纳米日，一个年度项目，将包括在加州科学中心和纳米周进行生物纳米技术演示。我们展示生物纳米技术内容的社交媒体平台。这些举措旨在激发、吸引和传播有关医学生物纳米材料及其解决流行病等全球公共卫生问题的潜力的知识给从K-12到公众的不同受众。将对每项活动的方案有效性进行评估。该奖项反映了美国国家科学基金会的法定使命，并通过使用基金会的知识价值和更广泛的影响审查标准进行评估，被认为值得支持。

NON-TECHNICAL ABSTRACT:Natural killer cell-derived extracellular vesicles (NKE) are natural nanoparticles that are inherently biocompatible, stable in vivo, and contain anti-viral components. As such, investigating the materials properties of NKE will be critical to informing the design of synthetic, biomimetic nanoparticles that can be scaled-up for anti-viral application including COVID-19. To meet these needs, the overall goal of this EAGER award by the Biomaterials Program is to study the nanomaterial properties, membrane composition, and cargo of NKE. Using this new knowledge, the PI and her lab will test how these material properties affect membrane fusion, as well as cell internalization and anti-viral function against SARS-CoV-2 infection. Since NK cell activity has been found to differ among various patient populations, the PI will also elucidate how the materials properties of extracellular vesicles vary with biological factors, and can be used to obtain new knowledge that is critical for compatibility, stability, and overall performance of drug delivery systems. These research efforts will be integrated with education and outreach initiatives for K-12, undergraduate, and graduate students and for the public. The activities at the college and graduate levels are centered on recruitment, mentorship, and training women, URM, and LGBTQ students through in-depth research experiences, while K-12 activities are based on classroom lectures and hands-on demos. In addition, the PI will interact with the public through an established collaboration with the California Science Center and a social media platform for K-12 students. The goal is to inspire, engage, and prepare the next generation of scientists and engineers from diverse backgrounds to develop innovative solutions that can benefit global public health.TECHNICAL ABSTRACT:Natural killer (NK) cells are the first line of defense against viral infections and helps eliminate unhealthy host cells. Although NK cells have been proposed as a cell-based therapy against diseases caused by viral infections such as COVID-19, cell therapy has challenges including limited cell survival in vivo, short shelf-life, and high cost. Instead, the cytotoxic, extracellular vesicles (EVs) that endow natural killing properties to NK cells have the potential to be scaled-up as long-term, off-the-shelf drug delivery carriers and can be used to inform the design of stable, biomimetic drug delivery carriers for anti-viral application. To that end, the PI and her lab will first 1) characterize the material properties, as well as lipid, protein, and RNA membrane and cargo composition of NK EVs (NKE) originating from several human sources. Then, the PI and her lab will 2) evaluate NKE cell internalization and anti-viral activity in cells infected with SARS-CoV-2 and assess how differences in NKE composition and materials properties influences their function. Success of the research plan is anticipated to advance fundamental knowledge of NKE, and elucidate their material properties as it relates to anti-viral function. Thus, the results from the research proposal may be significant during the current pandemic (e.g. COVID-19) in evaluating NKE as a novel source of anti-viral therapy, but will have potential benefits for immunotherapy in other viral infections. The research plan is integrated with the education and outreach plan which includes: 1) recruitment, training, and mentoring of undergraduate and graduate students from diverse backgrounds including women, URM, and LGBTQ groups through in-depth research experiences, 2) classroom lecture and hands-on activities regarding bio-nanomaterials including nanoparticle carriers in recent COVID-19 vaccines (32nd Street School, Los Angeles Unified School District), and 3) NanoDays, an annual program that will include bio-nanotechnology demos at the California Science Center, and NanoPeek, our social media platform that showcases bio-nanotechnology content. Together, these initiatives aim to excite, engage, and disseminate knowledge to a diverse audience ranging from K-12 to the public regarding bionanomaterials in medicine and its potential to solve global public health issues such as pandemics. The effectiveness of the programs will be evaluated for each activity.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.