CAREER: Nanoparticle mRNA and DNA Immunoengineering of Macrophages for Solid Tumor Targeting

信使RNA (mRNA) COVID-19脂质纳米颗粒（LNP）疫苗的成功为利用mRNA LNP技术设计免疫系统开辟了新的机会。一个令人兴奋的目标是设计免疫系统来治疗难以用细胞免疫疗法治疗的实体癌肿瘤。这个CAREER项目的目标是开发一种LNP技术来设计能够进入并杀死实体肿瘤的免疫细胞。该研究目标与教育目标相结合，以吸引女性，代表性不足的第一代STEM学生参与免疫工程。该项目的教育目标是为美国国家科学基金会资助的希腊国际暑期学校和宾夕法尼亚大学生物工程课程课程创建免疫工程模块，与费城富兰克林研究所科学博物馆一起开发K-12免疫工程演示，并通过研究实习吸引高中和本科STEM学生。该项目的具体目标是利用纳米技术平台，利用mRNA和质粒DNA （pDNA）来设计巨噬细胞，作为产生针对实体肿瘤的免疫反应的新手段。细胞免疫疗法，如嵌合抗原受体（CAR） T细胞，已经成功地治疗了血癌，但在浸润性实体瘤方面的成功有限。巨噬细胞被积极招募并容易浸润实体肿瘤。因此，工程CAR巨噬细胞刺激免疫系统治疗实体肿瘤具有巨大的潜力。将采用一种新的LNP平台，安全有效地将mRNA和pDNA传递到巨噬细胞中，作为设计CAR巨噬细胞并表征其与实体肿瘤细胞相互作用的新手段。本项目旨在确定LNP的物理化学性质如何影响mRNA和pDNA向巨噬细胞的传递，利用LNP设计和表征CAR巨噬细胞，评估其靶向和杀死实体肿瘤细胞的能力，并量化mRNA和pDNA CAR巨噬细胞浸润实体肿瘤并引发小鼠抗肿瘤反应的能力。拟议的研究将对治疗实体肿瘤的免疫疗法的发展产生变革性影响，所开发的纳米技术将使巨噬细胞免疫调节应用于一系列应用，包括传染病、植入物、自身免疫性疾病和mRNA COVID-19疫苗。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。

The success of messenger RNA (mRNA) COVID-19 lipid nanoparticle (LNP) vaccines has opened new opportunities to engineer the immune system using mRNA LNP technology. An exciting target is engineering the immune system to treat solid cancer tumors which are difficult to treat with cellular immunotherapies. The goal of this CAREER project is to develop an LNP technology to engineer immune cells that can enter and kill solid tumors. This research goal is integrated with educational objectives to engage women, underrepresented, and first-generation STEM students in immunoengineering. The educational objectives of the project are to create immunoengineering modules for the NSF-funded International Summer School in Greece and the University of Pennsylvania Bioengineering course curriculum, develop K-12 immunoengineering demos with the Franklin Institute Science Museum of Philadelphia, and engage high school and undergraduate STEM students through research internships.The specific goal of this project is to exploit a nanotechnology platform to engineer macrophages using mRNA and plasmid DNA (pDNA) as a new means to generate an immune response against solid tumors. Cellular immunotherapies, such as chimeric antigen receptor (CAR) T cells, have been successful at treating blood cancers, but have had limited success in infiltrating solid tumors. Macrophages are actively recruited and readily infiltrate solid tumors. Therefore, there is immense potential for engineering CAR macrophages to stimulate the immune system to treat solid tumors. A new LNP platform will be employed that safely and efficiently delivers mRNA and pDNA into macrophages, as a new means to engineer CAR macrophages and characterize their interactions with solid tumor cells. This project aims to determine how LNP physicochemical properties influence mRNA and pDNA delivery to macrophages, to engineer and characterize CAR macrophages using LNPs, to assess their ability to target and kill solid tumor cells, and to quantify the ability of mRNA and pDNA CAR macrophages to infiltrate solid tumors and elicit an antitumor response in mice. The proposed studies will have a transformative impact on the development of immunotherapies to treat solid tumors, and the nanotechnology developed will enable macrophage immunomodulation for a range of applications, including infectious diseases, implants, autoimmune disorders, and mRNA COVID-19 vaccines.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.