Direct imaging of RNA loaded lipid-nanoparticles’ self assembly and cellular expression for cancer immunotherapy and RNA-based vaccines
Direct imaging of RNA loaded lipid-nanoparticles’ self assembly and cellular expression for cancer immunotherapy and RNA-based vaccines
复制标题
用于癌症免疫治疗和基于 RNA 的疫苗的 RNA 负载脂质纳米颗粒的自组装和细胞表达的直接成像
DOI：
10.1016/j.bpj.2021.11.2866
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发表时间：
2022-02-11
影响因子：
3.4
通讯作者：
Guan J
中科院分区：
文献类型：
--
作者：
Ashbrook J;Chung MC;Mendez-Gomez H;Sayour E;Guan J
免疫学领域在新的癌症免疫疗法和基于RNA的疫苗方面迅速扩张。基于RNA的疫苗最近在应对COVID-19等公共卫生危机方面表现出巨大的前景。此外，RNA纳米颗粒（RNA-NP）技术可用于创建可行的癌症治疗选择。RNA-NP是通过将RNA添加到脂质体中制成的，脂质体作为递送载体将RNA安全地运输到身体的免疫细胞中，试图引发免疫应答。然而，对RNA-NP组装过程以及它们如何与下游免疫原性相关的理解仍然有限，阻碍了提高癌症免疫治疗疗效的进展。需要具有高时空分辨率的先进成像来了解RNA-NPs如何组装，细胞如何与RNA-NPs相互作用，以及哪些生物物理特性影响转染效率。在这里，我们操纵各种RNA-NP制备参数，如温度和浓度，同时使用自制的落射荧光显微镜观察对纳米颗粒聚集的影响。进行活细胞实验以通过RNA-NP摄取以及转染效率确定这些操作的下游效应。我们的研究结果表明，RNA-NPs孵育的温度极大地影响聚集体面积。他们还表明，较小的纳米颗粒聚集体在DC 2的转染中更有效。4（通过实验使用的小鼠树突细胞系）。我们的工作提供了对RNA-NP组装过程的深入了解，并为微调RNA-NP的生物物理特性以改善癌症免疫治疗开辟了新的机会。
The field of immunology has seen a rapid expansion in new cancer immunotherapy and RNA-based vaccines. RNA-based vaccines have recently demonstrated great promise in addressing public health crises such as COVID-19. Additionally, RNA nanoparticle (RNA-NP) technology may be used to create a viable cancer treatment option. RNA-NPs were made by adding RNAs to liposomes which act as a delivery vehicle to safely transport RNA to the body’s immune cells in an attempt to elicit an immune response. However, the understanding of RNA-NP assembly processes and how they relate to downstream immunogenicity is still limited, hindering progress on improving the efficacy of cancer immunotherapy. Advanced imaging with high spatiotemporal resolution is needed to understand how RNA-NPs assemble, how cells interact with RNA-NPs, and which biophysical properties affect transfection efficiency. Here, we manipulate various RNA-NP preparation parameters, such as temperature and concentration, while observing the effect on nanoparticle aggregation using a home-built epifluorescent microscope. Live cell experiments were conducted to determine the downstream effects of these manipulations through RNA-NP uptake as well as transfection efficiency. Our results show that the temperature at which RNA-NPs are incubated greatly affects aggregate area. They also suggest that smaller nanoparticle aggregates are more effective in transfection of DC2. 4 (the mouse dendritic cell line used through the experiment). Our work offers insight into the RNA-NP assembly process and opens up new opportunities to fine-tune the biophysical properties of RNA-NPs to improve cancer immunotherapy.