Dawei Liu , Xing Wang , Letao Xu , Zahraa Nima Saeed Al-Delfi , Zelalem Addis Mekonnen , Song Gao , Branka Grubor-Bauk , Chun-Xia Zhao
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引用次数: 0
Abstract
Quantifying the biodistribution of lipid nanoparticles (LNPs) is critical for optimizing mRNA delivery systems, yet current approaches have inherent limitations. This study introduces a cost-effective method utilizing double-stranded DNA (dsDNA) barcodes and quantitative polymerase chain reaction (qPCR) for rapid analysis of a small library of mRNA-LNPs biodistribution and functional delivery in vivo. Three unique 100-bp dsDNA barcodes were designed to represent for three FDA-approved LNP formulations. Concurrently, these three formulations carrying luciferase mRNA were mixed with DNA-barcoding LNPs as a pool. Following intravenous administration of the pooled LNPs in mice, qPCR analysis revealed the highest abundance of DNA barcodes and accumulation of luciferase mRNA in spleen, with positive correlation between barcodes presence and mRNA localization across organs, validating DNA barcodes as reliable indicators of mRNA-LNPs biodistribution in vivo. Bioluminescence imaging further confirmed successful delivery and protein translation of luciferase mRNA facilitated by the LNPs in vivo. Integrating DNA barcodes for biodistribution analysis and luciferase mRNA for assessing functional delivery enabled comprehensive evaluation of LNP performance. This robust methodology provides valuable insights into the localization patterns and mRNA delivery capabilities of different LNP formulations, paving the way for the development of more effective and targeted mRNA-based therapeutics.
定量脂质纳米颗粒（LNPs）的生物分布对于优化mRNA传递系统至关重要，但目前的方法存在固有的局限性。本研究介绍了一种利用双链DNA （dsDNA）条形码和定量聚合酶链反应（qPCR）快速分析mRNA-LNPs生物分布和体内功能传递的低成本方法。三个独特的100 bp dsDNA条形码被设计来代表三种fda批准的LNP配方。同时，将这三种携带荧光素酶mRNA的制剂与dna条形码LNPs混合作为一个池。经小鼠静脉给药后，qPCR分析显示，小鼠脾脏中DNA条形码的丰度和荧光素酶mRNA的积累量最高，条形码的存在与mRNA在各器官中的定位呈正相关，验证了DNA条形码是mRNA-LNPs在体内生物分布的可靠指标。生物发光成像进一步证实LNPs在体内促进了荧光素酶mRNA的成功传递和蛋白质翻译。整合DNA条形码用于生物分布分析和荧光素酶mRNA用于评估功能递送，可以全面评估LNP的性能。这种强大的方法为不同LNP配方的定位模式和mRNA传递能力提供了有价值的见解，为开发更有效和更有针对性的mRNA治疗方法铺平了道路。
期刊介绍：
Colloids and Surfaces B: Biointerfaces is an international journal devoted to fundamental and applied research on colloid and interfacial phenomena in relation to systems of biological origin, having particular relevance to the medical, pharmaceutical, biotechnological, food and cosmetic fields.
Submissions that: (1) deal solely with biological phenomena and do not describe the physico-chemical or colloid-chemical background and/or mechanism of the phenomena, and (2) deal solely with colloid/interfacial phenomena and do not have appropriate biological content or relevance, are outside the scope of the journal and will not be considered for publication.
The journal publishes regular research papers, reviews, short communications and invited perspective articles, called BioInterface Perspectives. The BioInterface Perspective provide researchers the opportunity to review their own work, as well as provide insight into the work of others that inspired and influenced the author. Regular articles should have a maximum total length of 6,000 words. In addition, a (combined) maximum of 8 normal-sized figures and/or tables is allowed (so for instance 3 tables and 5 figures). For multiple-panel figures each set of two panels equates to one figure. Short communications should not exceed half of the above. It is required to give on the article cover page a short statistical summary of the article listing the total number of words and tables/figures.
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