High-throughput and high-content bioassay enables tuning of polyester nanoparticles for cellular uptake, endosomal escape, and systemic in vivo delivery of mRNA

被引：114

作者：

Rui, Yuan ^{[1
,2
]}

Wilson, David R. ^{[1
,2
]}

Tzeng, Stephany Y. ^{[1
,2
]}

Yamagata, Hannah M. ^{[1
,2
]}

Sudhakar, Deepti ^{[1
,2
]}

Conge, Marranne ^{[1
,2
,3
]}

Berlinicke, Cynthia A. ^{[4
]}

Zack, Donald J. ^{[4
,5
,6
]}

Tuesca, Anthony ^{[7
]}

Green, Jordan J. ^{[1
,2
,4
,8
,9
,10
,11
]}

机构：

[1] Johns Hopkins Univ, Sch Med, Dept Biomed Engn, Inst NanoBioTechnol, Baltimore, MD 21218 USA

[2] Johns Hopkins Univ, Sch Med, Translat Tissue Engn Ctr, Baltimore, MD 21218 USA

[3] Berea Coll, Dept Biol, Berea, KY USA

[4] Johns Hopkins Univ, Sch Med, Dept Ophthalmol, Baltimore, MD 21205 USA

[5] Johns Hopkins Univ, Sch Med, Dept Neurosci Mol Biol & Genet, Baltimore, MD USA

[6] Johns Hopkins Univ, Sch Med, Dept Genet Med, Baltimore, MD USA

[7] AstraZeneca, BioPharmaceut Dev, Dosage Form & Design Dev, BioPharmaceut R&D, Gaithersburg, MD USA

[8] Johns Hopkins Univ, Dept Neurosurg & Oncol, Sch Med, Baltimore, MD 21218 USA

[9] Johns Hopkins Univ, Dept Mat Sci & Engn, Baltimore, MD 21218 USA

[10] Johns Hopkins Univ, Dept Chem & Biomol Engn, Baltimore, MD 21218 USA

[11] Johns Hopkins Univ, Bloomberg Kimmel Inst Canc Immunotherapy, Sch Med, Baltimore, MD 21218 USA

来源：

SCIENCE ADVANCES | 2022年 / 8卷 / 01期

关键词：

SIRNA DELIVERY; MECHANISM; CELLS; TRANSFECTION;

D O I：

10.1126/sciadv.abk2855

中图分类号：

O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];

学科分类号：

07 ; 0710 ; 09 ;

摘要：

Nanoparticle-based mRNA therapeutics hold great promise, but cellular internalization and endosomal escape remain key barriers for cytosolic delivery. We developed a dual nanoparticle uptake and endosomal disruption assay using high-throughput and high-content image-based screening. Using a genetically encoded Galectin 8 fluorescent fusion protein sensor, endosomal disruption could be detected via sensor clustering on damaged endosomal membranes. Simultaneously, nucleic acid endocytosis was quantified using fluorescently tagged mRNA. We used an array of biodegradable poly(beta-amino ester)s as well as Lipofectamine and PEI to demonstrate that this assay has higher predictive capacity for mRNA delivery compared to conventional polymer and nanoparticle physiochemical characteristics. Top nanoparticle formulations enabled safe and efficacious mRNA expression in multiple tissues following intravenous injection, demonstrating that the in vitro screening method is also predictive of in vivo performance. Efficacious nonviral systemic delivery of mRNA with biodegradable particles opens up new avenues for genetic medicine and human health.

引用

页数：14

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