Oncogene Knockdown via Active Loading of Small RNAs into Extracellular Vesicles by Sonication
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Extracellular vesicles (EVs), including exosomes and microvesicles, have emerged as promising drug delivery vehicles for small RNAs (siRNA and miRNA) due to their natural role in intercellular RNA transport. However, the application of EVs for therapeutic RNA delivery may be limited by loading approaches that can induce cargo aggregation or degradation. Here, we report the use of sonication as a means to actively load functional small RNAs into EVs. Conditions under which EVs could be loaded with small RNAs with minimal detectable aggregation were identified, and EVs loaded with therapeutic siRNA via sonication were observed to be taken up by recipient cells and capable of target mRNA knockdown leading to reduced protein expression. This system was ultimately applied to reduce expression of HER2, an oncogenic receptor tyrosine kinase that critically mediates breast cancer development and progression, and could be extended to other therapeutic targets. These results define important parameters informing the application of sonication as a small RNA loading method for EVs and demonstrate the potential utility of this approach for versatile cancer therapy.
KeywordsExosomes siRNA delivery microRNA HER2 Cancer nanoparticle Nanobiotechnology Biotherapeutic Biopharmaceutical
This work was supported by NIH R00 Grant HL112905, by an ORAU Ralph E. Powe Junior Faculty Enhancement Award, and by two University of Maryland Tier 1 seed Grants (all to SMJ).
TNL, AJ, DBP, BP, NKL, NA and JSS performed the research and analyzed data. TNL, AJ, DBP and SMJ contributed to conception and design of experiments and wrote the manuscript. All authors reviewed, edited and approved of the final manuscript.
Conflict of Interest
Authors Tek N. Lamichhane, Anjana Jeyaram, Divya B. Patel, Babita Parajuli, Natalie K. Livingston, Navein Arumugasaamy, John S. Schardt and Steven M. Jay declare that they have no conflicts of interest.
Ethics and Informed Consent
No human studies were carried out by the authors for this article. No animal studies were carried out by the authors for this article.
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