Abstract
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.
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Acknowledgments
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).
Author Contributions
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.
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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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Associate Editor Tejal Desai oversaw the review of this article.
Steven M. Jay
is an Assistant Professor in the Fischell Department of Bioengineering at the University of Maryland. Through generous support from the University and the National Institutes of Health (NIH), he has established a laboratory focused on biotherapeutic development and delivery, with specific interests in cancer therapy and vascular regeneration. Originally from the great state of Kentucky, Dr. Jay received a B.S.B.E. (Biological Engineering) from the University of Georgia before completing his Ph.D. in Biomedical Engineering under Dr. Mark Saltzman at Yale University. He then received postdoctoral training jointly from Dr. Richard T. Lee at the Brigham and Women’s Hospital and Dr. Linda Griffith at the Massachusetts Institute of Technology, and has also trained with Drs. Brian Rymond and Russ Mumper (University of Kentucky), Drs. Karen Burg and Thomas Jenkins (Clemson University), Dr. William Kisaalita (University of Georgia) and Drs. Themis Kyriakides and Jordan Pober (Yale University) among others. In addition to the NIH and the University of Maryland, Dr. Jay’s work has been supported by the National Science Foundation and the Oak Ridge Associated Universities. Dr. Jay’s research is enabled by the fortitude, diligence and intelligence of his trainees and by inspiration from his colleagues, friends and family.
This article is part of the 2016 Young Innovators Issue.
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Lamichhane, T.N., Jeyaram, A., Patel, D.B. et al. Oncogene Knockdown via Active Loading of Small RNAs into Extracellular Vesicles by Sonication. Cel. Mol. Bioeng. 9, 315–324 (2016). https://doi.org/10.1007/s12195-016-0457-4
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DOI: https://doi.org/10.1007/s12195-016-0457-4