title: Oncogene Knockdown via Active Loading of Small RNAs into Extracellular Vesicles by Sonication.
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.
authors: Fischell Department of Bioengineering, University of Maryland, College Park, MD 20742 USA.
authors: Tek N
authors: TN
authors: Lamichhane
authors: Fischell Department of Bioengineering, University of Maryland, College Park, MD 20742 USA.
authors: Anjana
authors: A
authors: Jeyaram
authors: Fischell Department of Bioengineering, University of Maryland, College Park, MD 20742 USA.
authors: Divya B
authors: DB
authors: Patel
authors: Fischell Department of Bioengineering, University of Maryland, College Park, MD 20742 USA.
authors: Babita
authors: B
authors: Parajuli
authors: Fischell Department of Bioengineering, University of Maryland, College Park, MD 20742 USA.
authors: Natalie K
authors: NK
authors: Livingston
authors: Fischell Department of Bioengineering, University of Maryland, College Park, MD 20742 USA.
authors: Navein
authors: N
authors: Arumugasaamy
authors: Fischell Department of Bioengineering, University of Maryland, College Park, MD 20742 USA.
authors: John S
authors: JS
authors: Schardt
authors: Fischell Department of Bioengineering, University of Maryland, College Park, MD 20742 USA; Program in Oncology, Marlene and Stewart Greenebaum Cancer Center, University of Maryland, College Park, MD 20742 USA; Program in Molecular and Cell Biology, University of Maryland, College Park, MD 20742 USA.
authors: Steven M
authors: SM
authors: Jay
keywords: Exosomes; HER2; biopharmaceutical; biotherapeutic; cancer nanoparticle; microRNA; nanobiotechnology; siRNA delivery