Reduction of the therapeutic dose of silencing RNA by packaging it in extracellular vesicles via a pre-microRNA backbone

Ryan Reshke; James A Taylor; Alexandre Savard; Huishan Guo; Luke H Rhym; Piotr S Kowalski; My Tran Trung; Charles Campbell; Wheaton Little; Daniel G Anderson; Derrick Gibbings

doi:10.1038/s41551-019-0502-4

Reduction of the therapeutic dose of silencing RNA by packaging it in extracellular vesicles via a pre-microRNA backbone

Nat Biomed Eng. 2020 Jan;4(1):52-68. doi: 10.1038/s41551-019-0502-4. Epub 2020 Jan 14.

Authors

Ryan Reshke^#^{1

2

3}, James A Taylor^#^{1

2}, Alexandre Savard^#^{1

2}, Huishan Guo^{1

2}, Luke H Rhym⁴, Piotr S Kowalski⁴, My Tran Trung^{1

2}, Charles Campbell^{1

2}, Wheaton Little⁵, Daniel G Anderson⁴, Derrick Gibbings^{6

7

8

9}

Affiliations

¹ Department of Cellular and Molecular Medicine, University of Ottawa, Ottawa, Canada.
² Ottawa Institute of Systems Biology, University of Ottawa, Ottawa, Canada.
³ Éric Poulin Center for Neuromuscular Disease, University of Ottawa, Ottawa, Canada.
⁴ The Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge, MA, USA.
⁵ Central Nervous System Disorders Drug Discovery Unit, Takeda Pharmaceuticals, Cambridge, MA, USA.
⁶ Department of Cellular and Molecular Medicine, University of Ottawa, Ottawa, Canada. gibbings@uottawa.ca.
⁷ Ottawa Institute of Systems Biology, University of Ottawa, Ottawa, Canada. gibbings@uottawa.ca.
⁸ Éric Poulin Center for Neuromuscular Disease, University of Ottawa, Ottawa, Canada. gibbings@uottawa.ca.
⁹ Brain and Mind Research Institute, Ottawa, Canada. gibbings@uottawa.ca.

^# Contributed equally.

PMID: 31937944
DOI: 10.1038/s41551-019-0502-4

Abstract

A small percentage of the short interfering RNA (siRNA) delivered via passive lipid nanoparticles and other delivery vehicles reaches the cytoplasm of cells. The high doses of siRNA and delivery vehicle that are thus required to achieve therapeutic outcomes can lead to toxicity. Here, we show that the integration of siRNA sequences into a Dicer-independent RNA stem-loop based on pre-miR-451 microRNA-which is highly enriched in small extracellular vesicles secreted by many cell types-reduces the expression of the genes targeted by the siRNA in the liver, intestine and kidney glomeruli of mice at siRNA doses that are at least tenfold lower than the siRNA doses typically delivered via lipid nanoparticles. Small extracellular vesicles that efficiently package siRNA can significantly reduce its therapeutic dose.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Cell Line, Tumor
Drug Delivery Systems / methods
Extracellular Vesicles / metabolism*
Gene Expression / drug effects
Humans
Mice
MicroRNAs / chemistry
MicroRNAs / metabolism*
Motor Neurons / drug effects
Nanoparticles / administration & dosage
RNA Interference
RNA, Small Interfering / administration & dosage*
RNA, Small Interfering / chemistry
RNA, Small Interfering / metabolism*

Substances

MIRN451 microRNA, human
MicroRNAs
RNA, Small Interfering