In vitro evolution of enhanced RNA replicons for immunotherapy
Name
s41598-019-43422-0(1).pdf
Description
Published version
Size
1.67 MB
Format
Adobe PDF
Checksum (MD5)
d2f91a80d9d48da59600847db0bb47a0
Author(s)
Li, Yingzhong
Teague, Brian Paul
Su, Zhijun
Porter, Ely
Dobosh, Brian S
Wagner, Tyler E
Irvine, Darrell J
Weiss, Ron
Date Issued
May 6, 2019
Journal
Scientific reports
Publisher
Springer Science and Business Media LLC
Citation
Li, Yingzhong, et al. "In vitro evolution of enhanced RNA replicons for immunotherapy." Scientific reports 9 (2019): 6932 © 2019, The Author(s)
Version
Final published version
Abstract
Self-replicating (replicon) RNA is a promising new platform for gene therapy, but applications are still limited by short persistence of expression in most cell types and low levels of transgene expression in vivo. To address these shortcomings, we developed an in vitro evolution strategy and identified six mutations in nonstructural proteins (nsPs) of Venezuelan equine encephalitis (VEE) replicon that promoted subgenome expression in cells. Two mutations in nsP2 and nsP3 enhanced transgene expression, while three mutations in nsP3 regulated this expression. Replicons containing the most effective mutation combinations showed enhanced duration and cargo gene expression in vivo. In comparison to wildtype replicon, mutants expressing IL-2 injected into murine B16F10 melanoma showed 5.5-fold increase in intratumoral IL-2 and 2.1-fold increase in infiltrating CD8 T cells, resulting in significantly slowed tumor growth. Thus, these mutant replicons may be useful for improving RNA therapeutics for vaccination, cancer immunotherapy, and gene therapy.
Subjects
Multidisciplinary
MIT Department
Massachusetts Institute of Technology. Department of Biological Engineering
Massachusetts Institute of Technology. Department of Materials Science and Engineering
Koch Institute for Integrative Cancer Research at MIT
Terms of Use
Creative Commons Attribution 4.0 International license
Persistent DSpace Link
DOI of Published Version
10.1038/s41598-019-43422-0
