In vitro evolution of enhanced RNA replicons for immunotherapy
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 identi...
| Main Authors: | , , , , , , , |
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| Format: | Article |
| Language: | English |
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Springer Science and Business Media LLC
2020
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| Online Access: | https://hdl.handle.net/1721.1/124309 |
| _version_ | 1810999165878009856 |
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| author | Li, Yingzhong Teague, Brian Paul Su, Zhijun Porter, Ely Dobosh, Brian S Wagner, Tyler E Irvine, Darrell J Weiss, Ron |
| author2 | Massachusetts Institute of Technology. Department of Biological Engineering |
| author_facet | Massachusetts Institute of Technology. Department of Biological Engineering Li, Yingzhong Teague, Brian Paul Su, Zhijun Porter, Ely Dobosh, Brian S Wagner, Tyler E Irvine, Darrell J Weiss, Ron |
| author_sort | Li, Yingzhong |
| collection | MIT |
| description | 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. |
| first_indexed | 2024-09-23T14:56:31Z |
| format | Article |
| id | mit-1721.1/124309 |
| institution | Massachusetts Institute of Technology |
| language | English |
| last_indexed | 2024-09-23T14:56:31Z |
| publishDate | 2020 |
| publisher | Springer Science and Business Media LLC |
| record_format | dspace |
| spelling | mit-1721.1/1243092022-09-29T11:35:21Z In vitro evolution of enhanced RNA replicons for immunotherapy Li, Yingzhong Teague, Brian Paul Su, Zhijun Porter, Ely Dobosh, Brian S Wagner, Tyler E Irvine, Darrell J Weiss, Ron 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 Multidisciplinary 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. National Institutes of Health (U.S.) (grant CA20618) National Cancer Institute (U.S.) (Grant P30-CA14051) 2020-03-25T12:15:04Z 2020-03-25T12:15:04Z 2019-05-06 2020-02-20T16:30:26Z Article http://purl.org/eprint/type/JournalArticle 2045-2322 https://hdl.handle.net/1721.1/124309 Li, Yingzhong, et al. "In vitro evolution of enhanced RNA replicons for immunotherapy." Scientific reports 9 (2019): 6932 © 2019, The Author(s) en 10.1038/s41598-019-43422-0 Scientific reports Creative Commons Attribution 4.0 International license https://creativecommons.org/licenses/by/4.0/ application/pdf Springer Science and Business Media LLC Scientific Reports |
| spellingShingle | Multidisciplinary Li, Yingzhong Teague, Brian Paul Su, Zhijun Porter, Ely Dobosh, Brian S Wagner, Tyler E Irvine, Darrell J Weiss, Ron In vitro evolution of enhanced RNA replicons for immunotherapy |
| title | In vitro evolution of enhanced RNA replicons for immunotherapy |
| title_full | In vitro evolution of enhanced RNA replicons for immunotherapy |
| title_fullStr | In vitro evolution of enhanced RNA replicons for immunotherapy |
| title_full_unstemmed | In vitro evolution of enhanced RNA replicons for immunotherapy |
| title_short | In vitro evolution of enhanced RNA replicons for immunotherapy |
| title_sort | in vitro evolution of enhanced rna replicons for immunotherapy |
| topic | Multidisciplinary |
| url | https://hdl.handle.net/1721.1/124309 |
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