Viral genome imaging of hepatitis C virus to probe heterogeneous viral infection and responses to antiviral therapies
Hepatitis C virus (HCV) is a positive single-stranded RNA virus of enormous global health importance, with direct-acting antiviral therapies replacing an immunostimulatory interferon-based regimen. The dynamics of HCV positive and negative-strand viral RNAs (vRNAs) under antiviral perturbations have...
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| Format: | Article |
| Language: | en_US |
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Elsevier
2017
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| Online Access: | http://hdl.handle.net/1721.1/110773 https://orcid.org/0000-0001-6214-4788 https://orcid.org/0000-0002-1293-2097 |
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| author | Luna, Joseph M. Hoffmann, Hans.-Heinrich Espiritu, Christine Sheahan, Timothy P. Rice, Charles M. Ramanan, Vyas Trehan, Kartik Ong, Mei Lyn Chandrasekar, Hamsika Christine, Kathleen van Oudenaarden, Alexander Bhatia, Sangeeta N Schwartz, Robert E. |
| author2 | Massachusetts Institute of Technology. Institute for Medical Engineering & Science |
| author_facet | Massachusetts Institute of Technology. Institute for Medical Engineering & Science Luna, Joseph M. Hoffmann, Hans.-Heinrich Espiritu, Christine Sheahan, Timothy P. Rice, Charles M. Ramanan, Vyas Trehan, Kartik Ong, Mei Lyn Chandrasekar, Hamsika Christine, Kathleen van Oudenaarden, Alexander Bhatia, Sangeeta N Schwartz, Robert E. |
| author_sort | Luna, Joseph M. |
| collection | MIT |
| description | Hepatitis C virus (HCV) is a positive single-stranded RNA virus of enormous global health importance, with direct-acting antiviral therapies replacing an immunostimulatory interferon-based regimen. The dynamics of HCV positive and negative-strand viral RNAs (vRNAs) under antiviral perturbations have not been studied at the single-cell level, leaving a gap in our understanding of antiviral kinetics and host–virus interactions. Here, we demonstrate quantitative imaging of HCV genomes in multiple infection models, and multiplexing of positive and negative strand vRNAs and host antiviral RNAs. We capture the varying kinetics with which antiviral drugs with different mechanisms of action clear HCV infection, finding the NS5A inhibitor daclatasvir to induce a rapid decline in negative-strand viral RNAs. We also find that the induction of host antiviral genes upon interferon treatment is positively correlated with viral load in single cells. This study adds smFISH to the toolbox available for analyzing the treatment of RNA virus infections. |
| first_indexed | 2024-09-23T15:11:54Z |
| format | Article |
| id | mit-1721.1/110773 |
| institution | Massachusetts Institute of Technology |
| language | en_US |
| last_indexed | 2024-09-23T15:11:54Z |
| publishDate | 2017 |
| publisher | Elsevier |
| record_format | dspace |
| spelling | mit-1721.1/1107732022-09-29T13:19:40Z Viral genome imaging of hepatitis C virus to probe heterogeneous viral infection and responses to antiviral therapies Luna, Joseph M. Hoffmann, Hans.-Heinrich Espiritu, Christine Sheahan, Timothy P. Rice, Charles M. Ramanan, Vyas Trehan, Kartik Ong, Mei Lyn Chandrasekar, Hamsika Christine, Kathleen van Oudenaarden, Alexander Bhatia, Sangeeta N Schwartz, Robert E. Massachusetts Institute of Technology. Institute for Medical Engineering & Science Harvard-MIT Program in Health Sciences and Technology Massachusetts Institute of Technology. Computational and Systems Biology Program Massachusetts Institute of Technology. Department of Biology Massachusetts Institute of Technology. Department of Electrical Engineering and Computer Science Massachusetts Institute of Technology. Department of Physics Koch Institute for Integrative Cancer Research at MIT Ramanan, Vyas Trehan, Kartik Ong, Mei Lyn Chandrasekar, Hamsika Schwartz, Robert E Christine, Kathleen van Oudenaarden, Alexander Bhatia, Sangeeta N Hepatitis C virus (HCV) is a positive single-stranded RNA virus of enormous global health importance, with direct-acting antiviral therapies replacing an immunostimulatory interferon-based regimen. The dynamics of HCV positive and negative-strand viral RNAs (vRNAs) under antiviral perturbations have not been studied at the single-cell level, leaving a gap in our understanding of antiviral kinetics and host–virus interactions. Here, we demonstrate quantitative imaging of HCV genomes in multiple infection models, and multiplexing of positive and negative strand vRNAs and host antiviral RNAs. We capture the varying kinetics with which antiviral drugs with different mechanisms of action clear HCV infection, finding the NS5A inhibitor daclatasvir to induce a rapid decline in negative-strand viral RNAs. We also find that the induction of host antiviral genes upon interferon treatment is positively correlated with viral load in single cells. This study adds smFISH to the toolbox available for analyzing the treatment of RNA virus infections. National Cancer Institute (U.S.) (Grant 1 R01 CA057973) National Institute of Allergy and Infectious Diseases (U.S.) (Grant 1 R01 AI099284) National Institute of Diabetes and Digestive and Kidney Diseases (U.S.) (NIH Roadmap for Medical Research Grant 1 R01 DK085713) National Cancer Institute (U.S.). Physical Sciences-Oncology Center (Grant U54 CA143874) National Institutes of Health (U.S.) (Pioneer Award 1DP1 OD003936) Starr Foundation Hertz Foundation National Science Foundation (U.S.). Graduate Research Fellowship Program 2017-07-19T14:03:18Z 2017-07-19T14:03:18Z 2016-04 2016-04 Article http://purl.org/eprint/type/JournalArticle 0042-6822 1096-0341 http://hdl.handle.net/1721.1/110773 Ramanan, Vyas et al. “Viral Genome Imaging of Hepatitis C Virus to Probe Heterogeneous Viral Infection and Responses to Antiviral Therapies.” Virology 494 (2016): 236–247. https://orcid.org/0000-0001-6214-4788 https://orcid.org/0000-0002-1293-2097 en_US http://dx.doi.org/10.1016/j.virol.2016.04.020 Virology Creative Commons Attribution-NonCommercial-NoDerivs License http://creativecommons.org/licenses/by-nc-nd/4.0/ application/pdf Elsevier PMC |
| spellingShingle | Luna, Joseph M. Hoffmann, Hans.-Heinrich Espiritu, Christine Sheahan, Timothy P. Rice, Charles M. Ramanan, Vyas Trehan, Kartik Ong, Mei Lyn Chandrasekar, Hamsika Christine, Kathleen van Oudenaarden, Alexander Bhatia, Sangeeta N Schwartz, Robert E. Viral genome imaging of hepatitis C virus to probe heterogeneous viral infection and responses to antiviral therapies |
| title | Viral genome imaging of hepatitis C virus to probe heterogeneous viral infection and responses to antiviral therapies |
| title_full | Viral genome imaging of hepatitis C virus to probe heterogeneous viral infection and responses to antiviral therapies |
| title_fullStr | Viral genome imaging of hepatitis C virus to probe heterogeneous viral infection and responses to antiviral therapies |
| title_full_unstemmed | Viral genome imaging of hepatitis C virus to probe heterogeneous viral infection and responses to antiviral therapies |
| title_short | Viral genome imaging of hepatitis C virus to probe heterogeneous viral infection and responses to antiviral therapies |
| title_sort | viral genome imaging of hepatitis c virus to probe heterogeneous viral infection and responses to antiviral therapies |
| url | http://hdl.handle.net/1721.1/110773 https://orcid.org/0000-0001-6214-4788 https://orcid.org/0000-0002-1293-2097 |
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