RNA-induced epigenetic silencing inhibits HIV-1 reactivation from latency
Abstract Background Current antiretroviral therapy is effective in controlling HIV-1 infection. However, cessation of therapy is associated with rapid return of viremia from the viral reservoir. Eradicating the HIV-1 reservoir has proven difficult with the limited success of latency reactivation str...
| Main Authors: | , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
BMC
2018-10-01
|
| Series: | Retrovirology |
| Subjects: | |
| Online Access: | http://link.springer.com/article/10.1186/s12977-018-0451-0 |
| _version_ | 1818923483709571072 |
|---|---|
| author | Catalina Méndez Scott Ledger Kathy Petoumenos Chantelle Ahlenstiel Anthony D. Kelleher |
| author_facet | Catalina Méndez Scott Ledger Kathy Petoumenos Chantelle Ahlenstiel Anthony D. Kelleher |
| author_sort | Catalina Méndez |
| collection | DOAJ |
| description | Abstract Background Current antiretroviral therapy is effective in controlling HIV-1 infection. However, cessation of therapy is associated with rapid return of viremia from the viral reservoir. Eradicating the HIV-1 reservoir has proven difficult with the limited success of latency reactivation strategies and reflects the complexity of HIV-1 latency. Consequently, there is a growing need for alternate strategies. Here we explore a “block and lock” approach for enforcing latency to render the provirus unable to restart transcription despite exposure to reactivation stimuli. Reactivation of transcription from latent HIV-1 proviruses can be epigenetically blocked using promoter-targeted shRNAs to prevent productive infection. We aimed to determine if independent and combined expression of shRNAs, PromA and 143, induce a repressive epigenetic profile that is sufficiently stable to protect latently infected cells from HIV-1 reactivation when treated with a range of latency reversing agents (LRAs). Results J-Lat 9.2 cells, a model of HIV-1 latency, expressing shRNAs PromA, 143, PromA/143 or controls were treated with LRAs to evaluate protection from HIV-1 reactivation as determined by levels of GFP expression. Cells expressing shRNA PromA, 143, or both, showed robust resistance to viral reactivation by: TNF, SAHA, SAHA/TNF, Bryostatin/TNF, DZNep, and Chaetocin. Given the physiological importance of TNF, HIV-1 reactivation was induced by TNF (5 ng/mL) and ChIP assays were performed to detect changes in expression of epigenetic markers within chromatin in both sorted GFP− and GFP+ cell populations, harboring latent or reactivated proviruses, respectively. Ordinary two-way ANOVA analysis used to identify interactions between shRNAs and chromatin marks associated with repressive or active chromatin in the integrated provirus revealed significant changes in the levels of H3K27me3, AGO1 and HDAC1 in the LTR, which correlated with the extent of reduced proviral reactivation. The cell line co-expressing shPromA and sh143 consistently showed the least reactivation and greatest enrichment of chromatin compaction indicators. Conclusion The active maintenance of epigenetic silencing by shRNAs acting on the HIV-1 LTR impedes HIV-1 reactivation from latency. Our “block and lock” approach constitutes a novel way of enforcing HIV-1 “super latency” through a closed chromatin architecture that renders the virus resistant to a range of latency reversing agents. |
| first_indexed | 2024-12-20T02:10:09Z |
| format | Article |
| id | doaj.art-9dca84304e8d494dbda576e744c43dfd |
| institution | Directory Open Access Journal |
| issn | 1742-4690 |
| language | English |
| last_indexed | 2024-12-20T02:10:09Z |
| publishDate | 2018-10-01 |
| publisher | BMC |
| record_format | Article |
| series | Retrovirology |
| spelling | doaj.art-9dca84304e8d494dbda576e744c43dfd2022-12-21T19:57:05ZengBMCRetrovirology1742-46902018-10-0115111810.1186/s12977-018-0451-0RNA-induced epigenetic silencing inhibits HIV-1 reactivation from latencyCatalina Méndez0Scott Ledger1Kathy Petoumenos2Chantelle Ahlenstiel3Anthony D. Kelleher4Department of Immunovirology and Pathogenesis, Level 5, Wallace Wurth Building, The Kirby Institute for Infection and Immunity, UNSW SydneyDepartment of Immunovirology and Pathogenesis, Level 5, Wallace Wurth Building, The Kirby Institute for Infection and Immunity, UNSW SydneyDepartment of Immunovirology and Pathogenesis, Level 5, Wallace Wurth Building, The Kirby Institute for Infection and Immunity, UNSW SydneyDepartment of Immunovirology and Pathogenesis, Level 5, Wallace Wurth Building, The Kirby Institute for Infection and Immunity, UNSW SydneyDepartment of Immunovirology and Pathogenesis, Level 5, Wallace Wurth Building, The Kirby Institute for Infection and Immunity, UNSW SydneyAbstract Background Current antiretroviral therapy is effective in controlling HIV-1 infection. However, cessation of therapy is associated with rapid return of viremia from the viral reservoir. Eradicating the HIV-1 reservoir has proven difficult with the limited success of latency reactivation strategies and reflects the complexity of HIV-1 latency. Consequently, there is a growing need for alternate strategies. Here we explore a “block and lock” approach for enforcing latency to render the provirus unable to restart transcription despite exposure to reactivation stimuli. Reactivation of transcription from latent HIV-1 proviruses can be epigenetically blocked using promoter-targeted shRNAs to prevent productive infection. We aimed to determine if independent and combined expression of shRNAs, PromA and 143, induce a repressive epigenetic profile that is sufficiently stable to protect latently infected cells from HIV-1 reactivation when treated with a range of latency reversing agents (LRAs). Results J-Lat 9.2 cells, a model of HIV-1 latency, expressing shRNAs PromA, 143, PromA/143 or controls were treated with LRAs to evaluate protection from HIV-1 reactivation as determined by levels of GFP expression. Cells expressing shRNA PromA, 143, or both, showed robust resistance to viral reactivation by: TNF, SAHA, SAHA/TNF, Bryostatin/TNF, DZNep, and Chaetocin. Given the physiological importance of TNF, HIV-1 reactivation was induced by TNF (5 ng/mL) and ChIP assays were performed to detect changes in expression of epigenetic markers within chromatin in both sorted GFP− and GFP+ cell populations, harboring latent or reactivated proviruses, respectively. Ordinary two-way ANOVA analysis used to identify interactions between shRNAs and chromatin marks associated with repressive or active chromatin in the integrated provirus revealed significant changes in the levels of H3K27me3, AGO1 and HDAC1 in the LTR, which correlated with the extent of reduced proviral reactivation. The cell line co-expressing shPromA and sh143 consistently showed the least reactivation and greatest enrichment of chromatin compaction indicators. Conclusion The active maintenance of epigenetic silencing by shRNAs acting on the HIV-1 LTR impedes HIV-1 reactivation from latency. Our “block and lock” approach constitutes a novel way of enforcing HIV-1 “super latency” through a closed chromatin architecture that renders the virus resistant to a range of latency reversing agents.http://link.springer.com/article/10.1186/s12977-018-0451-0HIV-1LatencyReactivationLatent reservoirEpigenetic silencingTranscriptional gene silencing |
| spellingShingle | Catalina Méndez Scott Ledger Kathy Petoumenos Chantelle Ahlenstiel Anthony D. Kelleher RNA-induced epigenetic silencing inhibits HIV-1 reactivation from latency Retrovirology HIV-1 Latency Reactivation Latent reservoir Epigenetic silencing Transcriptional gene silencing |
| title | RNA-induced epigenetic silencing inhibits HIV-1 reactivation from latency |
| title_full | RNA-induced epigenetic silencing inhibits HIV-1 reactivation from latency |
| title_fullStr | RNA-induced epigenetic silencing inhibits HIV-1 reactivation from latency |
| title_full_unstemmed | RNA-induced epigenetic silencing inhibits HIV-1 reactivation from latency |
| title_short | RNA-induced epigenetic silencing inhibits HIV-1 reactivation from latency |
| title_sort | rna induced epigenetic silencing inhibits hiv 1 reactivation from latency |
| topic | HIV-1 Latency Reactivation Latent reservoir Epigenetic silencing Transcriptional gene silencing |
| url | http://link.springer.com/article/10.1186/s12977-018-0451-0 |
| work_keys_str_mv | AT catalinamendez rnainducedepigeneticsilencinginhibitshiv1reactivationfromlatency AT scottledger rnainducedepigeneticsilencinginhibitshiv1reactivationfromlatency AT kathypetoumenos rnainducedepigeneticsilencinginhibitshiv1reactivationfromlatency AT chantelleahlenstiel rnainducedepigeneticsilencinginhibitshiv1reactivationfromlatency AT anthonydkelleher rnainducedepigeneticsilencinginhibitshiv1reactivationfromlatency |