Multiply spliced HIV RNA is a predictive measure of virus production ex vivo and in vivo following reversal of HIV latency.
Anti-Retroviral Agents
/ pharmacology
CD4-Positive T-Lymphocytes
/ cytology
Cell Proliferation
/ drug effects
HIV Infections
/ drug therapy
HIV-1
/ genetics
Histone Deacetylase Inhibitors
/ pharmacology
Humans
Polyhydroxyalkanoates
/ pharmacology
RNA Splicing
RNA, Viral
/ blood
Tetradecanoylphorbol Acetate
/ pharmacology
Virus Latency
/ physiology
Vorinostat
/ pharmacology
Biomarker
HIV
Latency reversal
Multiply-spliced HIV RNA
Reservoir
Shock and kill
Journal
EBioMedicine
ISSN: 2352-3964
Titre abrégé: EBioMedicine
Pays: Netherlands
ID NLM: 101647039
Informations de publication
Date de publication:
Mar 2021
Mar 2021
Historique:
received:
23
10
2020
revised:
11
01
2021
accepted:
27
01
2021
pubmed:
2
3
2021
medline:
3
11
2021
entrez:
1
3
2021
Statut:
ppublish
Résumé
One strategy being pursued to clear latently infected cells that persist in people living with HIV (PLWH) on antiretroviral therapy (ART) is to activate latent HIV infection with a latency reversing agent (LRA). Surrogate markers that accurately measure virus production following an LRA are needed. We quantified cell-associated unspliced (US), multiply spliced (MS) and supernatant (SN) HIV RNA by qPCR from total and resting CD4+ T cells isolated from seven PLWH on ART before and after treatment ex vivo with different LRAs, including histone deacetylase inhibitors (HDACi). MS and plasma HIV RNA were also quantified from PLWH on ART (n-11) who received the HDACi panobinostat. In total and resting CD4+ T cells from PLWH on ART, detection of US RNA was common while detection of MS RNA was infrequent. Primers used to detect MS RNA, in contrast to US RNA, bound sites of the viral genome that are commonly mutated or deleted in PLWH on ART. Following ex vivo stimulation with LRAs, we identified a strong correlation between the fold change increase in SN and MS RNA, but not the fold change increase in SN and US RNA. In PLWH on ART who received panobinostat, MS RNA was significantly higher in samples with detectable compared to non0detectable plasma HIV RNA. Following administration of an LRA, quantification of MS RNA is more likely to reflect an increase in virion production and is therefore a better indicator of meaningful latency reversal. NHMRC, NIH DARE collaboratory.
Sections du résumé
BACKGROUND
BACKGROUND
One strategy being pursued to clear latently infected cells that persist in people living with HIV (PLWH) on antiretroviral therapy (ART) is to activate latent HIV infection with a latency reversing agent (LRA). Surrogate markers that accurately measure virus production following an LRA are needed.
METHODS
METHODS
We quantified cell-associated unspliced (US), multiply spliced (MS) and supernatant (SN) HIV RNA by qPCR from total and resting CD4+ T cells isolated from seven PLWH on ART before and after treatment ex vivo with different LRAs, including histone deacetylase inhibitors (HDACi). MS and plasma HIV RNA were also quantified from PLWH on ART (n-11) who received the HDACi panobinostat.
FINDINGS
RESULTS
In total and resting CD4+ T cells from PLWH on ART, detection of US RNA was common while detection of MS RNA was infrequent. Primers used to detect MS RNA, in contrast to US RNA, bound sites of the viral genome that are commonly mutated or deleted in PLWH on ART. Following ex vivo stimulation with LRAs, we identified a strong correlation between the fold change increase in SN and MS RNA, but not the fold change increase in SN and US RNA. In PLWH on ART who received panobinostat, MS RNA was significantly higher in samples with detectable compared to non0detectable plasma HIV RNA.
INTERPRETATION
CONCLUSIONS
Following administration of an LRA, quantification of MS RNA is more likely to reflect an increase in virion production and is therefore a better indicator of meaningful latency reversal.
FUNDING
BACKGROUND
NHMRC, NIH DARE collaboratory.
Identifiants
pubmed: 33647768
pii: S2352-3964(21)00034-7
doi: 10.1016/j.ebiom.2021.103241
pmc: PMC7920823
pii:
doi:
Substances chimiques
Anti-Retroviral Agents
0
Histone Deacetylase Inhibitors
0
Polyhydroxyalkanoates
0
RNA, Viral
0
Vorinostat
58IFB293JI
Tetradecanoylphorbol Acetate
NI40JAQ945
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
103241Subventions
Organisme : NIAID NIH HHS
ID : P30 AI027763
Pays : United States
Commentaires et corrections
Type : CommentIn
Informations de copyright
Copyright © 2021 The Authors. Published by Elsevier B.V. All rights reserved.
Déclaration de conflit d'intérêts
Declaration of Interests SRL's institution has received funding from the National Health and Medical Research Council (NHMRC) of Australia, National Institutes for Health, American Foundation for AIDS Research; Merck, ViiV and Gilead for investigator-initiated research; Merck, ViiV and Gilead for educational activities. She is on the advisory board of Abivax and Innivirax.
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