Single-molecule RNA-FISH analysis reveals stochasticity in reactivation of latent HIV-1 regulated by Nuclear Orphan Receptors NR4A and cMYC

Res Sq [Preprint]. 2024 Apr 19:rs.3.rs-4166090. doi: 10.21203/rs.3.rs-4166090/v1.ABSTRACTHIV-1 eradication strategies require complete reactivation of HIV-1 latent cells by Latency Reversing Agents (LRA). Current methods lack effectiveness due to incomplete proviral reactivation. We employed a single-molecule RNA-FISH (smRNA-FISH) and FISH-Quant analysis and found that proviral reactivation is highly variable from cell-to-cell, stochastic, and occurs in bursts and waves, with different kinetics in response to diverse LRAs. Approximately 1-5% of latent cells exhibited stochastic reactivation without LRAs. Through single-cell RNA-seq analysis, we identified NR4A3 and cMYC as extrinsic factors associated with stochastic HIV-1 reactivation. Concomitant with HIV-1 reactivation cMYC was downregulated and NR4A3 was upregulated in both latent cell lines and primary CD4 + T-cells from aviremic patients. By inhibiting cMYC using SN-38, an active metabolite of irinotecan, we induced NR4A3 and HIV-1 expression. Our results suggest that inherent stochasticity in proviral reactivation contributes to cell-to-cell variability, which could potentially be modulated by drugs targeting cMYC and NR4A3.PMID:38699331 | PMC:PMC11065080 | DOI:10.21203/rs.3.rs-4166090/v1

Source: Cell Research - May 3, 2024 Category: Cytology Authors: Annalena LaPorte Rajiv Pathak Carolina Eliscovich Laura Martins Rachel Nell Adam Spivak Masako Suzuki Vicente Planelles Robert Singer Ganjam Kalpana Source Type: research