PP 1.32 – 00163 cGAS/STING Signaling Drives HIV-1 Replication in Acutely Infected Macrophages

Background: Macrophages play an important role in HIV-1 pathogenesis in vivo, including serving as a major viral reservoir that contributes to viral persistence during antiretroviral therapy.The determinants that govern HIV-1 replication in macrophages are not well understood.We hypothesize that recognition of double stranded DNA by cGAS/STING drives NF-kB-dependent HIV-1 replication at early time points after viral infection of macrophages.Methods: Monocyte-derived macrophages (MDMs) infected with HIV-1 were used to evaluate the effects of NF-kB and cGAS/ STING inhibition on viral replication and transcription.Western blot and ELISA analyses were used to evaluate NF-kB translocation to the nucleus in response to viral infection.Chromatin immunoprecipitation (ChIP) analysis was used to evaluate transcription factor recruitment to the viral promoter.Single cell RNA sequencing (scRNA-Seq) was used to evaluate transcriptomic changes in macrophages in response to HIV-1 infection.Results: We demonstrated that shortly following infection with HIV-1, the p65 subunit of NF-kB translocates to the nucleus and associates with the HIV-1 promoter.The inhibition of NF-kB signaling, but not NFAT or AP-1 signaling, was sufficient to prevent HIV-1 replication in MDMs.The vital role of NF-kB signaling in HIV-1 replication in MDMs was confirmed by scRNA-Seq that demonstrated a significant increase in NF-kB activity in infected cells.Inhibition of cGAS/STING signaling led to a significant decrease in HIV-1 replication in MDMs, whereas inhibition of other innate immune receptors did not.Conversely, activation of cGAS/STING signaling in infected MDMs led to an increase in viral replication in an interferon and NF-kBdependent manner.Finally, knockdown of STING using shRNA led to a significant decrease in HIV-1 replication in acutely infected MDMs.Together, our data suggest that cGAS/STINGmediated activation of NF-kB is necessary for viral replication in acutely infected macrophages.Conclusions: Our studies implicate the cGAS/STING/NF-kB signaling axis as a driver of HIV-1 replication in acutely infected macrophages and suggest that this pathway might serve as a target to prevent the establishment of the macrophage reservoir.

Abstract licence: CC BY-NC

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