Classical monocyte transcriptomes reveal significant anti-inflammatory statin effect in women with chronic HIV

Abstract

Aims During virally-suppressed chronic HIV infection, persistent inflammation contributes to the development of cardiovascular disease (CVD), a major comorbidity in people living with HIV (LWH). Classical blood monocytes (CMs) remain activated during antiretroviral therapy and are a major source of pro-inflammatory and pro-thrombotic factors that contribute to atherosclerotic plaque development and instability. Methods and Results Here we identify transcriptomic changes in circulating CMs in peripheral blood mononuclear cell samples from participants of the Women’s Interagency HIV Study, selected by HIV and subclinical CVD (sCVD) status. We flow-sorted CM from participants of the Women’s Interagency HIV Study and deep-sequenced their mRNA (n = 92). CMs of HIV+ participants showed elevated IL-6, IL-1β, and IL-12β, overlapping with many transcripts identified in sCVD+ participants. In sCVD+ participants LWH, those reporting statin use showed reduced pro-inflammatory gene expression to a level comparable with healthy (HIV-sCVD-) participants. Statin non-users maintained an elevated inflammatory profile and increased cytokine production. Conclusion Statin therapy has been associated with a lower risk of cardiac events, such as myocardial infarction in the general population, but not in those LWH. Our data suggest that women LWH may benefit from statin therapy even in the absence of overt CVD. Translational perspective Monocytes from women living with HIV express many more pro-inflammatory genes than uninfected controls. An overlapping list of genes is expressed in samples from women with ultrasound evidence of carotid plaque. The inflammatory burden is enhanced in women with both HIV and carotid plaque, and this is mitigated by statin treatment, almost to the level of healthy participants. Thus, the present monocyte transcriptome data from 92 women support the idea that participants with HIV may specifically benefit from statin treatment, perhaps more so than seronegative subjects.

Combined protein and transcript single cell RNA sequencing in human peripheral blood mononuclear cells

“…[34] As an example, we applied the classical monocyte transcriptomes (8 subsets) to bulk transcriptomes of sorted classical monocytes from 92 subjects. [7] We found 1 of the CM subsets in all subjects and others at varying proportions ( Figure 5 ).…”

“…Cell proportions of classical monocytes (CMs, CM1-8) from the present dataset in 92 samples of classical monocyte transcriptomes in women with HIV, CVD, both or neither. [7] CM, classical monocyte; CRT, cholesterol-reducing treatment.…”

“…The remaining 45,719 cells were analyzed using the Doublet Finder package on R (https://github.com/chris-mcginnis-ucsf/DoubletFinder) with the default doublet formation rate (7.5%). This removed another 3,322 doublets, leaving 42,397 Cells.Finally, we removed all cells that had less than 128 (27 ) antibody molecules sequenced. This removed 786 noisy cells, resulting in 41,611 cell transcriptomes.…”

Lack of Atorvastatin Effect on Monocyte Gene Expression and Inflammatory Markers in HIV-1- infected ART-suppressed Individuals at Risk of non-AIDS Comorbidities

“…Finally, when we focused specifically on gene functions and pathways that we previously found were dys-regulated in HIV/ART compared with control participants, there was no evidence that they were impacted as a group by atorvastatin. In contrast, a recent retrospective study of HIV+ ART-treated women with subclinical cardiovascular disease reported that treatment with statins for clinical indications was associated with downregulation of inflammatory genes in classical monocytes [ 25 ]. Thus, in HIV/ART participants without clinical indications for statin use, we find no evidence for monocyte gene expression effect of atorvastatin that exceeds random inter-participant and intra-participant variability, or that treatment shifts monocyte gene expression closer towards “normal” patterns.…”

“…Statins, widely used in people living with HIV for lipid disorders, have pleiotropic immunomodulatory effects beyond those related to cholesterol lowering, which are mediated by lipid modifications of small signaling molecules [ 20 ]. Statins have been the focus of considerable interest as adjunctive therapy in HIV/ART patients [ 21 – 25 ]. In vitro , we [ 24 ] and others [ 26 , 27 ] have shown that statins modulate monocyte activation by triggers that contribute to residual immune activation in HIV/ART patients.…”