Sex differences in beneficial and pathogenic bacteria in People With HIV (PWH) with a history of heavy alcohol drinking

Aakarsha V Rao¹Smita S Ghare^1,2Vasuk Gautam²Kristi L Hoffman³Joseph Petrosino³Kaku So-Armah⁴Jeffrey H Samet⁴Gregory J Patts⁵Debbie M Cheng⁶Elena Blokhina^7,8Evgeny Krupitsky^7,9Dmitry Lioznov⁷Edwin Zvartau⁷Craig James McClain¹Hillary Tindle¹⁰Matthew S Freiberg^10,11Shirish Barve^1,2*

• ¹University of Louisville, Louisville, United States
• ²Norton Research Institute, Louisville, United States
• ³Baylor College of Medicine, Houston, Texas, United States
• ⁴Chobanian & Avedisian School of Medicine, Boston University, Boston, Massachusetts, United States
• ⁵School of Public Health, Boston University, Boston, Massachusetts, United States
• ⁶Department of Biostatistics, School of Public Health, Boston University, Boston, Massachusetts, United States
• ⁷Pavlov First Saint Petersburg State Medical University, Saint Petersburg, Saint Petersburg, Russia
• ⁸VM Bekhterev National Medical Research Center for Psychiatry and Neurology, St. Petersburg, Russia
• ⁹VM Bekhterev National Medical Research Center of Psychiatry and Neurology, St. Petersburg, Russia
• ¹⁰Vanderbilt University School of Medicine, Nashville, United States
• ¹¹Geriatric Research Education and Clinical Center, Tennessee Valley Healthcare System, Veterans Health Administration, United States Department of Veterans Affairs, Nashville, Tennessee, United States

Background: HIV-1 infection and hazardous levels of alcohol consumption have been independently linked to gut dysbiosis affecting beneficial butyrate-producing bacteria. However, sex-based differences in the composition and function of gut microbiome of People With HIV (PWH) with a history of heavy alcohol drinking remain undetermined, which is the focus of this study. Methods: Cross-sectional study examining structural and functional features of the gut microbiome in PWH between men and women with a history of hazardous alcohol drinking recruited at St. Petersburg, Russia. 16S rDNA sequencing information was used for metataxonomic, Phylogenetic Investigation of Communities by Reconstruction of Unobserved States (PICRUSt2) and Linear Discriminant Analysis Effect Size (LEfSe) analyses. Group-wise comparisons were done using Mann-Whitney U-test. Further, linear and logistic regression models were used to evaluate the association between sex and measures of gut microbial dysbiosis and Firmicutes/Bacteroidota (F/B) ratio respectively. Data were adjusted for confounding covariates particularly, HIV-viral load, Anti-retroviral Therapy (ART) and alcohol usage. Results: Women consumed significantly less alcohol compared to men, although viral load was not different. Metataxonomic analysis demonstrated that women depicted significantly higher microbial diversity (Operational Taxonomic Units, OTUs and Shannon Index), higher percent relative abundance (%RA) of Firmicutes, lower %RA of Bacteroidota and higher F/B ratio. Importantly, logistic regression revealed that women had twice the odds of having F/B ratio > 1. Notably, women demonstrated significantly higher %RA of butyrate-producing bacterial families i.e. Lachnospiraceae, Oscillospiraceae, Rikenellaceae and Marinifilaceae and genera. Correspondingly, significantly greater expression of bacterial genes involved in butyrate synthesis in women was demonstrated by PICRUSt2 analysis. Additionally, women depicted lower %RA of pathobiont, Prevotellaceae particularly, Prevotella_9 genus. Conclusions: Overall, we observed significant sex-based differences in the relative abundances of beneficial bacterial communities such as butyrate producers and potential pathogenic Prevotella community in the gut microbiome of PWH with a history of heavy alcohol consumption. The observed sex-based differences are clinically relevant and could inform therapeutic strategies with evidence-based probiotics. In PWH with a history of heavy alcohol drinking, the observed sex-based differences in the gut microbiome composition are clinically relevant and could inform therapeutic strategies with evidence-based probiotics.