In‐Vitro Evaluation of HIV/SARS‐CoV‐2 Co‐Infection Mediated Proteomic Changes in Astrocytes and Pericytes Reveals Altered Signaling Pathways Associated With Neurodegenerative Disorders

ABSTRACT

Coronavirus disease 2019 (COVID‐19) survivors frequently experience a wide range of symptoms known as post‐acute sequelae of SARS‐CoV‐2 (PASC) or long COVID. Importantly, complications arising from microvascular dysfunction, blood‐brain barrier (BBB) disruption, and chronic neuroinflammation have been implicated in driving PASC within the central nervous system (CNS), known as neuro‐PASC. Notably, people with HIV (PWH), who suffer from chronic neuroinflammation, BBB impairment, and glial cell dysfunction, collectively known as neuro‐HIV, are generally at higher risk of neuro‐PASC. The overlap between neuro‐PASC and neuro‐HIV raises concerns that HIV and SARS‐CoV‐2 co‐infection may exacerbate neurological dysfunctions among PWH. In this study, using an in‐vitro cell culture model, we examine the effects of HIV and SARS‐CoV‐2 mono‐ and co‐infection in microglia, astrocytes, and pericytes. Our results demonstrated that majority of brain cell types support SARS‐CoV‐2 replication, in the presence and absence of HIV infection. Furthermore, in both mono‐ and co‐infected cells, there were varying degree of up‐ and downregulation of SARS‐CoV‐2 host cell entry factors, such as ACE2, TMPRSS2, NRP1, and TRIM28, and inflammatory cytokines including IL‐6, TNF‐α, and IL‐1β. Moreover, conditioned media collected from HIV, SARS‐CoV‐2, and HIV/SARS‐CoV‐2 co‐infected astrocytes and pericytes were shown to be neurotoxic. Additionally, proteomic analysis has revealed a unique set of proteins significantly up/down regulated in HIV/SARS‐CoV‐2 co‐infected astrocytes and pericytes. The gene set enrichment analysis of these proteins indicates dysregulation of lipid, energy, and immune metabolism pathways linked to neurodegenerative disorders like Alzheimer's, Parkinson's, Huntington's disease, and amyotrophic lateral sclerosis. These in‐vitro findings indicate that astrocytes and pericytes from HIV/SARS‐CoV‐2 co‐infection exhibit altered protein expression profiles, implicating dysregulated signaling pathways associated with neurodegenerative dysfunction.