Zika virus dysregulates human Sertoli cell proteins involved in spermatogenesis with little effect on tight junctions

Abstract

Zika virus (ZIKV), a neglected tropical disease until its re-emergence in 2007, causes microcephaly in infants and Guillain-Barré syndrome in adults. Its re-emergence and spread to more than 80 countries led the World Health Organization in 2016 to declare a Public Health Emergency. ZIKV is mainly transmitted by mosquitos, but can persist in infected human male semen for prolonged periods and may be sexually transmitted. Testicular Sertoli cells support ZIKV replication and may be a reservoir for persistent ZIKV infection. Electrical impedance analyses indicated ZIKV infection rapidly disrupted Vero cell monolayers but had little effect upon human Sertoli cells (HSerC). We determined ZIKV-induced proteomic changes in HSerC using an aptamer-based multiplexed technique (SOMAscan) targeting >1300 human proteins. ZIKV infection caused differential expression of 299 proteins during three different time points, including 5 days after infection. Dysregulated proteins are involved in different bio-functions, including cell death and survival, cell cycle, maintenance of cellular function, cell signaling, cellular assembly, morphology, movement, molecular transport, and immune response. Many signaling pathways important for maintenance of HSerC function and spermatogenesis were highly dysregulated. These included IL-6, IGF1, EGF, NF-κB, PPAR, ERK/MAPK, and growth hormone signaling. Down-regulation of the PPAR signaling pathway might impact cellular energy supplies. Upstream molecule analysis also indicated microRNAs involved in germ cell development were downregulated by infection. Overall, this study leads to a better understanding of Sertoli cellular mechanisms used by ZIKV during persistent infection and possible ZIKV impacts on spermatogenesis.

Paracrine IFN Response Limits ZIKV Infection in Human Sertoli Cells

“…However, it is also important to note that, in contrast to SC, ZIKV-infected NPC cultures undergo different degrees of cell death and experience impairments in cellular growth depending on their state of differentiation (Wells et al, 2016;Wen et al, 2017), and thus it is plausible that the antiviral responses in NPC may be similarly affected in this regard. In contrast to our LC-MS/MS proteomics analysis, Rashid and colleagues recently employed the SOMAscan aptamer-based multiplexed proteomics approach, that targets a predefined set of proteins, to characterize proteins dysregulated by ZIKV in SC (Rashid et al, 2020). However, although their study reported altered levels related to cell growth, death, and survival in infected SC (Rashid et al, 2020), they could not detect changes in ISGencoded proteins since they were not included in the predefined set of protein targets of the SOMAscan aptamer-based platform.…”

“…In contrast to our LC-MS/MS proteomics analysis, Rashid and colleagues recently employed the SOMAscan aptamer-based multiplexed proteomics approach, that targets a predefined set of proteins, to characterize proteins dysregulated by ZIKV in SC (Rashid et al, 2020). However, although their study reported altered levels related to cell growth, death, and survival in infected SC (Rashid et al, 2020), they could not detect changes in ISGencoded proteins since they were not included in the predefined set of protein targets of the SOMAscan aptamer-based platform.…”

ZIKV Infection Induces DNA Damage Response and Alters the Proteome of Gastrointestinal Cells

“…A total of 446 host proteins were significantly dysregulated at any time point, and 71 were dysregulated > 1.375-fold ( Table 1 ). The majority of dysregulated proteins were significantly upregulated at 12 hpi, irrespective of the fold-change cut-off used ( Table 1 ), a pattern that was not reflected by our previous analyses of ZIKV-infected Vero [ 15 ], U-251 astrocytoma [ 16 ] or Sertoli cells [ 17 ]. Bioinformatic analysis using Ingenuity Pathway analysis (IPA) predicted several pathways that were activated and inhibited by ZIKV infection.…”

“…We have been examining proteomic alterations induced by ZIKV in various cell types, including monkey kidney Vero [ 15 ], human U-251 astrocytoma [ 16 ] and human Sertoli [ 17 ]. Most protein dysregulation occurred at 48 hpi or later in these other cells.…”

BAF45b Is Required for Efficient Zika Virus Infection of HAP1 Cells

“…Recent studies have pointed out the importance of microRNA (miRNA), including miR-9, during ZIKV infection, leading to apoptosis-associated microcephaly, reduced numbers of NPCs, and newly formed neurons in embryonic mouse cortex [ 13 , 14 , 15 , 16 ]. ZIKV infection in neuronal stem cells also upregulates miR-124-3p, which has been predicted to be involved in microcephaly [ 17 ].…”