RESEARCH ARTICLE Aptamers www.proteomics-journal.com Vero Cell Proteomic Changes Induced by Zika Virus Infection Kathleen K. M. Glover, Ang Gao, Ali Zahedi-Amiri, and Kevin M. Coombs* The re-emergence and the recent spread of the Zika virus (ZIKV) has raised signiﬁcant global concerns due to lack of information in patient diagnosis and management. Thus, in addition to gaining more basic information about ZIKV biology, appropriate interventions and management strategies are being sought to control ZIKV-associated diseases and its spread. This study’s objective is to identify host cell proteins that are signiﬁcantly dysregulated during ZIKV infection. SOMAScan, a novel aptamer-based assay, is used to simultaneously screen >1300 host proteins to detect ZIKV-induced host protein dysregulation at multiple time points during infection. A total of 125 Vero cell host proteins, including cytokines such as CXCL11 and CCL5, interferon stimulated gene 15, and translation initiation factors EIF5A and EIF4G2, are signiﬁcantly dysregulated after ZIKV infection. Bioinformatic analyses of 77 host proteins, that are signiﬁcantly dysregulated 1.25-fold, identify several activated biological processes, including the JAK/STAT, Tec kinase, and complement cascade pathways. 1. Introduction Zika virus (ZIKV), which belongs to the family Flaviviridae, has a plus-sense non-segmented single-stranded RNA genome. ZIKV, like other Flaviviridae (West Nile virus, Dengue virus, Japanese encephalitis virus and yellow fever virus), is an ar- bovirus, transmitted mostly by the Aedes aegypti and A. albopic- tus species of mosquitos. The three primary cell types targeted by ﬂaviviruses, including ZIKV, are monocytes, macrophages, and dendritic cells. [4–7] However, CD14 + monocytes have been reported as the main targets of ZIKV during infection. [8–11] Trans- mission of ZIKV starts when an infected mosquito bites an an- imal. Cells of the skin (ﬁbroblasts and epidermal keratinocytes) are highly permissive to ZIKV infection. [12] Virus then migrates into the lymph nodes and undergoes further replication, [13] and migrates into the circulatory system where it crosses the blood– brain barrier and enters the central nervous system. [14] The ﬁrst K. K. M. Glover, A. Zahedi-Amiri, Prof. K. M. Coombs Department of Medical Microbiology and Infectious Diseases University of Manitoba Winnipeg, Manitoba, R3E OJ9, Canada E-mail: kevin.coombs@umanitoba.ca K. K.M. Glover, Dr. A. Gao, A. Zahedi-Amiri, Prof. K. M. Coombs Manitoba Centre for Proteomics & Systems Biology Winnipeg, Manitoba, R3E 3P4, Canada Prof. K. M. Coombs Children’s Hospital Research Institute of Manitoba Buhler Research Centre Winnipeg, Manitoba, R3E 3P4, Canada DOI: 10.1002/pmic.201800309 line of host cell defense to ZIKV, as well as to other viral infections, is the innate immune response which includes secretion of type I interferons (IFNs) and inﬂammatory cytokines, comple- ment responses, and Natural Killer cell immunity. [14] Arboviruses like ZIKV hijack other innate responses such as apoptosis, autophagy, and unfolded protein response during infection and replication. [15] After cell entry, ﬂaviviruses release their viral genetic material into the cell cytoplasm [16–18] and the plus-sense single-stranded RNA genome is im- mediately translated into a polyprotein. The polyprotein is cleaved by host and viral proteases into three structural and seven non-structural proteins. Structural proteins are: capsid protein (C) that binds viral RNA, the premembrane (prM) protein that blocks premature viral fusion, and the enve- lope (E) protein which mediates viral attachment, membrane fu- sion, and viral assembly. [16] The seven non-structural proteins (NS1, NS2A, NS2B, NS3, NS4A, NS4B, and NS5) regulate vi- ral transcription and replication and attenuate host antiviral responses. [13] Viral replication occurs in the rough endoplasmic reticulum. The immature viral particle is then transported to the Golgi apparatus where furin-mediated cleavage of prM to M gen- erates mature, infectious virions which acquire their envelopes as they bud through host cell membranes. [19–21] Approximately half of the global population is at risk for arbovirus infections. [22] The Brazilian 2015 ZIKV outbreak re- sulted in “a 20-fold increase in the number of infants born with microcephaly,” [23] and barely a year later, the Brazilian Ministry of Health reported 4500 cases of microcephaly, making ZIKV a pub- lic health problem. [23] This caused anxiety in expectant mothers as well as women of reproductive age who were living or had vis- ited ZIKV endemic countries. Some countries therefore recom- mended couples to delay child bearing and in the United States, the CDC advised couples to use condoms to reduce the spread of the virus as it is believed to also be sexually transmitted. [24] In ad- dition, pregnant women who have had a history of visit to ZIKV endemic regions are put under observation to detect any pathol- ogy within the developing fetus. [25,26] Gaining more basic information about ZIKV biology to iden- tify potential targets in its replicative cycle might help in the de- velopment of appropriate interventions and management strate- gies to control ZIKV-associated diseases as well as its spread. Given the inherent mutability of RNA genomes, one approach which is becoming more widespread is the identiﬁcation of host Proteomics 2019, 19, 1800309 C  2018 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim 1800309 (1 of 16)