Research Mentor Name
Dr. Ashok Kumar
Research Mentor Email Address
Institution / Department
Ophthalmology, visual and anatomical sciences and Kresge Eye Institute
Level of Research
Type of Post-Bachelor Degree
Masters in Medical Research
Modulation of mitochondrial dynamics and bioenergetics in Zika Virus infected retinal cells
Karim Dirani1, Sneha Singh1, Shailendra Giri2, Ashok Kumar1
1 Department of Ophthalmology, Visual and Anatomical Sciences, and Kresge Eye Institute, Wayne State University School of Medicine, Detroit, MI
2 Department of Neurology, Henry Ford Health Systems, Detroit, MI
Purpose: In addition to the powerhouse of the cells and regulation of apoptosis, mitochondria are being increasingly recognized in playing crucial role in modulating innate immune response in viral infections. In this study, we sought to determine the impact of mitochondrial dynamic (fission, fusion, and mitophagy) and bioenergetics status in regulating antiviral innate response to Zika virus (ZIKV) infection.
Methods: Human ARPE-19 cells were infected with ZIKV (strain PRVABC59) at MOI of 1. The mitochondria were visualized for fission and fusion process using TOM20 and mitotracker dyes while the mitochondrial membrane potential was measured using JC-1 staining. The temporal expression of key genes/molecules involved in mitochondrial dynamic was assessed by RT-qPCR and Western blot. ARPE-19 cells stably expressing individual ZIKV genes were used to assess the functional role of viral proteins in altering mitochondrial dynamics. The bioenergetics activity of the cells was assessed using Seahorse XF analyzer for monitoring oxygen consumption rate (OCR) and Extracellular acidification rate (ECAR) upon ZIKV infection and with individual viral proteins.
Results: All cells tested were permissive to ZIKV infection, as evidenced by a time dependent increase in virion production and immunostaining of viral antigen in the cells. Infected cells exhibited an increase in the expression of antiviral (e.g., IFNs, ISG15, MX1, OAS2) and inflammatory mediators (e.g., CCL5, TNFa). Amongst the molecules regulating the mitochondrial fission and fusion, levels of p-Drp1, mitofusins (MFN1, MFN2), and Opa1 were altered at both mRNA and protein levels. Similarly, molecules regulating mitophagy, PINK1/Parkin -dependent and independent pathways were modulated in ZIKV infected cells. Among individual viral proteins, the non-structural proteins exerted profound effects on mitochondrial dynamic. This correlated with a decrease in OCR response upon ZIKV infection. Interestingly, the pharmacological inhibition of Drp1 reduced ZIKV replication.
Conclusion: Collectively, our data indicates that ZIKV virus modulates mitochondrial dynamics and individual ZIKV proteins exert differential effects on bioenergetics and antiviral innate immune response of infected cells. Targeting mitochondrial dynamic may provide new therapeutic avenues for antiviral therapy against ZIKV infection.
Medicine and Health Sciences
Dirani, Karim; Singh, Sneha; Kumar, Ashok; and Giri, Shailendra, "Modulation of mitochondrial dynamics, bioenergetics and the innate antiviral immune response in Zika Virus infected Retinal cells" (2021). Medical Student Research Symposium. 60.