MECHANISMS REGULATING BREACH OF BLOOD RETINAL BARRIER UPON ZIKA VIRUS INFECTION - PROJECT SUMMARY/ABSTRACT: Since 2015-26 epidemic in Brazil, Zika virus (ZIKV) infection has been linked to cause microcephaly in newborn infants and World Health Organization declared it a global public health emergency. One third of these infants were also reported to have ocular anomalies, yet the ocular pathology of ZIKV remain poorly understood, with no available treatments. Being RNA virus, ZIKV can evolve rapidly, underscoring the need for continued research. My long-term goal is to investigate how flaviviruses affect the blood-retinal barrier (BRB) and cause ocular issues. During my postdoctoral training, I conducted high-throughput transcriptomics on ZIKV-infected RPE cells and identified potential role of Ang/Tie2, S1P, and AMPK pathways involved in barrier integrity. However, the role of these pathways in ZIKV-induced BRB disruption leading to ocular anomalies remains unexplored. My hypothesis is that ZIKV infection dysregulates these pathways, compromising barrier integrity, and enabling viral entry into the eye, resulting in ocular complications. Therefore, therapeutic strategies aimed at restoring BRB integrity may alleviate ZIKV-induced ocular pathology. To test this hypothesis, I propose three aims: 1) Investigate the role of Ang/Tie-2 and S1P signaling in ZIKV-induced BRB permeability; 2) Examine the impact of AMPK activation on BRB integrity during ZIKV infection; and 3) Study the effects of RPE-specific knockout of AMPK, Ang/Tie2, and S1P pathway signaling on ZIKV-induced chorioretinal atrophy in mice. During the mentored phase, I will establish a congenital ZIKV-infection model in immunocompetent mice with guidance from Dr. Kumar (mentor) and Dr. Mor (collaborator), both experts in host-pathogen interactions and ZIKV-pregnancy models. I will also learn to culture iPSC lines to generate retinal organoids and perform gene knockouts using the CRISPR/Cas approach, guided by Dr. Arumugaswami (co-mentor), an expert in stem cell and retinal organoid culture. The mentored phase will involve regular meetings with mentors, co-mentor, and advisory committee members, attendance at scientific conferences, and ongoing career development. In the independent phase, I will establish my own laboratory and complete the proposed aims by utilizing an RPE- specific knockout ZIKV-congenital infection model. Successful completion of these aims will elucidate (i) the roles of host Ang/Tie2 and S1P pathways, and AMPK in BRB regulation; and (ii) the feasibility of mitigating ZIKV-mediated ocular tissue damage via restoring barrier integrity. These findings will provide crucial insights into the pathobiology of ocular ZIKV infection. I am well-positioned for this research and my career development, thanks to Dr. Kumar's state-of-the-art facility at Wayne State University and collaborations with renowned scientists, Dr. Arumugaswami, Dr. Mor, Dr. Yu, Dr. Shukla, and Dr. Ljubimov, which will contribute significantly to my research efforts and facilitate the establishment of my independent laboratory at an academic institution in the US.
