Research Projects

Rationale Nipah virus (NiV), a zoonotic Henipavirus, is significant public health threat due to its high case fatality rate & neurotropism. NiV has been identified as priority pathogen by both WHO & ICMR and has been responsible for six outbreaks in India. NiV infection spreads rapidly often transitioning from flu-like symptoms to encephalitis seizures, coma, and brainstem dysfunction. Microglia, the resident immune cells of brain, are highly responsive to NiV due to abundant expression of entry receptors for NiV attachment glycoprotein (G) such as ephrin-B2. NiV-G–induced mitochondrial stress, as evidenced by Johnston et al. (2025), could compromise mitochondria’s functioning in antiviral defense through MAVS and induce a proinflammatory state. The molecular connection between NiV-G–mediated mitochondrial impairment and immune activation in human microglia is poorly understood. By combining high-resolution mitochondrial assays with transcriptomic and proteomic profiling, and integrative bioinformatics, we aim to define early molecular events linking mitochondrial dysfunction to immune activation. Objectives 1.To study the impact of attachment glycoprotein G of Nipah virus on structure and function of mitochondria in human microglia 2.To characterize transcriptomics and proteomic alterations linked to mitochondrial dysfunction and immune activation during NiV-G exposure on human microglia 3.To validate key molecular signatures of NiV-G mediated mitochondrial disruption and assess their mechanistic role in inflammatory responses of human microglia. Hypothesis We hypothesize that NiV-G binding to ephrin-B2 on human microglia can independently stimulate mitochondrial dysfunction, resulting in ROS production, inflammasome activation, and proinflammatory gene expression. This mitochondrial damage is an early, replication-independent signal that induces microglial immune activation and contributes to NiV neuropathogenesis. Main Experiments Objective 1: HMC3 microglia will be treated with recombinant NiV-G (0.1–1 µg/mL, 24 h), with PBS, rotenone, and LPS as controls. Mitochondrial structure and function will be assessed via confocal microscopy, ΔΨm, ATP, ROS, OCR/ECAR, and Western blotting for mitochondrial markers. Objective 2: RNA-seq and 16-plex TMT proteomics will identify DEGs and DEPs followed by pathway and network analysis using edgeR, clusterProfiler, STRING, WGCNA, and PathVisio. Objective 3: Key candidates will be validated by qPCR, Western blot, and immunofluorescence. Gene knockdown or overexpression will determine functional roles, and ephrin-B2 knockdown will confirm receptor specificity. Significance This research will determine whether NiV-G independently disrupts mitochondrial function and stimulates immune signaling in human microglia. Results will uncover function of ephrin-B2 in NiV induced neuroinflammation and may lay foundation for host-targeted therapy and vaccine development against Nipah and other neurotropic viruses.