Multi-omics Analysis of Bacteria-Host Interactions in Brugia malayi, a Medically Important Nematode with a Wolbachia Endosymbiont

SUMMARY Filarial nematodes that cause the most devastating filarial diseases all contain obligate bacterial Wolbachia endosymbionts. We will focus on Brugia malayi, the only filarial nematode that infects humans and that is zoonotic, enabling experimental interrogation. Efforts to eliminate disease caused by filarial nematodes are stymied by obstacles such as limited treatment options, drug resistance, and coendemicity with nematodes that prevent treatment, like Loa loa. However, targeting Wolbachia bacterial endosymbionts with antibiotics has emerged as a promising avenue for the development of novel therapeutics against disease caused by filarial nematodes. However, the molecular underpinnings of this symbiosis are not fully characterized. The independent loss of Wolbachia endosymbionts at least three times in filarial nematodes suggests that the symbiosis could be fragile. If fragile, the selective pressures placed by antibiotics have the potential to drive loss of the endosymbiont, creating antibiotic resistant populations of medically important filarial nematodes. Understanding the molecular underpinnings of the interactions between these obligate host-associated nematodes and their obligate Wolbachia endosymbionts is therefore important. We propose experiments that draw upon insights from research on Wolbachia-insect interactions to interrogate Wolbachia-nematode interactions, establishing similarities and differences. We will examine tissue-specific transcriptomic changes, chromatin state changes, and epitranscriptomic changes upon removal of the obligate mutualistic Wolbachia endosymbiont wBm in Brugia malayi with two different antibiotics. To do this, we propose using single nuclei RNA-seq (snRNA-seq) and single nuclei ATAC-seq (snATAC-seq) techniques to examine tissue-specific responses to Wolbachia depletion. We will also use direct RNA sequencing to study epitranscriptomics changes upon treatment with tetracycline and rifampicin to remove the Wolbachia endosymbiont. The direct RNA sequencing data will also be used to improve transcript annotation, which will improve data analysis and rigor for these experiments as well as future research by the entire filarial nematode research community. We will make these data and analyses readily available to other scientists to interrogate, enabling other scientists to develop and test their own hypotheses, further advancing research in this area. Project Number: 1R01AI190371-01A1 | Fiscal Year: 2026 | NIH Institute/Center: National Institute of Allergy and Infectious Diseases (NIAID) | Principal Investigator: Julie Dunning Hotopp | Institution: UNIVERSITY OF MARYLAND BALTIMORE, BALTIMORE, MD | Award Amount: $678,218 | Activity Code: R01 | Study Section: Interspecies Microbial Interactions and Infections Study Section[IMII] View on NIH RePORTER: https://reporter.nih.gov/project-details/1R01AI19037101A1