The global healthcare community faces escalating threats from antimicrobial resistance (AMR); therefore, the need for novel, non-traditional antimicrobials has become increasingly urgent. Antimicrobial peptides (AMPs), naturally produced by a wide array of organisms, including marine invertebrates, offer a promising alternative to conventional antibiotics. AMPs typically act by disrupting microbial membranes, a mechanism less prone to resistance development. Mussels, being sessile filter feeders in pathogen-rich marine environments, have evolved expansive and diverse innate immune systems. The genomic diversity and adaptive resilience of Mytilus spp. remain underexploited resources for drug discovery, particularly in the search for new AMPs with clinical and ecological relevance.
The central objective of the project is to identify, functionally validate, and characterize novel AMPs from Mytilus genomes. The research plan integrates in silico genomic screening with experimental in vitro validation. Initially, genome assemblies and transcriptomic datasets from multiple Mytilus species will be analyzed to identify gene families coding for AMP-like sequences. The bioinformatics strategy includes sequence motif detection, gene clustering, evolutionary analysis of presence/absence variation, and structural modeling of predicted peptides. These tasks will be supported by national HPC resources provided through the PLGrid infrastructure, which allows high-throughput data processing, molecular simulations, and structural predictions. Experimentally, we will establish short-term primary cultures from Mytilus hemocytes and digestive gland cells. These cultures will serve as models to assess immune gene expression and AMP induction under controlled stimulation. Culture viability and responsiveness will be monitored using fluorescein diacetate (FDA) assays and quantitative PCR. Peptides derived from highly expressed or computationally prioritized genes will be purified either from conditioned media or synthesized chemically when necessary. Functional testing will involve comprehensive antimicrobial profiling against a library of multidrug-resistant strains.
Project No. 2025/57/B/NZ8/03838 funded by the National Science Centre (Poland).
