Identification of compounds with novel MoA targeting Klebsiella pneumoniae

This proposal aims to expand the repertoire of drug targets and lead compounds by developing an optimized screening workflow to identify molecules with novel MoA against Klebsiella pneumoniae. The approach will leverage, but not restricted to, Japan's two largest open-access academic chemical libraries and integrate high-throughput transcriptomic perturbation analysis (Quartz-seq2) to differentiate the MoA of identified hits from existing antibiotics. Transcriptome profiling will be performed on a subset of active compounds identified through high-throughput screening (HTS), alongside a panel of known antibiotics for reference. The transcriptome will be conducted by Knowledge Palette, a leader in high-throughput RNA sequencing and the developer of Quartz-seq2 (Sasagawa et al., 2018, Genome Biol), which ranked first in accuracy and performance in an international benchmarking study by the Human Cell Atlas project (Mereu et al., 2020, Nat Biotechnol). The overarching objective is to discover compounds with distinct MoAs and their corresponding molecular targets, using transcriptomic signatures to guide prioritization and characterization of novel antibacterial candidates, and resistant generation combined with whole genome sequencing in order to identify their molecular targets.