SBIR-STTR Award

Environmental DNA from three extreme samples will be isolated, cloned into a fosmid library (5760 clones) for sequencing. The sequencing data will cover ~ 240 Megabases and will be assembled into contigs long enough (~ 40 Kilobases) to represent potential biosynthetic gene clusters (BGCs). This data will be compared to BGC databases looking for homology to antibiotic targets of DoD interest and with reasonable expression profiles. Subcloning and expression will be performed on BGC clones of interest followed by inhibitory growth assays to prove in vitro antibacterial activity. The Option period will be used to test the antibacterial cell extracts against DoD pathogens at the Core facility.

This project is a unique metagenomic evaluation of under-studied extremophilic microbiome natural products (NPs), with the goal of discovering novel antibiotic biowarfare agent (BWA) countermeasures. A metagenomic biosynthetic gene cluster library from extremophilic sources was created, sequenced, analyzed, and exploited to produce several broad-spectrum antibiotic extracts during the Phase I effort. A novel platform for extremophilic microbiome drug discovery (EMDD) was created as well and will be expanded during the Phase II effort. These extracts will be purified and biochemically characterized before testing in vitro against BWAs. Cytotoxicity and mechanism of action studies will be conducted using in vitro and in silico methods. The leads will be optimized by a structure-activity relationship expansion study, which will generate a series of improved analogues for further in vitro screening. The best lead compounds will be tested for in vivo efficacy against priority bacterial BWAs. The goal is to discover a novel antibiotic with efficacy of >50% survival in a lethal challenge mouse model for subsequent preclinical studies and FDA investigational new drug application.