The Convergence of Metabolism and Virulence Factor Regulation on Streptococcus pyogenes Pathogenesis

Nutrient acquisition is a strong driving force in the evolution of pathogenesis. For many pathogens such as Streptococcus pyogenes, the coordination of metabolism with the expression of virulence factors is necessary for colonization. Previous research examining the regulation of virulence factors in S. pyogenes identified a unique catabolite sensing regulatory pathway composed of the tagatose bis-phosphate aldolase LacD.1. Examination of LacD.1 has determined that while it still maintains its ancestral enzymatic activity its regulation occurs via a mechanism independent of catalysis. Our approach to understanding how S. pyogenes\u27 metabolism contributes to pathogenesis involves both understanding the adaptation of LacD.1 as a novel metabolic regulator as well as investigating the role of LacD.1-regulated genes in pathogenesis. Analysis of LacD.1\u27s ancestral enzymatic activity and comparison to a paralogous protein encoded in the genome, LacD.2, suggested that the adaptation of LacD.1 required a decrease in enzymatic activity as compared to LacD.2. This decrease in activity is largely due to an increase in Km, and suggests an optimization in affinity for the signaling substrate(s) necessary for LacD.1\u27s regulation. Additionally the regulatory function of LacD.1 was found to be essential for the fitness of bacteria when cultured in media that mimicked a soft tissue environment. The selective advantage observed for bacteria in this environment can likely be attributed to LacD.1\u27s positive regulation of the arc operon, which encodes genes necessary for arginine catabolism and contributes to both the production of ATP and protection from acid stress. To further investigate the arc operon\u27s contribution to the fitness of bacteria in a soft tissue environment we examined the influence of arginine catabolism on pathogenesis. Analysis of several mutants defective in catabolism revealed that both the utilization of arginine as well as the novel utilization of citrulline aids in S. pyogenes pathogenesis. This influence on pathogenesis included contributions to both the cellular metabolism of S. pyogenes as well as the modulation of the host immune response. Taken together this work demonstrates that LacD.1\u27s adaptation to a regulator and its coordination of metabolism and virulence factor expression was essential to increase the fitness of S. pyogenes as a pathogen

Antivirulence <i>C</i>‑Mannosides as Antibiotic-Sparing, Oral Therapeutics for Urinary Tract Infections

Gram-negative uropathogenic Escherichia coli (UPEC) bacteria are a causative pathogen of urinary tract infections (UTIs). Previously developed antivirulence inhibitors of the type 1 pilus adhesin, FimH, demonstrated oral activity in animal models of UTI but were found to have limited compound exposure due to the metabolic instability of the <i>O</i>-glycosidic bond (<i>O</i>-mannosides). Herein, we disclose that compounds having the <i>O</i>-glycosidic bond replaced with carbon linkages had improved stability and inhibitory activity against FimH. We report on the design, synthesis, and in vivo evaluation of this promising new class of carbon-linked <i>C</i>-mannosides that show improved pharmacokinetic (PK) properties relative to <i>O</i>-mannosides. Interestingly, we found that FimH binding is stereospecifically modulated by hydroxyl substitution on the methylene linker, where the <i>R</i>-hydroxy isomer has a 60-fold increase in potency. This new class of <i>C</i>-mannoside antagonists have significantly increased compound exposure and, as a result, enhanced efficacy in mouse models of acute and chronic UTI