Deep longitudinal multi-omics analysis of Bordetella pertussis cultivated in bioreactors highlights medium starvations and transitory metabolisms, associated to vaccine antigen biosynthesis variations and global virulence regulation

Abstract

Bordetella pertussis is the bacterial causative agent of whooping cough, a serious respiratory illness. An extensive knowledge on its virulence regulation and metabolism is a key factor to ensure pertussis vaccine manufacturing process robustness. The aim of this study was to refine our comprehension of B. pertussis physiology during in vitro cultures in bioreactors. A longitudinal multi-omics analysis was carried out over 26 h small-scale cultures of B. pertussis. Cultures were performed in batch mode and under culture conditions intending to mimic industrial processes. Putative cysteine and proline starvations were, respectively, observed at the beginning of the exponential phase (from 4 to 8 h) and during the exponential phase (18 h 45 min). As revealed by multi-omics analyses, the proline starvation induced major molecular changes, including a transient metabolism with internal stock consumption. In the meantime, growth and specific total PT, PRN, and Fim2 antigen productions were negatively affected. Interestingly, the master virulence-regulating two-component system of B. pertussis (BvgASR) was not evidenced as the sole virulence regulator in this in vitro growth condition. Indeed, novel intermediate regulators were identified as putatively involved in the expression of some virulence-activated genes (vags). Such longitudinal multi-omics analysis applied to B. pertussis culture process emerges as a powerful tool for characterization and incremental optimization of vaccine antigen production.

Integrating proteomic data with metabolic modeling provides insight into key pathways of Bordetella pertussis biofilms

“…A recent study (Anziani et al, 2023), performed a temporal multi-omics analysis on planktonic B. pertussis cells and identified that the genes and proteins involved in the TCA cycle from the conversion of ⍺-ketoglutarate to fumarate had relatively low activity until after 12 h. These reactions would have lower activity during activation of the glyoxylate shunt reinforcing the flux predicted in this study. Additionally, it was reported that the proteins and genes involved in glyoxylate metabolism decreased following 12 h before increasing again after 18 h and 45 min (Anziani et al, 2023). The extracellular metabolites that were measured also reinforced this trend (Anziani et al, 2023).…”

“…Additionally, it was reported that the proteins and genes involved in glyoxylate metabolism decreased following 12 h before increasing again after 18 h and 45 min (Anziani et al, 2023). The extracellular metabolites that were measured also reinforced this trend (Anziani et al, 2023). Variation in the glyoxylate shunt has been observed in the biofilms of other species.…”