Real-Time Monitoring of Host–Gut Microbial Interspecies Interaction in Anticancer Drug Metabolism

Abstract

Gut microbiome can affect drug metabolism considerably, leading to modified drug response. However, quantitative estimation of host vs. microbial contributions in a living host–gut microbiome system has been challenging. Using the interspecies system of Caenorhabditis elegans and gut bacteria, we developed a real-time approach for monitoring their metabolic interaction in vivo during anticancer drug 5-fluorouracil (5-FU) metabolism. The fluorine NMR-based approach yielded the quantitative contributions to the host 5-FU metabolism made by human gut-microbial species of variable genetic backgrounds. It also experimentally confirmed a bacterial gene–metabolism relationship. Differential 5-FU catabolism among bacterial substrains and the contributions to the host metabolism, unobservable by conventional 16S rRNA metagenomic sequencing, were also found. The metabolic contributions could be correlated with phenotypic developmental toxicity of 5-FU to the host fed with different substrains. Our convenient platform should help to reveal heterogeneity in host–gut microbiome interactions for many drugs in a living symbiotic system.

NMR Metabolomics Methods for Investigating Disease

“…However, studies in a living host−gut microbiome system have been challenging. Recently, development of a real-time monitoring method for metabolic interaction in vivo during the application of an anticancer drug, 5-fluorouracil (5-FU), using C. elegans and gut bacteria was reported 59 (Figure 4). 19 F NMR enabled investigation of 5-FU metabolism, quantitatively, and in real time within the host utilizing human gut-microbes with variable genetics.…”

A Structure-Guided Molecular Network Strategy for Global Untargeted Metabolomics Data Annotation

“…The network connectivity was evaluated by the clustering coefficient, network density, characteristic path length, network diameter, network radius, and connected components (Table 1). Both GCMN KEGG and the KEGG MRN exhibited small-world properties 57 with relatively small values of characteristic path length (3,9) and high values of the clustering coefficient (0.6, 0.076). The network density of GCMN KEGG (3.51 × 10 −2 ) was larger than that of KEGG MRN (3.29× 10 −4 ), indicating stronger connectivity of GCMN KEGG .…”

“…With the progress of analytical techniques, especially the rapid development of high-resolution mass spectrometry (HRMS), rich metabolome information can be achieved from the untargeted metabolomics analysis of biological samples. − Unfortunately, metabolite annotation remains a bottleneck of metabolomics. − …”

Real‐time monitoring of enzyme‐catalyzed phosphoribosylation of anti‐influenza prodrug favipiravir by time‐lapse nuclear magnetic resonance spectroscopy

“…[23][24][25] Given the many useful properties of 19 F-NMR, methods employing 19 F-NMR-based techniques are widely considered to offer robust assays for monitoring reaction processes. [26][27][28] Notably, the favipiravir molecule includes a fluorine atom (Figure 1 and Figure S1); therefore, 19 F-NMR is expected to be a preferable approach for direct observation of the metabolic behavior of favipiravir. Similarly, phosphorus nuclei 31 P also have several favorable characteristics as an NMR detection probe, including the following: (1) 31 P constitutes 100% of the natural abundance; and (2) 31 P has relatively high NMR sensitivity, given that this isotope's gyromagnetic ratio is approximately 0.40 relative to 1 H. These characteristics of 31 P also allow NMR to serve as a significant analytical method for elucidation of events involving biomolecules, including investigations of nucleic acid structure, characterization of lipid membranes, and structure-function correlation studies of the phosphorylation of proteins.…”

“…The fluorine atom 19 F has several favorable characteristics as a site‐ or target‐specific NMR detection probe, including the following: (1) 19 F represents 100% of this element's natural abundance; (2) 19 F has high NMR sensitivity because of the isotope's high gyromagnetic ratio (approximately 0.94 relative to 1 H); and (3) 19 F‐NMR signals derived from the target molecules can be observed unambiguously (with no background) because 19 F is not naturally present in most biomolecules 23–25 . Given the many useful properties of 19 F‐NMR, methods employing 19 F‐NMR–based techniques are widely considered to offer robust assays for monitoring reaction processes 26–28 . Notably, the favipiravir molecule includes a fluorine atom (Figure 1 and Figure S1 ); therefore, 19 F‐NMR is expected to be a preferable approach for direct observation of the metabolic behavior of favipiravir.…”