Gut microbial beta-glucuronidase and glycerol/diol dehydratase activity contribute to dietary heterocyclic amine biotransformation

Zhang, Jianbo; Lacroix, Christophe; Wortmann, Esther; Ruscheweyh, Hans‐Joachim; Sunagawa, Shinichi; Sturla, Shana J.; Schwab, Clarissa

doi:10.1186/s12866-019-1483-x

Cited by 54 publications

(66 citation statements)

References 53 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…54 It produces butyrate abundantly (>10 mM) in vitro. 12,55 To determine if F. prausnitzii cells actively produce butyrate when co-cultured with human colon epithelia, we compared the concentration of butyrate in the apical medium and the GuMI effluent in the absence (GuMI-NB) and presence of F. prausnitzii (GuMI-FP). As expected, the concentration of butyrate in the apical medium in GuMI-NB (0.08±0.04 mM) or the inlet (source) medium (0.10±0.04 mM) was close to or below the detection limit 0.08 mM (Figure 2k).…”

Section: The Gumi Physiome Platform Supports Fermentation By the Supementioning

confidence: 99%

“…Afterward, PCR was performed in triplicate to amplify 16s rDNA using DreamTaq Green PCR Master Mix (K1081, Thermo Fisher Scientific Inc.) with primers F8 (5'-AGTTTGATCCTGGCTCAG-3') and 1492R (5'-TACGGYTACCTTGTTACGACTT-3') by following the procedures described elsewhere. 55 PCR products were purified using DNA purification solid-phase reversible immobilization magnetic beads (G95, Applied Biological Materials Inc.) and the purified products were sent out for Sanger sequencing (Genewiz Inc.).…”

Section: Bacteria Co-culture With Colon Epithelial Monolayersmentioning

confidence: 99%

See 1 more Smart Citation

Primary human colonic mucosal barrier crosstalk with super oxygen-sensitiveFaecalibacterium prausnitziiin continuous culture

Zhang

Huang

Yoon

et al. 2020

Preprint

Self Cite

View full text Add to dashboard Cite

AbstractThe gut microbiome plays an important role in human health and disease. Gnotobiotic animal and in vitro cell-based models provide some informative insights into mechanistic crosstalk. However, there is no existing system for a chronic co-culture of a human colonic mucosal barrier with super oxygen-sensitive commensal microbes, hindering the study of human-microbe interactions in a controlled manner. Here, we investigated the effects of an abundant super oxygen-sensitive commensal anaerobe, Faecalibacterium prausnitzii, on a primary human mucosal barrier using a Gut-MIcrobiome (GuMI) physiome platform that we designed and fabricated. Chronic continuous co-culture of F. prausnitzii for two days with colon epithelia, enabled by continuous flow of completely anoxic apical media and aerobic basal media, resulted in a strictly anaerobic apical environment fostering growth of and butyrate production by F. prausnitzii, while maintaining a stable colon epithelial barrier. We identified elevated differentiation and hypoxia-responsive genes and pathways in the platform compared with conventional aerobic static culture of the colon epithelia, attributable to a combination of anaerobic environment and continuous medium replenishment. Furthermore, we demonstrated anti-inflammatory effects of F. prausnitzii through HDAC and the TLR-NFKB axis. Finally, we identified that butyrate largely contributes to the anti-inflammatory effects by downregulating TLR3 and TLR4. Our results are consistent with some clinical observations regarding F. prausnitzii, thus motivating further studies employing this platform with more complex engineered colon tissues for understanding the interaction between the human colonic mucosal barrier and microbiota, pathogens, or engineered bacteria.

show abstract

Section: The Gumi Physiome Platform Supports Fermentation By the Supementioning

confidence: 99%

Section: Bacteria Co-culture With Colon Epithelial Monolayersmentioning

confidence: 99%

Primary human colonic mucosal barrier crosstalk with super oxygen-sensitiveFaecalibacterium prausnitziiin continuous culture

Zhang

Huang

Yoon

et al. 2020

Preprint

Self Cite

View full text Add to dashboard Cite

show abstract

“…CRC-associated gut microbiota also exerts tumorigenic effect through the production of genotoxin, which induces DNA or Alter the abundance, composition, or metabolic activities of gut microbiota, accompanied by colonic inflammation (Khalil et al, 2010;Ribière et al, 2016;Defois et al, 2017;Defois et al, 2018) Bioactivate and transform HCAs and PAHs into compounds with lower toxicity (Van de Wiele et al, 2005;Fekry et al, 2016;Zhang et al, 2019) Trimethylamine-Noxide (TMAO) --Involved in TMAO synthesis; microbial composition could define body response toward TMAO intake (Koeth et al, 2013;Tang et al, 2013;Romano et al, 2015;Cho et al,…”

Section: Bacterial Toxinmentioning

confidence: 99%

Colon Carcinogenesis: The Interplay Between Diet and Gut Microbiota

Loke

Chew

Ngeow

et al. 2020

Front. Cell. Infect. Microbiol.

View full text Add to dashboard Cite

Colorectal cancer (CRC) incidence increases yearly, and is three to four times higher in developed countries compared to developing countries. The well-known risk factors have been attributed to low physical activity, overweight, obesity, dietary consumption including excessive consumption of red processed meats, alcohol, and low dietary fiber content. There is growing evidence of the interplay between diet and gut microbiota in CRC carcinogenesis. Although there appears to be a direct causal role for gut microbes in the development of CRC in some animal models, the link between diet, gut microbes, and colonic carcinogenesis has been established largely as an association rather than as a cause-and-effect relationship. This is especially true for human studies. As essential dietary factors influence CRC risk, the role of proteins, carbohydrates, fat, and their end products are considered as part of the interplay between diet and gut microbiota. The underlying molecular mechanisms of colon carcinogenesis mediated by gut microbiota are also discussed. Human biological responses such as inflammation, oxidative stress, deoxyribonucleic acid (DNA) damage can all influence dysbiosis and consequently CRC carcinogenesis. Dysbiosis could add to CRC risk by shifting the effect of dietary components toward promoting a colonic neoplasm together with interacting with gut microbiota. It follows that dietary intervention and gut microbiota modulation may play a vital role in reducing CRC risk.

show abstract

“…the colon. Gut microbial glucuronidases then cleave the glucose and reactivate the compound leading to genotoxicity in the colon [27]. An alternative notion is that IQ is bioactivated to a mutagen in situ in the colon.…”

Section: Introductionmentioning

confidence: 99%

Genotoxic Assays for Measuring P450 Activation of Chemical Mutagens

Fasullo¹

2021

Genotoxicity and Mutagenicity - Mechanisms and Test Methods

View full text Add to dashboard Cite

This review discusses using yeast as a model organism for studying the biological effects of P450-mediated metabolism of xenobiotics. We discuss the challenges of testing the safety of thousands of chemicals currently introduced into the market place, the limitations of the animal systems, the advantages of model organisms, and the humanization of the yeast cells by expressing human cytochrome P450 (CYP) genes. We discuss strategies in utilizing multiple genetic endpoints in screening chemicals and yeast strains that facilitate phenotyping CYP polymorphisms. In particular, we discuss yeast mutants that facilitate xenobiotic import and retention and particular DNA repair mutants that can facilitate in measuring genotoxic endpoints and elucidating genotoxic mechanisms. New directions in toxicogenetics suggest that particular DNA damaging agents may interact with chromatin and perturb gene silencing, which may also generate genetic instabilities. By introducing human CYP genes into yeast strains, new strategies can be explored for high-throughput testing of xenobiotics and identifying potent DNA damaging agents.Genotoxicity and Mutagenicity -Mechanisms and Test Methods 2 the Ames assays are carcinogenic, while others that test positive in the Ames assays are not carcinogens [8,9]. Many chemicals are not genotoxic per se but require metabolic bioactivation [10]. The bioactivated compound is generally a highly reactive intermediate in a pathway which renders hydrophobic compounds more hydrophilic to facilitate excretion. While bioactivation does occur in specific animal models, toxicity outcomes differ depending on the species [11]. Thus, there is a need for metabolic competent cell-based assays that can measure multiple genotoxic endpoints.Bioactivation occurs by phase I and phase II enzymes; phase I enzymes generally hydroxylate compounds so that phase II enzymes can conjugate larger molecules, facilitating the export and excretion of the modified compound. Phase I enzymes include cytochrome P450 monooxygenases (CYPs), which compose a superfamily of over 50 genes, and catalyze the formation of highly reactive electrophiles and epoxides, as intermediates in xenobiotic metabolism [12,13]. Up to 80% of all bioactivations require CYPs [14]. For catalytic efficiency, the CYP proteins must be reduced by oxidoreductases, which are colocalized with CYPs in the endoplasmic reticulum (ER [15]).Saccharomyces cerevisiae (budding yeast) is an excellent eukaryotic model organism for studying the genotoxicity of xenobiotics, including pharmaceuticals, pesticides, insecticides, and suspected carcinogens. Similar to bacterial organisms, yeast strains are easy to culture, grow rapidly, and can be manipulated genetically, rendering it possible to perform high-throughput analysis [16]. Many yeast genes are similar to human genes, and approximately 30% of can be functionally replaced by the human orthologue [17,18]. DNA repair pathways and genes are also similar [17,16]. Mitochondrial genotoxicity can also be measured [19]. Thus, identi...

show abstract

Gut microbial beta-glucuronidase and glycerol/diol dehydratase activity contribute to dietary heterocyclic amine biotransformation

Cited by 54 publications

References 53 publications

Primary human colonic mucosal barrier crosstalk with super oxygen-sensitiveFaecalibacterium prausnitziiin continuous culture

Primary human colonic mucosal barrier crosstalk with super oxygen-sensitiveFaecalibacterium prausnitziiin continuous culture

Colon Carcinogenesis: The Interplay Between Diet and Gut Microbiota

Genotoxic Assays for Measuring P450 Activation of Chemical Mutagens

Contact Info

Product

Resources

About