Structure, function, and inhibition of drug reactivating human gut microbial β-glucuronidases

Biernat, Kristen A.; Pellock, S.J.; Bhatt, Aadra P.; Bivins, Marissa M.; Walton, William G.; Tran, Bryant; Wei, Lianjie; Snider, Michael; Cesmat, Andrew P.; Tripathy, Ashutosh; Erie, Dorothy A.; Redinbo, Matthew R.

doi:10.1038/s41598-018-36069-w

Cited by 67 publications

(77 citation statements)

References 28 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Inhibition. The hundreds of unique gut microbial GUS enzymes mapped to date have been categorized into six functionally distinct groups based on active site architecture (13)(14)(15)(16)(17)(18)(19)(20). GUSs are encoded by Bacteroidetes, Firmicutes, and Proteobacteria; among these phyla, Proteobacteria are unique in encoding only Loop 1 (L1) GUS orthologs, which are most efficient with smaller glucuronidated substrates (14).…”

Section: Gus Loop Architecture Dictates Sn38-g Processing Efficiency Andmentioning

confidence: 99%

“…It has previously been established that inhibiting bacterial GUSs does not alter the pharmacokinetics of irinotecan or its key metabolites in mouse serum (9). To determine whether an L1 GUS enzyme (e.g., Escherichia coli GUS) can drive irinotecan-induced gut damage, we monoassociated germ-free mice with either wild-type (WT) E. coli or the isogenic gus gene deletion strain (ΔGUS) (13). One irinotecan dose (50 mg/kg i.p.)…”

Section: Gus Loop Architecture Dictates Sn38-g Processing Efficiency Andmentioning

confidence: 99%

See 1 more Smart Citation

Targeted inhibition of gut bacterial β-glucuronidase activity enhances anticancer drug efficacy

Bhatt

Pellock

Biernat

et al. 2020

Proc. Natl. Acad. Sci. U.S.A.

Self Cite

126

View full text Add to dashboard Cite

Irinotecan treats a range of solid tumors, but its effectiveness is severely limited by gastrointestinal (GI) tract toxicity caused by gut bacterial β-glucuronidase (GUS) enzymes. Targeted bacterial GUS inhibitors have been shown to partially alleviate irinotecan-induced GI tract damage and resultant diarrhea in mice. Here, we unravel the mechanistic basis for GI protection by gut microbial GUS inhibitors using in vivo models. We use in vitro, in fimo, and in vivo models to determine whether GUS inhibition alters the anticancer efficacy of irinotecan. We demonstrate that a single dose of irinotecan increases GI bacterial GUS activity in 1 d and reduces intestinal epithelial cell proliferation in 5 d, both blocked by a single dose of a GUS inhibitor. In a tumor xenograft model, GUS inhibition prevents intestinal toxicity and maintains the antitumor efficacy of irinotecan. Remarkably, GUS inhibitor also effectively blocks the striking irinotecan-induced bloom of Enterobacteriaceae in immunedeficient mice. In a genetically engineered mouse model of cancer, GUS inhibition alleviates gut damage, improves survival, and does not alter gut microbial composition; however, by allowing dose intensification, it dramatically improves irinotecan's effectiveness, reducing tumors to a fraction of that achieved by irinotecan alone, while simultaneously promoting epithelial regeneration. These results indicate that targeted gut microbial enzyme inhibitors can improve cancer chemotherapeutic outcomes by protecting the gut epithelium from microbial dysbiosis and proliferative crypt damage. microbiome | chemotherapy | cancer | gastrointestinal toxicity I rinotecan (CPT-11) is essential for treating colorectal and pancreatic cancers and is frequently administered as a mixture with 5-fluorouracil and/or oxaliplatin. Ongoing clinical trials seek to extend irinotecan to other forms of cancer. However, irinotecan causes both myelosuppression and gastrointestinal (GI) side effects, including mucositis and late-onset diarrhea, which are often dose limiting: 88% of irinotecan patients develop diarrhea, with 30% experiencing acute grade 3 to 4 diarrhea (1-3). This toxicity is frequently refractory to antimotility drugs. Thus, treating irinotecaninduced diarrhea is a significant clinical need (4, 5).The irinotecan prodrug is activated in vivo to SN38, a potent topoisomerase I inhibitor (6) that retards the growth of rapidly proliferating cells in tumors and the intestinal epithelium, which renews every 5 d. SN38 is detoxified through the addition of glucuronic acid (GlcA) to form SN38-G, a compound marked for elimination via the GI tract. Gut commensal bacterial β-glucuronidase (GUS) proteins remove GlcA from SN38-G. Reactivated SN38 inflicts epithelial damage, resulting in bleeding diarrhea and acute weight loss in animal models (7). We reduced irinotecan-induced gut toxicity using inhibitors that selectively, nonlethally, and potently block the actions of gut bacterial GUS enzymes (8,9). The utility of GUS inhibition also extends to drugs be...

show abstract

Section: Gus Loop Architecture Dictates Sn38-g Processing Efficiency Andmentioning

confidence: 99%

Section: Gus Loop Architecture Dictates Sn38-g Processing Efficiency Andmentioning

confidence: 99%

Targeted inhibition of gut bacterial β-glucuronidase activity enhances anticancer drug efficacy

Bhatt

Pellock

Biernat

et al. 2020

Proc. Natl. Acad. Sci. U.S.A.

Self Cite

126

View full text Add to dashboard Cite

show abstract

“…Beta-glucuronidase was another protein that we found to be downregulated in AgNP-treated samples. β-Glucuronidase and phospholipase are factors in creating colonization surface that facilitate bacteremia and severe infections (Stewart et al, 2007; Biernat et al, 2019).…”

Section: Resultsmentioning

confidence: 99%

Label-Free Proteomic Approach to Study the Non-lethal Effects of Silver Nanoparticles on a Gut Bacterium

et al. 2019

View full text Add to dashboard Cite

Among all the food-related nanoparticles consumed every day, silver nanoparticles (AgNPs) have become one of the most commonly utilized because of their antimicrobial properties. Despite their common use, the effects of sublethal concentrations of AgNPs, especially on gut biofilms, have been poorly investigated. To address this issue, we investigated in vitro the proteomic response of a monospecies Escherichia coli gut biofilm to chronic and acute exposures in sublethal concentrations of AgNPs. We used a new gel- and label-free proteomic approach based on shotgun nanoflow liquid chromatography–tandem mass spectrometry. This approach allows a quantification of the whole proteome at a dynamic range that is higher than the traditional proteomic investigation. To assess all different possible exposure scenarios, we compared the biofilm proteome of four treatments: (i) untreated cells for the control treatment, (ii) cells treated with 1 μg/ml AgNPs for 24 h for the acute treatment, (iii) cells grown with 1 μg/ml AgNPs for 96 h for the chronic treatment, and (iv) cells grown in the presence of 1 μg/ml AgNPs for 72 h and then further treated for 24 h with 10 μg/ml AgNPs for the chronic + acute treatment. Among the 1,917 proteins identified, 212 were significantly differentially expressed proteins. Several pathways were altered including biofilm formation, bacterial adhesion, stress response to reactive oxygen species, and glucose utilization.

show abstract

“…Another process that has been characterized mechanistically is drug reactivation following β-glucuronic acid conjugation in the liver and biliary secretion back into the gut. This process is catalyzed by gut bacterial β-glucuronidases, which are widely present in gut bacteria [54] and have broad substrate specificities [55,56] that allow them to reactivate toxic drugs like the chemotherapeutic irinotecan. This process may be circumvented by drugs that, in turn block, β-glucuronidases to halt drug retoxification.…”

Section: Metabolism Of Drugs and Other Xenobiotics By Gut Microbesmentioning

confidence: 99%

In sickness and health: Effects of gut microbial metabolites on human physiology

Glowacki

Martens

2020

PLoS Pathog

View full text Add to dashboard Cite

The connection between intestinal microbes and human health has been appreciated since the 1880s with Theodor Escherich's investigation of Escherichia coli and other fecal bacteria. Escherich hypothesized that indigenous micoorganisms play roles in both digestion and intestinal diseases [1]. In the last century, our understanding of the bacteria, viruses, archaea, and eukaryotes that normally inhabit the gut has expanded alongside the rest of the field of microbiology, and numerous fundamental roles have been established for this community, now termed the microbiome. As speculated by Escherich, these roles definitively include nutrient digestion [2, 3] and protection from invading pathogens [4] but also extend to short-and long-term instruction of the immune system [5-7] and production of a wide range of metabolites that are unable to be produced by human physiology. Although the gut microbiome is typically described as being composed of nonharmful or beneficial microorganisms, it is now appreciated that both individual species [8] or multiple community members acting together [9, 10] can exert pathogenic effects, which are often more subtle than those of classical pathogens. Indeed, the presence of common intestinal microorganisms with discrete virulence factors (e.g., enterotoxins, genotoxins) that may only manifest in diseases like colorectal cancer or inflammatory bowel disease (IBD) over long periods of time or in certain host genetic backgrounds obscures the definition of pathogen. Accelerated in the 2000s by the "-omics" revolution, along with a recent resurgence of cultivation [11-13], countless studies in the past 2 decades have implied or established connections between altered gut microbiomes and many diseases. These studies have demonstrated the malleability (or fragility) of the microbiome in the face of environmental and dietary perturbations encompassing antibiotic use [14], geography [15], immigration [16], and dietary changes, including fiber deprivation [17, 18]. Although Escherich's original ideas were logically predicted with respect to microbiome effects in the gut, less-anticipated connections between gut microbes and health have extended to neurobiology [19-22] and systemic immune responses that impact allergy [23]. Emerging studies, often extending from omics-based observations, are providing causal and mechanistic understanding of the relationships that connect host responses with changes in the microbiome and its metabolism. Here, we look at recent examples that illustrate how the gut microbiome can augment or perturb host physiology through complementary or novel metabolism often initiating or modifying disease trajectories. The studies we highlight provide details that underscore the importance of gut microbes in human health, which Escherich postulated long ago. The impact of gut bacterial metabolites on host physiology The collective diversity of microbial species that compose the gut microbiome harbor approximately 10 million unique, annotated genes [24]-probably many more [25]-that...

show abstract

Structure, function, and inhibition of drug reactivating human gut microbial β-glucuronidases

Cited by 67 publications

References 28 publications

Targeted inhibition of gut bacterial β-glucuronidase activity enhances anticancer drug efficacy

Targeted inhibition of gut bacterial β-glucuronidase activity enhances anticancer drug efficacy

Label-Free Proteomic Approach to Study the Non-lethal Effects of Silver Nanoparticles on a Gut Bacterium

In sickness and health: Effects of gut microbial metabolites on human physiology

Contact Info

Product

Resources

About