Cranberry-Derived Proanthocyanidin and Its Gut Microbial Metabolites Affect the Intestinal miRNome in a Distinct Manner In Vitro

Abstract

Objectives Polyphenols are emerging as novel prebiotic compounds. Cranberries are a rich source of polyphenols, such as proanthocyanidin (PAC), which has known benefits including anti-cancer properties. In the colon, PACs are catabolized by the gut microbiota into 3,4-dihydroxyphenylacetic acid (DHPAA) and 3-(4-hydroxyphenyl) propionic acid (HPPA), which may mediate prebiotic effects. Mechanisms are unknown but may involve host microRNA (miRNA). The objective of this study was to investigate the effects of nutritionally relevant doses of cranberry PAC, DHPAA, and HPPA on the human intestinal miRNome. Methods Differentiated Caco-2BBe1 colonic epithelial cells, a morphologically homogenous subclone of Caco-2 cells, were treated with cranberry extract containing 94% PAC (Ocean Spray Cranberries, 50 μg/ml), DHPAA, HPPA (5 μg/ml) or control vehicle (Dulbecco's Modified Eagle Medium) for 24 hours. Experiments were repeated 3 times. Cell viability was assessed by fluorescence microscopy. Cell RNA was extracted and used for miRNA profiling via NanoString Technology. Data were processed and normalized in nSolverTM 4.0, statistics and hierarchical clustering were done with R and ClustVis. Gene targets were predicted with miRNet and pathway enrichment analysis was done in PathDIP. Results The treatments had no effect on cell viability. Of the 829 miRNAs assessed, 248 were expressed. Five miRNAs were differentially expressed among groups (ANOVA, P < 0.01; FDR < 5%). Unsupervised hierarchical clustering with these miRNAs revealed perfect separation based on treatment. These miRNAs were found to target 686 genes, enriched in 116 KEGG pathways including “miRNAs in cancer” and “pathways in cancer”. Treatment-specific responses included increased miR-2116-5p, miR-6721-5p, and miR-1290 for PAC, DHPAA, and HPPA, respectively. Pathways in glucagon signaling and central carbon metabolism in cancer were enriched in response to DHPPA only. Conclusions Cranberry PAC and polyphenol metabolites at concentrations representing dietary intakes elicit different miRNA signatures in colonic cells, providing a novel mechanism to explain their effects on intestinal health. Cranberry-mediated miRNA modulation may represent a potential strategy for preventing chronic disease. Funding Sources NSERC, Ocean Spray Cranberries, Inc., NSERC Graduate Scholarship.

Cranberry polyphenols‐gut microbiota interactions and potential health benefits: An updated review

“…Recent studies have shown that cranberry polyphenols gut metabolites 3,4-dihydroxyphenylacetic acid and 3-(4-hydroxyphenyl) propionic acid may mediate prebiotic effects by modulating small noncoding endogenous microRNA (miRNA) profiles in colonic cells (Lofft, Taibi, & Comelli, 2020). However, extrapolating these in vitro results to explain cranberry effects on human health is difficult and warrant further preclinical studies, particularly pharmacokinetics, and metabolism studies of these metabolites in vivo.…”

“…Similarly, probiotic metabolites of cranberry polyphenols by Lactobacillus rhamnosus, such as 4-hydroxyphenylacetic acid, have cell growth inhibitory effects against hepatocellular carcinoma HepG2 cells in vitro (Rupasinghe, Parmar, & Neir, 2019). As for other polyphenol metabolites, structural changes during metabolic transformation of CPs may lead to increased or decreased activities (van Duynhoven et al, 2011).Recent studies have shown that cranberry polyphenols gut metabolites 3,4-dihydroxyphenylacetic acid and 3-(4-hydroxyphenyl) propionic acid may mediate prebiotic effects by modulating small noncoding endogenous microRNA (miRNA) profiles in colonic cells(Lofft, Taibi, & Comelli, 2020). However, extrapolating these in vitro results to explain cranberry effects on human health is difficult and warrant further preclinical studies, particularly pharmacokinetics, and metabolism studies of these metabolites in vivo.In recent years, it is becoming increasingly clear that the composition of gut microbiota is highly individualized and associated with the host health.…”

Interplay between gut microbiota and the master iron regulator, hepcidin, in the pathogenesis of liver fibrosis

“…Another possible factor is a mutation on TP53, the gene encoding P53, which activates HAMP transcription (Hussain et al 2007 , Shen et al 2014 ). Also, there may be an elevation of matriptase-2 expression, which acts as a negative regulator for hepcidin expression (Lofft et al 2020 ).…”