Fecal Metabolome Research Articles

The anti-cholestatic effects of Coptis chinensis Franch. alone and combined with Tetradium ruticarpum (A. Jussieu) T. G. Hartley: dual effects on fecal metabolism and microbial diversity.

Introduction: Drug dosages and combinations are the main factors that affect the efficacy of pleiotropic traditional Chinese medicine (TCM). Coptis chinensis Franch. (CF) is a representative TCM with multiple effects and is often combined with Tetradium ruticarpum (A. Jussieu) T. G. Hartley (TR) to treat cholestasis. The present study assessed the influence of CF dose and its combination with TR on the efficacy of CF in cholestasis treatment, including their effects on fecal metabolism and fecal microorganisms. Methods: Rats with α-naphthylisothiocyanate (ANIT, 50mg/kg)-induced cholestasis were administered low (0.3g/kg) and high (0.6g/kg) doses of CF, as well as CF combined with TR at doses of 0.6g/kg and 0.9g/kg, respectively. The anti-cholestatic effects of these treatments were assessed by determining their anti-inflammatory, hypolipidemic, and anti-oxidative stress properties. Additionally, fecal metabolomics and fecal microorganisms were analyzed. Results: Low dose CF had a more potent hypolipidemic effect than high dose CF, whereas high dose CF had more potent anti-inflammatory and anti-oxidative stress effects. Combination with TR enhanced the hypolipidemic effect, but antagonized the anti-inflammatory effect, of CF. Analyses of fecal metabolomics and fecal microorganisms showed differences in the regulation of lipid- and amino acid metabolism-related pathways, including pathways of linoleic acid, tyrosine, and arachidonic acid metabolism, and amino acid biosynthesis between different doses of CF as well as between different doses of CF in combination with TR. These differences may contribute to differences in the anti-cholestatic effects of these preparations. Conclusion: CF dose influences its anti-cholestatic efficacy. The combination with TR had synergistic or antagonistic effects on the properties of CF, perhaps by altering fecal metabolism and fecal microbial homeostasis.

Gut microbiota and metabolites in patients with COVID-19 are altered by the type of SARS-CoV-2 variant.

Patients with COVID-19 have dysbiosis of the intestinal microbiota with altered metabolites in the stool. However, it remains unclear whether the differences among SARS-CoV-2 variants lead to differences in intestinal microbiota and metabolites. Thus, we compared the microbiome and metabolome changes for each SARS-CoV-2 variant in patients with COVID-19. We conducted a multicenter observational study of patients with COVID-19 and performed fecal microbiome, metabolome, and calprotectin analyses and compared the results among the different SARS-CoV-2 variants. Twenty-one patients with COVID-19 were enrolled and stratified according to the SARS-CoV-2 strain: six with the Alpha, 10 with the Delta, and five with the Omicron variant. Fecal microbiome analysis showed that α-diversity was reduced in the order of the Omicron, Delta, and Alpha variants (p = 0.07). Linear discriminant analysis revealed differences in the abundance of short-chain fatty acid-producing gut microbiota for each SARS-CoV-2 variant. Fecal metabolome analysis showed that the Omicron and Delta variants had markedly reduced propionic and lactic acid levels compared to the Alpha strain (p < 0.05). The intestinal microbiota of patients with COVID-19 varies depending on the SARS-CoV-2 variant. Dysbiosis of the intestinal microbiota due to differences in SARS-CoV-2 variants causes a decrease in intestinal short-chain fatty acids.

Imbalances in the gut microbiota and metabolome are associated with functional constipation in pregnancy

Functional constipation in pregnancy (FCP) is a common gastrointestinal disorder unique to pregnant women. The gut microbiota supports gastrointestinal function and maintains immune homeostasis, suggesting gut microbiota can become the target for FCP prevention and treatment. However, it remains unclear whether and how the gut microbiota and its correlated metabolites change for FCP. In this study, we aimed to explore the role of the gut microbiota in the pathogenesis of FCP and provide a reference for the selection of probiotics in further clinical trials. We comprehensively profiled the gut microbiota, Bifdobacterium and Bacteroides communities via high throughput sequencing, and faecal metabolome based on untarged metabolomics and targeted metabolomics of short-chain fatty acids (SCFAs) in 100 patients with FCP and 100 matched pregnant controls. We found that gut dysbiosis occurs in FCP patients, characterised by a reduced abundance of SCFAs-producing bacteria (e.g., Faecaliberium) and the enrichment of several opportunistic bacteria (e.g., Escherichia-Shigella), which were highly correlated with the clinical indicators of FCP. Furthermore, a reduced abundance of anti-inflammatory metabolites was seen, particularly arachidonic acid (AA) and butyric acid. We observed a complex co-occurrence between FCP-associated microorganisms and discriminative metabolites in an integrated network, with AA and Ruminococcaceae UCG-002 being identified as the main contributors. Collectively, imbalances in the gut microbiota and metabolome are associated with functional constipation in pregnancy. These findings provide a greater understanding of the pathogenesis and targets for probiotic interventions in FCP.

Study on the intervention effect of Epimedium before and after suet-oil-processed on kidney yang deficiency rats based on intestinal flora and fecal metabolomics

Epimedium is a Chinese herbal medicine commonly used in clinical practice to reinforce yang. Previous studies have shown that Epimedium fried with suet oil based has the best effect on warming kidney and promoting yang. Evidence suggests a relationship between kidney yang deficiency syndrome (KYDS) and metabolic disorders of the intestinal microflora. However, the specific interaction between KYDS and the intestinal microbiome, as well as the internal regulatory mechanism of the KYDS intestinal microbiome regulated by Epimedium fried with suet oil, remain unclear. The purpose of this study was to investigate the regulatory effects of different processed products of Epimedium on intestinal microflora and metabolites in rats with kidney yang deficiency, and to reveal the processing mechanism of Epimedium fried with suet oil warming kidney and helping yang. 16S rRNA and LC-MS/MS technology were used to detect fecal samples. Combined with multivariate statistical analysis, differential intestinal flora and metabolites were screened. Then the content of differential bacteria was then quantified using quantitative real-time fluorescence PCR. Furthermore, the correlation between differential bacterial flora and metabolites was analyzed using Spearman's method. The study found that the composition of intestinal flora in rats with kidney yang deficiency changed compared to healthy rats. Epimedium fried with suet oil could increase the levels of beneficial bacteria, while significantly reducing the levels of harmful bacteria. Real-time quantitative PCR results were consistent with 16S rRNA gene sequencing analysis. Fecal metabolomics revealed that KYDS was associated with 30 different metabolites, involving metabolic pathways steroid hormone biosynthesis etc. Moreover, differential bacteria were closely correlated with potential biomarkers. Epimedium could improve metabolic disorders associated with KYDS by acting on the intestinal flora, with Epimedium fried with suet oil demonstrating the most effective regulatory effect. Its potential mechanism may involve the regulation of abnormal metabolism and the impact on the diversity and structure of the intestinal flora.

Integrated 16S rRNA sequencing and metabolomic analysis reveals the potential protective mechanism of Germacrone on diabetic nephropathy in mice.

Diabetic nephropathy (DN) is a severe complication of diabetes and the leading cause of end-stage renal disease and death. Germacrone (Ger) possesses anti-inflammatory, antioxidant and anti-DN properties. However, it is unclear whether the improvement in kidney damage caused by Ger in DN mice is related to abnormal compositions and metabolites of the gut microbiota. This study generates a mouse model of DN to explore the potent therapeutic ability and mechanism of Ger in renal function by 16S rRNA sequencing and untargeted fecal metabolomics. Although there is no significant change in microbiota diversity, the structure of the gut microbiota in the DN group is quite different. Serratia_marcescens and Lactobacillus_iners are elevated in the model group but significantly decreased after Ger intervention ( P<0.05). Under the treatment of Ger, no significant differences in the diversity and richness of the gut microbiota are observed. An imbalance in the intestinal flora leads to the dysregulation of metabolites, and non-targeted metabolomics data indicate high expression of stearic acid in the DN group, and oleic acid could serve as a potential marker of the therapeutic role of Ger in the DN model. Overall, Ger improves kidney injury in diabetic mice, in part potentially by reducing the abundance of Serratia_marcescens and Lactobacillus_iners, as well as regulating the associated increase in metabolites such as oleic acid, lithocholic acid and the decrease in stearic acid. Our research expands the understanding of the relationship between the gut microbiota and metabolites in Ger-treated DN. This contributes to the usage of natural products as a therapeutic approach for the treatment of DN via microbiota regulation.

Efficacy Assessment of the Co-Administration of Vancomycin and Metronidazole in Clostridioides difficile-Infected Mice Based on Changes in Intestinal Ecology.

Vancomycin (VAN) and metronidazole (MTR) remain the current drugs of choice for the treatment of non-severe Clostridioides difficile infection (CDI); however, while their co-administration has appeared in clinical treatment, the efficacy varies greatly and the mechanism is unknown. In this study, a CDI mouse model was constructed to evaluate the therapeutic effects of VAN and MTR alone or in combination. For a perspective on the intestinal ecology, 16S rRNA amplicon sequencing and non-targeted metabolomics techniques were used to investigate changes in the fecal microbiota and metabolome of mice under the co-administration treatment. As a result, the survival rate of mice under co-administration was not dramatically different compared to that of single antibiotics, and the former caused intestinal tissue hyperplasia and edema. Co-administration also significantly enhanced the activity of amino acid metabolic pathways represented by phenylalanine, arginine, proline, and histidine, decreased the level of deoxycholic acid (DCA), and downregulated the abundance of beneficial microbes, such as Bifidobacterium and Akkermansia. VAN plays a dominant role in microbiota regulation in co-administration. In addition, co-administration reduced or increased the relative abundance of antibiotic-sensitive bacteria, including beneficial and harmful microbes, without a difference. Taken together, there are some risks associated with the co-administration of VAN and MTR, and this combination mode should be used with caution in CDI treatment.

Multi-omic analysis of precocious puberty girls: pathway changes and metabolite validation.

Precocious puberty (PP) is a prevalent endocrine disorder affecting the physical and mental wellbeing of children. Identifying the triggering factors of PP has become a central issue. This study seeks to investigate the metabolomic and transcriptomic alterations in PP. First, 37 school-aged girls diagnosed with PP and 25 age-matched prepubertal control girls were recruited, and the fecal samples were collected for non-targeted metabolomic analysis to screen for differentially expressed metabolites (DEMs). Subsequently, an animal model of PP was constructed by danazol administration to neonatal female rats, and both fecal non-targeted metabolomics and serum next-generation transcriptomic sequencing were performed to screen DEMs and differentially expressed genes (DEGs) in PP. Moreover, the DEM co-existing in clinical and animal models was administrated to PP rats to explore the role of the target metabolite in PP. A total of 24 DEMs in PP clinical samples and 180 DEMs and 425 DEGs in PP animal samples were identified. The Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis showed that these DEMs and DEGs were enriched in disease-associated pathways, including fatty acid synthesis, glycerolipid metabolism, pyrimidine metabolism, steroid hormone biosynthesis, progesterone-mediated oocyte maturation, and gonadotropin-releasing hormone (GnRH) signaling pathway, forming a tight DEM-DEG pathway regulatory network. Further DEM validation demonstrated that thymine supplementation delayed the opening of the vagina and development of PP in model rats. This study reveals that the metabolomic and transcriptomic changes, along with enriched pathways, are implicated in PP based on clinical and animal analyses. The findings may provide new strategies and research avenues for PP treatment.

Altered gut mycobiome in patients with end-stage renal disease and its correlations with serum and fecal metabolomes

BackgroundThe relationship between the gut mycobiome and end-stage renal disease (ESRD) remains largely unexplored.MethodsIn this study, we compared the gut fungal populations of 223 ESRD patients and 69 healthy controls (HCs) based on shotgun metagenomic sequencing data, and analyzed their associations with host serum and fecal metabolites.ResultsOur findings revealed that ESRD patients had a higher diversity in the gut mycobiome compared to HCs. Dysbiosis of the gut mycobiome in ESRD patients was characterized by a decrease of Saccharomyces cerevisiae and an increase in various opportunistic pathogens, such as Aspergillus fumigatus, Cladophialophora immunda, Exophiala spinifera, Hortaea werneckii, Trichophyton rubrum, and others. Through multi-omics analysis, we observed a substantial contribution of the gut mycobiome to host serum and fecal metabolomes. The opportunistic pathogens enriched in ESRD patients were frequently and positively correlated with the levels of creatinine, homocysteine, and phenylacetylglycine in the serum. The populations of Saccharomyces, including the HC-enriched Saccharomyces cerevisiae, were frequently and negatively correlated with the levels of various toxic metabolites in the feces.ConclusionsOur results provided a comprehensive understanding of the associations between the gut mycobiome and the development of ESRD, which had important implications for guiding future therapeutic studies in this field.

Children with idiopathic short stature have significantly different gut microbiota than their normal height siblings: a case-control study.

To investigate the role of gut microbiota (GM) in pathogenesis of idiopathic short stature (ISS) by comparing GM of ISS children to their normal-height siblings. This case-control study, conducted at the Schneider Children's Medical Center's Institute for Endocrinology and Diabetes between 4/2018-11/2020, involved 30 pairs of healthy pre-pubertal siblings aged 3-10 years, each comprising one sibling with ISS and one with normal height. Outcome measures from fecal analysis of both siblings included GM composition analyzed by 16S rRNA sequencing, fecal metabolomics, and monitoring the growth of germ-free (GF) mice after fecal transplantation. Fecal analysis of ISS children identified higher predicted levels of genes encoding enzymes for pyrimidine, purine, flavin, coenzyme B, and thiamine biosynthesis, lower levels of several amino acids, and a significantly higher prevalence of the phylum Euryarchaeota compared to their normal-height siblings (p<0.001). ISS children with higher levels of Methanobrevibacter, the dominant species in the archaeal gut community, were significantly shorter in stature than those with lower levels (p=0.022). Mice receiving fecal transplants from ISS children did not experience stunted growth, probably due to the eradication of Methanobrevibacter caused by exposure to oxygen during fecal collection. Our findings suggest that different characteristics in the GM may explain variations in linear growth. The varying levels of Methanobrevibacter demonstrated within the ISS group reflect the multifactorial nature of ISS and the potential ability of the GM to partially explain growth variations. The targeting of specific microbiota could provide personalized therapies to improve growth in children with ISS.

The differential effect of two cereal foods on gut environment: a randomized, controlled, double-blind, parallel-group study.

Cereal-based foods such as fruit granola (FG) and corn flakes (CF) form part of a fiber-rich diet. Dietary fiber has a good effect on human health. However, changes in gut microbiota and intestinal immunity have not been investigated. We conducted a randomized, double-blind, placebo-controlled trial to investigate the effects of FG and CF intake on gut microbiota, metabolome, and the immune system. Subjects continuously consume CF or FG for 4 weeks. Stool samples, and questionnaires on defecation were collected before, 2 weeks after, and 4 weeks after intake. Gut microbiota was analyzed using 16S rRNA gene amplicon sequencing. Fecal metabolomes were analyzed using GC/MS and CE-TOF/MS. Fecal IgA was analyzed using ELISA. The defecation frequency after cereal based food intake was improved. The different cereal-based foods had different effects on gut microbiome. The increase in intestinal IgA levels was positively correlated with the relative abundance of Dialister and the Lachnospiraceae ND3007 group in CF and FG group, respectively. SCFAs showed a positive correlation with Prevotella 9 in the FG group. This study showed that the supplement in dietary fiber contained in CF and FG improves bowel movements. CF and FG each had different effects on gut microbes, metabolites and different relationships between fecal IgA or SCFAs and gut microbiota.

Electroacupuncture at ST25 corrected gut microbial dysbiosis and SNpc lipid peroxidation in Parkinson's disease rats.

Parkinson's disease (PD) remains one kind of a complex, progressive neurodegenerative disease. Levodopa and dopamine agonists as widely utilized PD therapeutics have not shown significant positive long-term outcomes. Emerging evidences indicate that electroacupuncture (EA) have potential effects on the therapy of nervous system disorders, particularly PD, but its specific underlying mechanism(s) remains poorly understood, leading to the great challenge of clinical application and management. Previous study has shown that acupuncture ameliorates PD motor symptoms and dopaminergic neuron damage by modulating intestinal dysbiosis, but its intermediate pathway has not been sufficiently investigated. A rat model of PD was induced using rotenone. The therapeutic effect of EA on PD was assessed using the pole and rotarod tests and immunohistostaining for tyrosine hydroxylase (TH) in the substantia nigra (SN) of brain. The role of gut microbiota was explored using 16S rRNA gene sequencing and metabonomic analysis. PICRUSt2 analysis, lipidomic analysis, LPS and inflammatory factor assays were used for subsequent exploration and validation. Correlation analysis was used to identify the key bacteria that EA regulates lipid metabolism to improve PD. The present study firstly reappeared the effects of EA on protecting motor function and dopaminergic neurons and modulation of gut microbial dysbiosis in rotenone-induced PD rat model. EA improved motor dysfunction (via the pole and rotarod tests) and protected TH+ neurons in PD rats. EA increased the abundance of beneficial bacteria such as Lactobacillus, Dubosiella and Bifidobacterium and decreased the abundance of Escherichia-Shigella and Morganella belonging to Pseudomonadota, suggesting that the modulation of gut microbiota by EA improving the symptoms of PD motility via alleviating LPS-induced inflammatory response and oxidative stress, which was also validated by various aspects such as microbial gene functional analysis, fecal metabolomics analysis, LPS and inflammatory factor assays and SNpc lipidomics analysis. Moreover, correlation analyses also verified strong correlations of Escherichia-Shigella and Morganella with motor symptoms and SNpc lipid peroxidation, explicating targets and intermediate pathways through which EA improve PD exercise symptom. Our results indicate that the improvement of motor function in PD model by EA may be mediated in part by restoring the gut microbiota, which intermediate processes involve circulating endotoxins and inflammatory mediators, SNpc oxidative stress and lipid peroxidation. The gut-microbiome - brain axis may be a potential mechanism of EA treatment for the PD.

Serum aryl hydrocarbon receptor activity is associated with survival in patients with alcohol-associated hepatitis.

Patients with alcohol-associated hepatitis (AH) have an altered fecal metabolome, including reduced microbiota-derived tryptophan metabolites, which function as ligands for aryl hydrocarbon receptor (AhR). The aim of this study was to assess serum AhR ligand activity in patients with AH. The study included 74 controls without AUD, 97 patients with AUD, and 330 patients with AH from 2 different multicenter cohorts (InTeam: 134, AlcHepNet: 196). Serum AhR activity was evaluated using an AhR reporter assay with HepG2-Lucia cells incubated with serum for 24 hours. Serum AhR activity was significantly higher in patients with AH compared with both controls (1.59 vs. 0.96-fold change, p < 0.001) and patients with AUD (1.59 vs. 0.93, p < 0.001). In both AH cohorts, patients with AhR activity ≥ 2.09 had significantly lower cumulative survival rates at 30, 60, 90, and 180 days compared to those with AhR activity < 2.09. When serum AhR activity was used to further stratify patients with severe AH, the cumulative 30, 60, 90, and 180-day survival rates for patients with severe AH and the AhR activity ≥ 2.09 group were all significantly lower than those with an AhR activity < 2.09 group. Serum AhR activity was significantly higher in patients with AH compared with controls and individuals with AUD, and this increased activity was associated with higher mortality. Consequently, serum AhR activity holds potential as a prognostic marker.

Effects of radiofrequency field from 5G communication on fecal microbiome and metabolome profiles in mice

With the rapid development of 5G networks, the influence of the radiofrequency field (RF) generated from 5G communication equipment on human health is drawing increasing attention in public. The study aimed at assessing the effects of long-term exposure to 4.9 GHz (one of the working frequencies of 5G communication) RF field on fecal microbiome and metabolome profiles in adult male C57BL/6 mice. The animals were divided into Sham group and radiofrequency group (RF group). For RF group, the mice were whole body exposed to 4.9 GHz RF field for three weeks, 1 h/d, at average power density (PD) of 50 W/m2. After RF exposure, the mice fecal samples were collected to detect gut microorganisms and metabolites by 16S rRNA gene sequencing and LC–MS method, respectively. The results showed that intestinal microbial compositions were altered in RF group, as evidenced by reduced microbial diversity and changed microbial community distribution. Metabolomics profiling identified 258 significantly differentially abundant metabolites in RF group, 57 of which can be classified to Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways. Besides, functional correlation analysis showed that changes in gut microbiota genera were significantly correlated with changes in fecal metabolites. In summary, the results suggested that altered gut microbiota and metabolic profile are associated with 4.9 GHz radiofrequency exposure.

Integrating fecal metabolomics and intestinal microbiota to study the mechanism of cannabidiol in the treatment of idiopathic pulmonary fibrosis.

Introduction: Idiopathic pulmonary fibrosis is a chronic interstitial lung disease characterized by excessive deposition of extracellular matrix. Cannabidiol, a natural component extracted from plant cannabis, has been shown to have therapeutic effects on lung diseases, but its exact mechanism of action is unknown, hindering its therapeutic effectiveness. Methods: To establish a pulmonary fibrosis model, combined with UPLC-Q-TOF/MS metabolomics and 16S rDNA sequencing, to explore cannabidiol's mechanism in treating pulmonary fibrosis. The rats were randomly divided into the control group, pulmonary fibrosis model group, prednisone treatment group, and cannabidiol low, medium, and high dose groups. The expression levels of HYP, SOD, and MDA in lung tissue and the expression levels of TNF-α, IL-1β, and IL-6 in serum were detected. Intestinal microbiota was detected using UPLC-QTOF/MS analysis of metabolomic properties and 16S rDNA sequencing. Results: Pathological studies and biochemical indexes showed that cannabidiol treatment could significantly alleviate IPF symptoms, significantly reduce the levels of TNF-α, IL-1β, IL-6, MDA, and HYP, and increase the expression level of SOD (p < 0.05). CBD-H can regulate Lachnospiraceae_NK4A136_group, Pseudomonas, Clostridia_UCG-014, Collinsella, Prevotella, [Eubacterium]_coprostanoligenes_group, Fusobacterium, Ruminococcus, and Streptococcus, it can restore intestinal microbiota function and reverse fecal metabolism trend. It also plays the role of fibrosis through the metabolism of linoleic acid, glycerol, linolenic acid, and sphingolipid. Discussion: Cannabidiol reverses intestinal microbiota imbalance and attenuates pulmonary fibrosis in rats through anti-inflammatory, antioxidant, and anti-fibrotic effects. This study lays the foundation for future research on the pathological mechanisms of IPF and the development of new drug candidates.

In-depth investigation of the therapeutic effect of Tribulus terrestris L. on type 2 diabetes based on intestinal microbiota and feces metabolomics

Ethnopharmacological relevanceThe fruit of Tribulus terrestris L. (TT) is extensively documented in the Tibetan medical literature 'Si Bu Yi Dian', has been used to treat diabetes mellitus for more than a thousand years. However, the underlying mechanisms and comprehensive effects of TT on diabetes have yet to be investigated. Aim of the studyThe aim of the study was to systemically elucidate the potential mechanisms of TT in treating diabetes mellitus, and further investigate the therapeutic effects of the water extract, small molecular components and saccharides from TT. Materials and methodsFecal metabolomics was employed to draw the metabolic profile based on UHPLC-Q-TOF-MS/MS. The V3–V4 hypervariable regions of the bacteria 16S rRNA gene were amplified to explore the structural changes of the intestinal microbiome after TT intervention and to analyze the differential microbiota. The microbial metabolites SCFAs were determined by GC-MS, and the BAs and tryptophan metabolites were quantified by UPLC-TQ-MS. Spearman correlation analysis was carried out to comprehensively investigate the relationship among the endogenous metabolites profile, intestinal microbiota and their metabolites. ResultsTT exhibited remarkably therapeutic effect on T2DM rats, as evidenced by improved glucolipid metabolism and intestinal barrier integrity, ameliorated inflammation and remission in insulin resistance. A total of 24 endogenous biomarkers were screened through fecal metabolomics studies, which were mainly related to tryptophan metabolism, fatty acid metabolism, bile acid metabolism, steroid hormone biosynthesis and arachidonic acid metabolism. Investigations on microbiomics revealed that TT significantly modulated 18 differential bacterial genera and reversed the disordered gut microbial in diabetes rats. Moreover, TT notably altered the content of gut microbiota metabolites, both in serum and fecal samples. Significant correlation among microbial community, metabolites and T2DM-related indicators was revealed. ConclusionsThe multiple components of TT regulate the metabolic homeostasis of the organism and the balance of intestinal microbiota and its metabolites, which might mediate the anti-diabetic capacity of TT.

Fibre-rich Foods to Treat Obesity and Prevent Colon Cancer trial study protocol: a randomised clinical trial of fibre-rich legumes targeting the gut microbiome, metabolome and gut transit time of overweight and obese patients with a history of noncancerous adenomatous polyps

IntroductionRecently published studies support the beneficial effects of consuming fibre-rich legumes, such as cooked dry beans, to improve metabolic health and reduce cancer risk. In participants with overweight/obesity and a...

Effect of Lactiplantibacillus plantarum CCFM8661 on serum metabolites and gut microbiota in a lead-exposed population

In this study, we investigated the impact of Lactiplantibacillus plantarum (L. plantarum) CCFM8661 on the gut microbiota, and the serum and fecal metabolomes in lead (Pb)-exposed individuals. The volunteers recruited for this study were divided into two treatment groups, (i) the placebo (control) and (ii) the L. plantarum CCFM8661 treatment groups. The analysis revealed that probiotic intervention reversed some of the changes in Pb exposure-induced intestinal bacterial abundance, including the abundance of Parabacteroides, Bacteroides, Clostridiaceae, and Erysipelotrichaceae. An analysis of the fecal metabolome identified 26 differential metabolites involved in purine metabolism, unsaturated fatty acid metabolism, and other pathways. Serum metabolite analysis showed that L. plantarum CCFM8661 treatment altered the serum metabolite levels of various metabolic pathways, such as the glycerophospholipid, amino acid, and glutathione metabolism pathways. These results suggest that L. plantarum CCFM8661 may have beneficial effects on Pb-exposed populations by modulating the gut microbiota, host serum metabolism, and the metabolism of the gut microbiota.

P030 Gut-host metabolites and microbiota interactions in a murine model of checkpoint inhibitor induced colitis

Abstract Background The advent of Immune Checkpoint Inhibitors (CPIs) has revolutionised cancer treatment by bolstering anti-tumour immune responses. However, the clinical success of CPIs is hampered by immune-related adverse events (irAEs), with colitis posing a common and formidable challenge. Despite this, the precise mechanisms triggering CPI-induced colitis remain elusive. Recognising the influence of gut microbiota on CPI therapy response and colitis development, this study seeks to unravel the intricate details of these mechanisms to facilitate the development of targeted therapies. Methods Utilising a murine model of CPI-induced colitis, we combined CPI therapy with faecal microbial transplant (FMT) of colitogenic bacteria. FMT induced colitogenic bacteria in wild-type mice, followed by weekly injections of anti-CTLA4 and anti-PD1 for 3 weeks. Metataxonomic data, obtained through 16S rRNA gene amplicon sequencing, and metabolomic profiles, captured by 1H-NMR spectroscopy, were analysed. Alpha and beta diversity, as well as differential expression of taxa, were assessed using 16S data. Metabolomics data underwent multivariate and univariate analyses to identify metabolic profiles and differential metabolites. Pathway analysis was conducted with MetaboAnalyst. Correlation analyses were performed between bacterial taxa at the family level and metabolite concentrations. Results After one week of FMT and CPI treatment, distinct profiles in faecal metabolome and microbiome were observed through Principal Component Analysis (PCA) and Principal Coordinates Analysis (PCoA) for beta-diversity, respectively. Altered metabolites included increased trimethylamine, methylamine, pyruvate, and cytidine, along with reduced tryptophan and proline. Changes in bacterial composition were noted, with increased Bacteroides, Faecalibaculum, Mucispirillum, and Roseburia, and decreased Muribaculum. Metabolic differences persisted after 3 weeks of treatment, revealing additionally decreased acetate, propionate and tyramine. Pathway analysis highlighted profound alterations in tyrosine, alanine, aspartate and glutamate metabolism, pyrimidine metabolism and glycolysis. Desulfovibrionaceae was positively correlated with dimethylamine, glycine and valine. Bacteroidaceae was positively correlated with propionate and butyrate. Conclusion This study provides novel mechanistic insights into the metabolite and microbiota-metabolism pathways associated with CPI-induced colitis. The alterations in short-chain fatty acids (SCFA), amino acids, and the SCFA-producing bacteria and colitogenic bacteria shed light on the metabolic changes underlying CPI colitis. Targeting these microbiomes and metabolites could offer promising avenues for enhancing clinical outcomes during CPI therapy.

DOP09 Models for predicting Crohn Disease (CD) exacerbation using serum and fecal metabolomics

Abstract Background Metabolites, produced endogenously or derived from the gut microbiome, are key mediators and effectors of many cellular functions, but there are only scant data on their temporal changes along CD course. We analyzed the dynamics of serum and fecal metabolome in a prospective cohort of quiescent CD patients over 2 years or until the occurrence of a clinical flare, aiming to identify whether changes in the metabolome precede and predict a clinical relapse. Methods Untargeted metabolomics was performed using LC/MS. Multivariate analyses in MaAsLin2 were applied to identify differentially abundant metabolites between CD cases who relapsed and those who remained in remission. Our models used the "Leave-One-Out" approach to compensate for over-fitting bias. Results We included 110 serum samples and 232 fecal samples from 44 CD participants recruited in remission (mean age 31.73 years, 59% males; Table 1). The median follow-up time was 630 days (IQR 500-808). Demographic, biomarkers, and treatment exposures were not significantly different between the 11 that experienced flare during follow-up and the 33 that remained in remission throughout. We compared the metabolomics between relapsers and non-relapsers using only preflare samples after excluding samples from a particular subject in the "Leave-One-Out" approach. The top 3 metabolites in each direction and comparison and their abundance preflare in the non-relapsers and relapsers are shown (Fig. 1A-B). We then calculated a flare index for that excluded subject, using the sum peak area ratio of all metabolites that were increased and decreased preflare. A receiver operating characteristic (ROC) area under the curve (AUC) using the "individual" flare index after z-score normalization was able to predict a subsequent flare with AUC=0.73, 95% CI 0.52-0.93 for serum metabolomics (Fig. 1C) compared with AUC=0.62 (95% CI 0.40-0.84) for CRP and AUC=0.74 (95% CI 0.54-0.94) for FCP, in the same sampling cohort. For the stool sampling cohort, metabolomics predictive accuracy was AUC=0.75 (95% CI 0.5-0.99; Fig. 1D) compared with AUC=0.67 (95% CI 0.49-0.84) for CRP and AUC=0.71 (95% CI 0.55-0.86) for FCP. Survival plot analyses after selecting Youden point based on ROC (Fig. 1C-D) showed moderately better performance for fecal flare index with log-rank Hazard Ratio (HR) of 8.20 (95% CI 1.98-34.20), than for the serum with HR of 3.45 (95% CI 1.5-11.37). Conclusion In a prospective cohort of quiescent CD patients, we identified metabolites in the serum and feces that were incorporated in models that can predict CD flare. These can be used to guide personalized preemptive therapy intensification and provide insight into the underlying mechanism of CD flares.

P1206 Mechanistic implications of the Mediterranean diet in patients with newly diagnosed Crohn’s disease- multi-omic analysis of a prospective cohort

Abstract Background The Mediterranean diet (MED) is hypothesized to play a protective role in Crohn’s disease (CD) etiology, yet the underlying mechanisms remain unclear. Here we explored the association between adherence to MED, CD course, clinical and inflammatory markers, and microbial and metabolite composition in patients with newly diagnosed CD. Methods Patients with newly diagnosed CD were recruited and prospectively assessed. Food frequency questionnaires (FFQ) were collected each visit and assessed for adherence to MED, using a predefined IBDMED score, alongside validated MED adherence screeners. The Crohn’s disease activity index (CDAI) was used to evaluate clinical activity and serum and fecal samples collected for fecal calprotectin, C-reactive protein (CRP), and microbial composition using 16S rRNA sequencing. Baseline serum and fecal metabolomics were analyzed using a targeted quantitative metabolomics approach. Patients were classified as having a complicated course if during follow-up, they were steroid dependent, switched ≥2 biologics, hospitalized or underwent CD-related surgery. Results We recruited and prospectively observed 271 consecutive patients with newly diagnosed CD (52% males, average age- 31±12 years, inflammatory phenotype, B1- 75%) and collected 636 FFQs (range 1-5 FFQ per patient). Adherence to MED was associated with a non-complicated course and was inversely correlated with CDAI, fecal calprotectin, CRP and microbial dysbiosis index (all p&amp;lt;0.05). Increasing adherence to MED over time was associated with decreasing CDAI and fecal calprotectin in patients with complicated and non-complicated courses (p&amp;lt;0.05). Diet microbial interactions revealed two microbial clusters, showing that adherence to MED and intake of its recommended foods positively correlated with commensals and short-chain fatty acid producers including Faecalibacterium. Conversely, adherence to MED inversely correlated with a cluster of members of the Proteobacteria and included Ruminococcus gnavus. Diet metabolomics interactions revealed two clusters showing that adherence to MED positively correlated with multiple serum and fecal organic compounds, vitamin derivatives, plant metabolites, amino acids and tryptophan metabolites and inversely associated with fecal triglycerides, ceramides and cholic acid, the major primary bile acid (FDR&amp;lt;0.1). Conclusion Adherence to MED is associated with lower levels of primary bile acids and microbial dysbiosis and with a beneficial microbial and metabolite composition in patients with newly diagnosed CD. This may drive the inverse association of MED with clinical activity, inflammatory markers and complicated course. Microbial and metabolite-targeted dietary approaches like MED, should play a role in CD management.