Introduction

Inflammatory bowel disease (IBD) is a chronic inflammatory disease of the gastrointestinal tract that results from various factors, including genetic factors, inappropriate immune response, changes in gut microbiota balance, and oxidative stress.^{1, 2}Bifidobacterium longum is a representative bifidobacterium that exists in the human gastrointestinal tract and plays a significant role in protecting against colitis.³ Oxidative stress elicits an unfavorable environment for the survival and colonization of beneficial probiotics as well as causing tissue damage in colitis.^{4, 5}

Aims & Methods

In this study, we aimed to identify the genome characteristics, ROS-resistant properties, colonization abilities, and anti-colitic effects of the newly isolated B. longum strain (B. longum S2) from the feces of a healthy adult in South Korea. Viability of B. longum S2 against hydrogen peroxide was assessed using H₂O₂-containing media. The antioxidant activity of B. longum S2 was measured using the ABTS and DPPH assays. Colonization in the colon was observed using a scanning electron microscope. Whole genome sequencing, data assembly, and gene analysis were performed. The anti-colitic effects were evaluated in mice using 2.5% DSS-induced colitis and adoptive T cell transfer models. Bifidobacterium was administrated by oral gavage 1 × 10⁹ CFU and the disease activity index (DAI) was scored. Colon length was measured and the histopathologic score was assessed with PAS-stained colon tissue. T cells in lamina propria mononuclear cells from the small intestine, lymphocytes from the spleen, and peritoneal cavity cells were analyzed using flow cytometry. Epithelial barrier function was evaluated using a FITC-dextran permeability assay on Caco-2 cells. Gene expression and cytokine profiles were analyzed using qRT-PCR.

Results

B. longum S2 colonies in H₂O₂-containing media were observed more frequently than those of other bifidobacterium strains. The antioxidant activity of B. longum S2 on ABTS and DPPH radicals was higher than that exhibited by other bifidobacterium strains. We identified that B. longum S2 possesses oxidative response genes, such as LexA and HrcA, through pan-genome and comparative genome analysis. However, the average nucleotide identity highlights a 94.92% value with respect to B. longum ATCC, 98.65% with respect to B. longum JCM1217 and 98.64% with respect to B. longum NCTC 11818, supporting the characteristics of B. longum S2 as a novel strain of bifidobacteria. B. longum S2-treated group showed alleviation of the colitis index, including a higher survival rate, restoration of body weight loss and colon shortening, and improvement of the DAI compared to the control group. B. longum S2 treatment significantly decreased the loss of goblet cell and the histopathological score in colon tissues. B. longum S2 induced the expression of markers of Treg cells and M2 macrophages in the isolated mouse tissues. Moreover, the B. longum S2-treated group showed decreased gene expression of pro-inflammatory cytokines, including mucin gene, compared to the control group. Pre-treatment with B. longum S2 significantly reduced the TNF-α-induced permeability of intestinal epithelial cells, suggesting that B. longum S2 improves epithelial barrier function.

Conclusion

Our study revealed that the newly isolated B. longum strain, B. longum S2, possesses unique anti-colitic effects and colonization abilities, as demonstrated in both in vitro experiments and in two different mouse colitis models. These findings strongly suggest that B. longum S2 holds great promise as a potential treatment for IBD.

References

1. Khor B, Gardet A, Xavier RJ. Genetics and pathogenesis of inflammatory bowel disease. Nature 2011;474:307-17.
2. Blumberg RS. Environment and Genes: What Is the Interaction? Dig Dis 2016;34:20-6.
3. Yao S, Zhao Z, Wang W, et al. Bifidobacterium Longum: Protection against Inflammatory Bowel Disease. J Immunol Res 2021;2021:8030297.
4. Sanz Y, De Palma G. Gut microbiota and probiotics in modulation of epithelium and gut-associated lymphoid tissue function. Int Rev Immunol 2009;28:397-413.
5. Xiao M, Xu P, Zhao J, et al. Oxidative stress-related responses of Bifidobacterium longum subsp. longum BBMN68 at the proteomic level after exposure to oxygen. Microbiology (Reading) 2011;157:1573-1588.

Introduction

Hericium erinaceus (H. erinaceus), berberine and quercetin showed anti-inflammatory potential in mouse models of experimental colitis. H. erinaceus (class Agaricomycetes, phylum Basidiomycota) is a mushroom belonging to traditional Chinese medicine that has been demonstrated in experimental colitis to be able to control pathways induced by Nuclear Factor kappa-light-chain-enhancer of activated B cells in an anti-inflammatory direction by suppressing the production of Tumor Necrosis Factor (TNF) and increasing the expression of Interleukin-10 (IL-10).

Aims & Methods

This ex vivo study aimed to evaluate the anti-inflammatory potential of a nutraceutical compound of H. erinaceus, berberine, quercetin, biotin and niacin (HBQ-Complex®) in Inflammatory bowel diseases (IBD) tissues obtained from both Normal-Appearing Mucosa (NAM) and Inflamed Mucosa (IM) tracts of patients with Crohn's Disease (CD) and patients with Ulcerative Colitis (CD). NAM tracts were identified from colic segments with Simple Endoscopic Score for CD = 0 or Mayo Endoscopic subscore = 0 (for CD and UC, respectively). NAM and one IM samples were frozen at -70° and then evaluated at T0. Two other IM samples were grown in DMEM medium in a temperature-controlled incubator and exposed to HBQ-Complex® for 120 minutes (T1) and 180 minutes (T2). RT-PCR and Western blot evaluations were performed for TNF, IL-10 and cyclooxygenase 2 (COX-2) at T0, T1, and T2. The HBQ-Complex® consisted of 525 mg of H. erinaceus powder (5 % polysaccharides from sporophorum) and 225 mg of H. erinaceus as an extract (30 % polysaccharides from sporophorum), 75 mg of quercetin titled to 98 %, 225 μg of biotin, 27 mg of niacin and, finally, 75 mg of Berberis vulgaris titled to 97 %.

Results

20 IBD treatment-naïve patients (50% CD, 50% UC) were enrolled. None of them had other comorbidities or was taking drug therapies within six months of the start of the study. According to the Harvey-Bradshaw index and partial Mayo score, four patients had mild CD and six moderate CD. The same was for UC patients. At T0, both CD and UC samples showed TNF and COX-2 genic expressions higher in IM than NAM (p < 0.0001). In IM samples, Incubation with HBQ-Complex® resulted in a progressive decrease in gene and protein COX-2 and TNF expression at T1/T2 in CD (p=0.002) and UC (p=0.003) samples. However, IL-10 showed an opposite trend, with a progressive genic expression increase from T0 to T1 (p=0.054) and T1 to T2 (p=0.002). The same was for UC samples (p < 0.01). These variations in PCR expressions of mRNAs were also recalculated from the same protein variations at Western blot analysis.

Conclusion

HBQ-Complex® might possess nutraceutical therapeutic potential in IBD. This could be explored in translational studies evaluating whether HBQ-Complex® can impact IBD's already validated clinical, endoscopic, and histological parameters.

References

- Diling C, Chaoqun Z, Jian Y, Jian L, Jiyan S, Yizhen X, et al. Immunomodulatory Activities of a Fungal Protein Extracted from Hericium erinaceus through Regulating the Gut Microbiota. Front Immunol. 2017;8:666.
- Ren Y, Geng Y, Du Y, Li W, Lu ZM, Xu HY, et al. Polysaccharide of Hericium erinaceus attenuates colitis in C57BL/6 mice via regulation of oxidative stress, inflammation-related signaling pathways and modulating the composition of the gut microbiota. J Nutr Biochem. luglio 2018;57:67–76.
- Qin M, Geng Y, Lu Z, Xu H, Shi JS, Xu X, et al. Anti-Inflammatory Effects of Ethanol Extract of Lion’s Mane Medicinal Mushroom, Hericium erinaceus (Agaricomycetes), in Mice with Ulcerative Colitis. Int J Med Mushrooms. 2016;18(3):227–34.

Introduction

Inflammatory bowel disease (IBD), comprising of Crohn’s disease and Ulcerative Colitis (UC), is a debilitating inflammatory disease of the gastrointestinal tract with no known cause. Although extensive work has linked numerous genetic loci to IBD, most of these associations remain poorly understood. In addition, some studies have associated IBD to metabolic diseases such as diabetes and non-alcoholic fatty liver disease (NAFLD), although the precise molecular and pathophysiological mechanisms underlying these associations are unknown.

Aims & Methods

The central objective of this study was to understand the molecular mechanisms underlying the associations between IBD and obesity related metabolic diseases. To address this research question, we used multiple mouse models of colitis (dextran sodium sulfate or DSS), DSS and Azoxymethane induced colitis associated colon cancer and Helicobacter hepaticus infection induced colitis in Il-10-/- mice) and high fat diet (HFD) and genetic models of obesity and obesity associated metabolic diseases. In addition, for mechanistic studies, we used chimeric mice generated through bone marrow transplantation as well as conditional knockout mouse models. Lastly, we employed CRISPR/Cas9 edited cells in vitro to understand the molecular mechanism.

Results

Employing multiple murine models of colitis, colitis-associated colon cancer (CAC) and obesity associated metabolic diseases, we have identified DUPD1, a phosphatase of unknown function, that links IBD with metabolic diseases. Specifically, Dupd1^-/-mice are protected from dextran sodium sulfate (DSS) and Helicobacter hepaticus induced colitis, and DSS/azoxymethane (AOM) induced CAC. Dupd1^-/- mice also exhibited protection against high fat diet (HFD) induced obesity and fatty liver disease and glucose intolerance. DUPD1 is highly expressed in the skeletal muscle. Indeed, using skeletal muscle specific conditional DUPD1 knockout mice (Dupd1^fl/flMyf6^Cre), we show that DUPD1 exerts its colitogenic effects from the skeletal muscle through modulation of autophagy, a pathway whose dysfunction is known to play a role in IBD. Importantly, a dual specificity phosphatase inhibitor (NSC-663284) inhibited DUPD1 enzyme activity and reduced DSS-induced colitis in wild type mice but not in Dupd1^-/-mice.

Conclusion

Our study identifies Dupd1 as a central gene that links IBD and obesity-associated metabolic diseases and suggests that it is a new therapeutic target in IBD, and obesity associated metabolic diseases.

Disclosure

Nothing to declare.

Introduction

Head-to-head (H2H) comparisons of biologics and small molecules in randomised clinical trials (RCTs) are considered the gold standard in inflammatory bowel disease (IBD). However, comparative real-world evidence (RWE) registry studies can provide important additional knowledge. Our aim is to perform a range of such H2H RWE studies, including subgroup analyses, to examine different treatments in patients with IBD. To do so, we draw upon data from a large data warehouse (UMBRELLA-IBD), which pools data from all eight prospective RWE registries in the Competence Network IBD. All patients were recruited to each registry in a similar manner.

Aims & Methods

Pooled analyses have a range of important methodological and logistical requirements, all of which are met in the eight prospective RWE registries that feed data into the UMBRELLA-IBD data warehouse: (1) The prospectively collected data are highly homogeneous, with each registry using similar inclusion and exclusion criteria, and the analyses of registry data using similar outcome parameters; (2) The quality of the data is high, and the data are comparable due to comprehensive remote and onsite monitoring; (3) The advanced statistical methods used to compensate for the absence of randomisation include propensity score adjustment with inverse probability of treatment weighting (IPTW) and are applied similarly in each analysis (Table 1). In concordance with a priori protocols, data and parameters from the eight Competence Network IBD registries were checked for consistency, merged and then migrated to a common server. Additionally, a data validation plan (DVP) was created to process all data, with individual items being adapted by appropriate recoding for the pooled evaluation.

Results

In the new UMBRELLA-IBD data warehouse of the Competence Network IBD, prospectively recorded data from a total of 6,689 IBD patients in eight registry studies in Germany are available for a planned pooled analysis (Table 1), albeit with some patients potentially having been recruited to more than one of these studies. As of 31 March 2023, 35% of the patients included in the data warehouse are biologic-naïve and started their first biologic/small molecule treatment at recruitment. Prospectively documented one-year records are available for 75% of patients, and supplementary biomaterials have been collected in 40% of patients. Recruitment and follow-up are ongoing in some registries.

Table 1: IBD patients in the prospective registries of the Competence Network IBD at a glance: Umbrella-IBD (as of 31 March 2023)
* Some patients may be recruited in more than one study

Conclusion

The large UMBRELLA-IBD RWE data warehouse from eight different prospective Competence Network IBD registries is a promising initiative for conducting further comparative studies that will provide important evidence in addition to that from existing RCTs in IBD patients.

Disclosure

BB has received consulting fees from: AbbVie, MSD, Shire, Ferring, Hospira, Takeda, Galapagos, Shield Therapeutics, Pfizer, Biogen, Janssen, Hexal, and Celgene, outside the submitted work. Financial support for lectures and teaching from: AbbVie, Ferring, MSD, Merckle, Falk, Galapagos, Shield Therapeutics, Pfizer, Celltrion, Takeda, and Janssen. Grant and financial support for research from AbbVie, Takeda, Jansen, Pfizer, Galapagos, and Ferring, outside the submitted work. SPD has no relevant financial or non-financial interests to disclose. TW has no relevant financial or non-financial interests to disclose. EG has no relevant financial or non-financial interests to disclose. FT receives scientific funding from Sanofi, received consulting fee's from LEK Consulting and speaker's fee from Lilly, Janssen, Falk, Abbvie, Celltrion, onkowissen.de, CED Service GmbH. AB received consulting and speaker's fee from Abbvie, Amgen, BMS, Falk, Ferring, Janssen, Pfizer, Sanofi and Takeda. SS has served as a consultant or advisory board member for AbbVie, Amgen, Arena, Bristol Myers Squibb, Biogen, Celltrion, Celgene, Ferring, Fresenius, Galapagos, Gilead, HIKMA, IMAB, Janssen, Lilly, MSD, Mylan, Pfizer, Protagonist, Provention Bio, Sandoz/Hexal, Takeda, Theravance and UCB.

Introduction

Liver transplant is the only definitive treatment for chronic liver disease (CLD). Referral to a hepatologist for consideration for liver transplant should be made when a patient with CLD experiences a hepatic decompensation or has a MELD score ≥ 15. Failure to refer in a timely manner can result needing a liver transplant evaluation (LTE) in the inpatient setting while a patient is experiencing acute decompensation.

Aims & Methods

We aimed to compare the clinical presentation of patients undergoing LTE in the inpatient setting versus the outpatient setting to identify potential risk factors for requiring urgent evaluation. A list of all LTEs performed at our institution between 10/2017-8/2021 was obtained. Medical records were preliminarily reviewed for evaluation setting (Inpatient vs. Outpatients); evaluations for re-transplantation and acute liver failure were excluded. After all Inpatients (n=159) were identified, Outpatients (n=159) were selected at random to create a sample matched for age, gender, race, and ethnicity (n=318). Data were collected using our institutions electronic medical record and outside health records as obtained during LTE process. “Prior decompensations” refers to the number of the following clinical decompensations experienced prior to the time of LTE: ascites, hepatic encephalopathy (HE), variceal/acute UGIB, SBP and HRS. Categorical variables and continuous were compared between using Fisher’s Exact test, and student’s T test, respectively.

Results

Alcohol was the most common CLD etiology (42.1% Inpatient, 48.4% Outpatient). Inpatients tended to live further from our center than Outpatients (p=0.079). Inpatients had significantly higher MELD-Na scores (27.1 ± 8 vs. 13.3 ± 6; p<0.0001). Malnutrition was more common in 5.6 times more common in inpatients than outpatients (OR 5.6, 95% CI 2.9-10.8, p<0.001).. A similar proportion of patients had 1 hospitalization. But inpatients were 3 times as likely to have had ≥ 1 liver-related hospital admission prior to LTE (p<0.0001). Inpatients had significantly more prior decompensations than Outpatients prior to inpatient LTE (3.2 ± 1.4 vs. 1.8 ± 1.1, p<0.001), with the greatest differences in SBP and HRS.

Conclusion

Multiple liver-related hospitalizations and malnutrition may be considered a risk factor for ultimately needing inpatient LTE and should be taken seriously by health care providers caring for patients with CLD. Episodes of SBP and HRS should prompt providers to consider immediate or urgent LTE referral regardless of other factors.

References

Multiple liver-related hospitalizations may be considered a risk factor for ultimately needing inpatient LTE and should be taken seriously by health care providers caring for patients with CLD. Episodes of SBP and HRS should prompt providers to consider immediate or urgent LTE referral regardless of other factors.

Disclosure

D Devuni has received grant funding from Sequana Medical and receives grant support from the NIAAA (AA017986-11), both for research unrelated to this work.