Introduction

Inflammatory bowel disease (IBD), are characterized by chronic inflammation of the intestine, and affects millions of people worldwide. There are two main classifications of IBD: Crohn's disease and ulcerative colitis with different mechanisms and etiology of disease progression. IBD is multifactorial; the incidence of this disease may be associated with a dysregulated immune response, genetics, environment, and interactions with the host microbiome. The gastrointestinal epithelium acts as a barrier against harmful stimuli, and defects in its integrity are a precursor for the development of IBD. Our laboratory has shown that the conditional deletion of the Hepatocyte nuclear factor 4A (HNF4A) in the mouse intestinal epithelium (Hnf4a) leads to chronic intestinal inflammation. However, the functional impact of HNF4A on epithelial barrier integrity is still controversial.

Aims & Methods

To evaluate the impact of Hnf4a deletion on barrier integrity during acute inflammation.
We used a tamoxifen-inducible Cre^ER-loxP system to delete Hnf4a in the adult intestinal epithelium. Intestinal permeability was assessed in Hnf4a^ΔIEC-ind and control littermate mice in the collected serum after oral gavage of FITC-dextran. Mice were infected with an invasion-attenuated strain of Salmonella Typhimurium (SB103 (invA::strep)). S. Typhimurium loads were scored in the feces, liver, and spleen by plating homogenates on LB agar supplemented with streptomycin. Histological assessment was performed by various staining. In addition, we used a model of T-cell-induced enteropathy triggered by a T-cell activating monoclonal anti-CD3 Ab. Control and Hnf4a^ΔIEC-ind mice received an intraperitoneal injection of 50 μg monoclonal antiCD3ε Ab (145-2C11). Mice were assessed for clinical signs of enteropathy and sacrificed 1-, 3-, and five days post-injection. Gene expression levels of selected targets were measured by RT-qPCR. Also, Ki67+ staining was performed in the small intestinal tissue

Results

FITC-dextran assay showed a significant increase of intestinal permeability in uninfected Hnf4a^ΔIEC-ind mice compared to controls. In contrast, an oral infection with an invasion-deficient Salmonella Typhimurium strain did not differentially impact tissue bacterial loads or epithelial damage between groups. Likewise, the loss of epithelial HNF4A led to increased Muc2 transcripts expression in the ileum of infected Hnf4a^ΔIEC-ind mice. Histological examinations at four days post-infection showed an elevated number and size of goblet cells in the ileal crypts of Hnf4a^ΔIEC-ind mice. Bacterial localization using the general bacterial probe EUB388-cy3 (red) staining showed that infected Hnf4a^ΔIEC-ind mice tended to enhance barrier mucus layer thickness compared to the controls. On the contrary, acute T cell activation with anti-CD3 mAb induced a significant increase of Cxcl1 transcripts in the small intestine of Hnf4a^ΔIEC-ind mice. H&E staining showed elongated ileal crypts in Hnf4a^ΔIEC-ind mice compared to controls after anti-CD3 injection. Moreover, five-days after inflammatory stimulus, the ki67+ staining showed a proliferative phenotype located in the hyperplastic crypts of Hnf4a^ΔIEC-ind mice.

Conclusion

Altogether, these results support that HNF4A could play a crucial adaptive role in modulating the small intestine epithelial barrier function and controlling the immune system during acute inflammatory stress.

Disclosure

No conflict of interest in the text

Introduction

Inflammatory bowel disease (IBD), encompassing Crohn’s disease (CD) and ulcerative colitis (UC), is characterized by chronic inflammation affecting the gastrointestinal tract and extraintestinal organs. The etiology of IBD is multifactorial, involving genetic, immunological, and environmental factors, with significant genetic predisposition indicated by over 200 identified loci associated with the disease. Despite advancements in understanding the genetic basis of IBD, clinical management remains challenging due to heterogeneity in disease presentation and variable treatment responses. Current therapies, including 5-aminosalicylates and biologics, are not universally effective, highlighting the urgent need for reliable biomarkers to predict therapeutic responses. This study investigates the potential role of interferon regulatory factor 5 (IRF5) in IBD pathogenesis, particularly in UC.

Aims & Methods

We conducted a systematic analysis of colon biopsies from adult patients diagnosed with UC and non-IBD controls. Immunostaining was performed to assess IRF5 expression in colonic tissues, and statistical analyses were conducted to evaluate the correlation between IRF5-positive cell counts, disease activity, and inflammatory markers such as calprotectin

Results

Our analysis revealed a significant increase in IRF5-positive macrophage-like cells in the inflamed mucosa of IBD patients compared to healthy controls. The number of IRF5-positive cells correlated positively with disease activity and calprotectin levels, indicating that higher IRF5 expression is associated with increased inflammation.

Conclusion

This study establishes a significant correlation between IRF5 expression and disease activity in UC, suggesting that IRF5 may play a crucial role in the inflammatory processes associated with the disease. The findings propose IRF5 as a novel biomarker for therapeutic intervention in IBD. Further research is warranted to elucidate the exact mechanisms by which IRF5 contributes to IBD pathogenesis and to evaluate the therapeutic potential of targeting this pathway in clinical settings.

References

Anderson CA, Boucher G, Lees CW, Franke A, D'Amato M, Taylor KD, et al. Meta-analysis identifies 29 additional ulcerative colitis risk loci, increasing the number of confirmed associations to 47. Nat Genet. 2011;43(3):246-52.
Dideberg V, Kristjansdottir G, Milani L, Libioulle C, Sigurdsson S, Louis E, et al. An insertion-deletion polymorphism in the interferon regulatory Factor 5 (IRF5) gene confers risk of inflammatory bowel diseases. Hum Mol Genet. 2007;16(24):3008-16.
Yang Y, Zhang C, Jing D, He H, Li X, Wang Y, et al. IRF5 Acts as a Potential Therapeutic Marker in Inflammatory Bowel Diseases. Inflammatory bowel diseases. 2021;27(3):407-17.
Troncone E, Laudisi F, Di Fusco V, Dinallo V, De Simone E, Monteleone I, et al. Interferon Regulatory Factor 5 Expressing Cells infiltrate Lamina Propria of IBD Patients and produce inflammatory Cytokines. Digestive and Liver Disease. 2017;49(s2):e73-e223.

Disclosure

Lecturing fees from Immndiagnostik, MSD, Janssen, Takeda, Abbvie, consultancy fees from Vifor Pharma and Takeda

Introduction

Inflammatory Bowel Diseases (IBD) patients are associated to a higher risk of developing colonic dysplasia and colorectal cancer due to long-standing colonic inflammation and immunosuppressant therapies. They seem to have also a higher risk of developing extra-intestinal malignancies, such as cholangiocarcinoma, skin cancer, and others. Different studies have been conducted to assess whether a correlation exists between IBD, medical treatments and cancers, but the results are controversial.

Aims & Methods

Our study aimed to evaluate the risk factors of malignancies in our IBD patients, particularly considering the medication history.
In this single-center retrospective study, IBD patients with a history of at least one malignancy post-IBD diagnosis followed at the Gastroenterology Unit of Azienda Ospedale Università Padova were enrolled (cases). Demographics and clinical data, including medication information, were collected before and at the time of malignancy diagnosis. A group of IBD patients, matched by sex and age, without a malignancy history were also included (controls), considering the medications taken before and at the time of the last visit for comparison.

Results

Among 2213 IBD patients followed at our center (1212 males, 1040 CD, and 1173 UC), 153 cases (80 UC and 73 CD, 46.4% males, mean age at diagnosis: 38.4±14.7) and 50 sex and matched controls were included (30 UC and 20 CD, 52% males, mean age at diagnosis: 38.9±0.8). Twelve patients had two malignancies, and 2 had three. For the scope of our study, we considered only the first tumor. Results are shown in the Table 1. The mean time from IBD diagnosis to the first tumor was 15 years (SD 10.5). Considering malignancies, the most frequent in our population were breast cancer (27/1001 females, 2.7%), colorectal cancer (23/2213, 1%), melanoma (24/2213, 1.1%), and non-melanoma skin cancer (15/2213, 0.7%). Patients with colon cancer were younger at the time of IBD diagnosis (31.0±15.8 vs. 38.9±0.8), while patients with prostate and urinary cancer (11/2213, 0.5%) were older compared to controls (49.6±13.7 vs. 38.9±0.8). A lower percentage of patients with colon (13%), breast (4%), and prostate cancers (9.1%) had taken biological therapy before cancer occurrence, as compared to controls (44%, p<0.01). Finally, we observed a significantly higher number of melanoma and breast cancer patients taking azathioprine (AZA) at the time of cancer diagnosis for more than 4 years. No difference in terms of duration of AZA and biological therapies before cancer diagnosis was observed.

Table 1: Type and frequency of Malignancies and characteristics of the patients

Conclusion

Our study found that biological therapy was not associated to malignancies, while azathioprine use was associated with breast and melanoma cancers. A younger age at diagnosis was associated with colon cancer, while an older age with prostate and urinary cancers.

Introduction

Inflammatory bowel disease (IBD) is a chronic multi-factorial disease characterized by inflammation of the gastrointestinal tract. Currently, the mechanisms underlying the disease are far from being elucidated. Thus, the study of proteins involved in its pathogenesis would allow further insights into the molecular mechanisms underlying IBD.

Aims & Methods

The serum and intestinal mucosa (ileum and left colon) proteomic profiles of patients with Crohn´s disease (CD) and ulcerative colitis (UC) and healthy controls (HC) were analyzed through label-free relative quantitative proteomic profiling using nLC MS/MS. Data mining and pattern recognition techniques were performed to identify hidden relationships that are not detectable using classical linear classifiers, and thus identify potential markers able to discriminate between the different study groups. Six candidate biomarkers were selected for further validation using enzyme-linked immunosorbent assay (ELISA) and immunohistochemistry (IHC) in serum and intestinal tissue samples, respectively.

Results

Comparison of serum proteins between UC patients and HC revealed statistically lower levels of protein 1 and 2 and higher levels of protein 3. The comparison of potential biomarkers between CD and HC group showed lower levels of protein 1 and higher levels of protein 3. In addition, protein 4 was differentially upregulated in CD patients compared to UC. Regarding the intestinal biopsies, protein 5 showed higher intensity in the IHC staining of ileum from CD patients compared to HC, whereas protein 6 was significantly decreased in left colon and ileum from CD and UC patients compared to HC. These proteins are involved in thyroxine transport, regulation of chemotactic chemokine activity, inflammatory processes, triglyceride homeostasis, oxidative stress and regulation of NF-kappa-B activation in the cytosol.

Conclusion

In this study, we have identified a panel of six potential biomarkers that could help to elucidate mechanisms involved in IBD pathogenesis. Their roles in the pathophysiology need to be clarified in further experiments.

Introduction

Inflammatory Bowel Diseases (IBD) can lead to various complications, with Acute Severe Colitis (ASC) being among the most critical. ASC is a medical and surgical emergency that poses a significant risk to life, necessitating prompt and effective diagnosis and management. The first-line treatment typically involves intravenous corticosteroids. However, some cases are resistant to corticosteroids and require second-line medical therapy or immediate surgical intervention.

Aims & Methods

The objective was to identify factors associated with the failure of first-line medical treatment in patients with ASC. A retrospective descriptive and analytical study was conducted over ten years (January 2014 to March 2024) in the gastroenterology department of Charles Nicolle Hospital. The study included patients with IBD complicated by ASC, defined by the Truelove and Witts criteria and a Lichtiger score greater than 10.Patients with bacterial superinfection were not included.

Results

Forty-five patients over 18 years old were included, with a mean age of 37.87 years (range: 18–74 years). There was a slight female predominance (53%), with a male-to-female ratio of 0.87. In 31% of cases, ASC was the initial presentation of the disease. Endoscopic signs of severity at admission were observed in 49% of patients. All patients received intravenous corticosteroid therapy with hydrocortisone hemisuccinate as the first-line treatment. The response rate to first-line treatment was 64.5%. Second-line medical therapy was administered to 24.4% of patients, consisting of cyclosporine in 11.1% and infliximab in 13.3%. Five patients (11.1%) underwent subtotal colectomy due to corticosteroid failure.
In univariate analysis, factors associated with corticosteroid failure included male gender (p=0.011), absence of extraintestinal manifestations (p=0.017), higher average number of stools on the first day (p=0.013), Lichtiger score >10 on the third (p=0.011) and fifth day (p<0.001), hemoglobin levels >11 g/dL on the first day (p<0.001), >10 g/dL on the third day (p=0.009), >9 g/dL on the seventh day (p=0.013), white blood cell count >10,000/mm³ on the first day (p=0.008), >12,000/mm³ on the third (p=0.001) and seventh day (p=0.004), C-reactive protein (CRP) level >30 mg/L on the seventh day (p=0.005), and persistent endoscopic signs of severity after three to five days of corticosteroid therapy (p=0.001).
Multivariate Analysis showed that only the presence of severe endoscopic signs on follow-up endoscopy after three to five days of corticosteroid therapy was predictive of corticosteroid failure (Odds Ratio = 42.2; p=0.028).

Conclusion

Acute Severe Colitis is a potentially life-threatening complication. The presence of severe endoscopic signs after five days of corticosteroid therapy should prompt the initiation of second-line therapy. Understanding the predictive factors for corticosteroid failure is essential for individualized management to improve both immediate and long-term outcomes.

Disclosure

The authors declare no conflicts of interest.

Introduction

Patients with inflammatory bowel disease (IBD) have an increased risk of developing intestinal and extra-intestinal cancers. Specific therapies used to control inflammation in IBD may have a different impact on cancer risk. However, there are no long-term studies investigating cancer risk in patients with IBD.

Aims & Methods

In the present study we aimed to quantify the overall risk for of developing cancer and to identify specific risk factors for the occurrence of intestinal and extra-intestinal cancers in 560 consecutive patients with Crohn's disease (CD, N=310) or ulcerative colitis (UC, N=250) referred to our outpatient department between January 2021 and February 2022. Demographic data, smoking status, alcohol consumption, age at IBD diagnosis, disease location and course, therapies, and cancer development after IBD diagnosis were recorded. Odds ratios (OR) were estimated by logistic regression analysis. Standardized Incidence Ratios (SIRs) were estimated compared with the general population.

Results

During a median follow-up interval or time of 10 years (range 1-26 years), 6.6% of patients (37/550, female 65 %, mean age 51 years) developed cancer (SIR 1.94, 95%CI 1.4-2.6). The most common cancers were colorectal (12.3%), skin (8.2%), and breast cancer (7.2%). Female patients had an increased risk for all cancers (SIR 3.11, 95%CI 2.06-4.3), melanoma (SIR 5.6, 95%CI 1.14-16.2) and colorectal cancer (SIR 7.59, 95%CI 3-15.4). Male patients had an increased risk of lymphoma (SIR 26.15, 95%CI 3.2-95.7). Younger age at diagnosis of CD (A1 Montreal classification, OR 37.9, 95%CI 10-14), immunosuppressive monotherapy (OR 5.5, 95%CI 2.65- 11) or in combination with anti-TNFα (OR 9.9, 95%CI 3.3-29.09) were associated with an increased risk of cancer in the study population.

Conclusion

We have identified subgroups of IBD patients who would particularly benefit from screening programs. Our findings need to be validated in independent cohorts.

Introduction

The true effect of having inflammatory bowel disease (IBD) in the development of infections has not been studied in depth since cohort studies are scarce and the results of studies are sometimes contradictory. Especially, this complication has not been addressed prospectively in an incident cohort of patients with IBD.

Aims & Methods

Aims: To assess, regarding infections in IBD: 1) incidence and type, 2) risk factors, 3) effect of type and the duration of IBD treatment, 4) impact in mortality 5) effect of IBD activity.
Methods: The INFEII registry (ClinicalTrials.gov: NCT02904590) is a prospective study promoted by GETECCU to determine the incidence and risk factors of infection in an inception cohort of patients with IBD and follow-up of at least 5 years. This longitudinal prospective study within the ENEIDA registry included incident cases with IBD. Patients were recruited between 2016 and 2020 with a planned follow-up of 5 years. All infections were detailed analysed. An infection was considered relevant when: 1) required hospital admission, 2) led to death or endangered the patient’s life, 3) was treated with specific antibiotics,4) occurred recurrently, and/or 5) required change/withdrawal of immunosuppressive treatment.

Results

The INFEII registry recruited 1456 patients from 28 centres throughout Spain. The median follow-up was 71 months (IQR 60-79 months). The description of the cohort is presented in Table 1. Of the 1456 patients, 580 (40%) presented at least one relevant infection. In total, 1321 infections were collected. One-hundred and seven infections (8%) required hospitalisation and four patients died (0.3%) over infection. Ninety-nine patients were lost to follow-up (22 died and 34 changed hospitals). The most prevalent infections were: ear-nose-throat in 238 cases, urinary tract infections in 237, respiratory in 234, COVID-19 in 133, skin infections in 127, dental in 64 and colitis/enteritis in 64. From the whole cohort and at some point, in time, 756 (52%) patients received 5ASA, 481 (33%) steroids (369 (25%) oral, 112 (7.7%) intravenous), 568 (39%) azathioprine, 72 (4.9%) methotrexate, 21 (1.4%) tofacitinib, 533 (37%) antiTNF, 141 (9.9%) ustekinumab and 77 (5.3%) vedolizumab. The effect of treatment and IBD activity in each infection is being analysed.

Table 1. Clinical characteristics of patients included in the INFEII study

Conclusion

Infections are frequent complications in patients with IBD, presented in 40% of them. Being a woman, having a history of past serious infections or occupational risk may be associated with infection occurrence. It has to be determined if IBD activity and treatment pose a greater risk for infections. This paradigmatic study implies the first use of the ENEIDA registry in prospective research.