Introduction

Celiac disease (CeD) is an immune-mediated disease with excess morbidity and mortality. CeD is associated with inflamed intestinal mucosa, which may be mediated IL-15. There are currently no established non-invasive biomarkers to monitor disease activity following treatment of CeD.

Aims & Methods

Thus, we aimed to characterize disease biomarkers of CeD, primarily IL-15 levels, in the gut and in the serum of non-responsive CeD (NRCD), Treated responsive CeD (TrCD), and non-CeD controls to evaluate the relationship between cytokine levels and CeD disease status.
Based on the existing archives of our Olmsted County Celiac Registry, we identified 29 NRCD, 30 TrCD, and 24 non CeD controls. NRCD was defined as patients with a diagnosis of CeD presenting with continued celiac symptoms and partial or total villous atrophy after 1 or more years of following a strict gluten-free diet (GFD). In contrast, TrCD was defined as patients with a diagnosis of CeD who were presenting with no celiac symptoms and normal duodenal histology after 1 year or more of a strict GFD. Control samples were matched by age, gender, and confirmed as negative celiac serology. We assessed duodenal biopsy and serum IL-15 protein expression levels in patients with NRCD, TrCD, and controls using a quantitative ligand binding assay with electrochemiluminescent detection. Furthermore, we conducted the proteomic profiling by utilizing targeted proteomics panels to assess the changes in serum and duodenal biopsy tissues samples of patients with NRCD, TrCD, or non-CeD controls.

Results

The measured levels of IL-15 in serum and duodenal biopsy samples are summarized according to disease status in Table 1. No apparent separation in median IL-15 levels (serum and gut tissue) was observed across the three groups (non-CeD controls, TrCD and NRCD). It was also difficult to draw a correlation between serum and gut tissue IL-15 levels. In contrast to the TrCD and control groups, high variability in IL-15 levels was seen with the NRCD group, potentially due to differences in histology and/or other clinical manifestations by the subjects within the group. Interestingly, the highest IL-15 level (serum or tissue) was noted in NRCD patients with total villous atrophy. However, small n (2 out of 29) precludes definitive association.

Table 1. Demographics and IL-15 protein levels in samples from patients with NRCD, RCD, and non-CeD controls.

Conclusion

Data generated from this retrospective study did not show clear difference in the mean IL-15 (serum or tissue) levels between the responsive and non-responsive CeD groups. However, for the limited number of NRCD samples with total villous atrophy, the IL-15 levels were substantially higher (either in serum or tissue) compared to partial or near normal. Testing of more samples representing the different villous atrophy status is warranted to confirm these results.

Disclosure

Study was conducted as part of a Teva funded research collaboration between the Mayo Clinic, which provided the gut biopsies and serum samples, and Teva, Pharmaceuticals Industries, Ltd., which developed and performed the assays and analyses.

Introduction

There is an ongoing paradigm shift toward serology-based diagnosis of celiac disease (CeD). It is essential to target potential endoscopies for those most likely to benefit. Anemia is a common symptom of CeD that may warrant further investigations, but the prevalence and clinical relevance of endoscopic findings in anemic CeD patients remain unclear.

Aims & Methods

The aim of this study was to compare the prevalence of upper and lower endoscopic findings between anemic patients and non-CeD controls, and to identify predictors of significant findings in CeD patients. Medical data, including upper and lower endoscopy findings, were collected from 162 CeD patients and 350 controls. Binary logistic regression analysis was used to identify predictors of findings.

Results

CeD patients were younger (median 49 vs 66 years, p<0.001) and had fewer risk factors such as smoking (14% vs 25%, p<0.001), alcohol abuse (4% vs 14%, p<0.001), chronic comorbidities (55% vs 83%, p<0.001), and previous malignancies (1% vs 8%, p=0.002).
In comparison of upper endoscopy findings, CeD patients had fewer significant findings (21% vs 53%, p<0.001), H. Pylori positive gastritis (6% vs 19%, p<0.001), gastric ulcers (0% vs 5%, p=0.006), and atrophic gastritis (3% vs 9%, p=0.005) compared to the controls.
Twenty-four patients and 208 controls had undergone lower endoscopy with celiac patients more frequently demonstrating significant findings (54% vs 28%, p=0.008), non-specific inflammation (21% vs 1%, p<0.001) and colorectal cancer (25% vs 9%, p=0.018).
Predictors of significant upper endoscopy findings in CeD patients were older age (OR 1.03, 95% CI 1.01-1.05), age >50 (2.44, 1.18-5.06), co-existing chronic conditions (3.45, 1.53-7.78), gastrointestinal symptoms (2.31, 1.07-4.98), and use of acetylsalicylic acid (2.38, 1.01-5.06), and of colonoscopy findings older age (1.01, 0.99-1.04), co-existing conditions (3.48, 1.12-10.8), and gastrointestinal symptoms (4.73, 1.34-16.7).

Conclusion

CeD patients had fewer findings on upper endoscopy but, despite being younger and having fewer risk factors, showed more findings on lower endoscopy compared to controls. In older CeD patients presenting with anemia, lower endoscopy should be strongly considered.

Introduction

Celiac disease, a chronic autoimmune enteropathy caused by exposure to gluten in genetically predisposed individuals, is characterized by a broad clinical spectrum often with multisystem involvement. Symptoms related to the reproductive system include delayed menarche, early menopause, fertility disorders, miscarriages, and prematurity.

Aims & Methods

To study the impact of celiac disease on fertility in a female Greek population. A retrospective review of all patient records from the celiac disease outpatient clinic at our hospital.

Results

Medical histories of 231 female patients were reviewed (69% of the total patient population) with a mean age of 44.3 years and a mean age at diagnosis of 35.3 years. The mean age of menarche was 12.7 years. Menstrual irregularities were reported in 27% (20% instability, 7% dysmenorrhea). A total of 28% (65/231) were in menopause, with an average age of menopause onset at 50.7 years. A history of miscarriages was recorded in 15% of all women (35/231), but in 29% of women in menopause (19/65). 82% (53/65) of women in menopause had children, with 98% (52/53) having been pregnant before the diagnosis of the disease. Among all women who gave birth, only 12/113 (11%) had been diagnosed before pregnancy. 56% (63/113) and 44% (50/113) had a vaginal delivery and cesarean section, respectively. Pregnancy complications were recorded in 24 women (18 preterm births, 15 low birth weight infants, 1 intrauterine death).

Conclusion

Women with celiac disease tend to have relatively late menarche and an increased rate of miscarriages. However, the disease does not seem to significantly affect fertility.

References

1.Ehud Galron MD, Rayna Boyarsky MD,Ido Sold MD, Chen Ben David MD,and Eyal Gal MD,Interactions between celiac disease and pregnancy:A literature review, Isr Med Assoc J.2023 Dec;25(12):830-835
2.Giovanni Casella et. Al. Celiac disease and obstetrical-gynecological contribution, Gastroenterol Hepatol Bed Bench. 2016 Autumn; 9(4): 241–249.
3.Stephanie M. Moleski et. Al., Increased rates of pregnancy complications in women with celiac disease, Annals of Gastroenterology (2015) 28, 236-240
4.Nafeesa N. Dhalwani et Al., Women With Celiac Disease Present With Fertility ProblemsNo More Often Than Women in the General Population, Gastroenterology 2014;147:1267–1274

Introduction

Obesity and metabolic dysfunction-associated steatotic liver disease (MASLD) have been linked to gut dysbiosis.

Aims & Methods

This study aimed to investigate the relationship between gut microbiota composition, hepatic fat content and stiffness, and body mass index (BMI) status. This cross-sectional study enrolled adolescents aged 12 to 18. Transient elastography with controlled attenuation parameter (CAP) was employed to diagnose hepatic fibrosis and steatosis. Fecal samples were analyzed by 16S rRNA sequencing. Relative abundance, as well as alpha- and beta-diversity of the microbiota, were compared across participants with different nutrition statuses and the presence of hepatic steatosis and fibrosis.

Results

A total of 76 participants (51 males) were recruited for this study, including 43 with normal weight, 19 overweight, and 14 obese. The clinical characteristics of the participants are presented in Table 1. Hepatic steatosis was present in 8 out of 14 obese participants (57.2%) and 2 out of 19 overweight (10.5%) participants. Hepatic fibrosis was detected in 4 out of 14 obese participants (28.6%) but none in the normal weight or overweight groups. There were no significant differences in alpha- or beta-diversity between the normal weight, overweight, and obese groups. The Firmicutes to Bacteroidetes (F/B) ratio was 1.2 across all three groups. Several genera, including Eubacterium_halli_group and Ruminococcus_gauvreauii_group were present in higher abundances in the gut microbiome of the obese group compared to the normal-weight group, with p values of 0.025 and 0.036, respectively. In contrast, Bifidobacterium was more abundant in the normal-weight participants (p=0.012). Obese individuals with hepatic fibrosis exhibited a significantly higher F/B ratio compared to those without hepatic fibrosis (p=0.04). There were significant differences in the relative abundance of specific genera between the fibrosis and non-fibrosis groups: Phascolarctobacterium (p=0.020), Dorea (p=0.040), and Ruminococcus_gauvreauii_group (p=0.045) were present in higher abundances in the fibrosis group, whereas Senegalimassilia was more abundant in the non-fibrosis group (p=0.037).

Conclusion

Hepatic steatosis and fibrosis are common among obese adolescents. The gut microbiota abundance in obese individuals with hepatic fibrosis differ from those without it. Further studies are crucial to explore the role of gut microbiota modification in the progression of hepatic steatosis and fibrosis.

Introduction

Barrett’s esophagus (BE), the major precursor of esophageal adenocarcinoma (EAC), is a condition where the normal stratified squamous esophageal epithelium undergoes transdifferentiation into the specialized, intestinal-like epithelium. This reprogramming, also termed ‘paligenosis’, is predominantly caused by gastroesophageal reflux disease (GERD)¹. On the other hand, hydrogen sulfide (H₂S) next to carbon monixide (CO) is an important endogenous gaseous mediator. H₂S is biosynthesized by cystathionine β-synthase (CBS) and cystathionine γ-lyase (CTH). Both, H₂S and CO have been shown to maintain gastrointestinal (GI) integrity^2,3,4. However, the contribution of gaseous mediators signaling to esophageal transdifferentiation process is overlooked.

Aims & Methods

Thus, in this study we aimed to explore the role of H₂S bioavailability in mTOR-dependent paligenosis induced by chronic epithelial injury. Furthermore, we evaluated the relationship between GERD, the barrier function of cultured cells, and the pattern of selected components involved in the formation of the tight junctions (TJs) and responsible for epithelial barrier integrity.
We implemented an animal model of GERD, based on Male Wistar rats with esophagogastroduodenal anastomosis (EGDA) treated i.g. for 8 weeks with vehicle or with H₂S- releasing chemicals: GYY4137 (5.0 mg/kg)/NaHS (1.0 mg/kg) or CTH/CBS inhibitors: PAG (15 mg/kg)/AOAA (11 mg/kg). In parallel, we employed optimized in-vitro GERD model based on human-derived esophageal epithelial (EPC2) cells +/- CRISPR-Cas9-induced knockout of H₂S-producing enzymes that were treated with acidified bile mixture (BM) or with GYY4137/NaHS (100 µM). We evaluated activation of AKT/mTOR pathway by multiplex microbeads fluorescent assay and an expression of TJ-associated protein (ZO-1) and occludin by capillary immunoblotting. The barrier function of cultured cells was evaluated by measurement of transepithelial electrical resistance (TER).

Results

We observed that either chronic reflux in EGDA-rats and/or treatment of human esophageal cells with BM led to a slight or a significant activation of AKT/mTOR signaling pathway and its down-stream effector - p70S6K. Additionally, we observed that pharmacologically-derived H₂S reversed this effect, while inhibition of endogenous H₂S biosynthesis (pharmacological or induced by knockout of H₂S-producing enzymes) had either no effect or increased GERD-induced phosphorylation of AKT/mTOR pathway components. Moreover, we showed that ZO-1 is highly expressed in BM treated EPC2 cells and this was reflected by the increase in TER measurment. Level of occludin remained unchanged. We did not observe any changes in expression of ZO-1 and occludin after GYY4137 treatment; however, TER measurment decreased in comparison to BM treated cells. Selective knockout of H₂S-producing enzymes led to a decrease in ZO-1 protein content in comparison to wild-type cells.

Conclusion

We conclude that interventional enhancing or decreasing of H₂S bioavailability in esophageal epithelium respectively delay or accelerate the GERD-induced paligenosis process via regulation of the mTOR pathway. H₂S-prodrugs may provide a protective effect on the esophageal epithelium by substituting the phisiological defensive response of the tissue against GERD, which is reflected by an increase in the expression of ZO-1 and tightening of the cellular barrier.

References

1. Souza RF, Spechler SJ. Mechanisms and pathophysiology of Barrett oesophagus. Nat Rev Gastroenterol Hepatol 2022.
2. Fiorucci S, Distrutti E, Cirino G, Wallace JL. The emerging roles of hydrogen sulfide in the gastrointestinal tract and liver. Gastroenterology 2006.
3. Magierowska K, Korbut E, Wójcik-Grzybek D, Bakalarz D, Sliwowski Z, Cieszkowski J, Szetela M, Torregrossa R, Whiteman M, Magierowski M. Mitochondria-targeted hydrogen sulfide donors versus acute oxidative gastric mucosal injury. J Control Release 2022.
4. Magierowska K, Brzozowski T , Magierowski M. Emerging role of carbon monoxide in regulation of cellular pathways and in the maintenance of gastric mucosal integrity. Pharmacol Res 2018.

[Funding: National Science Centre, Poland (UMO-2016/23/D/NZ4/01913, UMO-2019/33/B/NZ4/00616) and Statutory Grant (N41/DBS/000874), Jagiellonian University Medical College].