Introduction

In patients with colorectal cancer (CRC), intestinal dysbiosis has been observed, with abnormal colonization of the colonic mucosa by pathogenic Escherichia coli strains producing a cyclomodulin named cytotoxic necrotizing factor (CNF). Cyclomodulins are bacterial toxins capable of altering the cell cycle of the infected cell. One of the mechanisms involved in host defence against pathogens and in carcinogenesis is autophagy. However, the role of autophagy in the host response to the infection by CNF-producing E. coli, designated as CyPEC, has not yet been described.

Aims & Methods

Thus, the aim of this study was to investigate the role of autophagy in colorectal carcinogenesis in the context of CyPEC infection.
Mice predisposed to CRC development with autophagy deficiency specifically intestinal epithelial cells (Apc^Min/+/Atg16l1^ΔIEC mice) and their control littermates with functional autophagy (Apc^Min/+ mice) were infected with the clinical CyPEC strain 21F8. Tumor size and number were determined using a dissecting microscope. The mouse colons were collected and used for histological score assessment, and to analyse the levels of markers of autophagy, cell proliferation, apoptosis and inflammation by immunoblot, immunohistochemical staining and ELISA.

Results

We showed that in Apc^Min/+/Atg16l1^ΔIEC mice, infection with the CyPEC 21F8 strain led to increases in the number and the size of colonic tumours, compared with the uninfected condition. However, this was not observed in Apc^Min/+ mice. The increase in tumorigenesis in Apc^Min/+/Atg16l1^ΔIEC mice upon 21F8 infection versus uninfected condition was associated with increased pro-inflammatory cytokine production, enhanced cellular proliferation and decreased apoptosis in colonic epithelial cells.

Conclusion

Our results suggest that autophagy is necessary to limit the pro-carcinogenesis effects of the CyPEC 21F8 strain, thereby decreasing colorectal tumorigenesis. In another word, autophagy deficiency could be a susceptibility factor for the development of CRC in a host with colonization by CyPEC strains.

Introduction

The aging with weight gain induces pathological changes in the liver, exacerbating non-alcoholic fatty liver disease (NAFLD). In the reproductive age, NAFLD is more prevalent and severe in males, however, NAFLD occurs at a higher rate among postmenopausal females. The liver and small bowel are closely interrelated through enterohepatic circulation, such that damage to the small bowel can significantly impact hepatic function. In small intestine, the expression of tight junction protein (TJP) in nearly associated with the development of NAFLD. The aim of this study was to investigate the effects of age and sex on hepatic steatosis and the small bowel under conditions of a high fructose, high-fat diet (HFHFD) in rats.

Aims & Methods

6-week-old male and female F344 rats divided into control and HFHFD groups, along with 2-year-old F344 rats. Over an 8-week period, the high fat diet (Raonbio, 60 kcal% fat) provided twice weekly and high fructose water (20%) gave once weekly for HFHFD groups. After 8 weeks, all animals were sacrificed, and both liver and small intestinal tissue were collected. All tissue samples were subjected to hematoxylin and eosin staining for histological analysis. RNA extraction from the jejunum mucosa was followed by qRT-PCR analysis to assess claudin-2 (cldn-2, one of TJP) gene expression.

Results

The body weight change (%) and the hepatic steatosis scores were significantly higher in the HFHFD groups compared to both male and female control groups (Table). Histological scores of old rat HFHFD groups were significantly higher than those of 6 week HFHFD groups, regardless of sex (Table). In terms of histological inflammation scores of old control group small bowel were higher than those of 6 week control group regardless of sex (Table). The jejunum villus/crypt ratio of 6-week-old rats significantly decreased only among males (Table). In contrast, in the 2-year-old groups, the villus/crypt ratio resulted in a reduction only among females and the mRNA expression of cldn-2 observed in females with significant elevation (Table).

Conclusion

Our data showed that high fructose, high-fat diet induced hepatic steatosis and inflammation of small bowel, especially in old rats. This indicates that NAFLD is related with damage of small bowel brush border membrane. The upregulated gene expression of cldn-2 (p = 0.005) and the decreased tendency of jejunum villus/crypt ratio (p = 0.076) in the aged females rats suggests a potential role of TJP in advancing NAFLD in postmenopausal females.

References

Lee, Sun Min, et al. "Microbial changes and host response in F344 rat colon depending on sex and age following a high-fat diet." Frontiers in microbiology 9 (2018): 386521.
He, Yongyuan, et al. "Emerging role of aging in the progression of NAFLD to HCC." Ageing Research Reviews 84 (2023): 101833.
Hollander, Daniel, and Jonathan D. Kaunitz. "The “leaky gut”: tight junctions but loose associations?." Digestive diseases and sciences 65.5 (2020): 1277-1287.
Ahmad, Rizwan, et al. "Obesity-induces organ and tissue specific tight junction restructuring and barrier deregulation by claudin switching." Scientific reports 7.1 (2017): 1-16.
Zhou, Xiaoqiong, et al. "Gut microbiota induces hepatic steatosis by modulating the T cells balance in high fructose diet mice." Scientific Reports 13.1 (2023): 6701.
Amasheh, Salah, et al. "Claudin-2 expression induces cation-selective channels in tight junctions of epithelial cells." Journal of cell science 115.24 (2002): 4969-4976.
Lonardo, Amedeo, et al. "Sex differences in nonalcoholic fatty liver disease: state of the art and identification of research gaps." Hepatology 70.4 (2019): 1457-1469.

Introduction

Capsule endoscopy (VCE) is generally safe, however a subset of patients require confirmation of bowel patency before undergoing the procedure, using a self-dissolvable patency capsule of similar calibre to the video capsule, with interval imaging to confirm excretion. Patency capsule absence on imaging confirms bowel patency. Prucalopride, a 5-HT4 agonist that enhances bowel motility, has been used in VCE to improve completion rates¹, but its use in the context of patency testing remains unknown. Similarly, the role of fasting in improving pass rate is unclear.

Aims & Methods

This study aims to determine whether Prucalopride and fasting enhance the pass rate of patency tests. Data from procedures performed in 3 centers in the Republic of Ireland was reviewed retrospectively. We compared the pass rate in the context of Prucalopride (Group A), fasting (B), and non-fasting (C) states. All patients had an abdominal x-ray between 24-30 hours post-patency capsule ingestion to confirm passage. ANOVA test was used to compare means between all groups, and student t-test between individual groups.

Results

A total of 127 patients were included. Group A (Pruculopride): 59 patients, Group B (fasting, no Prucalopride): 22 patients, Group C (non-fasting, no Prucalopride): 46 patients.
Demographics:

• Group A: 47.5% males and 52.5% females, Group B: 45.5% males and 54.5% females, Group C: 32.6% males and 67.4% females
• Mean age – Group A: 50.3 years (range 18-82), Group B: 48.4 years (24-82), Group C: 47.9 (17-79).

• Appropriate: 80 (63%)
• Inappropriate: 35 (27.6%)
• Undocumented: 12 (9.4%)

• Group A: 36/59 (61%), Group B: 17/22 (77%), Group C: 19/46 (67%).
• ANOVA test f-ratio 4.55, p= 0.012.
• T-test: Group A vs B (61% vs 77%, p=0.087). A vs C (61%vs 67%, p=0.023), B vs C (77% vs 67%, p=0.002).

Conclusion

This study shows that fasting improves patency tests pass rates, but Prucalopride does not. Fasting and subsequent patency capsule ingestion promoting the gastro-ileal reflex is a possible explanation. It is unclear in this study why Prucalopride, a prokinetic and laxative, failed to show an improvement in pass rates despite being shown to improve VCE completion rates in other studies. Longer duration of Prucalopride at split intervals is a potential area for further studies.

References

1. Deding U, Kaalby L, Baatrup G, Kobaek-Larsen M, Thygesen MK, Epstein O, et al. The effect of prucalopride on the completion rate and polyp detection rate of colon capsule endoscopies. Clinical Epidemiology 2022;Volume 14:437–44. doi:10.2147/clep.s353527.

Disclosure

None

Introduction

As no direct comparison between IBD drugs to prevent endoscopic postoperative recurrence (POR) in Crohn’s disease (CD) are available, hierarchizing these therapeutic options remains challenging.

Aims & Methods

The objective of this network meta-analysis was to compare the effectiveness of treatments to prevent endoscopic POR of CD.
We performed a systematic review and network meta-analysis of comparative studies conducted in adults, evaluating a treatment to prevent endoscopic POR after ileocecal resection. The selection of studies was made according to PRISMA guidelines. The primary endpoint was endoscopic POR at 6 months defined as Rutgeerts score ≥ i2.
The network meta-analysis was carried out according to a random effects model. The results are presented as odds ratio (OR) with 95% confidence interval, adjusted for risk factors of POR. The risk of bias of the studies was assessed according to the Cochrane risk-of-bias tool for randomized trials and overall statistical heterogeneity between studies was assessed using τ.
The SUCRA ranking method (surface under the cumulative ranking) was used to rank the treatments.

Results

Twenty studies were included in this network meta-analysis, including a total of 2414 patients. Overall heterogeneity was moderate (τ =0.34). After adjustment for POR risk factors, ustekinumab (OR = 0.23 [0.07-0.70]), vedolizumab (OR = 0.17 [0.05 -0.59]), infliximab (OR = (0.18 [0.36-0.88]) and adalimumab (OR = 0.17 [0.07-0.42]) were superior to placebo to prevent endoscopic POR, contrary to 5-ASA (OR = 0.79 [0.42 -1.48]) and thiopurines (RR = 0.52 [0.22-1.24]) (Table 1). Ustekinumab (OR = 0.29 [0.08-0.99]), vedolizumab (OR = 0.22 [0.06-0.85]), infliximab (OR = (0.23 [0.09-0.54]) and adalimumab (OR = 0.22 [0.08-0.59]) were more effective than 5-ASA in preventing endoscopic POR. Adalimumab (OR = 0.33 [0.15-0.74]) and infliximab (OR = 0.34 [0.13-0.87]) were also more effective than thiopurines.
We observed no difference in efficacy between the 4 biologics (infliximab, adalimumab, ustekinumab and vedolizumab). The SUCRA ranking identified adalimumab (SUCRA = 0.81), infliximab (SUCRA = 0.80), vedolizumab (SUCRA = 0.79) and ustekinumab (SUCRA = 0.72) as the most effective treatments. The probability that thiopurines (SUCRA = 0.41), 5-ASA (SUCRA = 0.24), placebo (SUCRA = 0.16) and antibiotics (SUCRA = 0.08) were significantly higher weak.

Conclusion

This network meta-analysis confirms the efficacy of anti-TNF agents, vedolizumab and ustekinumab in preventing endoscopic CD POR without any difference between them. When prophylactic therapy is indicated, biologics appear as the best therapeutic option to prevent of endoscopic POR. Due to lower effectiveness, the use of 5-ASA and thiopurines should be limited in this situation.

Disclosure

Anthony BUISSON: Consulting fees for Abbvie, Amgen, Arena, Biogen, Celltrion Healthcare, CTMA, Galapagos/Alfa Sigma, GutyCare/Resilience, Janssen, MSD, Nexbiome, Pfizer, Roche, Sandoz, Takeda, Tillotts. And Viatris. Lecture fees for Abbvie, Amgen, Biogen, Celltrion Healthcare, Galapagos/Alfa Sigma, Janssen, Mayoli-Spindler, MSD, Nordic Pharma, Norgine, Pfizer, Roche, Takeda, Tillotts and Vifor Pharma. Research grants from: Abbvie, Celltrion Healthcare, Galapagos/Alfa Sigma, Janssen, Lessaffre, Lilly, Pfizer, Takeda.

Introduction

Colorectal cancer (CRC) is one of the most common neoplasms world widely. G-protein coupled receptors (GPCR) affect crucial elements of oncogenesis, such as responses to various endo- and exogenous stimuli (e.g. medications), angiogenesis and forming metastases. The magnitude of GPCR signaling is terminated by regulators of G-protein signaling (RGS), which typically possess GTPase activity. AXIN is an atypical RGS without GTPase function. Its main role is stabilization of β-catenin destruction complex in which it cooperates with adenomatous polyposis coli (APC) and glycogen synthase kinase3 (GSK3). Through this complex, AXIN antagonizes prooncogenic canonical Wnt signaling pathway that depends on β-catenin. On the other hand, recently attention has been paid to the potential tumor-suppressive properties of cannabinoids (CB) which are ligands of Gαi/o-type GPCR – CB1 and CB2.

Aims & Methods

In this project we aim to test the therapeutic potential of simultaneous targeting of AXIN and CB receptors with KYA1797K and WIN 55,212-2 in CRC cell lines (SW480, Caco-2) and non-cancerous enterocytes CCD 841 CoN as comparator. We hypothesize that KYA1797K is effective in monotherapy and in combination with cannabinoids in CRC. Moreover, we expect that stabilization of AXIN does not to weaken the effect of CB receptor ligands.
Cytotoxicity of KYA1797K (1; 10; 100, 300 µM) and WIN 55,212-2 (1; 10; 100 µM) in monotherapies and combined treatment (KYA1797K(10µM)+WIN 55,212-2(10µM) [K+W10] and KYA1797K(100µM)+WIN 55,212-2(10µM)) was assessed using MTT test. Relative genes’ expression was evaluated using quantitative real-time PCR and Western Blot. Statistical analysis included Shapiro-Wilk test, one-way ANOVA with post-hoc Dunnett’s and Tukey’s tests.

Results

Caco-2 cells were insensitive to KYA1797K in monotherapy excluding concentration of 300 µM, for which we observed significant cytotoxicity. Treatment with WIN 55,212-2 in concentration of at least 10 µM in both mono- and combined therapy significantly reduced viability of this cell line. Similar profile of changes was observed in SW480 line, however the strength of observed effect and sensitivity to KYA1797K were higher. In CCD 841 CoN cells significant additive effect of KYA1797K and WIN 55,212-2 was observed for 100 µM with no changes observed for 10 µM. Moreover, we found that treatment with K+W10 causes significant cytotoxicity in cancerous SW480 cells but not in normal enterocytes CCD 841 CoN. Moreover, K+W10 treatment resulted in increased expression of AXIN 1 and 2 in each cell line except AXIN 2 in Caco-2 line, in which opposite effect was observed. The combined treatment decreased CB1 receptors’ expression in all examined cell lines, whereas opposing tendency was observed for CB2. Furthermore, K+W10 treatment led to increased expression of RGS2, 11, APC and GSK-3. At the protein level K+W10 treatment surprisingly tended to decrease β-catenin relative expression. CB2 receptors were significantly more expressed in SW480 than in CCD 841 CoN that was consistent with cytotoxicity results.

Conclusion

AXIN stabilization is a promising new approach in CRC treatment. AXIN stabilization does not weaken CB1 and 2-dependent effects. The combined treatment of AXIN stabilizer and CB agonist may be especially important as life-prolonging therapy for patients with advanced CRC.

Introduction

Esophageal Achalasia (EA) is a rare esophageal motility disorder, characterized by impaired peristalsis and incomplete relaxation of the Lower Esophageal Sphincter (LES) in response to swallowing​​​​​. Key symptoms of the disease include dysphagia, food regurgitation, and retrosternal pain, significantly impacting the quality of life (QoL) and often leading patients to restrict their food intake. Consequently, EA patients often adopt self-managed dietary practices that may result in nutritional imbalances and deficiencies, given the lack of specific dietary guidelines.

Aims & Methods

This retrospective study aimed to describe and evaluate the dietary patterns, eating habits, and nutritional status of achalasic patients. Additionally, the secondary aim was to explore potential associations between food rheology and symptomatology. Patients with a manometric diagnosis of EA were recruited from our outpatient gastrointestinal pathophysiology center. Disease severity was assessed with Eckardt Symptoms Score (ESS), symptoms were evaluated with Modular Questionnaire Roma IV and QoL with the MD Anderson Dysphagia Inventory (MDADI). The MDADI outcomes were stratified into severe, moderate, mild and not perceived disability. Patients also filled out a food diary for 15 days, recording ingested food for each meal and symptoms onset. Estimated caloric intake and macronutrient composition were compared to those recommended by the Italian Society of Human Nutrition (SINU).

Results

Forty-three achalasic patients were included (M 24, F 19, mean age 56.7 +- 15.8 yrs). The median BMI was 23.7 (18.8-32.2), 34 patients (79%) had an active disease (ESS ≥ 3). Most patients reported mild to moderate disability (respectively 42% and 35%), while 21% reported severe disability and 2% perceived no disability. Disease activity, measured by ESS, strongly correlated with perceived disability (r= 0.59; p <0.0001), while no significant correlation was found between ESS and BMI. The diary analysis showed a mean caloric intake of 1572±373 kcal versus the estimated needs of 2293,71±374 kcal based on anthropometric parameters and physical activity, resulting in a mean caloric deficit of 720 ± 416 kcal (p<0.0001). Macronutrient analysis also reported an unbalanced distribution: carbohydrates 43.3%±7.3 vs recommended 55-59%, protein 18.2%±4.5 vs 15-18%, total fat 37.6%±7.7 vs 26–27% and fibre 14g±5.2 vs 25-30g. Solid food caused significantly more symptoms than semiliquid and liquid ones: dysphagia (35.5% vs 3.5% vs 5.7%, p<0.0001), chest pain (10% vs 12.1% vs 0.2%, p<0.0001), regurgitation (8.9% vs 1.4% vs 1.9%, p=0.001), cough (5% vs 0.2% vs 0.6%, p=0.0002). Bread, pasta, and pizza (50-60%), fresh fruit (44%), lean meat (39.5%), crackers, snacks, and vegetables (30-35%) were the most commonly reported triggers of dysphagia. Additionally, 60% of patients reported worsened symptoms with cold foods, while 53% reported improvement with hot foods.

Conclusion

This study underscores the significant impact of EA on QoL, influenced by disease activity. Achalasic patients tend to modify their diets to minimize symptoms, favoring easily swallowable soft and fatty foods while reducing intake of solid foods rich in carbohydrates and fiber. Thus, comprehensive nutritional assessment is strongly recommended for EA patients to mitigate dietary imbalances and manage possible deficiencies promptly.