Introduction

Anastomotic leakage (AL) is a major complication in colorectal cancer (CRC) surgery. Its impact on survival and recurrence remains debated.

Aims & Methods

This meta-analysis aimed to provide a comprehensive assessment of the effect of AL on oncological outcomes in patients undergoing CRC surgery. A systematic search was conducted in three databases on the 17th of November, 2023 (PROSPERO: CRD42023480523). Studies examining the impact of AL after CRC surgery were included. Our outcomes were overall survival (OS), disease-free survival (DFS), cancer-specific survival (CSS), local (LR) and distant recurrence (DR). Hazard ratios (HR) with a 95% confidence interval (CI) were calculated using a random-effect model. Additionally, restricted mean survival time (RMST) was calculated for individual patients using Kaplan-Meier curves.

Results

A total of 66 studies with 235,626 patients were analyzed. AL was associated with worse OS, DFS, and CSS across all cancer types. For rectal cancer, the HRs for OS, DFS, and CSS were 1.45 (CI: 1.14–1.84), 1.65 (CI: 1.30–2.09), and 1.91 (CI: 0.95–3.85), respectively. Similar trends were observed for colorectal and colon cancer. AL was also associated with significantly higher rates of LR for rectal and colorectal cancer (HR: 1.69, CI: 1.03-2.79 and HR: 3.80, CI: 1.13-12.82, respectively), but no significant impact was found on DR. The RMST for OS was significantly shorter for patients with AL: 54.05 months (CI: 48.52-58.78) vs without AL: 114.56 months (CI: 101.74-121.29).

Conclusion

Anastomotic leakage was significantly associated with poorer survival outcomes and an increased risk of local recurrence in patients who underwent colorectal cancer surgery.

Introduction

Patient-derived organoid culture is a powerful and versatile system for investigating the molecular mechanisms underlying a wide range of gastrointestinal diseases, including inflammatory bowel disease (IBD) [1]. In the context of IBD, intestinal organoids derived from patients retain key genetic, epigenetic, and disease-specific features, offering a unique platform to model individual responses to inflammatory triggers [2, 3]. This approach also shows promise in predicting personalized responses to biologic and small-molecule therapies, advancing precision medicine strategies for IBD [4]. Although intestinal organoids represent a major advancement in personalized ex vivo cellular modeling, the detailed effects of differentiation and inflammation on the cellular landscape remain poorly understood.

Aims & Methods

In this study, we analyzed differentiated and undifferentiated intestinal epithelial organoids derived from both an IBD patient with active ulcerative colitis and a healthy donor (non-IBD) (n = 4). Inflammation was induced by treating organoids with the pro-inflammatory cytokines TNF-α (10 ng/mL) and IFN-γ (10 ng/mL). Organoids were dissociated using TrypLE Express Enzyme supplemented with Y27632, and viable cells were enriched by fluorescence-activated cell sorting (FACS) using 7-AAD staining. Cell viability and concentration were assessed using the Countess 3 automated cell counter and Trypan Blue staining. Single-cell suspensions were then processed using the 10x Genomics GEM-X Universal 3’ Gene Expression application, and libraries were sequenced on the NovaSeq X Plus platform. Primary data processing was performed using Cell Ranger (10X Genomics), followed by downstream visualization in Loupe Browser (10X Genomics) and an in-house pipeline.

Results

The mean number of recovered cells per sample was 1,957, with an average of 142,015 reads per cell and 27,127 genes detected across all samples. Single-cell transcriptome analysis revealed five distinct clusters in undifferentiated organoids (both treated and untreated), and six to seven clusters in differentiated organoids (treated and untreated). The majority of cells expressed the epithelial marker EPCAM, confirming their epithelial origin. Inflammatory stimulation led to a significant upregulation of TNF-α and IFN-γ gene expression (p < 0.05) in both differentiated and undifferentiated samples. Furthermore, differentiated organoids displayed a significant increase in specific epithelial subtypes, including Paneth cells, enteroendocrine cells, and proliferative cells (p < 0.05), compared to their undifferentiated counterparts.

Conclusion

To conclude, single-cell transcriptomic profiling of patient-derived intestinal organoids revealed a dynamic and heterogeneous cellular landscape shaped by both differentiation and inflammatory signals. The observed upregulation of inflammatory markers underscores the organoid model's utility in mimicking inflammatory responses. Additionally, the potential expansion of distinct epithelial subpopulations in differentiated conditions highlights the model's ability to faithfully recapitulate the complexity of the intestinal epithelium.

References

1. Günther, C., Winner, B., Neurath, M. F., & Stappenbeck, T. S. (2022). Organoids in gastrointestinal diseases: from experimental models to clinical translation. Gut, 71(9), 1892-1908.\
2. Dotti, I., Mora-Buch, R., Ferrer-Picón, E., Planell, N., Jung, P., Masamunt, M. C., ... & Salas, A. (2017). Alterations in the epithelial stem cell compartment could contribute to permanent changes in the mucosa of patients with ulcerative colitis. Gut, 66(12), 2069-2079.
3. Sato, T., Stange, D. E., Ferrante, M., Vries, R. G., Van Es, J. H., Van Den Brink, S., ... & Clevers, H. (2011). Long-term expansion of epithelial organoids from human colon, adenoma, adenocarcinoma, and Barrett's epithelium. Gastroenterology, 141(5), 1762-1772.
4. Arnauts, K., Verstockt, B., Santo Ramalho, A., Vermeire, S., Verfaillie, C., & Ferrante, M. (2020). Ex vivo mimicking of inflammation in organoids derived from patients with ulcerative colitis. Gastroenterology, 159(4), 1564-1567.

Introduction

Liver cirrhosis is characterized by microbial dysbiosis, intestinal inflammation and a weakened gut barrier and associated with an increased risk of complications. We previously observed a positive influence of L-ornithine-L-aspartate (LOLA) on gut microbiome composition in cirrhosis (1). The current study aimed to investigate the effect of LOLA on microbiome dysbiosis, quality of life, sarcopenia, intestinal permeability, inflammation and metabolite composition.

Aims & Methods

In this monocentric, open-label, phase 4 study, LOLA (18g/day) was administered orally for three months to liver cirrhosis patients with hepatic encephalopathy grades (0-2). We investigated changes in their gut microbiome composition (16s rRNA sequencing, alpha and beta diversity, differential abundance), quality of life (SF36), blood ammonia concentrations, measurements of sarcopenia and frailty (appendicular lean muscle mass, anthropometric measures, liver frailty index, SARC-F), biomarkers of gut permeability, inflammation and sarcopenia as well as metabolic alterations in stool, serum, and urine.

Results

65 patients were enrolled, and 52 patients completed the 3-month study (40.4% female, age 62 (58; 65)). While LOLA reduced the abundance of the genus Romboutsia, it did not significantly change overall microbiome diversity. LOLA led to a significant improvement in the vitality dimension of the SF36 questionnaire (45 (35; 60) to 50 (45; 60), p=0.019) and decreased plasma ammonia levels in patients with elevated baseline ammonia (69.5 (54; 225) to 46.0 (22; 66) µmol/L, p=0.004) Although muscle mass generally declined across the study, this was not observed in patients whose elevated ammonia levels improved. Sarcopenia and frailty scores and the muscle biomarkers myostatin, irisin, fibroblast growth factor 21, insulin-like growth factor 1 were unaffected by LOLA treatment. Diamine oxidase, a marker of gut barrier dysfunction, significantly decreased (from 12.75 U/mL (11.25; 17.45) to 11.15 U/mL (9.9; 15.65), p=0.016), while LPS binding protein, a marker of innate immune response to bacterial translocation, significantly increased (from 17.05 µg/mg (14.7; 19.8) to 17.39 µg/mg (16.2; 21.9), p=0.006). Calprotectin, zonulin and sCD14 did not show significant changes. Metabolomic analysis revealed an increase in serum alanine levels (pFDR=0.014) with additional subtle shifts in lipoprotein and amino acid profiles.

Conclusion

In summary, LOLA was associated with an improvement in vitality, an important patient-reported outcome accompanied by alterations in gut microbial composition and prevented muscle loss in patients with hyperammonaemia. Furthermore, LOLA was associated with improvements in gut permeability and endotoxin handling. The slight increase in alanine observed in the metabolomics analysis may be a consequence of LOLA metabolism. LOLA may be a valuable supportive treatment for enhancing quality of life in cirrhosis by targeting the gut liver axis and may be particularly beneficial in preventing muscle loss in patients with hyperammonaemia.

References

1. Horvath A, Traub J, Aliwa B, Bourgeois B, Madl T, Stadlbauer V. Oral Intake of L-Ornithine-L-Aspartate Is Associated with Distinct Microbiome and Metabolome Changes in Cirrhosis. Nutrients. 2022 Feb 10;14(4):748.

Introduction

Eosinophilic esophagitis (EoE) is a chronic immune-mediated disorder marked by type-2 inflammation mediated eosinophil infiltration in the esophagus. Oxylipins derived from polyunsaturated fatty acids (PUFAs) are key players in inflammation, yet their role in EoE remains unclear. Activation of ALOX15 expression in esophageal tissue has been reported for the development of EoE. Lu et al. suggest ALOX15 derived 15-HETE as an activator of a type-2 inflammation response in EoE patients. In this context an approximately twofold increase in the concentrations of 15-HETE in blood plasma of active EoE patients have been reported earlier. Moreover, COX derived PGD2, released from activated mast cells, supports eosinophilic infiltration into the esophagus and PGD2 receptor antagonists have been suggested to offer therapeutic options besides topicals steroids. Since most studies have focused on esophageal tissue, little is known about the formation of oxylipins in the blood of patients with active EoE.

Aims & Methods

This study aims to investigate oxylipin profiles in plasma from patients with active EoE to improve understanding of EoE pathogenesis. Here we analyzed blood samples of 13 EoE patients and 15 healthy controls. Red blood cells (RBC) fatty acid (FA) compositions were analyzed by using gas chromatography (GC) and oxylipin concentrations in the plasma were analyzed by liquid chromatography-tandem mass spectrometry (LC-MS/MS). Prism 10 software (GraphPad, San Diego, CA, USA) was used for statistical analyses of the data.

Results

In order to assess FA composition of the two study groups, we performed GC analyses from the lipid fractions of processed RBC samples. In patients with active EoE, we found a small but significant increase in saturated fatty acids, in particular palmitic acid (16:0) When comparing the omega-6 (n-6) and omega-3 (n-3) PUFAs, ratio of n-6/n-3 PUFA and the EPA + DHA content did not differ between the groups. ALOX5-derived 5-HETE was significantly lower in plasma of patients with active EoE. The ALOX12 products 12-HETE, 12-HEPE and 14-HDHA, were also significantly reduced in the plasma of EoE patients. For ALOX15 derivend 15-HETE we could only detect tendencies of an increase in plasma without reaching a significance level. 17-HDHA was below the limit of quantification and not detectable in plasma samples. Concering COX metabolites, only PGF2α and TxB2 were detectable in plasma, with no significant group differences.

Conclusion

Our study provides new insights into the relationship between FA composition and plasma oxylipin formation in active EoE. Interestingly, while we measured significantly reduced concentrations of 5LOX and 12LOX metabolites in plasma, we could not detect significant changes for 15LOX and COX metabolites. This differs from previous data describing an increase in these 15LOX and COX metabolites in esophageal tissue under the condition of active EoE. This observations suggests that the the systemic circulation may reflect a different inflammatory profile compared to local esophageal tissue inflammation in EOE.

References

Greuter, T., Straumann, A., Fernandez-Marrero, et al., (2024). A Multicenter Long-Term Cohort Study of Eosinophilic Esophagitis Variants and Their Progression to Eosinophilic Esophagitis Over Time. Clinical and translational gastroenterology, 15(4), e00664
Lu S, Herzlinger M, Cao W, et al. (2018). Utility of 15(S)-HETE as a Serological Marker for Eosinophilic Esophagitis. Sci Rep. 2018;8(1):14498. Epub 2018/09/30.
Zhang S, Wu X, Yu S. (2014). Prostaglandin D2 receptor D-type prostanoid receptor 2 mediates eosinophil trafficking into the esophagus. Dis Esophagus. 2014;27(6):601-6. Epub 2013/10/30