Introduction

While robotic surgery has enhanced the precision of laparoscopy, its rigid instrumentation presents limitations. Flexible endoscopic instruments offer potential advantages in reach and access, yet have historically lacked the stability and force needed for human application. The advent of a flexible robotic platform holds promise in providing the necessary visualization, stability, and instrumentation for safe and effective procedures in humans. This video demonstrates the proof of concept for cholecystectomy using a novel flexible robotic platform.

Aims & Methods

In a porcine in-vivo model, a flexible robotic platform was used to perform a cholecystectomy. The platform is a master-slave system with a physician console, patient cart, an 18mm wide, 45cm long flexible shaft, and two 6mm robotic effector arms with 7 degrees of freedom. A 2cm incision was made at the level of the umbilicus to introduce the platform. Once in position, an atraumatic grasper and scissors were introduced under endoscopic control of the cobra camera. Cholecystectomy was performed in a hybrid format, using flexible instruments with one additional 5mm laparoscopic grasper to optimize exposure of the operating field. The triangle of Calot and gallbladder were successfully dissected, and the cystic duct and artery were clipped and divided with the flexible endoscopic instruments. A 5mm laparoscopic camera was used to enhance lighting in the peritoneal cavity beyond the cholecystectomy site, facilitating comprehensive monitoring of the entire procedure and platform in action.

Results

Total operative time was 22 minutes. There were no intraoperative complications. Cholecystectomy was performed safely and completely with the flexible endoscopic instruments.

Conclusion

Single incision cholecystectomy with this novel flexible robotic platform is safe and feasible. The approach may combine the best technical aspects of laparoscopic, robotic, and endoscopic surgery, minimizing trauma and external scars although improvement in exposure and lighting are desirable. Clinical investigation with this promising technology is ongoing.

Disclosure

Silvana Perratta is a member of EndoQuest's Clinical Adivisory Board
Jacques Marescaux is a member of EndoQuest's Scientific Adivisory Board

Introduction

Body composition parameters in patients with celiac disease differ from the normal population, and in some cases, they do not completely normalize even with a gluten-free diet. Despite weight gain on the diet, the change in body composition is generally unfavorable (less increase in fat-free body mass, significantly higher increase in fat content). One reason for this is improved absorption, but the high calorie, sugar, and fat content of the gluten-free diet can also lead to undesirable weight gain and unfavorable metabolic changes (dyslipidemia, fatty liver, insulin resistance) without dietary control.

Aims & Methods

In our prospective multicenter, case-control study (ARCTIC study), we examined body composition parameters and blood lipid levels of patients with celiac disease who had been on a gluten-free diet for at least 1 year and compared them to those of healthy control individuals.
Patients from three university centers with celiac disease participated in the study. Body composition analysis was performed using an Inbody 770 device, measuring the following parameters: body mass index (BMI), waist-hip ratio, Inbody score, Skeletal muscle mass (SMM), Percent body fat (PBF), Visceral fat area (VFA). Blood lipids including total cholesterol, LDL, and triglyceride levels were measured. Abdominal ultrasound was performed to assess the degree of hepatic steatosis. Fischer's exact test and Chi-Square test were used for categorical variables, and the Welch Two Sample t-test was used to assess continuous variables. A p-value less than 0.05 is considered statistically significant.

Results

97 patients with celiac disease (78 female, 19 male) and 47 (40 female, 7 male) control individuals were included. The mean age of patients with celiac disease (36.2 years) was slightly higher than that of controls (32.5 years), but the difference was not significant. The mean waist and hip circumference were higher in patients with celiac disease but did not reach the significance threshold (p=0.051 and p=0.10). Among the body composition parameters, higher mean BMI, Skeletal muscle mass, Percent body fat, Visceral fat area, and lower Inbody score were observed, but the differences were not significant (p=0.31, 0.16, 0.72, 0.39, 0.62). Based on BMI, 9.3% of celiac patients were underweight, 54.7% had normal weight, 18.5% were overweight, and 17.5% were obese. These proportions for controls were: 8.5%, 66%, 19%, 6.5%. The mean total cholesterol level was lower in patients with celiac disease (p=0.043), while the mean triglyceride level was higher (p=0.85). We found significantly more cases of hepatic steatosis in celiac patients than in controls (p=0.012). Among them, 83% met the Metabolic Dysfunction–Associated Liver Disease (MASLD) criteria, while 60% of controls with hepatic steatosis did.

Conclusion

The body composition parameters of celiac patients on a gluten-free diet are less favorable compared to control cases, albeit not significantly. Our data suggest that for most of these patients, the problem is not caused by malabsorption and malnutrition. Preventing cardiovascular and metabolic complications poses a new challenge in patient care.

Disclosure

Funding: National Research, Development and Innovation Office (grant FK142942); ÚNKP-23-3-II-PTE-1855; PTE-KITEP-2023-311

Introduction

Colorectal cancer (CRC) is the third-most common malignancy worldwide. Lysyl oxidase-like protein 2 (LOXL2), which promotes crosslinking of collagen and elastin, contributes to tumor progression in different tumour entities, but its role in CRC has not been evaluated. Here, we unraveled the role of LOXL2 in regulating tumour immune microenvironment of CRC and identified LOXL2 as a potential target for immunotherapy.

Aims & Methods

We analyzed the expression of LOXL2 in the Cancer Genome Atlas (TCGA) data. The effect of LOXL2 on CRC cell growth was determined by cell viability and colony formation assays. In vivo models were established by subcutaneous injection of colorectal cancer cell lines into immunodeficient NOD-SCID-γ (NSG) mice and immunocompetent wild-type BALB/c mice. Flow cytometry and immunohistochemistry were used to analyze immune cell composition. The collagen fibers in the extracellular matrix (ECM) were observed by masson staining, second harmonic generation imaging (SHG), and two-photon fluorescence imaging.

Results

Survival prognosis analysis revealed that high LOXL2 expression was correlated with poor survival of patients with CRC in TCGA cohort. However, we found that LOXL2 overexpression in human (HCT116 and DLD1) and mouse (CT26 and MC38) CRC cell lines had little effect on tumor growth in vitro and in immunodeficient mice. But its overexpression in mouse CRC cell lines could significantly promote tumor growth in immunocompetent mice (p<0.05). Overexpression of LOXL2 led to CXCL1 upregulation in the supernatant of CRC cells and CRC tumor tissues; subsequently, upregulation of CXCL1 induced more migration of myeloid-derived suppressor cells (MDSCs) in vitro and recruited more MDSCs in the tumor microenvironment. On the other hand, LOXL2 increased the crosslinking of collagen fibers in vitro and in the tumor microenvironment, which subsequently activated the function of MDSCs (up-expression of Arg1, IDO1, and INOS) and increased the oncostatin M (OSM) secreted by MDSCs. In turn, OSM secreted by MDSCs induced the expression of LOXL2 in CRC cells. The accumulation and activation of MDSCs in the tumor microenvironment inhibited the infiltration and function of CD8+ T cells, which suppressed the anti-tumour immunity of the host. Targeting of LOXL2 with a neutralizing antibody or inhibitor (PAT-1251) significantly impaired tumor growth (p<0.05) and MDSC infiltration and activation (p<0.05) in immunodeficient mice. Finally, PAT-1251 boosted anti-PD1 efficacy in colorectal cancer, suggesting that it could be repurposed to improve immunotherapy efficacy.

Conclusion

Our findings revealed that LOXL2 exerted a pro-tumorigenic role in CRC. LOXL2 recruited and activated MDSCs through CXCL1 and crosslinking of collagen fibers, respectively, subsequently reducing infiltration and activation of anti-tumor CD8⁺ T cells in the tumor microenvironment. Targeting this axis with PAT-1251 improved the efficacy of immunotherapy in colorectal cancer.

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

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

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.