scholarly journals P073 An increased autophagy and decreased apoptosis is detected in intestinal fibroblasts from Crohn’s Disease patients

2021 ◽  
Vol 15 (Supplement_1) ◽  
pp. S176-S177
Author(s):  
L Gisbert-Ferrandiz ◽  
M Queralt ◽  
J Cosín-Roger ◽  
S Coll ◽  
C Bauset ◽  
...  
Keyword(s):  
Crohn’S Disease ◽  
Crohn's Disease ◽  
Lamina Propria ◽  
Caspase 3 ◽  
Statistical Significance ◽  
Intestinal Tissue ◽  
Protein Ratio ◽  
Protein Levels ◽  
Mtor Protein ◽  
Significant Increment

Abstract Background Fibrosis is a complication commonly present in Crohn’s disease (CD) patients with a structuring (B2) or penetrating (B3) phenotype, with no effective treatment. This process is characterized by a disequilibrium between the production and degradation of the extracellular matrix (ECM), mainly regulated by myofibroblasts. We aim to analyse here, the expression of markers of autophagy, apoptosis and proliferation in intestinal fibroblasts from CD patients. Methods Fibroblasts were isolated from the damaged intestinal mucosa of CD patients with a penetrating and stenotic behaviour. Control cells were obtained from the non-damaged intestine of patients with colorectal cancer. Protein levels of markers of autophagy and apoptosis were determined by Western Blot in isolated fibroblasts. The proliferation marker Ki67 was analysed by immunohistochemistry (IHC) in 5 µm slides of intestinal tissue from control or CD patients. Statistical significance was measured by t-test. Results In fibroblasts from CD patients, we detected a significant decrease in the ratio phospho-mTOR / mTOR (Fig. A) in parallel with a non-significant increment in the LC3 II / LC3 I protein ratio (174% ± 46.5), and a decrease in p62 protein levels (84.8% ± 5.5). When compared between CD behaviours, a significant decreased in the phospho-mTOR / mTOR protein ratio was detected in fibroblasts from B2- compared to that obtained in cells from B3-CD patients (Fig. B). The analysis of the expression of an apoptosis marker, Caspase 3, revealed a decreased of cleaved caspase 3 protein levels in CD fibroblasts compared to levels detected in control cells (Fig C). Finally, we observed in the lamina propria of the intestine from CD patients an increased number of Ki67 positive cells, compared to that detected in control tissue. Conclusion Our data show an increased autophagy and decreased apoptosis in isolated intestinal fibroblasts from CD patients; the high number of cells proliferating in the lamina propria of the intestinal tissue of these patients, strongly suggests a higher viability of these cells in the fibrotic context.

scholarly journals Anti-TNF antibodies do not induce the apoptosis of lamina propria mononuclear cells in uninflamed intestinal tissue in patients with Crohn’s disease

2013 ◽  
Vol 51 (3) ◽  
pp. 239-243 ◽  
Author(s):  
Piotr Eder ◽  
Liliana Lykowska-Szuber ◽  
Iwona Krela-Kazmierczak ◽  
Kamila Stawczyk-Eder ◽  
Karolina Sterzynska ◽  
...  
Keyword(s):  
Crohn’S Disease ◽  
Crohn's Disease ◽  
Lamina Propria ◽  
Mononuclear Cells ◽  
Intestinal Tissue ◽  
Lamina Propria Mononuclear Cells

scholarly journals P046 Vitamin D decreases PDIA3 and prevents the enhanced migration of fibroblasts from stricturing Crohn’s disease

2020 ◽  
Vol 14 (Supplement_1) ◽  
pp. S156-S156
Author(s):  
L Gisbert-Ferrandiz ◽  
J Cosín-Roger ◽  
S Coll ◽  
C Bauset ◽  
D Ortiz-Masiá ◽  
...  
Keyword(s):  
Vitamin D ◽  
Wound Healing ◽  
Crohn’S Disease ◽  
Crohn's Disease ◽  
Protein Disulfide Isomerase ◽  
Statistical Significance ◽  
Protein Levels ◽  
And Migration ◽  
Protein Disulfide ◽  
Wounded Area

Abstract Background Fibrosis is a common complication in Crohn’s disease (CD) patients and fibroblasts play an important role in the fibrogenic process. Low vitamin D (VD) levels and a defective VD-signalling pathway have been reported in CD. VD signals through both vitamin D receptor (VDR) and protein disulfide-isomerase A3 (PDIA3) and we have previously demonstrated that VDR protein levels are reduced in fibroblasts isolated from CD patients and that VD increased VDR expression in these cells (A-2080; ECCO 2019). We aim to analyse here the effect of VD on both PDIA3 protein levels and migration in CD fibroblasts. Methods We used intestinal fibroblasts isolated from surgical resections of the damaged mucosa of CD patients with stricturing behaviour (B2). Control fibroblasts were obtained from the non-damaged intestine of patients with colorectal cancer. Fibroblasts were treated with VD (100 nM) or its vehicle for 24 h and PDIA3 protein levels were measured by Western Blot. In the wound healing analysis, a single scraping was done in the centre of the fibroblasts monolayer and FBS-free medium was added to the cells, which allows us to determine the ability of fibroblasts to migrate and close the wound. Photos were taken at 0, 24 and 48 h. Results of wound healing were expressed as the percentage of the wound at each time point for the maximal wounded area (time 0, 100%). Statistical significance was measured by ANOVA or t-test. Results No significant differences in PDIA3 protein levels were detected between control and CD fibroblasts but VD significantly decreased PDIA3 expression in CD fibroblasts (Figure 1A). In the wound healing assay, we detected that CD-B2 fibroblasts migrate faster than control cells, resulting in a reduced wounding area, 48 h later (Figure 1B). Treatment of these CD-B2 cells with VD decreased their migration rate, and 48 h later cells exhibited a higher and significant wounding area than vehicle-treated cells (Figure 1C). Conclusion Vitamin D decreased PDIA3 expression and prevented the accelerated migration detected in intestinal fibroblasts from B2-CD patients which suggest an anti-fibrotic effect of VD mediated by a direct effect of this hormone on intestinal fibroblasts.

ACTIVATED INTESTINAL MUSCLE CELLS PROMOTE PREADIPOCYTE MIGRATION: A NOVEL MECHANISM OF CREEPING FAT FORMATION IN CROHN’S DISEASE

2021 ◽  
Vol 27 (Supplement_1) ◽  
pp. S34-S34
Author(s):  
Ren Mao ◽  
Genevieve Doyon ◽  
Ilyssa Gordon ◽  
Jiannan Li ◽  
Sinan Lin ◽  
...  
Keyword(s):  
Extracellular Matrix ◽  
Crohn’S Disease ◽  
Crohn's Disease ◽  
Ex Vivo ◽  
Muscularis Propria ◽  
Dominant Factor ◽  
Stricture Formation ◽  
Intestinal Tissue ◽  
Mesenteric Fat

Abstract Background and Aims Creeping fat, the wrapping of mesenteric fat around the bowel wall, is a typical feature of Crohn’s disease, and is associated with stricture formation and bowel obstruction. How creeping fat forms is unknown, and we interrogated potential mechanisms using novel intestinal tissue and cell interaction systems. Methods Tissues from normal, ulcerative colitis, non-strictured and strictured Crohn’s disease intestinal specimens were obtained. Fresh and decellularized tissue, mesenteric fat explants, primary human adipocytes, pre-adipocytes, muscularis propria cells, and native extracellular matrix were used in multiple ex vivo and in vitro systems involving cell growth, differentiation and migration, proteomics, and integrin expression. Results Crohn’s disease muscularis propria cells produced an extracellular matrix scaffold which is in direct spatial and functional contact with the immediately overlaid creeping fat. The scaffold contained multiple proteins, but only fibronectin production was singularly upregulated by TGF-b1. The muscle cell-derived matrix triggered migration of pre-adipocytes out of mesenteric fat, fibronectin being the dominant factor responsible for their migration. Blockade of α5β1 on the pre-adipocyte surface inhibited their migration out of mesenteric fat and on 3D decellularized intestinal tissue extracellular matrix. Conclusion Crohn’s disease creeping fat appears to result from the migration of pre-adipocytes out of mesenteric fat and differentiation into adipocytes in response to an increased production of fibronectin by activated muscularis propria cells. These new mechanistic insights may lead to novel approaches for prevention of creeping fat-associated stricture formation.

scholarly journals Lamina propria macrophage phenotypes in relation to Escherichia coli in Crohn’s disease

2015 ◽  
Vol 15 (1) ◽  
Author(s):  
Timothy R. Elliott ◽  
Neil B. Rayment ◽  
Barry N. Hudspith ◽  
Rebecca E. Hands ◽  
Kirstin Taylor ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Crohn’S Disease ◽  
Crohn's Disease ◽  
Lamina Propria ◽  
Macrophage Phenotypes

scholarly journals The association between inflammatory potential of diet and disease activity: results from a cross-sectional study in patients with inflammatory bowel disease

2020 ◽  
Vol 20 (1) ◽  
Author(s):  
Carlijn R. Lamers ◽  
Nicole M. de Roos ◽  
Ben J. M. Witteman
Keyword(s):  
Ulcerative Colitis ◽  
Inflammatory Bowel Disease ◽  
Crohn’S Disease ◽  
Crohn's Disease ◽  
Disease Activity ◽  
Bowel Disease ◽  
Activity Index ◽  
Statistical Significance ◽  
Inflammatory Bowel ◽  
Anti Inflammatory

Abstract Background Diet may play a role in disease status in patients with inflammatory bowel disease. We tested whether the inflammatory potential of diet, based on a summation of pro- and anti-inflammatory nutrients, is associated with disease activity in patients with Crohn’s disease and ulcerative colitis. Methods Participants completed a disease activity questionnaire (short Crohn’s Disease Activity (sCDAI) or Patient Simple Clinical Colitis Activity Index (P-SCCAI)) and a Food Frequency Questionnaire (FFQ). FFQ data were used to calculate the Dietary Inflammatory Index (DII) which enables categorization of individuals’ diets according to their inflammatory potential on a continuum from pro- to anti-inflammatory. Associations with disease activity were investigated by multiple linear regression. Results The analysis included 329 participants; 168 with Crohn’s disease (median sCDAI score 93 [IQR 47–156]), and 161 with ulcerative colitis (median P-SCCAI score 1 [IQR 1–3]). Mean DII was 0.71 ± 1.33, suggesting a slightly pro-inflammatory diet. In Crohn’s disease, the DII was positively associated with disease activity, even after adjustment for confounders (p = 0.008). The mean DII was significantly different between participants in remission and with mild and moderately active disease (0.64, 0.97 and 1.52 respectively, p = 0.027). In ulcerative colitis, the association was not significant. Conclusions Disease activity was higher in IBD participants with a more pro-inflammatory diet with statistical significance in Crohn’s disease. Although the direction of causality is not clear, this association strengthens the role for diet in medical treatment, which should be tested in an intervention study.

scholarly journals Trends in Inflammatory Bowel Disease Therapy: A Meta-Analytic Approach

1995 ◽  
Vol 9 (7) ◽  
pp. 405-411 ◽  
Author(s):  
Gregory W Whiting ◽  
Joseph Lau ◽  
Bruce Kupelnick ◽  
Thomas C Chalmers
Keyword(s):  
Crohn’S Disease ◽  
Crohn's Disease ◽  
Meta Analysis ◽  
Statistical Significance ◽  
Analytic Approach ◽  
Disease Therapy ◽  
The Matrix ◽  
Inflammatory Bowel ◽  
Acute Attacks ◽  
New Treatments

OBJECTIVE: To illustrate the current meta-analytic approach to extracting clinically useful information from the glut of randomized controlled trials (RCTs) of the treatment of acute attacks of Crohn's disease.PROCEDURE: Acceptable RCTs were classified in a matrical format so that the feasibility of a meta-analysis of each treatment could be quickly determined. Each potential inclusion was then photocopied after blinding the source and outcome to minimize bias by the investigators.RESULTS AND CONCLUSIONS: The search was started in 1990, when a previous meta-analysis was completed. Over 100 citations in MEDLINE contained 21 RCTs of the treatment of acute attacks of Crohn's disease, and 11 more were culled from the references of recent papers. The meta-analysis most indicated by the matrix was a comparison of several different immunosuppressive regimens with a placebo. The previously demonstrated statistical significance was confirmed and the point made that it is no longer justified to compare any new treatments with a placebo. Cumulative meta-analysis, the process of performing a new analysis every time a new trial is published, revealed the importance of employing both random and fixed effect models when heterogeneity of treatment results is appreciable, as in this case.

scholarly journals P303 Role of parietal healing by ultrasound in the evolution of patients with Crohn’s disease

2020 ◽  
Vol 14 (Supplement_1) ◽  
pp. S304-S305
Author(s):  
P Mayor Delgado ◽  
C J Suarez Ferrer ◽  
J Poza Cordon ◽  
E Martin Arranz ◽  
J L Rueda Garcia ◽  
...  
Keyword(s):  
Crohn’S Disease ◽  
Survival Analysis ◽  
Crohn's Disease ◽  
Median Survival ◽  
Statistical Significance ◽  
Inflammatory Activity ◽  
First Year ◽  
Ultrasound Findings

Abstract Background The objective of this study is to assess the degree of parietal involvement with ultrasound and see how they influence the prognosis according to the findings. Methods It was defined as ‘parietal healing’ the normalisation of ultrasound findings in a previously affected segment (parietal thickness less than 3 mm and absence of parietal hyperaemia). Therefore, patients with absence of ultrasound activity but without reaching parietal healing were considered to be those without significant inflammatory signs (parietal thickness <4.5 mm and absence of hyperaemia or other inflammatory data) but who had not ‘normalised’ the previously affected segment. Results A total of 111 patients without ultrasound inflammatory activity were included. It was studied whether normalisation of parameters in a previously affected segment, i.e. parietal healing gave a better prognosis during follow-up. It was detected that 74 patients achieved ‘parietal healing’ and this subgroup of patients generally had a flare in 12% of cases (9/74 patients) compared with 27% of cases (10/37) among patients without parietal healing, these differences were statistically significant (p = 0.05). Similar, it was identified that among patients without ultrasound activity who had a subsequent outbreak during follow-up, the median survival was lower (1,09 years) in patients without parietal healing vs. those with parietal healing (1.4 years), the differences reaching statistical significance (p = 0.03). Likewise, during the follow-up, it was objectived, by means of survival analysis, that the evolution of patients without ultrasound activity was more favourable in those with ‘parietal healing’ with remission 95% vs. 90% at first year, 89% vs. 81% at 2 years and 85% vs. 66% at 3 years of follow-up. The results are outlined in Figure 1. Conclusion To conclude, detecting parietal healing by ultrasound improves prognosis by decreasing the number of outbreaks and increasing the median survival until outbreaks occur.

Sign in / Sign up

Export Citation Format

Share Document