bluuebunny/crossref_metadata_2025 huggingface dataset:
Transforming growth factor-β1 disrupts angiogenesis during the follicular–luteal transition through the Smad–serpin family E member 1 (SERPINE1)/serpin family B member 5 (SERPINB5) signalling pathway in the cow
Authors: Yan Leyan , Qu Xiaolu , Yu Jianning , Robinson Robert S. , Woad Kathryn J. , Shi Zhendan
Published: May 2021

Intense angiogenesis is critical for the development of the corpus luteum and is tightly regulated by numerous factors. However, the exact role transforming growth factor-β1 (TGFB1) plays during this follicular–luteal transition remains unclear. This study hypothesised that TGFB1, acting through TGFB receptor 1 (TGFBR1) and Smad2/3 signalling, would suppress angiogenesis during the follicular–luteal transition. Using a serum-free luteinising follicular angiogenesis culture system, TGFB1 (1 and 10 ng mL–1) markedly disrupted the formation of capillary-like structures, reducing the endothelial cell network area and the number of branch points (P < 0.001 compared with control). Furthermore, TGFB1 activated canonical Smad signalling and inhibited endothelial nitric oxide synthase (NOS3) mRNA expression, but upregulated latent TGFB-binding protein and TGFBR1, serpin family E member 1 (SERPINE1) and serpin family B member 5 (SERPINB5) mRNA expression. SB431542, a TGFBR1 inhibitor, reversed the TGFB1-induced upregulation of SERPINE1 and SERPINB5. In addition, TGFB1 reduced progesterone synthesis by decreasing the expression of steroidogenic acute regulatory protein (STAR), cytochrome P450 family 11 subfamily A member 1 (CYP11A1) and 3β-hydroxysteroid dehydrogenase (HSD3B1) expression. These results show that TGFB1 regulates NOS3, SERPINE1 and SERPINB5 expression via TGFBR1 and Smad2/3 signalling and this could be the mechanism by which TGFB1 suppresses endothelial networks. Thereby, TGFB1 may provide critical homeostatic control of angiogenesis during the follicular–luteal transition. The findings of this study reveal the molecular mechanisms underlying the actions of TGFB1 in early luteinisation, which may lead to novel therapeutic strategies to reverse luteal inadequacy.
Experimental study of CO2 adsorption using activated carbon
Authors: Almoneef M M , Jedli H , Mbarek M
Published: May 2021

The current study aimed to investigate the CO2 adsorption isotherms of activated carbon (AC), prepared from olive waste via chemical activation by H3PO4. The textural parameters, the microstructural and morphological characterization of the AC were characterized by N2 adsorption-desorption, SEM and XRD. The AC presented the best textural properties with a BET surface area of 1565 m2 g−1 and a micropore volume of 0.649 cm3 g−1. The CO2 adsorption was measured with a volumetric method at three various temperatures: 298 K, 308 K and 318 K. Different models, such as the Langmuir and Freundlich isotherms, were applied to model the CO2 adsorption. Based on the correlation coefficient (R2), the Langmuir model provided a the best fit with the experimental data. Different thermodynamic functions, such as Gibbs free energy and entropy, were calculated using Van Hoff’s formulation. These thermodynamic parameters indicated that the CO2 adsorption is an exothermic and spontaneous process. The results obtained in this study can serve as a benchmark while designing an adsorption-based gas storage system. the Freundlich
DOP10 Persistent sonographic Inflammation as a predictor of clinical complications in patients with Crohn’s Disease
Authors: Vaughan R , Tjandra D , Patwardhan A , Mingos N , Gibson R , Boussioutas A , Christensen B
Published: May 2021

Background

Transmural healing has emerged as a new treatment target and can be assessed non-invasively with intestinal ultrasound (IUS). We investigated whether the presence of persistent sonographic changes were associated with clinical complications including medication escalation, corticosteroid use, hospitalisation and surgery in Crohn’s disease (CD) patients in clinical remission at baseline.

Methods

A retrospective study on 212 patients (50% male) with CD who had IUS between August 2017- June 2020 was performed at a tertiary centre. Our analysis included patients in clinical remission at baseline IUS (HBI≤2 or CDAI<150). Patients were excluded if they had disease confined to the rectum, stoma in situ, suboptimal IUS assessment or < 6 months of follow up available. We compared time of medical escalation, corticosteroid use, IBD related hospitalisation or related surgery in patients with and without sonographic inflammation defined as bowel wall thickness (BWT) >3mm and/or hyperaemia on colour doppler imaging, followed for a median of 19 months. We identified factors associated with survival using Kaplan Meier analysis and Cox proportional hazard model.

Results

At baseline IUS, 61% of patients had sonographic inflammation. During the follow-up period medical escalation occurred in 51% of patients, corticosteroid use in 23%, hospitalisation in 20% and IBD related surgery in 13% of patients. The presence of sonographic inflammation at baseline was associated with increased risk of medical escalation (p=0.0057), corticosteroid use (p=0.037), hospitalisation (P=0.0085) and surgery (P=0.0062) (Figure 1 A-D). On multivariable analysis only maximal BWT significantly predicted medical escalation (HR 1.22 (1.02–1.46), P=0.0268). The presence of hyperaemia at baseline IUS was significantly associated with corticosteroid use (HR 2.20 (1.13–4.28), P= 0.02). No sonographic parameter predicted the need for IBD related hospitalisation or surgery. Baseline immunomodulator use and stricturing (B3) phenotype was significantly associated with surgery (HR 3.59 (1.22–10.54), P=0.02, HR 4.01 (1.14–14.03), P=0.03, respectively).

Figure 1: Kaplan-Meier analysis of the effect of sonographic healing vs sonographic inflammation. A) Medication escalation-free survival. B) Corticosteroid-free survival. C) Hospitalisation-free survival. D) Surgery-free survival.

Conclusion

In CD patients in clinical remission, persistent sonographic inflammation can be used as a non-invasive marker of increased risk for clinical complications.
DOP21 Role of P2X7 in intestinal fibrosis and inflammasome activation: Relevance in Crohn′s Disease
Authors: Lis-López L , Bauset C , Ortiz-Masia D , Gisbert-Ferrándiz L , Coll S , Calatayud S , Barrachina M D , Cosín-Roger J
Published: May 2021

Background

Crohn’s disease (CD) is a chronic inflammatory disorder of the gastrointestinal tract whose etiology is unknown. CD is associated with complications such as fibrosis or fistula, which cannot be pharmacologically reversed, requiring repeated surgery. Although a profibrotic effect of the P2X7 receptor has been described in some scenarios such as lung, heart and liver, its role in intestinal fibrosis has not been analysed yet. Given the crosstalk between fibrosis and inflammasome, we aim to analyze the relevance of P2X7 in intestinal fibrosis and inflammasome activation.

Methods

Surgical intestinal resections of CD patients and healthy ileum of carcinoma patients were obtained. Murine chronic colitis was induced by 4 cycles of DSS in wild-type (WT) or P2X7-/- (KO) mice. HT29 cells were treated 24 hours with an inflammasome activator cocktail (LPS, TNF-α and IFN-γ) and the P2X7 antagonist A-80. Gene and protein expression of P2X7, inflammasome markers (NLRP3, ASC, CASPASE1, IL1β and IL18) and alternative inflammasome pathways (APIs) (NLRP1, NLRC4 and AIM2) were analysed by qPCR and Western Blot. The collagen layer was analysed by Sirius Red Staining. Results are expressed by mean±SEM. Statistical analysis was performed with one-way ANOVA and correlations were analysed with Spearman coefficient.

Results

In CD patients, the expression of P2X7 (2.97±0.50), Nlrp3 (2.53±0.41), Asc (5.61±0.76), Caspase1 (6.90±1.41), IL18 (4.17±0.89) and APIs Nlrp1 (3.07±0.40), Nlrc4 (6.99±1.19) is significantly increased vs non-IBD patients. Moreover, P2X7 expression positively and significantly correlates with the expression of the inflammasome markers NLRP3 (r=0.51), ASC (r=0.38), CASPASE1 (r=0.46), IL18 (r=0.36) and API such as NLRP1 (r=0.73), NRLC4 (r=0.67) and AIM2 (r=0.51) in CD patients (n≥45). The chronic murine model of DSS revealed that: a) KO-DSS showed more aggravated colitis with lower survival and greater weight loss compared with WT-DSS; b) the expression of NLRP3, IL18, IL1β and NLRP1 were significantly increased in KO-DSS (101.00±16.33, 3.28±1.49, 327.50±113.90, 4.92±1.00 respectively) vs WT-DSS; c) the thickness of the collagen layer in KO-DSS was increased vs WT-DSS. As expected, HT29 cells treated with the inflammasome cocktail increased protein expression of caspase-1 and the treatment with the P2X7 antagonist A-80 impaired the inflammasome activation since it significantly reduced the protein expression of caspase-1.

Conclusion

An increased expression of P2X7 receptor, the inflammasome and its APIs is detected in CD patients. Lack of P2X7 worsens chronic colitis associated with an increased activation of the inflammasome. Additional studies are needed in order to elucidate this dual role of P2X7 in intestinal fibrosis.
DOP38 An observational study to evaluate the burden of illness in patients with Crohn’s Disease with and without perianal fistulas in the USA
Authors: Jiang J , Cazzetta S E , Athavale A , Kuharic M , Fan T , Silber A , Vijay A , Hadker N , Sharpe E , Nazarey P P
Published: May 2021

Background

Crohn’s disease (CD) is a chronic inflammatory condition of the gastrointestinal tract that can lead to complications such as perianal fistulas (PAFs). This study compared disease burden, experiences and health-related quality of life between patients who have CD with PAFs (CPF) and those who have CD without PAFs (non-PAF CD).

Methods

This cross-sectional, observational study was conducted in three cohorts of US patients aged 18–89 years with self-reported physician-diagnosed CD: (1) non-PAF CD; (2) CPF without PAF-related surgery; and (3) CPF with PAF-related surgery. Data on medical and surgical interventions, CD-specific symptoms and Fecal Incontinence Quality of Life (FIQL) were collected via a web-enabled questionnaire. Statistical comparisons were assessed at the 0.05 level.

Results

The mean (standard deviation) age of patients in cohorts 1 (n = 300), 2 (n = 51) and 3 (n = 52) was 47 (16.4), 40 (12.2) and 39 (13.1) years, respectively. In patients with CPF (cohort 2 + 3), 59 (57%) reported multiple fistulas and 48 (47%) reported fistula recurrence/persistence. Compared with cohort 1 (non-PAF CD), more patients with CPF reported currently receiving biologic treatment or immunomodulators for CD (58% vs 43% and 23% vs 15%, respectively; both p = 0.01). More patients with CPF also reported undergoing ≥ 1 CD-related surgery and experiencing ≥ 1 failures of CD-related surgery (79% and 20%, respectively) versus cohort 1 (53% and 9%, respectively; both p < 0.001). In cohort 3, 63% of patients had ≥ 3 PAF-related surgeries, and in those receiving seton placement (n = 37), 8% reported placement failure. Post-surgical/seton placement complications were common; the most frequently reported were worsening of pain and swelling around the anus (33%) and fever/infection (29%).

CD-specific symptom frequency and severity results indicated a high symptom burden across cohorts; fatigue was reported with the highest frequency and severity. Faecal incontinence (FI) and leakage-related symptoms affected greater proportions of patients in cohorts 2 and 3, often with greater severity and frequency, compared with cohort 1 (Fig.1). Across all cohorts, 58% of patients (cohort 1/2/3, n = 158/35/40) reported experiencing FI and completed the FIQL questionnaire: cohorts 2 and 3 reported lower (worse) FIQL scores across domains than cohort 1 (Fig.2).

Conclusion

The burden of illness in patients with CPF is substantial, with medical and surgical intervention rates, and symptom severity and frequency greater than for those with non-PAF CD. Higher disease burden, including FI, was noted in patients with CPF compared with the non-PAF CD cohort. An unmet need remains for improved management and outcomes of CPF.

Sponsor: Takeda Pharmaceuticals USA, Inc.
DOP45 Utilization and sustainability of biologics in CD - A nationwide study from the epi-IIRN
Authors: Atia O , Yogev D , Chagit F , Gili F , Orlanski-Meyer E , Magen Rimon R , Natan L , Shira G , Kariv R , Yisca L W , Gabay H , Nevo D , Matz E , Gorelik Y , Chowers Y , Iris D , Turner D
Published: May 2021

Background

Real-world studies demonstrate that ~33% of CD patients fail to respond to induction therapy with biologics. In this nationwide study we aimed to evaluate trends in biologics utilization and sustainability in CD during the last 15 years.

Methods

This study was performed on data from the four Israeli Health Maintenance Organizations, covering 98% of the population. Sustainability was defined as continuous treatment without IBD-related surgeries and at most one short steroid course. Sustainability was compared across different biologics utilizing a propensity score (PS) weighted analysis, estimated by generalized boosted modeling (GBM).

Results

16,936 patients were diagnosed with CD in Israel since 2005 (2,932 [17%] pediatric-onset, 14,004 [83%] adult-onset), of whom 5,804 (34%) were ever treated with biologics (1,659 [57%] pediatric-onset, 4,145 adults [30%], OR 3.1 [95%CI 2.9–3.4]) with a median of 6.4 years follow up (IQR 3.4–9.9). Infliximab was the most common first-line treatment in children (58%, p<0.001), while adalimumab was the most common in adults (63%, p<0.001). However, in recent years there was an increase in adalimumab and vedolizumab utilization in parallel with a decrease in infliximab (Figure). The rate of initiating biologics in the first year of diagnosis increased from 20% among all biologics users during 2005–2010 through 36% during 2011–2014 and 74% since 2015 (p<0.001). The use of combination therapy with immunomodulators is becoming less common in last years and decreased in infliximab from 38% until 2010 to 21% in 2018 (P<0.001) and in adalimumab from 22% to 7%, respectively (p<0.001). The sustainability rate in those treated with infliximab was 65% at one year from initiation of biologics, and 46% and 42% at three and five years, thereafter; compared to 72%, 61% and 57% with adalimumab and 84% and 80% after one and two years in vedolizumab. The rate of primary non-response was 21% with infliximab, 17% with adalimumab and 12% with vedolizumab.

Sustainability was associated with male sex (HR 0.9 [95%CI 0.8–0.95]) and earlier initiation of biologics (HR 0.92 [95%CI 0.9–0.95]). In the PS-adjusted model the sustainability of both infliximab (HR 0.7 [95%CI 0.6–0.8]) and vedolizumab (HR 0.8 [95%CI 0.7–0.95]) were lower than adalimumab. Compared with monotherapy, combination therapy did not change significantly the sustainability rate of all three biologics (adalimumab: HR 1.3 [95%CI 0.8–2.1], infliximab: HR 1.2 [95%CI 0.7–1.9], and vedolizumab: HR 0.8 [95%CI 0.5–1.3]).

Conclusion

Biologics are being increasingly used in CD and earlier during the disease course, but most patients do not sustain treatment at five years. Sustainability rate was higher with adalimumab compared with either infliximab or vedolizumab
DOP51 De-escalation of biological therapy in Inflammatory Bowel Disease patients following prior escalation
Authors: Thomas P , Smits L , Te Groen M , West R , Russel M , Jansen J , Römkens T , Hoentjen F
Published: May 2021

Background

There are limited data available on de-escalation of biological therapy after prior escalation in inflammatory bowel disease (IBD) patients. The aim of this study was to assess the frequency and success rate of de-escalation of biological therapy in IBD patients after prior dose escalation and evaluate which measures are used prior to de-escalation.

Methods

This multicentre, prospective, cohort study enrolled IBD patients treated with infliximab (IFX), adalimumab (ADA) or vedolizumab (VEDO) in whom therapy was de-escalated at least once after prior biological escalation. Objective disease measures for de-escalation were defined as faecal calprotectin ≤ 200 µg/g and/or therapeutic or supratherapeutic trough levels and/or radiologic or endoscopic remission. Successful de-escalation was defined as remaining on the same or lower biological dose for ≥6 months after de-escalation.

Results

In total, 206 IFX users, 85 ADA users and 55 VEDO users underwent therapy escalation. Of these, 34 (17%) patients on IFX, 18 (21%) patients on ADA and 8 (15%) patients on VEDO had received at least one subsequent de-escalation. De-escalation was successful in 91% of IFX patients, 89% of ADA patients and 100% of VEDO patients. The probability of remaining on the de-escalated regimen or further de-escalation after 1 year was 85% for IFX, 62% for ADA and 100% for VEDO. De-escalation based on objective disease measures was performed in 67% of all de-escalations. Objective de-escalations were successful in 98% versus 80% of subjective de-escalations.

Conclusion

De-escalation after biological escalation is successful in the majority of patients. Objective markers of remission increase the likelihood of successful de-escalation.
DOP88 Effect of risankizumab on patient-reported outcomes in patients with Crohn’s Disease who had an inadequate response or intolerance to conventional and/or biologic treatments: Results from phase 3 MOTIVATE and ADVANCE trials
Authors: Peyrin-Biroulet L , Louis E , Ghosh S , Lee S D , Griffith J , Wallace K , Berg S , Loftus E V
Published: May 2021

Background

Risankizumab (RZB), an IL-23 inhibitor, is being investigated for Crohn’s disease (CD). MOTIVATE (NCT03105128) and ADVANCE (NCT03104413) were two double-blind, randomised, placebo (PBO)-controlled phase 3 trials evaluating the efficacy and safety of RZB as induction therapy for CD after inadequate response or intolerance to biologic treatment, or conventional or biologic treatment. We investigated the effect of RZB on patient-reported outcomes (PROs) in these two trials.

Methods

Patients with moderately to severely active CD (CD activity index of 220–450, Simple Endoscopic Score for CD excluding the narrowing component ≥ 6 or ≥ 4 for isolated ileal disease, and average daily stool frequency ≥ 4 and/or average daily abdominal pain score ≥ 2) were randomised 1:1:1 (MOTIVATE; N=569) or 2:2:1 (ADVANCE; N=850) to intravenous RZB 600 mg, RZB 1200 mg or PBO at Weeks 0, 4 and 8. PROs assessed were Inflammatory Bowel Disease Questionnaire (IBDQ), Functional Assessment of Chronic Illness Therapy-Fatigue (FACIT-F), and 36-Item Short Form Health Survey (SF 36). Least-squares mean change from baseline to Week 4 and 12 were assessed for each PRO. Additionally, the proportion of patients achieving IBDQ remission (IBDQ total score ≥ 170 points) was evaluated. Comparisons between RZB dosage groups and PBO were based on Cochran-Mantel-Haenszel or chi-square/Fisher’s exact tests; missing data were reported using non-responder imputation.

Results

In the MOTIVATE (100% biologic inadequate responder [bio-IR]) and ADVANCE (57% bio-IR) trials, significant (P<0.05) improvements in nearly all PROs evaluated were seen as early as Week 4 (Figures 1–4). At Week 12, significant improvements in all PROs were reported for both doses of RZB compared with PBO in both trials, except for the SF-36 Mental Component Summary score (RZB 1200 mg in MOTIVATE), which improved, but not significantly. In MOTIVATE, at Week 4 and 12, IBDQ remission was achieved by significantly (P<0.05) more patients taking RZB (600 mg, 25.1%/38.3%; RZB 1200 mg, 31.9%/44.6%) than PBO (15.5%/26.8%). Results were similar in ADVANCE: RZB 600 mg 30.9%/44.5%, RZB 1200 mg 33.7%/51.5%, and PBO 18.4%/29.8% at weeks 4 and 12, respectively (P<0.001 for both RZB groups compared to PBO).

Conclusion

Patients with active CD and an inadequate response to conventional or biologic treatment who received RZB (600 mg or 1200 mg) induction therapy reported significant improvements in disease-specific (IBDQ) and general health-related PROs (FACIT and SF-36) compared with PBO, with improvements seen as early as Week 4.

Funding statement: Study was funded by AbbVie.

Acknowledgement: Medical writing support was provided by Joann Hettasch of Fishawack Facilitate Ltd, and was funded by AbbVie.
N04 Being an IBD Clinical Nurse Specialist in the UK National Health Service: Challenges, Stressors and Coping Mechanisms
Authors: Dibley L , Kemp K , Duncan J , Mason I , Lisa Y
Published: May 2021

Background

Inflammatory Bowel Disease Clinical Nurse Specialists (IBD-CNSs) provide essential therapeutic and emotional support services to patients with often complex disease. The ever-increasing role expectations, treatment modalities and rising patient population places significant demands on IBD-CNSs (Stansfield, 2019), many of whom enter the role soon after qualifying. Posts in the UK-based RCN IBD Nurse Facebook© page suggest stress is high amongst members. Sustaining IBD-CNSs wellbeing is essential for ensuring sustainability of services. Our aim was to explore experiences of being an IBD-CNS in the UK, in order to inform the wider dialogue around workforce and the wellbeing of specialist nurses.

Methods

This hermeneutic phenomenological study used purposive sampling with maximum variation to recruit Band 6–8 IBD-CNSs who currently, or until recently, worked in NHS Trusts across the UK. Data were collected between July and September 2020, via online or telephone interviews. Focussing on personal perceptions, participants were invited to ‘Tell me what it means to be an IBD-CNS’ with follow-up prompts and probes as needed. Diekelmann et al’s (1989) team analysis method was used.

Results

Twenty-five IBD-CNSs participated [92% Female; Band 6 (n=3), Band 7 (n=10), Band 8 (n=13); 0.5–19 (mean 5.8) years in post; ex-NHS (n=4)]. Two constitutive patterns: ‘Giving and receiving support’, and ‘Developing potential’ were revealed, informed by relational themes addressing patients’ needs, service design/delivery challenges, workload, professional development opportunities, and peer support. Junior IBD-CNSs worried more about workload and learning the role, whilst senior nurses often struggled with the shift towards management responsibilities which they were expected to take on often without any managerial skills or training. The IBD-CNS role was often poorly understood by senior service managers, whose level of support varied widely. Some participants had techniques to mitigate against stressors, but there were also limited professional development opportunities especially in senior roles.

Conclusion

IBD-CNSs are committed to patients, despite an often-burdensome workload. Structural, institutional, and hierarchical issues undermine confidence. Many IBD-CNSs are stressed by mixed messages of delivering an excellent service yet being criticised/under-appreciated by the system. Robust senior support structures lead to better experiences for IBD-CNSs. This work contributes to the UK and global data evidencing the need to pursue strategies which promote wellbeing of all specialist nurses.
N08 Construction of an explanatory model for quality of life in outpatients with ulcerative colitis
Authors: Takahashi M , Nunotani M , Aoyama N
Published: May 2021

Background

Previous studies have reported explanatory models of health-related quality of life (HRQoL) in patients with Crohn disease or inflammatory bowel disease. However, no model for HRQoL has been developed that is specialised for patients with ulcerative colitis (UC). In this study, we aimed to develop and evaluate a predictive explanatory model for HRQoL among outpatients with UC in Japan.

Methods

We conducted a cross-sectional survey between December 2019 and July 2020 at a clinic in Japan. HRQoL was evaluated using the 32-item Inflammatory Bowel Disease Questionnaire (IBDQ-32). We extracted explanatory variables of HRQoL, including disease activity, psychological symptoms, and social support, from previous studies and created an explanatory model based on the conceptual model of Wilson & Cleary (1995). The relationship between explanatory variables and the IBDQ-32 total score was examined using the Spearman’s rank correlation coefficient, Mann–Whitney test, or Kruskal–Wallis test. We conducted multiple regression analysis and path analysis to examine the effect of explanatory variables on IBDQ-32 total score.

Results

We included a total of 203 patients with UC. Variables that were significantly associated with the IBDQ-32 total score were partial Mayo Score, presence or absence of treatment side effects, Hospital Anxiety and Depression Scale (HADS) score, and having/not having an adviser when patients were severely ill-conditioned. HADS had the largest negative effect on IBDQ-32 total score (β = −0.474), followed by partial Mayo Score (β = −0.408), presence/absence of treatment side effects (β = −0.116), having/not having an adviser when patients were ill-conditioned (β = 0.081). Having an adviser or not when ill-conditioned had an indirect effect on patients’ IBDQ-32 total score via HADS (β = −0.111). We verified the final model, which included IBDQ-32 total score and the above four explanatory variables (adjusted R² = 0.501, GFI = 0.996, AGFI = 0.970, CFI = 1.000, RMSEA = 0.010, AIC = 28.043).

Conclusion

Psychological symptoms had the most direct effect on HRQoL in patients with UC and acted as a mediator in the relationship between social support and HRQoL. Nursing interventions to improve HRQoL in patients with UC should consider the effect of psychological symptoms and support when patients are severely ill-conditioned.
N09 Sexuality as lived incompleteness: exploring intimacy and sexuality experiences of people living with inflammatory bowel disease
Authors: Fourie S , Norton C , Jackson D , Czuber-Dochan W
Published: May 2021

Background

Sexual function in those living with Inflammatory Bowel Disease (IBD) is known to be affected by their condition, with impact on their psychosocial well-being. Little is known about the causality of low sexual functions scores and their contribution to low quality of life reports. The aim of this study was to explore how intimacy and sexuality is experienced in the context of IBD.

Methods

This was a phenomenological study guided by van Manen’s methodology. Data were collected from 43 participants from interviews and narrative accounts submitted via Google Forms. Reflection on four existential domains (body, relationships, space and time) and thematic analysis were used to interpret the data.

Results

Four themes were generated from data analysis: Otherness of the sick body, Interrupted connectedness, Missing out on life fullness and Fragmented openness. These led to the overarching theme Sexuality as lived incompleteness, which reflected the essence of the experiences described by participants. Their experiences were deeply embedded in everyday life, and the IBD posed an obstacle to intimacy and sexuality, negatively affecting their life, relationships, family planning and their social position, imposing limitations and isolation.

Conclusion

IBD has a significant impact on sexual well-being with negative effects on overall quality of life, which may be unknown to healthcare professionals, therefore remain unaddressed. A better understanding of the potential issues could help healthcare professionals identify and address concerns and worries related to sexual well-being, and approach them in a reflective, holistic manner in situations arising in practice.
P013 Potential Role of Epithelial Protein Disulphide Isomerases in Crohn’s Disease Fibrosis
Authors: Vieujean S , Hu S , Bequet E , Salée C , Massot C , Bletard N , Pierre N , Quesada Calvo F , Baiwir D , Mazzucchelli G , De Pauw E , Coimbra Marques C , Delvenne P , Edouard L , Meuwis M A
Published: May 2021

Background

Intestinal fibrosis is a common complication of Crohn’s disease (CD) characterized by an accumulation of fibroblasts differentiating into activated myofibroblasts secreting excessive extracellular matrix. In in-vitro experiments, this myofibroblastic differentiation is elicited by a whole series of factors among which transforming growth factor-β1 (TGF-β1) seems to play a key role. The potential role of the intestinal epithelium in this fibrotic process remains poorly defined.

Methods

We performed a pilot proteomic study comparing the proteome of surface epithelium isolated by laser-capture microdissection in normal and fibrotic zones of resected ileal CD strictures (13 zones collected in 5 patients). The pro-fibrotic role of selected epithelial proteins was investigated through in-vitro experiments using HT-29 epithelial cells and a CCD-18Co fibroblast to myofibroblast differentiation model.

Results

Proteomic study revealed an endoplasmic reticulum (ER) stress proteins increase in the epithelium of CD ileal fibrotic strictures, including Anterior gradient protein 2 homolog (AGR2), Protein disulphide isomerase A6 (PDIA6) and Endoplasmic reticulum resident protein 44 (ERP44) which are 3 protein disulphide isomerases. In HT-29 cells, tunicamycin-induced ER stress triggered AGR2, PDIA6, ERP44 as well as TGF-β1 intracellular expression and their secretion. Supernatant of these HT-29 cells, pre-conditioned by tunicamycin (Tm), led to a myofibroblastic differentiation when applied on CCD-18Co fibroblasts. The application of blocking agents for AGR2, PDIA6, ERP44 or TGF-β1 in the supernatant of these Tm-pre-conditioned HT-29 cells, attenuated the myofibroblastic differentiation induced by this supernatant, suggesting a pro-fibrotic role of these secreted epithelial proteins.

Conclusion

The development of CD fibrotic strictures may involve ER stress in epithelial cells, releasing a whole set of proteins into their environment, including AGR2, PDIA6, ERP44 as well as TGF-β1, which could exercise a pro-fibrotic role through a paracrine action.
P018 Myeloid diversity in inflammatory bowel disease revealed by single-cell transcriptomics
Authors: Garrido Trigo A , Corraliza A M , Melón-Ardanaz E , Veny M , Ordás I , Ricart E , Masamunt M C , Panès J , Hemberg M , Mereu E , Heyn H , Azucena S
Published: May 2021

Background

Recent studies have applied single-cell RNA sequencing (sc-RNAseq) of the human intestine to resolve the cellular complexity and heterogeneity characteristic of inflammatory bowel disease (IBD). Nonetheless, an in-depth analysis of the myeloid mucosal compartment, a crucial player in gut immune responses, has not yet been conducted.

Methods

Colonic biopsies from healthy controls (HC, n=6), endoscopically active Crohn’s disease (CD, n=5) or ulcerative colitis (UC, n=6) patients were collected and immediately disaggregated. Single-cell suspensions were processed using the 10x Genomics Next-GEM Single Cell kit assays (Chromium 3’ v3.0) and the resulting libraries sequenced (NovaSeq 6000 S1, Illumina). Data from 3452 myeloid cells was separated in silico and analyzed using Cell Ranger v3.1.0 and the R package Seurat.

Results

In HC, mast cells, CD1c+ dendritic cells (DCs) and resident macrophages (RMΦ) in different transcriptional states were identified. All RMΦ clusters were HLA-IIhi and expressed C1Q genes, SELENOP, CD74 and LYZ. Within RMΦ, a distinct cluster of M2-like cells characterized by high expression of CD209, CD163L1, LILRB5, AXL and MRC1 formed an independent cluster. IBD showed a marked increase in both the number of cells and heterogeneity within the myeloid compartment compared to HC. While RMΦ and M2 macrophages were decreased, we observed an infiltration of inflammatory monocytes and M1-like macrophages expressing the monocyte markers VCAN and CD300E. A cluster of TNIP3-expressing M1 macrophages that co-expressed ACOD1, KYNU, TRAF1, MMP19, IRAK2, TNF and IL1B was present in most UC and CD samples. Additional M1-like macrophage subsets, including a cluster characterized by high expression of CXCL5, HTRA1, SPP1, INHBA, MMP9, MMP14 and MMP10, or a cluster of NRG1, RETN, EREG, HBEGF, TGFB1 and VEGFA-expressing macrophages were identified in UC. Furthermore, IBD samples presented an additional cluster of mature DCs expressing CCL22, CCL17, CCL19 and CCR7. Importantly, our study is the first to capture infiltrating granulocytes in IBD patients by scRNA-seq; specifically, eosinophils expressing CLC, IL4 and IL13, and 3 neutrophil states, expressing FCGR2B, CMTM2, S100A8 and CXCL8.

Conclusion

We describe an unbiased approach for the identification of intestinal myeloid cells that revealed previously undescribed macrophage subpopulations. In addition, we discovered a marked heterogeneity within intestinal inflammatory macrophages that showed high patient-to-patient variability. We suggest that the diverse macrophage states in mucosal lesions could explain important differences in disease pathophysiology and contribute to the observed diversity in patient behavior.
P023 SZN-1326, a Wnt agonist, improved epithelial healing and ameliorated colitis in a chronic DSS model, in stark contrast to anti-TNF and anti-IL-12/23p40 antibodies
Authors: Xie L , Handa P , Tun C , Liu A , Lu C , Baribault H , Newman M , Le L , Shah D , Deshmukh S , Mutha D , Le T , Fletcher R , Chen H , Ye J , Tibbitts J , Srinivasan U , Li Y , Vanhove G , Yeh W C
Published: May 2021

Background

All currently approved treatments for Inflammatory bowel disease (IBD) and the majority of drugs in the pipeline are anti-inflammatory agents. Clinical remission rates have reached a plateau, and there is an unmet need for agents that directly repair and regenerate the intestinal epithelial barrier as mucosal healing has been associated with improved clinical outcomes. Wnt/β-catenin signaling promotes intestinal stem cell renewal and epithelial regeneration. We have engineered a Wnt activator, SZN-1326, which increased Wnt target gene expression in the colon of mice treated with Dextran Sulfate Sodium (DSS), an animal model of IBD and demonstrated efficacy in the acute and chronic forms of this model. The goal of this study was to compare the efficacy of SZN-1326 to the anti-inflammatory agents anti-TNFα or anti-IL-12/23p40 antibodies in a chronic DSS model.

Methods

7- to 8-week old female C57BL6/J mice were administered 3% DSS for three 7-day cycles separated by 7-days off, followed by a 3-day 1% DSS wash-out period. SZN-1326 was injected intraperitoneally (I.P.) at 1, 3, or 10 mg/kg for 2, 4 or 6 doses. Anti-TNFa (MP6-XT22, Biolegend) was injected I.P. at 5 or 25 mg/kg for 4 or 7 doses, and the mice were euthanized on day 38. A separate study examined the efficacy of SZN-1326 relative to anti-IL-12/23p40 (C17.8, Bioxcell) in the same model. Anti-IL12/23p40 antibody at 3 or 10 mg/kg for 4 or 8 doses was given on days 23 or 7 respectively.

Results

During each cycle of DSS treatment, animals exhibited a marked increase in disease activity index (DAI), characterized by body weight loss, diarrhea and bleeding. SZN-1326 at various dosing regimen (1 mg/kg for 4 doses to 10 mg/kg for 2, 4 or 6 doses) significantly improved body weights and decreased DAI compared to isotype control, while anti-TNFa or anti-IL12/23p40 had no efficacy on body weight restoration or DAI. The antibodies could be detected in the serum in accordance with their doses. Treatment with SZN-1326, but not with anti-TNFα significantly decreased colon inflammation, mucosal erosion and overall histopathological score. Further, SZN-1326 treatment significantly reduced serum levels of IL-6 and lipocalin-2, while anti-TNFa decreased only lipocalin-2 levels and anti-IL12/23p40 reduced serum IL-6 level.

Conclusion

In a chronic mouse IBD model, SZN-1326 at various dosing regimens stimulated intestinal epithelial regeneration, induced mucosal healing and restored the epithelial barrier resulting in reduced inflammation, improved body weight and reduced disease activity. In contrast, treatment with anti-TNFα or anti-IL-12/23p40 antibodies led to reduction only in certain serum inflammatory cytokines but had no efficacy on DAI or colon histology in this model.
P033 Dendritic cells from patients with active IBD imprint pro-inflammatory α4β7+CLA+ T cells with potential for gut and skin homing
Authors: Gordon H , Rodger B , Hoti I , Lindsay J O , Stagg A
Published: May 2021

Background

T cell trafficking contributes to inflammation and is a therapeutic target in IBD. Exposure of T cells to retinoic acid during activation induces expression of the gut trafficking integrin α4β7 and suppresses expression of cutaneous leukocyte antigen (CLA) which is required for entry into the skin. Although α4β7+ (gut homing) and CLA+ (skin homing) T cells are generally non-overlapping populations, we have recently observed a low frequency of α4β7+CLA+ (dual tropic) T cells in human blood; these cells may provide an immunological link between gut and skin. Here we investigate the generation of these population by DC in vitro, analyse the function of cells with different tissue tropism and examine the impact of intestinal and skin inflammation.

Methods

Using flow cytometry, expression of α4β7 and CLA was assessed in whole blood, gut tissues, and on proliferating cells generated by stimulation of naïve CD4+ T cells with monoclonal antibodies (anti-CD3/28/2), or with allogeneic colonic or monocyte-derived DC (moDC). Production of TNFα and IFNγ was measured by intracellular cytokine staining.

Results

α4β7+CLA+ T cells were found at low frequency in ileal lamina propria tissue as well as peripheral blood. Naïve CD4 T cells activated in vitro with antibodies expressed α4β7 but not CLA. In contrast, activation with DC resulted in the expression of both α4β7 and CLA and generated a dual tropic a4b7+CLA+ population. Colonic DC generated significantly more α4β7+ cells (p<0.05) and higher levels of α4β7 (p<0.05) than did moDC; colonic DC generated significantly fewer CLA+ cells (p<0.01). Nonetheless, a similar frequency of dual tropic cells was generated by both types of DC. moDC from healthy controls and patients with active IBD or skin inflammation all generated dual tropic cells. moDC from patients with active IBD, but not from patients with active inflammatory skin disease generated more gut tropic (α4β7+) T cells (p<0.05) compared with moDC from healthy controls. Following activation with moDC, cytokine production varied with homing profile. The frequency of TNFα+ and IFNg+ cells was significantly greater in CLA+ expressing populations (p<0.05 and p<0.01, respectively).

Conclusion

CLA expression is induced by gut DC even though they potently upregulate α4β7. The dual tropic population generated is enriched for cells that produce inflammatory cytokines and it may provide a link between gut and skin inflammation. Monocyte-derived cells may contribute to the imprinting of T cell homing to the intestine and upregulation of this activity, specifically in active IBD, supports the concept that intestinal inflammation reprogrammes circulating monocytes.
P049 Development of human embryonic stem cell-derived intestinal organoids for in vitro studies on intestinal inflammation and fibrosis
Authors: Kandilogiannakis L , Filidou E , Drygiannakis I , Tarapatzi G , Arvanitidis K , Bamias G , Valatas V , Paspaliaris V , Kolios G
Published: May 2021

Background

Organoids are self-renewing, 3D structures, consisting of different cell types, with histology and physiology features very close to the physiology of the studied organ. Specifically, human Intestinal Organoids (HIOs) develop epithelial crypts consisting of all subtypes of intestinal epithelial cells which are surrounded by mesenchymal cells. Our aim was to develop 3D HIOs from human embryonic stem cells (hESCs) and examine the expression of fibrotic and mesenchymal factors during their maturation process. Additionally, we investigated the effect of the pro-inflammatory cytokines, IL-1α and TNF-α on the expression of fibrotic and inflammatory mediators in HIOs.

Methods

The human ESC line (H1) was cultured and then differentiated towards HIOs using commercially available kit. HIOs were characterized by immunofluorescence in all differentiation stages. In order to examine their maturation process, we compared the mRNA expression of fibrotic and mesenchymal markers from passages 1–10. In order to examine their functionality, HIOs from different passages were stimulated with 5ng/ml IL-1α and 50ng/ml TNF-α for 12 hours, total RNA was collected and the fibrotic and inflammatory mRNA expression was examined. The mRNA transcripts of CD90, collagen type I, III, fibronectin, CXCL8, CXCL10 and CXCL11 were measured by reverse transcription quantitative PCR.

Results

HIOs were successfully developed as they were stained positive for all tested markers throughout their developmental process. Regarding their maturation process, we observed high expression of CD90, collagen type I, type III and fibronectin that was gradually decreased during passages. As for the fibrotic and inflammatory responses from HIOs, we found that the IL-1α and TNF-α stimulation resulted in statistically significant upregulation of the fibrotic factors, fibronectin, collagen type I and type III in culture passages 2 and 4, but had no effect in culture passages 8 and 10. Similarly, IL-1α and TNF-α stimulation led to the statistically significant induction of the inflammatory chemokines CXCL8, CXCL10 and CXCL11 in culture passages 2 and 4, while no effect was observed in culture passages 8 and 10.

Conclusion

Our findings indicate that HIOs contain a functional mesenchymal component that is gradually diminished during passages. Inflammatory and fibrotic responses of HIOs seem to depend on the fitness of their mesenchyme. IBD studies using HIOs as in vitro models should be performed on early passages, when HIO’s mesenchymal component is still functional.
P050 Effect of biological treatments (anti-TNFs) in the creeping fat of Crohn’s disease patients
Authors: Montfort-Ferré D , Serena C , Millan M , Boronat-Toscano A , Maymó-Masip E , Caro A , Espina B , Vendrell J , Fernández-Veledo S
Published: May 2021

Background

Crohn’s disease (CD) is characterized by persistent inflammation and ulcerations at the small or large bowel, provoking chronic low-grade systemic inflammation. Adipose tissue (AT) is believed to play an active role in the pathogenesis of CD, as the expansion of mesenteric fat attached to the inflamed segments of the intestine, also known as “creeping fat,” is a hallmark of the disease that seems to be directly related to disease activity. We demonstrated that adipose-stem cells (ASC) isolated from the creeping fat of CD patients showed a proinflammatory phenotype and increased the proliferation, migration, and phagocytic capacities of these cells. Taking into account the widely described effects of TNFalpha on the biology and functionality of adipocytes, we believe that biological therapies based on anti-TNF agents modify the inflammatory status of creeping fat. In this context, the effect of anti-TNF treatment on mesenteric fat is poorly studied, and the results are divergent.

Methods

Creeping fat biopsies were obtained from active CD patients that underwent surgery for symptomatic complications: 10 patients were on anti-TNF therapy (at least 6-months prior to surgery) and 10 patients never received any biological therapy. The groups were comparable in age, sex, and body mass index. We isolated from AT biopsies: AT explants, ASC, and adipose tissue macrophages (ATM). Adipose tissue was fixed in 10% phosphate-buffered formalin and embedded in paraffin for histological studies. The proliferation of ASC was performed using the CellTraceTM Violet Cell proliferation kit using flow cytometry and the cell migration of ASC was analyzed using a Tranwell system (8 mmpore polycarbonate membrane).

Results

Histological studies revealed that AT of patients treated with anti-TNF therapy recovered adipocytes morphology and showed lower infiltration of immune cells. Interestingly, we found a significant decrease in the gene expression of pro-inflammatory cytokines (IL1B, IL6, TNFA) in the creeping fat of CD patients treated with anti-TNF (Figure 1A). Furthermore, ATMs isolated from patients treated with anti-TNF showed a significant decrease in the gene expression of antigen-presenting markers (CD74, CIITa, HLA-DPB and HLA-DM) (Figure 1B). To note, ASC isolated from patients with anti-TNF therapy has significantly decreased their proliferation and migration capacities as well as the pro-inflammatory gene expression. Moreover, the anti-inflammatory gene expression and secretion were significantly increased in these cells (Figure 1C).

Conclusion

Anti-TNF therapies impact on the creeping fat of CD patients improving the phenotype of this tissue and this may cause a beneficial effect on CD.
P084 C-Reactive Protein Correlates with Tumor Necrosis Factor-α in Ulcerative Colitis
Authors: McDonagh P , O’Connell F , O’Connell J , Argue R , Corcoran R , Kevans D , O’Sullivan J
Published: May 2021

Background

Monitoring disease activity in Ulcerative colitis (UC) is essential. Inflammatory markers like C-Reactive Protein (CRP) and faecal calprotectin have been shown to correlate with clinical and endoscopic disease activity. To date the relationship between CRP and the inflammatory analytes within the colonic microenvironment have not been analysed.

Methods

Our primary aim was to evaluate the relationship between CRP and key inflammatory cytokine, TNF-α, in UC patients. We also sought to investigate whether CRP correlates with clinical parameters. Ethical approval was granted by our Research Ethics Committee. Patients over the age of 17 with a confirmed diagnosis of UC presenting for colonoscopy were offered the opportunity to participate. Basic patient demographics and Mayo score were recorded. At endoscopy colonic biopsies were taken and cultured in dimethyl sulfoxide (DMSO). Multiplex inflammatory and angiogenic enzyme-linked immunosorbent assay (ELISAs) were performed to evaluate the colonic microenvironment and assess real time secretion of TNF-α. Correlations were carried out on SPSS 24 and plotted using R Studio corrplot.

Results

26 patients with UC participated in the study. The mean Mayo score was 4 (range 0–10). At endoscopy 15% had severe colitis (Mayo endoscopy sub-score 3), 31% had moderate disease, 54% had mild disease. 35% of patients were receiving biologic therapy. The mean CRP was 6.19mg/L (range 1–48.35mg/L), albumin 43.5g/L (range 31-49g/L). CRP had a negative correlation with albumin (r=-0.585, p-value 0.002). CRP had a statistically significant positive correlation with clinical, endoscopic and total Mayo scores (r= 0.469, 0.543, 0.526. p-values 0.016, 0.004, 0.006, respectively). Bleeding at colonoscopy also correlated very strongly with an elevated CRP (r=0.859, p-value=0.00). CRP had a moderate positive correlation with TNF-α (r=0.603, p-value=0.001) and VEGF receptor (r=0.492, p-value=0.011) levels.

Conclusion

CRP displayed a statistically significant positive correlation with the Mayo score. There was a significant correlation between the CRP and TNF-α expression within the colonic micro-environment. TNF-α is a key cytokine and therapeutic target in UC, its relationship with CRP has not been previously assessed. CRP is produced primarily by hepatocytes and is activated by TNF-α, interleukin 6 and interleukin 1β. It therefore stands to reason that there would be correlation between the two. The importance of TNF-α in UC is well known, this study supports and provides new evidence for the use of CRP as a non-invasive marker of disease activity.