bluuebunny/crossref_metadata_2025_split huggingface dataset:
P020 Differential expression of miR-424-5p and miR-378c in stenotic and penetrating lessions of Crohn’s Disease associates with altered transcription of genes involved in vascular regulation
Authors: Coll S , Seco M , Bauset C , Lis L , Cosín-Roger J , Ortiz-Masia D , Gisbert-Ferrandiz L , Barrachina D , Calatayud S 
Published: January 2022

Abstract

Background
Crohn′s disease (CD) is often complicated by the appearance of intestinal stenosis (B2 behaviour) or fistulas (B3 behaviour), and the factors determining the pattern of disease and the etiology of these lesions, both associated with fibrosis and requiring surgery, are poorly defined. Our aim has been to detect transcriptomic alterations in intestinal tissues affected by stenosis or fistulas, and have centred our attention in genes affecting vessel formation and function as potential contributors to these lesions.

Methods
We conducted a small RNA sequencing profiling on ileal surgical resections from CD patients with B2 (n=10) or B3 (n=10) behaviour. We obtained a sample from affected tissue and the paired non-affected ileum from each patient.

Results
Tissues affected by stenotic or penetrating lesions present a differential expression of two pro-angiogenic miRNAs (Fig1A) and an increased expression of genes associated with vascular regulation (Fig1B), and both alterations are more pronounced in penetrating lesions. The expression of this set of genes shows a generalized and significant correlation with that of the pro-angiogenic miRNAs (negative with the down-regulated miR-378c, positive with the up-regulated miR-424-5p, Fig1C), especially in the affected tissue.

Fig1. Expression of pro-angiogenic miRNAs (A) and of genes associated with vascular regulation (B) in tissues from stenotic (B2) and penetrating (B3) lesions of CD patients. Data represent normalized counts of each RNA and are expressed as mean±SEM (A) or mean (B). Data in (C) correspond to Pearson’s R. Significant differences vs corresponding non-affected tissue are represented by *P&lt;0.05 (paired t-test) and #P&lt;0.05 (ANOVA+Tukey test)

Conclusion
Ileal tissues from CD patients affected by stenosis or penetrating injuries present a local alteration in the expression of miRNAs with pro-angiogenic functions that probably contributes to the up-regulation of genes associated with vessel formation and to the modification of the vascular compartment in these lesions.
P036 Identification and functional analysis of stromal subsets in experimental IBD mouse models
Authors: Zhou Z , Plug L G , de Jonge-Muller E S M , van de Beek L M , Brands L , van der Meulen-de Jong A E , Hawinkels L J A C , Barnhoorn M C 
Published: January 2022

Abstract

Background
Single cell RNA sequencing data from the inflamed colon of inflammatory bowel disease (IBD) patients revealed the presence of different stromal cell/fibroblast subsets and showed their importance in IBD pathology. Up to now, it is unknown if and how the stroma cells affect disease progression. In this study we compared the stromal cell subsets in three IBD mouse models and explored the interaction between these stromal subsets and epithelial cells.

Methods
Stromal cell subset markers, including CD55, C-X-C motif chemokine 12 (CXCL12), podoplanin (PDPN), CD90 and CD73, were analyzed in the inflamed murine colon by flow cytometry in interleukin (IL)-10 knockout (KO) mice, dextran sulfate sodium(DSS)-induced colitis and the T cell transfer model. In order to explore the changes in stromal subsets upon colitis induction in vitro, short hairpin RNAs were used to silence target gene expression of these cellular markers in murine fibroblasts. Fibroblast proliferation and migration was studied in addition to the effects on immune cell trafficking and tissue regeneration.

Results
In all three colitis models, an increase in the total amount of stromal cells was observed. The percentages of PDPN+ and CD73+ stromal cells significantly increased during colitis in the IL-10 KO mice, while the percentage of CD55+, CD90+ and CXCL12+ stromal cells decreased. For the T cell transfer model, the abundance of CD73+, CD90+ and CD55+ stromal cells was enriched, while the percentage of CXCL12+ stromal cells was reduced. Interestingly, in contrast to the two other, in the DSS-mice the percentage of CXCL12+ stromal cells was significantly increased. In vitro experiments showed the importance of CD55 and CXCL12 for fibroblast proliferation and CD55 for their migration. Furthermore, the fibroblast supernatant obtained after the knockdown of CXCL12 decreased epithelial cell wound healing. On the contrary, knock-down of CD90 in the fibroblasts improved epithelial migration. Current experiments explore the immune regulatory properties of the fibroblast subsets.

Conclusion
The composition of stromal cell subsets is significantly changed in experimental colitis and differs between three established colitis mouse models. Therefore they may possibly reflect different human IBD subtypes observed in the clinic. Finally the defined subsets influence fibroblast behavior and epithelial regeneration.
P042 Succinate promotes the conversion from white to beige adipose tissue in Crohn’s Disease
Authors: Montfort-Ferré D , Menacho M , Caro A , Espina B , Boronat-Toscano A , Nuñez-Roa C , Martí M , Espin E , Vendrell J , Fernández-Veledo S , Serena C 
Published: January 2022

Abstract

Background
Crohn’s disease (CD) is characterized by severe transmural inflammation with subsequent destruction of the intestinal barrier. Bacterial infiltration across this leaky gut facilitates access to the mesenteric fat and the development of a subsequent inflammatory reaction in the surrounding adipose tissue named creeping fat (CF). Dysbiosis in CD patients has been associated with an increase in succinate-producing bacteria and a decrease in succinate-consuming bacteria. In fact, elevated succinate levels have been found in the intestine and feces of CD patients. Furthermore, succinate is an inductor of beige adipogenesis in white adipose tissue progenitors.

Methods
Visceral and subcutaneous adipose tissue biopsies (VAT and SAT), serum, and plasma were obtained from a cohort of active CD (n=20), inactive CD (n=12), and control group (n=15). From adipose tissue biopsies, adipose-derived stem cells (ASCs) and adipose tissue macrophages (ATMs) were isolated. Different experiments were performed such as protein uncoupling protein 1 (UCP1)-immunohistochemistry and immunofluorescence, gene expression studies, adipose tissue explants, proteomic analysis of ASCs, succinate determinations, bio-impedancemetry, and infrared photographs.

Results
Succinate treatment increased markers of beige adipose tissue in ASCs from controls and inactive CD patients (Fig. A). Surprisingly, the markers of beige adipose tissue were already high in ASCs isolated from active CD, due to being exposed to an environment with high levels of succinate in vivo. Proteomic analysis of isolated CD-ASCs revealed that these cells secrete beige adipose tissue-related proteins compared to Control-ASCs (Fig. B). Furthermore, bacterial translocation increases the conversion of pre-adipocytes from white to beige adipose tissue in CD individuals (Fig. C). Immunohistochemical study of UCP-1, confirms increased staining in adipose samples from CD patients from both depots (SAT and VAT) (Fig. D). Altogether, adipose tissue depots from CD patients had a conversion from white to beige adipose tissue with a high protein UCP1-gene expression. Thermic images show an increase of temperature in the intestinal area of active CD patients that is not observed in inactive CD patients or the control group.

Conclusion
Understanding the role of beige adipose tissue in CD could help develop new therapeutic or diagnostic strategies in these patients.
P047 Effect of infliximab and vedolizumab on M1 and M2 macrophages derived from THP-1 cells
Authors: Notararigo S , Viñuela-Roldán J , Zurita-Gonzalez M J , Dominguez Medina E , Dominguez-Munos J E , Barreiro-de Acosta M 
Published: January 2022

Abstract

Background
Inflammatory bowel diseases encompass two pathologies affecting the digestive tract, Crohn’s disease (CD) and ulcerative colitis (UC). They are characterized by a high and chronic inflammatory rate, resulting from activation of the intestinal immune system and increased permeability of the intestinal barrier leading to lesions in the intestinal tract. Non-classical CD14- CD16+ monocytes, are recruited to the focus of inflammation from the peripheral blood and preferentially differentiate to anti-inflammatory macrophages CD206+ CD163+ HLA-DR- in the intestine, playing a key role in the repair of such lesions. In some cases, treatment with biologics, such as vedolizumab, could interfere with this biological process leading to a modification of the innate immune response. Previous results showed that CD and UC had a different distribution of non-classicalmonocytes, leading to the hypothesis that treatment with biologic could modulate macrophage wound healing response.

Methods
Macrophages derived by THP-1 cells were polarized to M0, after PMA treatment, followed by LPS and IFN-γ for M1 macrophages, or IL-4 and IL-13 for M2 macrophages. To assess the effect of biologics on macrophage immune response, Infliximab (IFX) and Vedolizumab (VDZ) were added independently to the cell cultures until polarization process finalized. Immunophenotyping of M0, M1 and M2, and transcription factor expression profiling of pSTAT-1, pSTAT-6 and pNFκB, were carried out by flow cytometry. Cytokine production of IL-6, TGF-β and IL-10, was performed by ELISA immunoblot. Macrophages immune response was assessed with wound healing assay with Caco-2.

Results
Immunophenotyping revealed that M1 macrophages increased the expression of surface markers like HLA-DR, CD38 and reduced CD11c, vs. M0 macrophages, indeed transcription factor expression of pSTAT-6 and pNFκB were modulated. M2 macrophages showed the decrease of CD163 and HLA-DR vs. M0 macrophages, while no significant effect was observed for CD38 and CD206. Transcription factor expression revealed that pSTAT 6 incremented vs. M0. Cytokine profiling showed a clear difference between the tree macrophages types. Finally wound healing assay with Caco-2 revealed how biologic interfered with innate immune response.

Conclusion
Our results demonstrated that the treatment administration could be responsible of macrophage differential immune response, especially upon biologic treatment. These findings could lead to support the establishment of precision medicine in order to modulate a suitable macrophage response.
P070 Novel long-acting lipidated interleukin-22 normalises colonic inflammation in an acute DSS colitis model
Authors: Van De Bunt M , Norrild J C , Sass-Ørum K , Kjølbye A L , Jorgensen R 
Published: January 2022

Abstract

Background
Most current treatments and new agents in clinical development for Inflammatory Bowel Disease (IBD) focus on inhibition of excessive inflammation. However, clinical remission rates for IBD have reached a plateau. New approaches directly improving mucosal healing, a process associated with reduced hospitalizations and long-term remission, therefore have great potential for addressing the unmet need. The cytokine interleukin-22 (IL-22) plays a key role in epithelial healing and has shown early proof-of-concept as a potential drug class in a phase 1 trial in Ulcerative Colitis. Here we present results from a mouse model of colitis on the efficacy of lipidated IL-22 analogues, which maintain more native-like molecular properties than current IL-22 modalities in development.

Methods
Chow-fed female C57Bl/6JRj mice were randomized to 4 groups (n=8 per group) based on body weight. Three groups received 2.5% (w/v) Dextran Sulfate Sodium (DSS) in the drinking water for 7 days from the start of the study. Animals receiving water without DSS served as healthy controls. Mice in the DSS groups received vehicle or lipidated IL-22 intraperitoneally at two dose levels for the entire duration of the study (10 days). At termination, plasma was sampled for assessment of target engagement biomarkers and intestines were removed and measured for weight and length before stereological analysis.

Results
Mice subjected to DSS developed colitis characterized by sustained weight loss and bloody diarrhoea. Vehicle dosed DSS mice showed clear histological features of colitis, with significantly (P&lt;0.05) increased weight/length ratio of the colon as well as colonic mucosal, submucosal and inflammation volumes compared to healthy controls. Treatment with lipidated IL-22 significantly (P&lt;0.05) improved all these parameters except mucosal volume compared to vehicle dosed DSS mice; in particular colonic inflammation volume was normalized to healthy control levels. Dose-dependent biomarker responses (e.g. Peptide YY levels) confirmed target engagement of the lipidated IL-22.

Conclusion
Novel long-acting lipidated IL-22, which acts on epithelial repair mechanisms, demonstrated strong improvements in histological parameters with normalisation of colonic inflammation in a DSS model of colitis. These beneficial effects were accompanied by dose-dependent increases in target engagement biomarkers. The underlying lipidation technology used for engineering these compounds has validated best in class potential in marketed treatments due to its molecular properties (e.g. size and potency). Clinical trials to assess safety and efficacy of lipidated IL-22 are expected to commence early 2023.
P074 SZN-1326 promotes colonic mucosal healing in an acute injury model of IBD by first accelerating epithelial regeneration and secondarily reducing inflammation
Authors: Fletcher R , Xie L , Phipps J , Sanman L , Deshmukh S , Shah D , Hill N , Liu A , Gupta S , Sura A , Chen H , Ye J , Yeh W C , Li Y , Lu C 
Published: January 2022

Abstract

Background
Wnt signaling plays critical roles in regulating intestinal stem cell maintenance and differentiation. The inflammatory bowel disease, ulcerative colitis (UC), is characterized by epithelial lesions, and there is a need for therapies that promote mucosal healing in addition to limiting inflammation. The Surrozen proprietary Wnt mimetic, SZN-1326, is a FZD5,8 and LRP6-specific, full-length, effectorless, bi-specific IgG1 molecule that is well-suited for impacting the intestinal epithelium where FZD5 is highly expressed. Previously, we have shown that in an acute dextran sodium sulfate (DSS) epithelial injury model, where 4% DSS was provided in the drinking water for seven days followed by 1% DSS until termination at day 10, a single dose of SZN-1326 at 1 mg/kg or higher on Day 4 robustly induced intestinal epithelial healing and reduced inflammation, and we wanted to understand more thoroughly the mechanism underlying this activity.

Methods
Here, we report on additional studies in the same DSS model including scRNA-seq and tissue analysis timepoints at days 3, 4, 5, 6, 7, and 10 (6-days post treatment).

Results
DSS impacted all tissue layers by day 3. The direct effect of SZN-1326 on the epithelium occurred as early as 24-hours after treatment (day 5 of the experiment). SZN-1326 induced and/or increased Wnt target and cell cycle gene expression in the epithelium leading to progenitor cell expansion, and following this wave of enhanced progenitor proliferation, the cells transited toward colonocyte differentiation more quickly than the progenitor cells in control treated samples. Even in the presence of 1% DSS, relative to the anti-GFP treatment samples, the absorptive cell lineage in the SZN-1326-treated epithelium was more similar at a transcriptomic level to the uninjured condition. In addition, when comparing SZN-1326 treatment to anti-GFP at 24-hours, there were no differentially expressed genes in either the stroma or immune cells. However, analysis of the later timepoints revealed a robust reduction in cytokine/inflammatory signaling in the stromal and immune cells and a drastic reduction of infiltrating monocytes and neutrophils, which we validated in the tissue by immunohistochemistry. We detected this reduction by 72-hours after treatment, and it was more pronounced by day 10, 6-days after treatment, implicating a secondary effect.

Conclusion
In this acute damage context, SZN-1326 promoted mucosal healing by transiently inducing epithelial progenitor cell expansion and accelerating epithelial repair. Secondary to its initial impact on the epithelium, SZN-1326 reduced stromal and immune cytokine signaling and immune cell infiltration.
P108 Identifying Patient-Centered Future Research Priorities: A Report from the Patient Reported Outcomes after Pouch Surgery (PROPS) Delphi Consensus Study
Authors: Cavallaro P , Bordeianou L 
Published: January 2022

Abstract

Background
The Patient Reported Outcomes after Pouch Surgery (PROPS) Delphi Consensus Study recently described Ileoanal Pouch Syndome – the constellation of functional outcomes that should be the focus of future research. Importantly, several additional items not related to bowel function were identified by patients as critical to their quality of life. The aim of this abstract is to highlight these additional items.

Methods
Three panels of expert stakeholders were chosen to correlate with the clinical scenario of the multidisciplinary team that cares for pouch patients: (A) patients, (B) colorectal surgeons, (C) gastroenterologists/other clinicians. IPAA patients were recruited by advertisements on the social media pages of a national patient advocacy groups. Clinicians were recruited by a 14 member scientific committee with the aim of selecting experts in inflammatory bowel disease and IPAAs. A starting list of items was generated by systematic literature review. Three rounds of Delphi surveys were employed to select high priority items based on an a priori threshold. Participants had the opportunity to suggest additional symptoms not included in the starting list. A final online consensus meeting with representation from all three expert panels was held to finalize a consensus statement.

Results
After three rounds of voting, 195 patients, 62 colorectal surgeons, and 48 gastroenterologists/other specialists completed all three rounds. In the patient group, 95% had UC and J-pouch anatomy. In the early Delphi rounds, approximately 74% of patients identified impact on sexual function/intimacy as a high priority outcomes, while 70% of patients identified impact on ability to conceive as a high priority. Similarly, 78% of patients believed that risk of pouchitis was a high priority, compared to 47% of surgeons and 66% of gastroenterologists. Furthermore, patients participating in the subsequent online focus groups and final consensus meeting felt strongly that future research on delivery of care, long term follow up, and proper consent before surgery (including accurate counseling on function and perception of being “cured”) were critical areas for future research.

Conclusion
PROPS was the first study to identify key functional outcomes after pouch surgery with direct input from a large panel of IPAA patients. The inclusion of patients in all stages of the consensus process allowed for a true patient-centered approach in defining the core domains that should be focused on in future studies of pouch function. In addition to the identification of key bowel function outcomes, the study highlighted additional patient-centered core outcomes that clinicians should focus future research efforts on.
P128 Accuracy of the combination of transabdominal and transperineal ultrasound for the evaluation of Ulcerative Colitis activity
Authors: Terracciano F , Bossa F , Marra A , Valvano M R , Carparelli S , Ippolito M A , Perri F 
Published: January 2022

Abstract

Background
Transabdominal Bowel Ultrasound (TBUS) is useful technique to assess disease activity and extent in patients with ulcerative colitis (UC); however, the evaluation of the rectum by TBUS is difficult. Recently, transperineal ultrasound (TPUS) has been proposed as a useful diagnostic tool in the evaluation of rectal disease. The aim of the study is to to assess the concordance between the combination of TPUS/TBUS and endoscopy in adult patients with UC.

Methods
Fifteen consecutive adult patients with UC were prospectively enrolled from June to November 2021. Disease activity was determined using Partial Mayo Score (PMS) and Mayo Endoscopic Subscore (MES). TBUS and TPUS were performed within a week from endoscopy. MES ≤ 1, bowel wall thickness ≤ 3mm in the colon and ≤ 4mm in the rectum were defined as remission. A concordance analysis comparing endoscopy and US was performed by kappa statistics

Results
Ten patients were male (66%) and 8 female (34%), with a median age of 39± 13,9. Two patients had a pancolitis, 6 a distal colitis, and 3 a proctitis. 4 patients were in remission. The agreement between endoscopy and rectal evaluation with TBUS was fair (k =0.40), while the agreement with TPUS was excellent (k =0.86). The evaluation of the colon with TBUS showed good concordance in the right colon (k= 0.63), left colon (k=0.65), and sigmoid colon (k=0.72), and excellent agreement in the transverse colon (k= 1).

Conclusion
These preliminary data showed that the combination of TBUS and TPUS allows to easily and accurately study all the segments of the large intestine in patients with UC, overcoming the limitation related to the reduced accessibility to the rectum with only trans abdominal approach.