bluuebunny/crossref_metadata_2025_split huggingface dataset:
P034 GB004, a novel gut-targeted prolyl hydroxylase inhibitor, induces HIF-1α target genes and is efficacious in mouse colitis
Authors: Taylor Meadows K , Kennedy M , Naiman Z , Sergeeva M , Paulson B , Opiteck G , Slee D , Carter L , Salter-Cid L 
Published: January 2020

Abstract

Background
GB004 is a small molecule prolyl hydroxylase inhibitor (PHDi) that stabilises hypoxia inducible factor (HIF-α), key transcription factors involved in the protective cellular responses at the intersection of hypoxia and inflammation. GB004 was selected based on its gut-targeted profile to limit systemic on-target effects associated with HIF-α stabilisation. GB004 is in clinical development for the treatment of ulcerative colitis.

Methods
Female wild-type C57BL/6 and immunodeficient B6.129S6-Rag2tm1FwaN12 ko/ko mice (Rag2) mice received either a single oral dose of GB004 or daily oral dosing for 7 days. Colitis was induced by administering 100 µg of anti-CD40 monoclonal antibody interperitoneally in Rag2 mice. GB004 oral dosing started on day 0 and continued once daily for 7 days. Body weight was measured daily. At termination, plasma was isolated for compound exposure and colons were excised, measured and weighed. Colon tissue was preserved in formalin for histology and 1 cm colon sections were snap-frozen for compound exposure and preserved with RNA for gene expression.

Results
A single oral administration or 7 days dosing of 10 mg/kg GB004 in healthy mice resulted in significantly higher exposure in the gastrointestinal tract (GI) (pylorus, duodenum, ileum and colon) than in plasma with colon to plasma ratio of ≥500. Target engagement in colon was demonstrated by induction of HIF-dependent genes (HIF1A), epithelial barrier genes (AKAP12, TFF3, NT5E) and anti-apoptotic genes (BNIP3). In colitis mice, GB004 plasma exposure was elevated compared with plasma concentrations in healthy mice, even so, GB004 exposure was still higher in the colon than plasma at a ratio of 90. Target engagement was demonstrated by an increase in NT5E, CLND1, BNIP3 and CA9 and a reduction in IL12B. In colitis mice, 1, 3 and 10 mg/kg GB004 demonstrated a dose-dependent reduction in colon inflammation and gland loss and 10 mg/kg GB004 showed a significant reduction in neutrophil score. Gene expression 4 h after the final GB004 dose demonstrated an increase in barrier genes AKAP12 and CLDN1 and a decrease in IL12B.

Conclusion
These studies demonstrate that oral GB004 has significantly higher exposure in the GI tract than in the plasma and induces HIF-1α-dependent genes in colon. In mouse colitis, GB004 has high colon exposure, induces target genes involved in barrier function and repair and demonstrates efficacy by reducing inflammation, gland loss and neutrophil inflammation. A clinical study of GB004 is ongoing in patients with ulcerative colitis to explore safety, PK and PD both systemically and in colon (NCT03860896). Sponsored by GB004, Inc., a wholly owned subsidiary of Gossamer Bio, Inc.
P044 Milk derived exosomes has a therapeutic effect on experimental colitis
Authors: Golan-Gerstl R , Koroukhov N , Elbaum Shiff Y , Shilo I , Reif S 
Published: January 2020

Abstract

Background
Inflammatory bowel disease (IBD) is a complex disorder that results from a dysregulated immune response in the gut. Emerging therapy for IBD treatment is mainly focused on regulation of the immune response. Exosomes are nanovesicle packing different molecules such as miRNAs that transfer their cargo to recipient cells. We and others found that mammalian milk contain high concentration of exosomes (milk-derived exosomes, MDE) carrying beneficial miRNAs such as miRNA-148. Furthermore, MDE are taken up by different cell type such as intestinal epithelial cells, modify target gene expression, promote proliferation and differentiation of colon epithelial cells. The aim of this study is to explore the therapeutic effect of MDE on colitis.

Methods
We used gavage administration of MDE labelled with DiR dye to track their localisation patterns in vivo. The therapeutic effect of MDE on colitis was study in mice model of SDS induced colitis, colon length, histopathological scoring grade, cytokine expression, stool consistency and miRNA expression were analysed.

Results
Imaging of mouse that have receive labelled MDE revealed an accumulation of fluorescent signal in the intestine. Moreover, fluorescent signal in the intestine and liver is time dependent. MDE reduced the histopathological scoring grade from 5.83 ± 1.47 to 0.6 ± 0.6, p &lt; 0.05. The length of the colon of MDE-treated animals was 7.9 ± 0.19 in comparison to 6.92 ± 0.3 p &lt; 0.05 of the untreated. The weight loss as a result of the colitis was reverted in MDE-treated mice. Likewise, MDE treatment reduced IL-6, TNF-α and caveolin expression from 3.83 to 0.78, 1.59 to 0.86 and 3.52 to 1.1, respectively. Highly expressed miRNAs (miRNA-320, 375, Let-7a and 6073) were found to be more abundant in colon of MDE treatment mice compared with the untreated.

Conclusion
This study demonstrated that MDE have a therapeutic effect on colitis in vivo. Proving the effect of MDE on colitis will have implications for the potential of adding MDE as a therapeutic nutrient to be included in the formulas for IBD patients.
P045 Profiling of human circulating dendritic cells and monocytes subsets discriminates type and mucosal status in patients with inflammatory bowel disease
Authors: Ortega Moreno L , Fernández Tomé S , Chaparro M , Marin A , Mora Gutiérrez I , Santander C , Baldán M , Gisbert J , Bernardo D 
Published: January 2020

Abstract

Background
Intestinal dendritic cells (DCs) and macrophages govern the mechanisms of immune homeostasis having a role in inflammatory bowel disease (IBD) onset. However, the profile of their circulating precursors (DC and monocytes) in IBD has not been previously described in depth. Our aim was to characterise blood DC and monocyte subsets in healthy controls (HCs) and IBD patients in order to understand their potential implication in IBD pathogenesis.

Methods
18 HC and 64 IBD patients were recruited. IBD patients were categorised into Crohn’s disease (CD) and ulcerative colitis (UC), either endoscopically active (aCD and aUC) or quiescent (qCD and qUC), based on the SES-CD or the Mayo index endoscopic subscore. Blood circulating type 1 conventional DC (cDC1), type 2 conventional DC (cDC2), plasmacytoid DC (pDC), classical monocytes, non-classical monocytes and intermediate monocytes were identified by flow cytometry and characterised for the expression of 18 homing and activation markers (β7, CCR1, CCR2, CCR3, CCR5, CCR6, CCR7, CCR9, CCRL1, CD40, CD86, CD137L, CD274 (PD-L1), CLA, CXCR1, CXCR3, ICOSL and HLA-DR). Association between markers and the presence, type or activity of IBD was tested by logistic regression. Discriminant canonical analysis was also performed to classify the patients on their own endoscopy category.

Results
All groups (HC, aCD, qCD, aUC and qUC) were separated from the others based on the discriminant canonical analyses of the 18 markers applied over all DC and monocytes subsets (Figure 1). Specifically, CCRL1, CCR3 and CCR5 expression on cDC1, CCRL1 on non-classical monocytes and CCR9 and b7 on classical monocytes were highly associated to IBD. CCR3 displayed an odds ratio (OR) of 2.29 along with its 95% confidential interval (CI) between 1.11 and 4.75, showing a strong association with activity in CD; whereas the other markers displayed an inverse association with IBD. Hence, expression of CCRL1 on cDC1 and non-classical monocytes from aUC showed an OR (95% CI) of 0.23 (0.08–0.66) and 0.52 (0.28–0.95), respectively. In the case of qUC, CCR5 on cDC1 and β7 on classical monocytes displayed an OR (95% CI) of 0.10 (0.01–0.83) and 0.56 (0.34–0.90), respectively. CCR9 was inversely associated to qCD with an OR (95% CI) of 0.64 (0.46–0.89) in the classical monocytes subset. Indeed, the same markers (excluding β7) were also associated with IBD when all DC and monocyte subsets were considered at the same time.

Conclusion
Differences on the expression of migration markers CCR3, CCR5, CCR9, b7 and decoy receptor CCRL1 on circulating DC and monocyte subsets from IBD groups suggest the presence of constitutive migratory differences underlying IBD pathogenesis in CD or UC and its condition (inflamed or non-inflamed).
P048 Mucosal macrophages express elevated levels of HDAC9 in inflamed and uninflamed mucosa of Crohn’s disease, but not ulcerative colitis
Authors: Ghiboub M , de Bruyn J , Reedquist K , Radstake T , Wichers C , Wildenberg M , Broen J , D’Haens G , de Jonge W 
Published: January 2020

Abstract

Background
Histone deacetylases (HDACs) are a group of enzymes that control histone and non-histone deacetylation and influence inflammatory gene transcription. Certain members of the HDAC family control the function of macrophages and play an important role in immune response. In this study, we aimed to study the expression of HDACs in mucosal macrophages isolated from inflammatory bowel diseases (IBD) patients.

Methods
Both macroscopically inflamed and non-inflamed colon resection tissue were collected from 15 Crohn’s disease (CD) and nine ulcerative colitis (UC) patients operated on for therapy refractory disease. Of the CD patients, 53% had ileal and 47% ileocolonic disease. Of the UC patients, 44% had left-sided colitis and 56% pancolitis. Lamina propria was separated from the muscularis externa, and a targeted array for epigenetic enzymes was performed. To assess the relevance of HDAC9 gene expression in terms of protein level, immunofluorescence staining of HDAC9 protein was undertaken in tissue sections from inflamed and non-inflamed mucosa. CD68 was used as a pan-macrophage marker.

Results
From our array, expression of HDAC9 was significantly higher in the inflamed mucosa of CD patients compared with UC patients (p = 0.005). Gene expression of HDAC9 in non-inflamed mucosa from CD was elevated compared with non-inflamed mucosa from UC. In addition, in CD, HDAC9 mRNA level was increased in inflamed tissue in comparison to non-inflamed tissue (p = 0.046). In conjunction with the expression data, HDAC9 protein was found highly expressed in inflamed tissue. HDAC9 was predominantly localised in the cytoplasmic compartment of macrophages in non-inflamed tissue whilst HDAC9 localised to the nucleus of macrophages in inflamed tissue.

Conclusion
HDAC9 is member of class IIA HDAC superfamily that exerts pro-inflammatory properties. The inhibition of HDAC9 in experimental murine colitis clearly enhances regulatory T-cell function, suggesting a critical role for HDAC9 in breaching immune homeostasis (de Zoeten EF et al, 2009). We suggest here that HDAC9 can serve as an additional marker to distinguish CD from UC in tissue biopsies. Furthermore, we show for the first time that HDAC9 protein is expressed in mucosal macrophages of CD patients, indicating its potential in mediating macrophage inflammatory function in IBD. Further studies are currently being undertaken to elucidate the role of HDAC9 in CD pathogenesis.
P049 A new preclinical rationale for first-line therapy of ulcerative colitis
Authors: Thorlacius H , Bjoerk A , Nordle Ö , Hedlund G 
Published: January 2020

Abstract

Background
Ulcerative colitis (UC) is a chronic inflammatory condition with no known medical cure. 5-Aminosalicylic acid (5-ASA [mesalazine]) represents the cornerstone of first-line therapy for mild-to-moderate UC. Sulfasalazine (SASP) is the original agent in this class of drugs. Meta-analyses of patients with mild-to-moderately active UC comparing 5-ASA to placebo showed 5-ASA to be significantly superior to placebo. However, about two-thirds of patients treated with 5-ASA fail to enter clinical remission. It is therefore most important to identify strategies to accelerate and maximise the therapeutic effects of 5-ASA. Therapeutic intervention against NFκB activation is a useful strategy for treatment of UC. The 4-alkanoylaminobenzamide PM0503 inhibits the breakdown of the NFκB inhibitor IκBβ, and SASP/5-ASA inhibits the breakdown of IκBα. This elicited a hypothesis of a possible synergistic action and converging effect on NFκB signalling. In the present study, we investigated the effect of combining SASP/5-ASA with PM0503 in experimental colitis.

Methods
SASP and PM0503 alone or in combination were administered for 5 days to Balb/c mice with colitis triggered by 5% dextran sulphate sodium (DSS). Blood in the stool, stool consistency and body weight loss were evaluated daily on a 0–4 point scale. The disease activity index (DAI) was calculated by summarising the total score of these three parameters.

Results
Addition of 5% DSS in the drinking water for 5 days produced reproducible symptoms of colitis. PM0503 was shown to inhibit DSS induced colitis by reducing mean DAI at day 5 from 6.9 in controls to 1.7 (a 75% decrease). Mean DAI recorded with SASP treatment at optimal doses in the same series of experiments was 4.4 (a 36% decrease). Furthermore, and most important, lower doses of PM0503 acted synergistically with SASP in ameliorating DSS-induced disease severity. The combination of PM0503 and SASP using suboptimal doses having minimal beneficial effects as monotherapies, showed more than 50% disease inhibition at day 5. In addition, no toxicity was observed with PM0503 alone or in combination with SASP.

Conclusion
Our findings offer a preclinical rationale for simultaneous coadministration of PM0503 and a 5-ASA agent such as SASP or 5-ASA as first-line treatment for patients with UC.
P051 Quantifying inflammation and fibrosis through a surgical histopathological score can differentiate ileal Crohn’s disease phenotypes and predict postoperative progressive disease
Authors: Tavares de Sousa H , Gullo I , Castelli C , Dias C C , Magro F 
Published: January 2020

Abstract

Background
The quantification of fibrosis in Crohn’s disease (CD) still relies on surgical specimens’ pathology. We aimed to correlate quantification of inflammation and fibrosis of CD ileal resection specimens with postoperative progressive disease.

Methods
All patients (patients) having primary ileal resection for CD complications with a follow-up &gt;3 years (n = 262) were considered. Unavailability of specimen excluded 72 patients and 22 for absence of the three study sections: (1) proximal ileal margin; (2) most affected area (B2: narrower calibre of stricture; B3: area with fistulas/deep ulcers); (3) inflamed area (inflamed bowel, no lesions as in 2). Of the 168 patients with the three studied phenotypes (B2 and B3 with stenosis (B3s) and without stenosis (B3o), we randomly excluded 65 B3s for overrepresentation. We analysed three sections per patient (3 × 103 patients = 309 sections), stained with haematoxylin and eosin and Masson’s trichrome. Chiorean et al. histopathological score grades inflammation 1–3 and fibrosis 0–2. We add a ‘0’ category to inflammation and observed adipose tissue and muscularisation in submucosa. Progressive disease was previously defined as one of eight post-operative outcomes (reoperation, hospitalisation, steroids, start or change of immunosupressives or biologics, new stricturing/penetrating/anal event). Statistics: Continuous variables were described by mean(standard deviation), median(interquartile range), minimum and maximum, and categorical variables by absolute(n) and relative(%) frequencies. Chi-square, Mann–Whitney and Kruskal–Wallis compared groups. Hypotheses were tested at 5% level of significance, using IBM SPSS Statistics for Mac, vs. 24.0.

Results
We assessed 103 patients (B2–29; B3o–20; B3s–54), 55% males, mean age at diagnosis 30(12) years, followed for a mean time of 10(4) years. Median time from surgery to reoperation was 8.0 (7.0, 12.0) years. B3 patients have significantly more inflammation than B2 patients [score 3: 78%vs.55% and 96%vs.76% in most affected (p = 0.027) and inflamed (p = 0.005) sections, respectively]. B3s patients had significantly more fibrosis than B3o [score 1 + 2: 90%vs.60% in most affected (p = 0.011) and 99%vs.85% in inflamed (p = 0.048) sections] and significantly more inflammation than B2 patients [score 3: 81%vs.55% in most affected (p = 0.020) and 94%vs.76% in inflamed (p = 0.019) sections]. B3s had higher total score than B3o and B2 [score 4–5: 78%vs.45%vs.52%, p = 0.019] and more new penetrating events (p = 0.043). Of the 25 patients changing biologic after surgery, 88% had inflammation at the margins [score 3: 55%vs.12%, p = 0.035].

Conclusion
B3s stood out as a distinctive phenotype, with significantly more fibrosis than B3o but significantly more inflammation than B2. It displayed the highest total score and was associated to progressive disease.
P057 IL-17 regulates expression of chemotactic chemokines in human colonic subepithelial myofibroblasts
Authors: Filidou E , Tarapatzi G , Boulkou M , Arvanitidis K , Vradelis S , Valatas V , Bamias G , Kolios G , Koutroubakis I , Drygiannakis I 
Published: January 2020

Abstract

Background
Crohn’s disease (CD) and ulcerative colitis (UC), the two main entities of inflammatory bowel disease (IBD), are characterised by chronic and relapsing/remitting inflammation of the gastrointestinal tract, and occasionally ultimately result in debilitating intestinal fibrosis. Apart from their key role in fibrosis, there is evidence that subepithelial myofibroblasts (SEMFs) participate in the IBD inflammatory cascade, as they express various pro-inflammatory cytokine receptors. We examined the effect of pro-inflammatory IL-17—the hallmark cytokine of T-lymphocytes differentiated to Th17—on the expression of lymphocyte-chemotactic chemokines in SEMFs.

Methods
SEMFs were isolated from endoscopically obtained colonic biopsies from healthy controls, set to culture and stimulated with 100 ng/ml IL-17 for 6 h. Total RNA was extracted and mRNA expression of CCL5, CXCL1 and CXCL11 was assessed with reverse transcription quantitative (RT-q) PCR. Changes in cytokine mRNA are provided as medians (IQR).

Results
Untreated SEMFs had a basal expression of chemokines of the CCL and CXCL family groups. So far, our study has shown that the IL-17 stimulation leads to a statistically significant upregulation of CCL and CXCL chemokines in SEMFs (p &lt; 0.001). In detail, CCL5 was upregulated 5.7-fold (4.9–7.5), CXCL1 72.9-fold (63.1–90.7) and CXCL11 25.4-fold (17.3–37.3).

Conclusion
IL-17 induced the expression of chemotactic factors in SEMFs. Our results further support a potential role of SEMFs in the shaping of intestinal mucosal immunity by serving as immunological intermediates that respond to cytokines of adaptive immunity and amplify the recruitment of immune cells via chemokine production.
P069 CD73 promotes colitis-associated tumourigenesis in mice
Authors: Wu X , Liu X , Lan N , Zheng X , Chen Y , Cai Z , Lan P , Wu X 
Published: January 2020

Abstract

Background
Patients with inflammatory bowel disease (IBD) are at a higher risk of developing colitis-associated colorectal cancer. The aim of the present study was to investigate the role of CD73 in IBD-associated tumourigenesis.

Methods
A mouse model of colitis-associated tumourigenesis (CAT) induced by azoxymethane and dextran sulphate sodium (AOM/DSS) was successfully constructed. Model mice were injected with CD73 inhibitor or adenosine receptor agonist. Colon length, body weight loss and tumour formation were assessed macroscopically. Measurement of inflammatory cytokines and RNA sequencing on colon tissues were performed.

Results
Inhibition of CD73 by adenosine 5′-(α,β-methylene) diphosphate (APCP) suppressed the severity of CAT with attenuated weight loss, longer colons, lower tumour number and smaller tumour size when compared with the model group. On the other hand, activation of adenosine receptors using 1-(6-amino-9H-purin-9-yl)-1-deoxy-N-ethyl-β-d-ribofuranuronamide (NECA) exacerbated CAT. Histological assessment indicated that inhibition of CD73 reduced while activation of adenosine receptors exacerbated the histological damage of the colon compared with the model group. Increased expression of pro-inflammatory cytokines (tumour necrosis factor-α and interleukin-6) in colonic tissue was detected in the NECA group. According to the results of RNA sequencing, potential oncogenes such as ALOX15, Bcl2l15 and Nat8l were found to be downregulated in the APCP group and upregulated in the NECA group compared with the model group.

Conclusion
Therefore, inhibition of CD73 attenuated IBD-associated tumourigenesis, while activation of adenosine receptors exacerbated tumourigenesis in a C57BL/6J mouse model. This effect may be associated with the expression of pro-inflammatory cytokines and the regulation of ALOX15, Bcl2l15 and Nat8l.
P079 High-dimensional mass cytometry after local MSC treatment in patients with refractory proctitis shows an effect on the myeloid compartment: preliminary data
Authors: Ouboter L , Barnhoorn M C , Hawinkels L J A C , van Pel M , Zwaginga J J , Verspaget H W , Koning F , Pascutti M F , van der Meulen-de Jong A E 
Published: January 2020

Abstract

Background
Ulcerative proctitis (UP) causes a chronic rectal mucosal inflammation. A subset of patients does not respond to one or more medication options. Mesenchymal stromal cells (MSCs) show beneficial effects in animal models and patients with immune diseases. However, the effects of MSCs on the mucosal immune composition in patients with inflammatory bowel disease (IBD) are not unravelled yet.

Methods
Patients with UP (N = 7) were locally treated with allogeneic bone marrow-derived MSCs. A total of 20 × 106 MSCs was injected if the inflammation was restricted to 7 cm and 40 × 106 if inflammation exceeds 7 cm. Medication was stable in all patients from at least 2 weeks before treatment up to week 6 after treatment. Biopsies were taken from the affected and unaffected colon before (t = 0) and 6 weeks (t = 6) after treatment from the same locations and faecal calprotectin (FCP) was determined. Single-cell suspensions from the biopsies were stained with a 42 antibody panel and analysed with a mass cytometer to identify and characterise possible effects of MSCs on immune subsets. The generated dataset was analysed with Hierarchical SNE (HSNE) in the Cytosplore analysis and visualisation tool.

Results
FCP decreased significantly in four out of seven patients (419→191; 365→40; 1744→346; 3270→291), ‘responders’, while in three patients FCP increased, ‘non-responders’. Unbiased hierarchical clustering of the subsets showed no clear differences in the major immune lineages before and after treatment. In some patients; however, a prominent myeloid population was present in the affected colon, whereas the CD8+− and CD3−CD7+ populations in affected tissue were in general less frequent compared with unaffected tissue from the same patients. We then zoomed into every major lineage separately, and observed changes in the frequencies of two myeloid clusters in the affected tissue of ‘responders’ compared with the ‘non-responders’ (see figure, Cluster 1=CD11b+CD11c+CD66b+CD15intCD16+CD45RA−CD45RO+ cells, Cluster 2= CD11b+CD11c+CD66b+ CD15−CD16−CD69+CD45RA+CD45RO+ cells).

Conclusion
These preliminary data show differences between ‘responders’ and ‘non-responders’ in the myeloid compartment in the inflamed tissue after MSC therapy. Since CD11b+ and CD11c+ populations mediate the tolerogenic effect of MSC in mouse models, we will explore the regulatory potential of the observed populations in MSC-treated UP patients in future studies. The inclusion of patients in a second cohort (N = 7) is currently ongoing.