bluuebunny/crossref_metadata_2025 huggingface dataset:
P032 Host transcriptome signatures in human fecal-washes predict histological remission in IBD patients
Authors: Ungar B , Yavzori M , Fudim E , Picard O , Kopylov U , Eliakim R , Shouval D , Levin Y , Savidor A , Ben-Moshe S , Manco R , Dan S , Egozi A , Bahar Halpern K , Mayer C , Barshak I , Ben-Horin S , Itzkovitz S
Published: January 2022

Background

Colonoscopy is the gold standard for evaluation of inflammation in inflammatory bowel diseases (IBD), yet entails cumbersome preparations and risks of injury. Existing non-invasive prognostic tools are limited in their diagnostic power. Moreover, transcriptomics of colonic biopsies have been inconclusive in their association with clinical features. Our aim was to assess the utility of host transcriptomics of fecal wash samples of IBD patients compared to controls.

Methods

In this prospective cohort study, we obtained biopsies and fecal-wash samples from IBD patients and controls undergoing lower endoscopy. We performed RNAseq of biopsies and matching fecal-washes, and associated them with endoscopic and histological inflammation status. We also performed fecal mass-spectrometry proteomics on a subset of samples. We inferred cell compositions using computational deconvolution and used classification algorithms to identify informative genes.

Results

We analyzed biopsies and fecal washes from 39 patients (19 IBD, 20 controls). Host fecal-transcriptome carried information that was distinct from biopsy RNAseq and fecal proteomics. Transcriptomics of fecal washes, yet not of biopsies, from patients with histological inflammation were significantly correlated to one another (p=5.3*10–12). Fecal-transcriptome was significantly more powerful in identifying histological inflammation compared to transcriptome of intestinal biopsies (150 genes with area-under-the-curve >0.9 in fecal samples versus 10 genes in biopsy RNAseq). Fecal samples were enriched in inflammatory monocytes, regulatory T cells, natural killer-cells and innate lymphoid cells.

Figure 1 - Fecal-wash host transcriptome predicts histological inflammation. A) Study layout, B) Clustergram of fecal-wash host cell mRNA signatures, demonstrating that patients with histological inflammation (red) are clustered when measuring fecal wash transcriptome yet not biopsy transcriptomes. C-D) Principle Component Analysis demonstrating improved separation of inflamed patients based on fecal host transcriptomes. E, F) Expression of host genes in fecal washes has higher statistical power (Area under the Curve, AUC) in classifying histological inflammation compared to biopsies. D shows NFKBIA as an example, E shows the AUC of the 5% best classifying genes, F shows the overall AUC based on biopsies or washes. Gray areas have AUC>0.9. G) UMAP of cells obtained from scRNAseq of mouse small intestine fecal washes.

Conclusion

Fecal wash host transcriptome is a powerful biomarker reflecting histological inflammation. Furthermore, it opens the way to identifying important correlates and therapeutic targets that may be obscure using biopsy transctriptomics.
P035 An examination of resting state functional connectivity in patients with active Crohn’s disease
Authors: Thapaliya G , Eldeghaidy S , Radford S , Francis S , Moran G
Published: January 2022

Background

Crohn’s disease (CD) and its comorbidities may be associated with a specific intrinsic neural phenotype that can be measured using resting state functional connectivity (RSFC). RSFC alterations have been reported in CD in remission but have not been investigated in the active phase of the disease. Here, the primary aim was to study the effects of active CD on resting state networks (RSNs), hence identifying RSFC alterations specific to active CD status. A second aim was to investigate associations between disease duration and abdominal pain with RSFC in active CD.

Methods

25 active CD patients and 25 age-, BMI- and gender-matched healthy controls (HC) were recruited. Active disease was defined as CRP > 5 mg/dl, or faecal calprotectin (FCP) >250 μg/g or through ileocolonoscopy or MRE. RsfMRI datasets were acquired on a 3T Philips Achieva scanner, with data corrected for physiological noise and motion. ICA analysis was carried out using MELODIC (FSL software). A multi-session temporal concatenation was used to generate 30 independent component (IC) maps of RSNs. A dual regression analysis with variance normalization, using group IC maps from HC group as the network template, was performed. Differences in RSN between HCs and CD patients and associations with covariates of interest were assessed.

Results

RSNs comprising the visual, cerebellar, frontal parietal and frontal RSN, default mode network (DMN), salience, temporal and dorsal attention RSN were identified. Enhancement of activity and increase connectivity in the medial visual RSN (orbital frontal cortex), in the frontal parietal network (occipital fusiform gyrus, thalamus, caudate, posterior cingulate cortex, postcentral gyrus) were observed in CD relative to HC. Further, decreased activity was seen in the salience network (cerebellum, postcentral gyrus), cerebellar network (occipital fusiform gyrus, cerebellum) and the DMN (parahippocampal gyrus, cerebellum) in CD relative to HC. Greater abdominal pain scores were associated with lower connectivity in the precuneus in the medial visual RSN, in the parietal operculum in the salience RSN, as well as reduced connectivity in the cerebellum in the frontal RSN. Finally, greater disease duration was associated with greater connectivity in the visual RSN in the middle temporal gyrus and planum temporale.

Conclusion

Alterations in RSFC between active CD and HC in RSNs implicated in cognition, attention, emotion and pain may reflect neuroplasticity in response to chronic systemic inflammation, abdominal pain, disease duration and severity. Identification of a neural phenotype specific to active CD status, may help guide intervention constituting a neurobehavioural precision medicine approach to treatment.
P048 Bridging the Gap Between Body and Soul: A Pilot Study Examining Inflammation and Depression / Anxiety in Inflammatory Bowel Disease Patients
Authors: . Mekori - Domachevsky E , Avinir A , Dar S , Haj O , Ben - Horin S , Gothelf D , Kopylov U , Taler M
Published: January 2022

Background

Inflammatory bowel diseases (IBD) have a varied phenotype, among which are increased rates of emotional difficultiesthat were proven to worsen patients’ prognosis. Common to IBD and depression / anxiety is the postulated role of inflammation in their pathogenesis. The aim of this study was to examine whether inflammation per se is linked to depressive and anxious symptoms is a cohort of IBD patients (and is not merely an emotional response to a medical condition).

Methods

Fifty patients with Crohn’s disease (CD) and 30 with ulcerative colitis (UC) were recruited at the gastroenterology department, Sheba Medical Center. They completed a set of questionaries that measured depressive symptoms (BDI), state and trait anxiety symptoms (STAI), stress levels (PSS), IBD quality of life (IBDQ) and sleep quality (PSQI). After which blood samples were drawn and analyzed for C – reactive protein (CRP) and complete blood count. In addition, the ratios between several leukocyte sub-population (that were previously proven to be correlated with IBD as well as depression / anxiety) were calculated, and medical data was obtained from patients’ records. Finally, backwards linear regression was used to determine which factors are associated with the depressive and anxious symptoms.

Results

For the Crohn’s group, predictive factors for depressive symptoms were gender (β = -0.17, p = 0.034), systemic symptoms (β = -0.59, p < 0.001), anxiety levels (β = 0.33, p < 0.001) and platelets to lymphocytes ratio (β = -0.33, p = 0.001). Predictive factors for anxiety were anxiety as a trait (β = 0.89, p < 0.001) and monocytes to lymphocytes ratio (β = 0.20, p = 0.010).

For the UC group, predictive factors for depressive symptoms were emotional functioning (β = -0.77, p < 0.001), systemic symptoms (β = 0.53, p = 0.025) and sleep quality (β = 0.52, p = 0.005). As for anxiety, predictive factors were gender (β = 0.21, p = 0.039), anxiety as a trait (β = 0.59, p < 0.001), depressive symptoms (β = 0.38, p = 0.004) and neutrophil to lymphocytes ratio (β = 0.25, p = 0.017). Intriguingly, CRP only had a single marginal contribution to depressive symptoms in the UC group (β = 0.20, p = 0.098).

Conclusion

Our results suggest that after controlling for demographics, disease activity, medication type and psychological factors inflammation by itself does play a role in the evolution of depression and anxiety in IBD patients. Furthermore, as the leukocytes’ ratios but not CRP had significant predictive values, we postulate that an imbalance between the innate and adaptive arms of the immune system mediates these phenomena, rather than an unspecified immune activation.
P049 The treatment effect of tonsil-derived mesenchymal stem cells (T-MSCs) via regulating programmed death-1/programmed death ligand-1 (PD-1/PD-L1) in ulcerative colitis
Authors: Song E M , Joo Y H , Park Y , Choe A R , Tae C H , Moon C M , Kim S E , Jung H K , Shim K N , Jung S A
Published: January 2022

Background

The programmed death 1 (PD-1)/programmed death-ligand 1 (PD-L1) pathway has not been fully evaluated in inflammatory bowel disease. We evaluated the level of PD-1/PD-L1 in the mucosa of patients with ulcerative colitis and revealed the role of PD-1/PD-L1 in the immunomodulatory mechanism of tonsil-derived mesenchymal stem cells (T-MSCs).

Methods

We used chronic murine colitis model induced by dextran sodium sulfate (DSS). We measured the PD-1 and PD-L1 levels in inflamed colonic tissues before and after the treatment with T-MSC. We also measured the levels of PD-1 and PD-L1 in the colonic tissues from patients with ulcerative colitis and compared with those from normal controls.

Results

In chronic colitis model, the level of PD-L1 was decreased than normal controls (fold change) (1.0 vs. 0.46±0.08, P=0.05). However, the level of PD-1 was increased than normal controls (1.0 vs. 6.84±8.16, P=0.142). After treatment with T-MSC which showed significant improvement in body weight, disease activity index and colon length, the levels of PD-1 and PD-L1 were recovered; PD-L1 was significantly increased (1.04±0.77 vs. 0.46±0.08, P=0.031) and the level of PD-1 was decreased (3.63±1.94 vs. 6.84±8.16, P=0.537). When measuring the level of PD-1 and PD-L1 in both soluble medium and cell lysate of T-MSC, we found that the PD-1 and PD-L1 were expressed in both forms (182.63 pg/ml in soluble medium and 11.85 pg/protein(ug) in cell lysate after 7 days of differentiation period). In the analysis using human colonic tissues, a significantly increase in the levels of PD-1 and PD-L1 was observed in the colonic mucosa of patients with UC compared with normal controls (PD-1: 2.09±3.95 vs. 7.93±14.21, P=0.034 and PD-L1: 2.24±2.91 vs. 10.08±18.13, P=0.044).

Conclusion

The altered expression of PD-1 and PD-L1 in colonic mucosa may a possible mechanism of ulcerative colitis and T-MSC-derived PD-L1 could repress the colitis.
P056 Local nanocarrier delivery of JAK1 siRNA for IBD treatment: an in vivo mouse model
Authors: Garrido Trigo A , Veny M , Dotti I , Esteller M , Sulpice E , Jary D , Clément F , Gidrol X , Navarro F , Salas A
Published: January 2022

Background

Janus Kinases (JAK), including JAK1, JAK2, JAK3 and TYK2, have garnered increasing interest as therapeutic targets for several chronic inflammatory diseases such as inflammatory bowel disease (IBD). Although oral small molecule JAK inhibitors are being used in the clinics as an effective treatment for moderate-to-severe ulcerative colitis, nonethelessthey can lead to severe systemic immunosuppression, which is associated with an increased risk of infections. Our project aimed to develop a nanomedicine that induced JAK1 mRNA degradation within the intestinal mucosa with little systemic distribution.

Methods

The NEWDEAL project (H2020) developed both potent small interfering RNAs (siRNA) specific for JAK1 (siJAK1) or TNF (siTNF), and lipid nanoparticles (CL40) as nano-vectors. Their biodistribution in a mice colitis model was analyzed using labeled AF488-siJAK1 complexed to CL40 and given intrarectally (IR) for 3 consecutive days. In vivo cellular uptake was determined by FACS analysis. To induce colitis, C57/Bl6 mice received 2.5% dextrate sulfate sodium (DSS) in drinking water for 5 days. To determine efficacy of siRNAs, mice started DSS on day 0 and received IR administrations of siRNAs alone or complexed with CL40 nanoparticles on days -3, -2, -1, 1, 3, 5 and 7. On day 9, mice were sacrificed and disease activity index (DAI), histological score, fecal inflammatory markers and qPCR of targeted genes were analyzed.

Results

Colonic phagocytic cells - such as macrophages, neutrophils and monocytes - are the main targets cells of the siRNA-CL40 nanocomplexes following IR administration. A small percentage of cells in the draining lymph nodes captured siRNA CL40 nanocomplexes, remaining almost undetectable in spleen. This suggests a localized effect of the siRNA nanocomplexes after IR administration. While siJAK1-alone effectively reduced JAK1 mRNA expression in the distal colon, it did not improve intestinal inflammation induced by administering DSS. siTNF-CL40 treatment, though not siTNF-alone, significantly decreased intestinal inflammation in the DSS colitis model of treated mice, revealing significant protection from weight loss, lower histological scores, and reduced production of fecal lipocalin.

Conclusion

In summary, these results show that despite effectively reducing JAK1 mRNA expression, siJAK1 administration does not ameliorate colitis in a DSS mice model. However, we showed that a locally delivered siTNF-nanocomplex has the capacity to improve clinical, macroscopic and histological symptoms as well as fecal biomarkers in DSS colitic mice. We conclude not only that siRNA-CL40 target delivery is a good therapeutic strategy, but also that siTNF-CL40 is effective in locally treating intestinal inflammation.
P057 Mucosal microbiota adapts to ATF6-induced alterations in host lipid metabolism with prognostic value in colorectal cancer
Authors: Bierwirth S , Sorbie A , Coleman O , Reuß E , Weber P , Köhler N , Kacprowski T , Dunkel A , Pauling J , Ecker J , Baumbach J , Rosenstiel P , Steiger K , Wirbel J , Zeller G , Janssen K P , Haller D
Published: January 2022

Background

Colorectal cancer (CRC) is a leading cause of cancer deaths worldwide. The endoplasmic reticulum unfolded protein response (UPRER) signal transducer activating transcription factor 6 (ATF6) is a clinically relevant pre-cancerous marker in CRC and colitis-associated CRC. We established the interrelated role of the microbiota and ATF6 signalling as a novel tumor-promoting mechanism in our transgenic mouse model of spontaneous microbiota-dependent ATF6-driven CRC (nATF6IEC).

Methods

To elucidate the transcriptional program initiated by acute and chronic ATF6 signalling, mRNA sequencing analyses of murine colonic intestinal epithelial cells were performed 4 days or 5 weeks after induction of ATF6, respectively. Germfree (GF) mice served to dissect microbiota contribution to the transcriptional response. CRC patient datasets (TCGA) were used to estimate ATF6 activity and validate ATF6-driven signatures. To investigate the impact of ATF6 signalling on metabolites, untargeted metabolomics of faeces and targeted lipidomics of tissue were performed. Mucosal microbiota was spatially characterised by 16S rRNA profiling at mm resolution along the colonic longitudinal axis. Based on 16S rRNA data, Phylogenetic Investigation of Communities Reconstruction of Unobserved States (PICRUSt2) was used to infer microbiota lipid-specific functional content.

Results

We identified an ATF6-UPR core of 368 differentially expressed genes fully activated by acute ATF6 signalling. Functional analysis using KEGG pathways showed that chronic ATF6 signalling predominantly alters UPR-related and metabolic pathways, with 22% of metabolic pathway genes classified as lipid metabolism. GF mice confirmed that the microbiota enhances ATF6-induced metabolic changes. Kaplan-Meier analyses significantly associate our microbiota-dependent ATF6-driven and lipid-specific ATF6-driven gene signatures with decreased disease-free survival in CRC patients since primary therapy. Moreover, ATF6 activity correlates with the presence of CRC-associated bacteria in TCGA samples. Tumor-susceptible mice show alterations in lipid metabolites, particularly long-chain fatty acids (FA) and elongation of saturated FA. PICRUSt2 revealed bacterial lipid detoxification mechanisms, with an increased total abundance of oleate hydratase-positive species.

Conclusion

Chronic ATF6-signalling alters host lipid metabolism and the lipid milieu in tumor-developing nATF6IEC mice. ATF6-driven microbiota changes are concomitant with bacterial lipid detoxification mechanisms in the tumor niche. We postulate that chronic ATF6 signalling represents a clinically relevant pathologic response that alters the intestinal lipid milieu and thus selects for a tumor-promoting microbiota.
P062 31P MRS and MRI phenotyping of muscle metabolic quality in IBD fatigue
Authors: Mcging J , Nicholas R , Serres S , Greenhaff P , Francis S , Moran G W
Published: January 2022

Background

Fatigue perception and exercise intolerance are common in inflammatory bowel disease (IBD). The mechanisms are unknown, although physical deconditioning is greater in fatigued relative to non-fatigued patients, suggesting an aetiological role for deconditioning. To assess peripheral muscle quality in Crohn’s disease (CD), we quantified the time-course of muscle phosphocreatine (PCr) recovery, which is wholly mitochondrial dependent, following ischaemic high-intensity exercise in fatigued CD and HV.

Methods

Participant body mass and regional body composition was assessed by DEXA. All MR data was collected using a 3T Philips Achieva scanner and a 14cm 31P coil on the medial gastrocnemius muscle. 1H mDIXON scans were acquired to image the calf prior to ischaemic plantar flexion exercise at 50% maximum voluntary contraction (MVC) using an MR-compatible ergometer (Trispect, Ergospect) during non-localized pulse-acquire 31P-MRS. 31P spectra were analysed using jMRUI Beta 6.0 using the AMARES function with prior knowledge. Cytosolic pH was estimated using the chemical shift difference between Pi and PCr peaks. Post exercise PCr recovery was quantified after reinstating limb blood flow. PCr recovery was expressed relative to baseline (non-ischaemic) signal amplitude and fit to a mono-exponential function in GraphPad prism. (PCr(t) = PCrinitial+(PCrend – PCrinitial)(1-exp(-k.t)) where t is the time from the start of non-ischaemic recovery, PCrinitial and PCrend is the PCr signal at the initial and end of recovery phases. An FSL and Matlab pipeline was used to obtain calf muscle volume and fat fraction from 1H mDIXON images.

Results

13 quiescent CD patients (7 female, HBI 1±2, CRP <5) and 9 HVs (4 female) were matched for age (P = 0.986) and BMI (P = 0.845). General fatigue score was greater in CD (13 ± 4) relative to HV (8 ± 3) (t19=3.08, P = 0.006) (Table 1.) End-exercise muscle PCr depletion (%) (85.25 ± 10.12 vs 83.28 ± 7.8%, P = 0.639) and estimated end-exercise muscle pH (6.6 ± 0.2 vs 6.5 ± 0.2, P = 0.320) was consistent between HV and CD. The mean rate constant of PCr resynthesis (kPCr min-1) in CD (0.76 ± 0.23) was significantly less (t20 = 2.6, P = 0.016) than HV (1.03 ± 0.23). PCr recovery curves were well described using a mono-exponential function in CD (R2=0.98 ± 0.01, R2value) and HV (R2=0.96 ± 0.03) (Table 3).

Conclusion

The PCr resynthesis rate following intensity matched ischemic exercise was less in fatigued CD patients relative to HV. This points to peripheral muscle deconditioning and associated loss of mitochondrial mass as a driver of exercise intolerance in IBD. This may be amenable to exercise training intervention.
P068 Discovery of IBD3540: A Novel Gut-Restricted Glutamate Carboxypeptidase II Inhibitor with Preclinical Anti-Colitis Activity
Authors: Peters D , Norris L , Tenora L , Snajdr I , Zhu X , Sakamoto S , Veeravalli V , Thomas A , Majer P , Rais R , Slusher B
Published: January 2022

Background

Glutamate carboxypeptidase II (GCPII) is robustly overexpressed in both Crohn’s disease and ulcerative colitis and is a promising therapeutic target for IBD. We have previously published that systemically or enema administered small molecule GCPII inhibitors have efficacy in multiple mouse colitis models. While this is a highly encouraging activity profile for a candidate IBD drug, an oral dose route would be strongly preferred. Thus, here we develop and characterize a new class of GCPII inhibitors designed to be gut-restricted for oral administration, with the intention of selecting a lead inhibitor for further clinical development.

Methods

Anti-inflammatory secondary bile acids lithocholic acid, deoxycholic acid and ursodeoxycholic acid were conjugated to GCPII inhibitor, 2-PMPA, yielding three novel constructs. These conjugates were screened for oral activity in murine dextran sulfate sodium (DSS) colitis, followed by detailed characterization of the most active inhibitor to: measure GCPII inhibition (IC50), confirm colon target engagement, evaluate plasma and colon pharmacokinetic profiles, compare efficacy versus standard-of-care agents, evaluate mechanisms of anti-inflammatory activity in vivo, confirm efficacy in a second colitis model (IL10-/-), and assess safety in standard preclinical assays.

Results

The deoxycholic acid 2-PMPA conjugate, IBD3540, was identified as the lead inhibitor (IC50 = 4nM). Oral IBD3540 was robustly efficacious in acute DSS colitis, where it dose-dependently inhibited colon GCPII, attenuated both gross disease activity index and blinded colon histopathology scores, and had improved efficacy relative to both sulfasalazine and tofacitinib when compared head-to-head. Mechanistically, we determined that IBD3540 attenuated pathogenic monocytic inflammation in the colon, as measured by flow cytometry, and also decreased colon pro-inflammatory cytokine content. We confirmed efficacy of IBD3540 in spontaneously occurring, chronic, IL10-/- colitis, where treatment initiated 4 weeks after the onset of colon inflammation improved multiple disease endpoints. Further, in preclinical safety screens, inclusive of CYP inhibition/induction assays, Eurofins SafetyScreen44TM, and AMES/hERG mutagenicity testing, no concerns were identified.

Conclusion

IBD3540 is a novel, gut-restricted, GCPII inhibitor with potent oral anti-colitis efficacy in both acute (DSS) and chronic (IL10-/-) mouse colitis models and a promising preclinical safety profile. Further development of this mechanistically unique candidate IBD drug is warranted.
P078 Vedolizumab Treatment of ex-vivo Human Ulcerative Colitis (UC) Explants Results in Altered Inflammatory Protein Secretion Profiles
Authors: Corcoran R , O’Connell F , O’Sullivan J , Kevans D
Published: January 2022

Background

Patient-derived inflammatory bowel disease (IBD) ex-plants have potential for biomarker and therapy discovery. Vedolizumab (VDZ) is a monoclonal antibody targeting a4b47 integrin whose mechanism of action is the reduction of inflammatory immune cell trafficking to the intestinal tract. The association between VDZ exposure and treatment response is unclear and appears insufficiently explained by serum levels. For this reason it is hypothesised that VDZ may also have effects at the tissue level.

Methods

We aimed to evaluate the effect of VDZ on inflammatory protein secretion profiles in ex-vivo human ulcerative colitis ex-plants (UC ex-plants).

Patients with UC, undergoing endoscopy, were prospectively recruited. Endoscopic biopsies were collected and UC ex-plants generated as per previously described methods. UC ex-plants were then co-cultured for 24 hours with an IgG control vehicle or VDZ. After 24 hours tissue conditioned media (TCM) from UC ex-plants was collected. TCM secreted protein profiles were quantified using 54 V-plex ELISA (Meso Scale Diagnostics, USA). Secreted cytokine profile were compared between IgG vehicle (control) and VDZ treated ex-plants. P values < 0.05 were considered significant in analyses.

Results

Thirteen patients with UC were included; age (mean, [range]) 45.8 years [30–78], 54% male; disease duration (mean, [range]) 8 [1–24] years; 62% of patients were anti-TNF naïve. Baseline total Mayo score (median [range]) was 6 [0–9]; endoscopic Mayo score (median [range]) was 2 [0–3]. Comparing VDZ with control treatment, 5 of 54 ex-plant secreted proteins differed significantly. GMCSF, IL-16, IL-22, IL-23 and sVCAM-1 secretion were significantly decreased comparing VDZ and control treatments, p < 0.03 for all comparisons.

Conclusion

The predominant mechanism of action of VDZ is the blocking of gut homing pro-inflammatory immune cell trafficking from the peripheral circulation to the intestinal tract. These data demonstrated that VDZ reduces pro-inflammatory protein secretions from UC ex-plants suggesting an additional local effect of this therapy. Further evaluation is required to determine whether a component of VDZ therapeutic effect occurs at the tissue level.
P079 The expression of some circulating micro RNAs and serum Vitamin D levels in IBD patients
Authors: Atanassova A , Georgieva PhD A C
Published: January 2022

Background

Serum miRNAs are secreted during the inflammation in the gastrointestinal tract and therefore could affect the regulatory functions of vitamin D and its signal pathways negative feedback 1,2,3. Additional tests are therefore needed to discover the correlation between the levels of vitamin D and the circulating miRNAs in patients with IBD.

Methods

The aim of this study is to assess the correlation between the serum levels of vitamin D and the expression of some miRNAs (Hs_miR-28_1, Hs_miR- 29c_1, Hs_miR-96_1, Hs_miR-191_1, Hs_miR-451_1, Hs_miR-142-5p_1, Hs_miR-199a_1, Hs_miR-363_1, Ce_miR39_1, Hs_miR-144_4, Hs_miR-142-3p_2, Hs_miR-155_2, Hs_miR-16_2, Hs_RNU6-2_11, Hs_miR-1228-2p_1) in a total of 50 patients with IBD: 27 with CD and 23 with UC. Serum levels of 25(OH)D were measured and correlated with serum CRP, FCP, IBD activity and the expression of different miRNAs. Correlation of levels of miRNAs with some clinical factors as 25(OH)D deficiency, treatment with corticosteroids or anti-TNF antibodies was also done.

Results

There is a significant difference (р=0.007) between the serum Vitamin D levels in patients with active disease (39.83 nmol/L ± 15.32 nmol/L) and those in remission (54.19 nmol/L ± 18.54 nmol/L). IBD activity is a risk factor for a decrease in the serum Vitamin D levels (OR=5.77 (1.552–21.503; p<0.01).

We found a weak inverse proportional correlation between levels of CRP and Vitamin D (r=-0.114;p<0.05). The correlative analysis found an inverse proportional weak to moderate correlation between the levels of FCP and Vitamin D (r=-0.259; p<0.01). The expression of miRNA-28_1 and miRNA-1228-3p_1 is increased in patients who normalized their Vitamin D levels. All others miRNAs have a higher expression in Vitamin D (25(OH)D) deficiency.

In Vitamin D (25(OH)D) deficit, the expression levels of all tested miRNAs were elevated if the patients received treatment with corticosteroids or monoclonal anti-TNF antibodies.

Conclusion

There is a correlation between the serum expression of different miRNAs and the Vitamin D levels in patients with IBD. Prospective cohort trials may identify the precise role of vitamin D as immune regulator as well as its relation to the expression of some miRNAs, participating in the processes of the pathogenesis, the optimization of the treatment and monitoring of the effect of different therapeutic regimens as a part of the personalized management.
P081 Therapeutic Drug Monitoring of infliximab in Crohn’s Disease: Cut-off points during maintenance therapy
Authors: Valdes T , Vilches Arenas A , Maldonado Perez B , Castro Laria L , Merino Bohorquez V , Argüelles Arias F
Published: January 2022

Background

Therapeutic drug monitoring of infliximab in Crohn’s disease is necessary to optimize treatments. However, infliximab serum levels are not well defined.

The aim of the study was to find our cut-off of infliximab serum levels in Crohn’s disease patients in remission in clinical practice. Furthermore, we tried to identify other predictors of long-term benefit of infliximab therapy.

Methods

Observational, retrospective and single-center study of patients diagnosed with Crohn’s disease on maintenance therapy with infliximab from January 2019 to July 2020.

Infliximab trough levels and antibodies to infliximab were measured at least three times, after 6 months of treatment. The tests were performed using enzyme linked immunosorbent assay. Clinical remission was defined using the Harvey Bradshaw index ≤4. The interpretation of data was by ROC analysis.

Results

105 Crohn’s disease patients were included in the study, 57.1% men, with a median age at diagnosis of 26 years. The rest of the characteristics are in table 1.

The best infliximab level cut-off point classifying patients in clinical remission was 4 μg/ml with an area under the curve of 0.801 (figure 1 and 2).

Median infliximab trough levels were significantly higher when patients achieved clinical remission [4.8 (3–8) μg/ml] versus [0.59 (0.1–2.4) μg/ml].

In the multivariate analysis we observed that the variables age, time on infliximab treatment and time of disease evolution were associated with non-remission (table 2).

Conclusion

Infliximab levels are an objective parameter related to clinical remission in maintenance therapy in patients with Crohn’s disease. Our cut-off point associated with clinical remission was 4 μg/ml.
P083 An integrative analysis of DNA methylation and RNA-Seq data in human adipose-stem cells of Crohn’s Disease patients with different clinical activity
Authors: Serena C , Montfort-Ferré D , Caro A , Menacho M , Espina B , Boronat-Toscano A , Martí M , Espín E , Millán M , Vendrell J , Fernández-Veledo S
Published: January 2022

Background

Crohn’s disease (CD) is characterized by the expansion of mesenteric fat attached to the inflamed segments of the intestine, also known as “creeping fat” which seems to be directly related to disease activity. Our group revealed that adipose-stem cells (hASCs) isolated from the creeping fat of CD subjects are dysfunctional (showing a high inflammatory profile, high invasive and phagocytic capacities, and worse immunosuppressive properties); and this dysfunction is maintained even in hASCs isolated from CD subjects in remission of the disease.1

Methods

1) Culture and characterization of hASCs isolated from adipose tissue biopsies of active CD, inactive CD, and healthy subjects (n=25). Visceral origin: creeping fat in active CD subjects and mesenteric fat in CD subjects in remission and healthy subjects. 2) Comparative DNA methylome analysis in hASCs isolated from the included population (Infinium Human-Methylation EPIC (850K) BeadChip; Illumina, Inc). 3) Transcriptomic study in hASCs isolated from the included population (Illumina HiSeq 2500 system). 4) Bioinformatic analysis & data integration.

Results

Methylation and transcriptomic studies revealed a multi-omic profile of hASCs isolated from CD subjects compared to control individuals (Heatmaps Fig 1A). Indeed, the integration of genome-wide methylation changes and gene expression differences revealed the main candidate’s genes (Figure 1B). The most significant upregulated gene in CD was Mab-21 like 2 (MAB21L2) (Fig 1B-C). This gene encodes a downstream target of transforming growth factor-beta signaling. MAB21L2 was validated in an independent cohort of hASC isolated from control, active, and inactive CD subjects (n=7/each group) using qPCR. In agreement, MAB21L2 gene expression was significantly higher in hASCs isolated from the active and inactive group than in the control group (Fig 1D). Another interesting candidate gene found was tyrosine kinase 2 (TYK2) which is significantly upregulated in active CD subjects. This gene promulgates cytokine signals by phosphorylating receptor subunits. It is also a component of both the type I and III interferon signaling pathways.2 Finally, another gene upregulated in remission of the disease is the calcium voltage-gated channel subunit alpha 1H (CACNA1H), which has been involved in low-grade inflammation and has been proposed as an important player in IBD during the remission phase.3

Conclusion

Integration of data revealed genes involved in the dysregulation of hASCs associated with CD and genes related to disease remission.

Reference

1. Serena C Stem Cell Rep 2017 9(4):1109–23; 2De Vries LCS JCC 2021 617-30; 3Picard E Br J Pharmacol 2019;176:950–63
P084 Secreted inflammatory protein profiles following treatment of ex-vivo human Ulcerative Colitis and Crohn’s Disease explants with licenced biologic therapies
Authors: Corcoran R , O’Connell F , O’Sullivan J , Kevans D
Published: January 2022

Background

Patient-derived inflammatory bowel disease (IBD) ex-plants have potential for biomarker and therapy discovery. Treatment of IBD ex-plants ex-vivo with licensed biologic therapies may result in further insights into their mechanism of action.

Methods

We aimed to evaluate the effect of Infliximab (IFX), Ustekinumab (USTK) and Vedolizumab (VDZ) on inflammatory protein secretion profiles in ex-vivo human IBD ex-plants.

Patients with IBD, undergoing endoscopy, were prospectively recruited. Endoscopic biopsies were collected from the sigmoid colon and IBD ex-plants generated as per previously described methods. IBD ex-plants were then co-cultured for 24 hours with an IgG control vehicle, IFX, USTK and VDZ. After 24 hours, tissue conditioned media (TCM) from IBD ex-plants was collected. TCM secreted inflammatory protein profiles were quantified using 54 V-plex ELISA (Meso Scale Diagnostics, USA). Secreted inflammatory protein profiles were compared between IgG vehicle (control) and USTK, IFX, VDZ treated ex-plants. All continuous variables are presented as median [interquartile range (IQR)]. P values < 0.05 were considered significant in analyses.

Results

18 patients with Ulcerative Colitis (UC) and 19 patients with Crohn’s Disease (CD) were included; age (median, [IQR]) 41[33–53], years, 49% male; disease duration (median, [IQR]) 9 [5–14] years; 54% of patients were anti-TNF naïve. IL-22 secretion was significantly lower in UC explants treated with IFX compared with IgG control; 4.5 [3.1 – 39.9] versus 17.5 [10.3 – 98.3] pg/ml per ug of protein respectively, p=0.004. IL-22 secretion was significantly lower in UC explants treated with USTK compared with IgG control; 7.8 [1.9 – 26.5] versus 27.9 [10.3 – 110.7] pg/ml per ug of protein respectively, p=0.004. IL-22 secretion was significantly lower in UC ex-plants treated with VDZ compared with IgG control; 10.6 [3.2 – 35.7] versus 17.5 [10.2 – 73.5] pg/ml per ug of protein respectively, p=0.01. For CD explants, IL-22 secretion was significantly lower in USTK compared with IgG control; 24.4 [17.6 – 57.7] versus 211.7 [75.9 – 588.1] pg/ml per ug of protein respectively, p < 0.001. In UC and CD explants, IL-23 secretion was significantly decreased comparing USTK with IgG control treatments, p < 0.004 for all comparisons.

Conclusion

IBD ex-plant IL-22 secretion is reduced following biologic therapy particularly in patients with UC. IL-22 is a pleotropic cytokine considered an essential factor in intestinal immune-epithelial cross talk and is capable of both pro- and anti-inflammatory functions depending on tissue microenvironment including cytokine milieu. Further evaluation of the contribution of IL-22 to IBD disease biology is warranted.
P085 Paneth cells translate dietary lipid exposure into gut inflammation
Authors: Grabherr F , Meyer M , Schmitz J J , Schwärzler J , Mayr L , Philipp M , Jukic A , Grander C , Tilg H , Adolph T E
Published: January 2022

Background

Dietary constituents impact the development and course of IBD (1). Especially a Western style diet is associated with an increased incidence and prevalence of IBD (2). The role of polyunsaturated fatty acids (PUFA) in the development of IBD is broadly discussed. Recently we could show that PUFAs elicit a pro-inflammatory response in intestinal epithelial cells (IEC) in a Glutathione peroxidase 4 (GPX4) deficient setting, which resembled phenotypic hallmarks of human CD (3). GPX4, an anti-oxidative enzyme, was described to be a risk gene for the development of CD (4) and is deficient in IECs derived from inflamed areas of the small intestine of CD patients (3). Stress responses in Paneth cells (PC) trigger small intestinal gut inflammation (5). In this study, we describe PC as originators of GPX4-restricted and PUFA-induced intestinal inflammation.

Methods

We analyzed PC morphology in mice lacking one allele of GPX4 specifically in IECs (GPX4+/-IEC) by electron microscopy. We generated mice lacking both alleles of Gpx4 specifically in PC (GPX4ΔPC) and exposed these mice (and wildtype controls) to a Western diet enriched with PUFAs for 12 weeks. Intestinal inflammation was assessed by histology and immunohistochemistry. Further we analyzed small intestinal whole tissue cultures and evaluated cytokine production by ELISA.

Results

Paneth cells of mice lacking one allele of Gpx4 showed morphological signs of ER-stress at baseline and after exposure to a PUFA-WD for 12 weeks when compared to WT mice (Fig 1a). Based on these results, we dissected a role for PC in GPX4-restricted gut inflammation. When fed a PUFA WD for 12 weeks GPX4ΔPC mice developed a CD like enteritis while WT controls did not (Fig 1b). A WD did not induce this inflammatory response in GPX4ΔPC or WT mice. The infiltrating leukocytes contained mainly neutrophils and macrophages (indicated by positive F4/80 staining, Fib 1c). Whole tissue cultures derived from GPX4ΔPC mice fed a PUFA WD produced more Interleukin-6 and CXCL-1, the murine homologue of the human Interleukin-8, compared to whole tissue cultures derived from WT mice fed the same diet (Fig 2).

Figure legend 1a) % of PC showing massive, medium or no ER dilatation 1b&c) enteritis score and representative H&E and F4/80 staining 2) IL-6 and CXCL-1 in supernatant of whole tissue jejunal samples

Conclusion

The presented data identify Paneth cells to be crucially involved in the development of PUFA induced enteritis in genetically susceptible hosts.

References

1. Roda G, NG S, et al Nat Rev Dis Primers 2020

2. Khalili H, Chan SSM, et al Nat Rev Gastroenterol Hepatol 2018

3. Mayr L, Grabherr F, et al Nat Com 2020

4. Jostins L, Ripke S, et al Nature 2012

5. Adolph TE, Tomczak M, Nature 2013
P089 Long ncRNA Landscape in the Rectum of Treatment Naïve Early Onset Ulcerative Colitis Highlights Association with Severity and Early and Late Disease Outcome, with Potential Role in Epithelial Metabolic Functions
Authors: Haberman Ziv Y , Braun T , Sosnovski K , Amir A , VanDussen K , Igor Ulitsky I U , Griffiths A , Walters T , Mack D , Boyle B , Kugathasan S , Jegga A , Hyams J , Denson L
Published: January 2022

Background

Long non-coding RNAs (lncRNAs) are tissue-specific and regulate cellular functions. We previously reported a dramatic inhibition of epithelial metabolic mitochondrial functions in mucosal biopsies from the UC PROTECT cohort. We aimed to define lncRNAs that are dysregulated in UC, linked with attenuated epithelial metabolic functions, and associated with worse outcome.

Methods

Transcriptomics of pre-treatment rectal biopsies in PROTECT UC [206 UC and 20 controls (Ctl)]. Weighted gene co-expression network analysis (WGCNA) to identify modules and gens associated with UC severity, week 4 remission after 5-ASA/steroids (W4R), week 52 steroid-free remission (W52SFR), and colectomy within 3 years. Epithelial mechanistic data of prioritized lncRNA.

Results

PROTECT transriptomics included 2,826 lncRNA genes with TPM>1 in 20% of samples. Principal components analysis (PCA, Fig 1) using these lncRNA showed distinct clusters of UC and Ctl, and PC2 values were linked (p<0.01) with endoscopic (Mayo score) and clinical (PUCAI) disease severities. Random forest classifiers using the 2,826 lncRNA trained to distinguish UC from Ctl using PROTECT, predicted the correct diagnosis in PROTECT (AUC=1) and in RISK (AUC=0.88) connecting lncRNA to UC pathogenesis in two independent cohorts. WGCNA on the 2,826 lncRNA resulted in 6 modules (M1-M6) associated with UC (p<0.01, Fig 2). M1, which contained lncRNA reduced in UC, was associated with lessW4R (p=0.01) less W52SFR (p=0.04) and increased colectomy (p=0.02). In contrast, M6, which contained lncRNA elevated in UC, was linked with W4R (p=0.008). The hub genes in the M1 and M6 modules included GATA6-AS1 and LINC01272 lncRNAs, respectively. Re-analysis of single cell datasets supported epithelial expression of GATA6-AS1 and myeloid expression of LINC01272. GATA6-AS1 co-expression with protein-coding genes in PROTECT showed significant enrichment for mitochondrial functions (p<1E-40). Reduction of GATA6-AS1 expression with 2 shRNAs resulted in reduced mitochondrial complex IV MT-CO2 mRNA and protein and reduction of LGR5, linking mitochondrial genes and function with epithelial renewal. We recently showed pronounced reduction of the mitochondrial membrane potential (MMP) by JC-1 staining in UC epithelia. Consistent with this finding, GATA6-AS1 inhibition reduced MMP levels from 0.22 in control to 0.08 (p=0.01), similarly to MMP of cells treated with CCCP that is known to depolarize and reduce MMP.

Conclusion

LncRNA are significantly linked with UC and disease severity, and the M1 epithelial enriched module shows significant association with worse outcome. GATA6-AS1 within the M1 hub genes show potential role in epithelial metabolic functions and renewal.
P101 Isosteviol Attenuates Dextran Sulfate Sodium-Induced Colitis by Maintaining the Intestinal Barrier Through PDK1/AKT/NF-κB Signaling
Authors: Yao L , Cao Q
Published: January 2022

Background

Inflammatory bowel disease (IBD) is a chronic debilitating inflammatory disorder of the gastrointestinal tract that is characterized by intestinal epithelial barrier dysfunction and excessive activation of the mucosal immune system. Isosteviol (IS) has been reported to possess anti-inﬂammatory properties. Thus, in this study, we aimed to investigate the therapeutic effects and detailed mechanisms of IS use against intestinal inflammation using both in vivo and in vitro models.

Methods

C57BL/6 mice were randomly divided into sham, dextran sodium sulfate (DSS), and DSS+IS groups. Colitis was established using 3.0% DSS through oral gavage for five consecutive days. In vitro, tumor necrosis factor-α (TNF-α)-treated Caco-2 cells were used as an inflammatory model. Clinical characteristics, histological performance, proinflammatory cytokine expression, and intestinal barrier function were measured. In addition, activation of the pyruvate dehydrogenase kinase 1/protein kinase B/nuclear factor-κB (PDK1/AKT/NF-κB) signaling pathway was determined by western blotting, quantitative polymerase chain reaction, and immunofluorescence assays.

Results

IS mitigated DSS-induced colitis by significantly reducing body weight loss, colonic shortening, and the disease activity index score via inhibition of colonic proinflammatory cytokine production of IL-1β, IL-6, and TNF-α. IS restored impaired barrier function by regulating tight junctions and intestinal epithelial barrier permeability. Furthermore, we found that IS ameliorated intestinal barrier injury by regulating the PDK1-dependent AKT/NF-κB signaling pathway.

Conclusion

Our results demonstrate that IS attenuates experimental colitis by preserving intestinal barrier function, probably mediated by the PDK1/AKT/NF-κB signaling pathway. These findings highlight the potential of IS as a novel therapeutic agent for IBD.
P117 point-of-care intestinal ultrasound for the detection of postoperative Crohn’s disease endoscopic recurrence
Authors: Dolinger M , Stauber Z , Spencer E , Kayal M , Pittman N , Colombel J F , Dubinsky M
Published: January 2022

Background

In patients with Crohn’s disease (CD), following an ileocolic resection (ICR), colonoscopy is the gold standard for the detection of endoscopic recurrence (ER). Colonoscopy, however, is invasive and not easily accepted by patients, particularly for repeated monitoring. In contrast, intestinal ultrasound (IUS) has the advantage of being non-irradiating, non-invasive, well-tolerated, and easy to repeat. The goal of this study was to assess the accuracy of IUS for ER in CD.

Methods

This was a cross-sectional study of CD patients who underwent IUS during a postoperative clinic visit within 30 days of a planned colonoscopy. Parameters on IUS included bowel wall thickness (BWT), bowel wall hyperemia (BWH; modified Limberg score 0-III), layer stratification, inflammatory fat, and complications. C-reactive protein (CRP), fecal calprotectin (FC), endoscopic healing index (EHI; Prometheus Labs, CA) and Harvey Bradshaw Index (HBI) were also measured. ER was defined as a Rutgeerts Score (RS) > i2. Primary outcome was the association between IUS parameters and ER. Secondary outcomes were the association of IUS parameters with other markers of disease activity.

Results

Eighteen CD patients (9 female; 29 [19–40] years old), underwent IUS examination 45 [29–99] months post-ICR during a routine clinic visit. Seven (39%) patients were on ustekinumab, four (22%) on adalimumab, two (11%) on infliximab, one (6%) on vedolizumab, and four (22%) on no therapy. ER was found in eight (44%) patients. BWT and BWH in the neo-terminal ileum and BWH at the ileocolic anastomosis were the only IUS parameters associated with ER (Table 1). Neo-terminal ileum BWT was 4.0 [3.2–4.8] mm in patients with ER compared to 2.0 [1.5–2.6] mm without (p=0.04). Neo-terminal ileum BWH was abnormal in six (75%) patients with ER compared to 0% without (p=0.007). BWT of 3.2 mm was the optimal cut point for predicting ER with an: AUROC of 0.82, positive predictive value of 100%, negative predictive value of 97.3%, sensitivity of 75%, and specificity of 100% (Figure 1) vs. a CRP of 10.4 mg/L (AUROC = 0.54) or FC of 1146 µg/g (AUROC = 0.56). Significant correlations were observed between neo-terminal ileum BWT and RS (ρ=0.51, p=0.04) and between BWT and CRP (ρ=-0.56, p=0.023). No correlation was observed between BWT and FC (ρ=-0.04, p=0.91), BWT and EHI (ρ=-0.04, p=0.91), or BWT and HBI (ρ=0.09, p=0.75).

Conclusion

IUS is a feasible, accurate, non-invasive monitoring tool for detection of postoperative CD recurrence. Larger prospective studies are needed to determine how IUS can be integrated in the monitoring of CD patients after surgery.