bluuebunny/crossref_metadata_2025_split huggingface dataset:
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

Abstract

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 &lt;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

Abstract

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

Abstract

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 &lt; 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 &lt; 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

Abstract

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&lt;0.01).
We found a weak inverse proportional correlation between levels of CRP and Vitamin D (r=-0.114;p&lt;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&lt;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

Abstract

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

Abstract

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

Abstract

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 &lt; 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 &lt; 0.001. In UC and CD explants, IL-23 secretion was significantly decreased comparing USTK with IgG control treatments, p &lt; 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

Abstract

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