bluuebunny/crossref_metadata_2025_split huggingface dataset:
DOP79 Dose escalation of upadacitinib in Crohn’s disease patients with an inadequate response: Data from the randomised CELEST study
Authors: Sandborn W , Panés J , Peyrin-Biroulet L , Louis E , Greenbloom S , Zhou W , Zhou Q , Liu J , Lacerda A P 
Published: January 2020

Abstract

Background
Patients with Crohn’s disease (CD) may lose response to treatment, requiring dose adjustments. This analysis from the CELEST study evaluated efficacy and safety in patients with CD who required an escalated dose of upadacitinib (UPA), an oral selective JAK1 inhibitor.

Methods
CELEST (NCT02365649) was a placebo-controlled phase 2 study of adults with moderate-to-severe CD refractory to immunosuppressants/biologics. Patients were randomised to placebo or UPA 3, 6, 12, or 24 mg twice daily (BID) or 24mg once daily (QD) for 16 weeks (weeks), followed by a 36-week double-blind maintenance period to receive 3, 6 or 12mg BID or 24mg QD (Figure 1). Patients who met criteria for inadequate response (average daily liquid/soft stool frequency [SF] &gt;2.2 or average daily abdominal pain score &gt;1.8, and increase in hs-CRP &gt;1 mg/l from baseline [BL] or hs-CRP ≥5 mg/l at previous or current visit) at or after week 20 were eligible to receive open-label (OL) therapy with UPA 12mg BID. Patients with inadequate response at or after 4 weeks of OL 12 mg BID could further escalate to OL 24 mg BID. Data were analysed using descriptive statistics of clinical/endoscopic improvement, change from BL in hs-CRP and faecal calprotectin (FC), and AEs at week 52.

Results
Among 220 randomised patients, 180 were re-randomised into the maintenance period; 60 had inadequate response and received OL 12 mg BID. Of the 60 patients, 25 (42%) were in clinical response at week 16. After receiving OL 12 mg, 27 patients (45%) required escalation to OL 24 mg BID. A higher proportion of patients who received OL 24 mg received 2 or more prior TNF antagonists, and had higher BL FC, SF, and Simplified Endoscopic Score for CD (SES-CD) compared with patients who remained on OL 12 mg BID (Table 1). At week 52, 15% of patients who received OL 12 mg achieved clinical remission and 10% achieved endoscopic response. Of the patients who escalated to OL 24 mg, 39% achieved clinical remission and 41% were in endoscopic response at week 52 (Table 2). At week 52, change from BL in FC and hs-CRP was −2123 ± 2475 µg/g and −5.1 mg/l, respectively, for patients who received OL 24 mg vs. −423 ± 2953 and −6.4 mg/l for OL 12 mg. Serious infection was reported in 3 patients (2 receiving OL 12 mg and 1 receiving OL 24 mg). Two nonserious herpes zoster events were reported (1 on each dose), but neither event led to discontinuation of UPA. No malignancies, cardiovascular events, thromboembolic events, intestinal perforations, tuberculosis, or deaths were reported for patients who dose-escalated.

Conclusion
Patients with non-response or loss of response during CELEST gained clinical remission and endoscopic response with UPA 12 mg BID or further escalation to 24 mg BID. No new safety risks were observed for either dose of UPA, consistent with double-blind CELEST results.
DOP80 Effectiveness of ustekinumab and vedolizumab in patients with Crohn’s disease refractory to anti-tumour necrosis factor: A multi-centre comparative study
Authors: Alric H , Amiot A , Kirchgesner J , Tréton X , Allez M , Bouhnik Y , Beaugerie L , Carbonnel F , Meyer A 
Published: January 2020

Abstract

Background
There is no head-to-head trial comparing ustekinumab and vedolizumab in patients with Crohn’s disease (CD) refractory to anti-TNF. In France between May 2014 and November 2016, vedolizumab (and not ustekinumab) was reimbursed for patients who had failed anti-TNF. Then, between December 2016 and August 2018, ustekinumab (and not vedolizumab) was reimbursed for this category of patients. Since September 2018, both ustekinumab and vedolizumab are reimbursed in patients who are refractory to anti-TNF. The aim of this study was to compare effectiveness and safety of ustekinumab and vedolizumab in patients with CD refractory to anti-TNF.

Methods
We studied all consecutive patients with active CD who were refractory to at least one anti-TNF, and were treated either with vedolizumab or ustekinumab, in five university hospitals of the Paris area, between May 2014 and August 2018. The primary endpoint was clinical remission rate at week 48. Adjustment according to propensity scores with inverse probability of treatment weighting was performed.

Results
239 patients were included, 107 received ustekinumab and 132 received vedolizumab. After propensity scoring with IPTW, there was no difference between the two groups (Figure 1). At week 48, the clinical remission rate was higher with ustekinumab than with vedolizumab (54.4% vs. 38.3%; OR =1.92, 95% CI [1.09–3.39]). At week 48, corticosteroid-free remission rate tended to be numerically higher with ustekinumab than with vedolizumab (44.7% vs. 34.0%; OR = 1.57, 95% CI [0.88–2.79]). Treatment persistence was significantly more frequent in the ustekinumab group (71.5% vs. 49.7%; OR = 2.54, 95% CI [1.40–4.62]). The dose optimisation rate at week 48 was higher with vedolizumab than with ustekinumab (53.5% vs. 30.1%; OR = 0.37, 95% Cl [0.21–0.67]). Subgroup analyses showed that ustekinumab was associated with higher clinical remission rates at week 48 in patients with ileal CD (OR = 3.49; 95% CI [1.33–9.17]), a penetrating phenotype (OR = 6.58; 95% CI [1.91–22.68]) and a history of perianal disease (OR = 2.48; 95% CI [1.04–5.93]). Regardless of treatment group, combotherapy was associated with a higher clinical remission rate at week 48 (OR = 1.93; 95% CI [1.09–3.43]).

Conclusion
This study suggests that, after 1 year of follow-up, ustekinumab is associated with a higher rate of clinical remission than vedolizumab in CD patients refractory to anti-TNF, particularly in those with ileal and penetrating disease.
DOP81 Time to loss of efficacy among patients in remission following induction treatment with tofacitinib 10 mg BID who reduced tofacitinib dose or discontinued tofacitinib in OCTAVE sustain
Authors: Dubinsky M C , Chou J W , Su C , Wang W , Modesto I , Guo X , Vermeire S 
Published: January 2020

Abstract

Background
Tofacitinib is an oral, small-molecule JAK inhibitor for the treatment of ulcerative colitis (UC). Safety and efficacy of tofacitinib were evaluated in two Phase 3 induction studies (OCTAVE Induction 1 and 2; NCT01465763, NCT01458951), a 52-week, Phase 3 maintenance study (OCTAVE Sustain, NCT01458574) and an ongoing, open-label, long-term extension study (NCT01470612).1,2 Here, we assess time to treatment failure among patients in remission at the end of OCTAVE Induction 1 and 2 who enrolled in OCTAVE Sustain.

Methods
In OCTAVE Induction 1 and 2, patients received placebo or tofacitinib 10 mg twice daily (BID) for 8 weeks; clinical responders were re-randomised to placebo, tofacitinib 5 or 10 mg BID in OCTAVE Sustain for 52 weeks. Kaplan–Meier method was used to estimate median time to treatment failure (withdrawal due to insufficient clinical response) in patients who received 10 mg BID in induction studies and entered OCTAVE Sustain in remission (total Mayo score ≤2 with no subscore &gt;1, and rectal bleeding subscore [RB] 0). Treatment failure: ≥3-point increase from baseline total Mayo score, plus ≥1-point increase in RB and endoscopic subscore (ES; centrally read) and absolute ES ≥2 after ≥8 weeks of maintenance therapy.

Results
Following induction, 156 patients in remission entered OCTAVE Sustain and received tofacitinib 5 mg BID (N = 57) or 10 mg BID (N = 50), or placebo (N = 49). Estimated treatment failure rates are reported in the table. At Week 52, Kaplan–Meier rates of treatment failure were higher in patients who switched to placebo (81.8% [95% confidence interval 67.0, 90.4]) vs. those who continued to receive tofacitinib 10 mg BID (25.6% [14.2, 38.6]) or reduced their dose to 5 mg BID (34.4% [21.8, 47.3]). Median time to treatment failure was 169 days for placebo and &gt;52 weeks for tofacitinib 5 and 10 mg BID (due to the low number of treatment failure events, exact median time to treatment failure was not available for tofacitinib 5 or 10 mg BID).

Conclusion
For patients in remission following 8 weeks of induction treatment with tofacitinib 10 mg BID, median time to treatment failure for those who continued tofacitinib treatment (5 or 10 mg BID) was &gt;52 weeks. After treatment interruption (remission patients who were re-randomised to placebo), median time to treatment failure was 169 days; this was similar to previously published time-to-treatment-failure data for patients with clinical response (including remission) following 8 weeks of induction treatment with tofacitinib who were re-randomised to placebo.3.

References
DOP86 An IFN-STAT1-MLKL axis drives programmed necrosis of Paneth cells in Crohn’s ileitis
Authors: HARTMANN L , Siegmund B , Weidinger C , Becker C , Neurath M F , Günther C 
Published: January 2020

Abstract

Background
Interferons (IFNs) are immune-modulatory cytokines expressed by epithelial and mucosal cells in response to viral and bacterial infection. Just recently, we discovered a correlation between IFN-λ expression and disease activity, including small intestinal inflammation and Paneth cell dysfunction, in human Crohn’s disease patients. On a molecular level, we uncovered that IFN-λ mediates epithelial cell death, in particular, Paneth cell death by a programmed necrosis, dependent on STAT1 activation and controlled by caspase-8. These results suggested that IFN-λ can be considered as a pathogenic cytokine in Crohn′s ileitis and should be considered as a new and promising target for future therapeutic intervention for this particular subtype of IBD. Our central question is now by which pathways interferon-regulated programmed necrosis of epithelial cells contributes to intestinal inflammation and how these mechanisms could be targeted for future therapeutic intervention.

Methods
We use a mouse model for Crohn’s Disease like inflammation and Paneth cell death that has a specific deletion of Caspase-8 in intestinal epithelial cells (Casp8∆IEC). We stimulate small intestinal organoids derived from Casp8∆IEC mice with IFNs in vitro and we overexpress IFN-λ in these mice in vivo by hydrodynamic tail vein injection of an IFN-λ expression vector. Furthermore, we use JAK-inhibitors to impede pharmacologically cell death pathways in the pathogenesis of intestinal inflammation in vitro and in vivo.

Results
We uncovered that gene expression of the cell death mediators Mlkl and Caspase-8 is dependent on IFN-λ-mediated JAK-STAT1 signalling. The non-specific pan JAK-inhibitor Tofacitinib is able to attenuate gene expression of Mlkl and Caspase-8 in vitro as well as in vivo. It prevents non-apoptotic as well as apoptotic cell death of small intestinal organoids stimulated with IFN-λ and is sufficient to prevent small intestinal tissue destruction in Casp8∆IEC mice challenged with IFN-λ. Additionally, we use the selective JAK1-inhibitor Filgotinib to limit the targeted JAK-STAT signalling pathways to only JAK1-STAT1 signalling and thus reduce side effects of the inhibitor on other signalling pathways. This had a similar effect as Tofacitinib suggesting that IFN controls MLKL-mediated cell death via JAK1.

Conclusion
In summary, our results indicate that targeting IFN-λ-mediated JAK-STAT1 signalling by the small-molecules Tofacitinib and Filgotinib impedes induction of Mlkl and Caspase-8-mediated cell death pathways. Therefore, JAK1 inhibitors such as Filgotinib might represent a promising novel therapy that may be sufficient to achieve efficacy particularly in Crohn′s ileitis patients who display elevated IFN-l serum levels.
DOP90 The role of Nlrp9b, a novel NLR member in regulating experimental colitis
Authors: Zheng D , Levy M , Elinav E 
Published: January 2020

Abstract

Background
The NOD-like receptors (NLR) are important family members of the intracellular pathogen recognition receptors, which recognise pathogen- or danger-associated molecular patterns in the cell. The NLR signalling pathways are increasingly recognised to play critical roles in the pathogenesis of inflammatory bowel diseases. We have identified an NLR member, Nlrp9b, whose expression is restricted to the epithelium of the gastrointestinal tract. However, the role of Nlrp9b in regulating intestinal inflammation and the underlying mechanism remains to be clarified.

Methods
We generated constitutive Nlrp9b-deficient (Nlrp9b−/−) mice to study the function of Nlrp9b. We induced experimental colonic inflammation by using dextran sulphate sodium (DSS). Bone marrow transfer experiments were conducted to clarify the cell type in which Nlrp9b exerts its function. We then performed RNA-sequencing of intestinal tissues from Nlrp9b−/− mice and WT controls to investigate the potential mechanisms.

Results
Nlrp9b−/− mice were more susceptible to DSS-induced colitis compared with cohousing wild-type (WT) mice, manifesting as exacerbated weight loss, more severe inflammation in colonoscopy and histopathology. Non-haematopoietic expression of Nlrp9b was responsible for protection against intestinal inflammation. Inflammasome activation was not impaired in Nlrp9b−/− mice, as we detected no defects in both cleavage of caspase 1 and secretion of IL18 in Nlrp9b−/− mice. Through RNA sequencing, we identified 300 differentially expressed genes in the intestinal tissue between Nlrp9b−/− and the co-housed WT controls, which primarily belongs to pathways involved in intestinal IgA production, as well as antigen processing and presentation. Indeed, IgA levels were globally reduced in Nlrp9b−/− mice, on the levels of mRNA expression, secreted antibodies measured in the intestinal lumen, and serum IgA. Furthermore, the total numbers of immune cells in the Peyer's patches were significantly reduced in Nlrp9b−/− mice, including leukocytes, B cells and B cells in the germinal centres, where they class switch to IgA producing cells.

Conclusions
Nlrp9b functions in the non-haematopoietic compartments of intestine and is protective against experimental intestinal inflammation, through potential mechanisms involving intestinal IgA production.
P002 Specific genome editing to model hypermethylation of the SP140 gene that associates to Crohn’s disease
Authors: Hageman I , Li Yim A , Joustra V , Ghiboub M , Gecse K , Te Velde A , D’Haens G , Paulusma C , Henneman P , De Jonge W 
Published: January 2020

Abstract

Background
SP140 (Speckled 140 KDa) encodes an epigenetic reader protein with an immune restricted expression, that binds to epigenetically modified (acetylated and methylated) histones and thereby regulates expression of large gene sets, including pro-inflammatory cytokines, in innate immune cells. SP140 is implicated in CD because single nucleotide polymorphisms, as well as defective protein function are associated with CD and marks anti-TNF response. Through a genome-wide methylation screen of Crohn’s disease (CD) patients peripheral blood, we identified two hypermethylated positions in SP140 locus associated with CD patients. We hypothesise that this DNA hypermethylation at the SP140 locus controls SP140 expression in CD patients contributing to their colitis development.

Methods
To address the role of SP140 DNA methylation, we used CRISPR ‘dead’ Cas9 (dCas9) epigenome-editing for specifically adding methyl groups (dCas9-DNMT) or removing methyl groups (dCas9-TET) in monocyte cell line THP1. We developed guide RNAs complementary to the gene expression regulatory region of the SP140 gene. With lentiviral delivery, we transduced THP-1 cells with guide RNA-lentiviruses, and with dCas9-DNMT or dCas9-TET lentiviruses. We assessed the level of SP140 methylation using bisulphite Sanger sequencing and the effect of methylation intervention of SP140 using qPCR and ELISA for SP140, IL-6, TNFα, IL-1β.

Results
We observed that SP140 gene in THP-1 cells under control conditions contained little methylated CpG sites. We induced sp140 hypermethylation through transduction of dCas9-DNMT. We validated hypermethylation of the two SP140 CpGs in transduced THP1 cells, thus mimicking the observed hypermethylation in CD patients cells. SP140 hypermethylation in THP1 cells polarised into M1 macrophages and stimulated with lipoteichoic acid (TLR-2 ligand), displayed a decrease of TNFα and (p = 0.042) protein levels. Similarly, we showed a decrease of TNFα (p = 0.02) and IL-6 (p = 0.03) protein release after transduction dCas9-DNMT and stimulation with LPS or Zymosan (TLR-2/4 ligand).

Conclusion
In this study, we demonstrated that editing SP140 gene methylation through CRISPR-dCAS9 technology allows modelling of the relevance of epigenetic marks for CD aetiology. Through methylome editing, we could affect the expression of CD-associated pro-inflammatory genes. Our dCas9 technique will allow us to investigate the role of DNA-methylation in the aetiology of CD.
P006 3D Multicellular intestine-on-a-chip model for disease modelling and drug discovery
Authors: Lo Sasso G , Gijzen L , Marescotti D , Naumovska E , Raineri E , Nicolas A , Lanz H , Guerrera D , van Vught R , Joore J , Vulto P , Peitsch M C , Hoeng J , Kurek D 
Published: January 2020

Abstract

Background
One of the major functions of the human intestine is to provide a protective epithelial barrier between the body and digestive environment. Additionally, the interplay of commensal microbes of the gut microbiome with the gut tissue and host immune system significantly contributes to intestinal homeostasis. Crohn’s disease and ulcerative colitis, collectively referred to as inflammatory bowel diseases, are both associated with increased permeability of the epithelial barrier and dysregulated immune response. Great efforts have been made to develop both in vitro and in vivo models of the human intestine in order to understand the development and underlying pathogenesis of IBD. These efforts have provided valuable insights into multiple aspects of the disease. However, none of these models has been able to capture the complexity and multifactorial nature of IBD. Animal models generally fail to accurately predict the efficacy and toxicity of novel compounds in human tissues, while in vitro human intestinal models developed on porous membranes within Transwell inserts fail to accurately recapitulate and mimic key aspects of the in vivo situation.

Methods
Here, we present the development and characterisation of a 3D multicellular perfused intestine-on-a-chip model in a microfluidic platform, the OrganoPlate®, and its application for investigating intestinal inflammation. The model described here comprises a coculture of Caco-2 and mucus-secreting HT29-MTX cells in the top compartment of the chip and a coculture of immune-competent cells THP-1 and MUTZ-3 in the bottom compartment, lining a collagen-I ECM in the middle.

Results
We show that the Caco-2 and HT29-MTX coculture form confluent and polarised tubular structures against the collagen-I ECM in the OrganoPlate®, with a stable barrier function over time as well as the capability to secrete mucus. By exposing the cultures to TNFα and/or IL-1β, we were able to induce an inflammatory state, characterised by cytokine release (IL-8) and a decrease in trans-epithelial electrical resistance. Finally, we proved the applicability of the model in screening anti-inflammatory compounds by its reversibility. Using a well-known anti-inflammatory drug, TPCA-1, we were able to prevent cytokine-induced inflammation. This result was evident from the decreased secretion of IL-8 and retention of barrier function in treated cultures, similar to that observed in untreated cultures.

Conclusion
Overall, this complex 3D multicellular perfused intestine-on-a-chip model provides the versatile modularity of mimicking key features of intestinal inflammation and can, therefore, further support drug screening efforts and provide a platform for personalised medicine.
P007 GC1qR driven oxidative phosphorylation is essential for intestinal goblet cell differentiation
Authors: SÜNDERHAUF A , Hicken M , Skibbe K , Schlichting H , Hirose M , Perner S , Ibrahim S , Derer S , Sina C 
Published: January 2020

Abstract

Background
Induction of goblet cell differentiation during inflammation has been shown to be impaired in ulcerative colitis (UC) but not Crohn’s disease (CD), possibly explaining the intestinal goblet cell and mucus reduction observed in active UC. A metabolic switch from glycolysis to mitochondrial oxidative phosphorylation (OXPHOS) is necessary for terminal differentiation of intestinal stem cells towards goblet cells. Interestingly, intestinal energy deficiency in general and reduced level of OXPHOS in specific have been attributed to UC pathogenesis more than 30 years ago. The c1q binding protein (C1QBP; gC1qR) is indispensable for the maintenance of OXPHOS. Nevertheless, experimental evidence linking mitochondrial dysfunction with goblet cell depletion and C1QBP expression are still missing.

Methods
Goblet cell differentiation was studied in human biopsies from UC patients in remission, in mucus-producing HT29MTX cells and in a conplastic mouse strain with diminished mitochondrial OXPHOS activity. Furthermore, mice were fed an experimental diet to shift cellular energy production from glycolysis to OXPHOS and mucosal cell differentiation was compared with mice on an isocaloric control diet.

Results
In vitro, siRNA experiments in HT29MTX cells showed that butyrate-induced expression of goblet cell differentiation factor KLF4 is highly dependent on gC1qR expression. Interestingly, the latter was significantly reduced in human ileal and colonic sections of UC patients in remission compared with HN. OXPHOS-deficient conplastic B6-mt FVB mice further confirmed these findings by depicting diminished klf4 expression, lowered goblet cell numbers, a thinned intestinal mucus layer and signs of intestinal inflammation. Finally, via nutritional intervention in C57BL/6 mice we were able to increase gC1qR level and to induce goblet cell differentiation via hath1 and klf4 compared with controls.

Conclusion
Taken together, we here describe a new pathway linking low intestinal expression of OXPHOS-regulating gC1qR to impaired goblet cell differentiation, mucus reduction and mucosal inflammation, which could be possibly reversed by nutritional intervention.