A. MERLI (1), F. QUESADA CALVO (1), C. MASSOT (1), N. BLETARD (2), N. SMARGIASSO (3), D. BAIWIR (4), G. MAZZUCCHELLI (3), M. DE PAUW-GILLET (5), M. MALAISE (6), E. DE PAUW (7), P. DELVENNE (2), M. MEUWIS (1), E. LOUIS (1) / [1] ULg and CHU de Liège, Liège, Belgium, Translational Gastroenterology, GIGA-R and Hepato-Gastroenterology and Digestive Oncology unit, [2] CHU de Liège, Liège, Belgium, Department of Preclinical and Biomedical Sciences - Laboratory of Experimental Pathology, [3] Université de Liège, Liège, Belgium, Department of Chemistry - Mass Spectrometry Laboratory, [4] Université de Liège, Liège, Belgium, GIGA-Technology platforms : Protemics Platform, [5] Université de Liège, Liège, Belgium, Department of Preclinical and Biomedical Sciences - Mammalian Cell Culture Laboratory, [6] CHU de Liège, Liège, Belgium, Department of Clinical Sciences - Rheumatology, [7] université de Liège, Liège, Belgium, Department of Chemistry - Mass Spectrometry Laboratory
Introduction: Chronic colonic inflammation in ulcerative colitis (UC) may induce dysplasia, which can itself progress and transform into neoplasia. Diagnosis of dysplasia in UC remains difficult particularly when tissue inflammation is present.
Aim: The aim of this retrospective pilot study was to highlight proteins specifically associated with inflammation induced dysplasia in UC.
Methods: We performed a pilot experiment on 15 Formalin-Fixed, Paraffin-Embedded (FFPE) samples isolated from 5 cases of UC patients with an Adenoma-Like Mass (ALM). We compared the proteomes of the ALM, the inflammatory (I) and the normal (NL) tissues of each patient. We performed Laser Capture Microdissection (LCM) in order to collect only epithelial cells, avoiding inflammatory infiltrating ones. Label free proteomic analysis using a 2D-nanoUPLC coupled with a hybrid Quadrupole-Orbitrap was applied, as well as differential analysis on the paired samples. Immunohistochemistry (IHC) characterisation of one of the selected proteins of interest was used for validation.
Results: Out of 985 quantified proteins, 7 were found significantly more abundant in ALM compared to I tissues, with 6 being only detected in ALM using proteomics. One of these is Solute Carrier Family 12 member 2 (SLC12A2), also known as Na-K-2Cl co-transporter 1 (NKCC1), a protein involved in ionic balance, in T-cell migration promotion and in some features involved in cancer development like proliferation, migration or invasion. IHC results obtained were in correlation with proteomic results and showed that SLC12A2 was more abundant in ALM tissue than in I and NL tissues, with a signal clearly delimiting the dysplastic region from the surrounding inflammatory tissue.
Conclusions: This pilot experiment shows a different proteomic profile in inflammation-associated dysplasia and simple inflammation. This should be replicated using other types of dysplasia in IBD (DALM and flat dysplasia). SLC12A2 could be a potential biomarker of inflammation-associated dysplasia.
M. NOBEN (1), N. HENDRIKS (2), S. VERMEIRE (3), G. VAN ASSCHE (3), C. VERFAILLIE (4), M. FERRANTE (3) / [1] KU Leuven, , Belgium, Translational Research Center for Gastrointestinal Disorders (TARGID), [2] KU Leuven, Leuven, Belgium, Translational Research Center for Gastrointestinal Disorders (TARGID), [3] University Hospitals Leuven, Leuven, Belgium, Department of Gastroenterology and Hepatology, [4] KU Leuven, , Belgium, Stem Cell Institute Leuven
Introduction: The intestinal epithelium is the first line of contact between the host and microbiota, and other luminal particles which may have a pathogenic role in patients with inflammatory bowel disease (IBD). Previously we showed that culturing organoids from healthy controls (HC) and patients with Crohn’s disease (CD) or ulcerative colitis (UC) works with equal efficiency, and that the transcriptional profiles are largely comparable (LGR5, MUC2, among others).
Aim: Here we investigated how the inflammatory burden affects cultured organoids.
Methods: Biopsies were derived from both the inflamed and non-inflamed mucosa of patients with IBD, including 5 patients with UC (3 males, median disease duration 4.6 years), and 7 patients with colonic Crohn’s disease (CDc, 4 males, median disease duration 10.1 years), and 3 female HC. Next, crypts were isolated and colon organoids were derived following previous described methods (Sato et al., Gastroenterology, 2011). RNA was isolated both from the original biopsies, as well as from organoids at the end of passage 1 of culture (14 days after isolation, kept in expansion medium with Wnt3a, EGF, Noggin, RspoI, B27, nicotinamide, p38-inhibitor, A83-01). We used RTqPCR to assess expression levels of the following genes: LGR5, CXCL8 (Interleukin-8), CXCL3, IL1β, IFNgamma , and TNFα. Ct values were normalized on the geometric mean of 3 reference genes (RPS14, HPRT1, and B2M). DeltaCt values were used for statistical analysis for: IBD vs HC for biopsy and organoid data and inflamed vs non-inflamed in the IBD groups.
Results: The intestinal stem cell marker LGR5 was equally expressed in the different groups and was enriched in organoids compared to biopsies (HC vs UC vs CD). A significant decreased in expression of TNFα was observed in organoids compared to biopsies for the CDc groups (in organoids derived from normal as well as inflamed tissue), while expression levels of TNFα in organoids were equal between the groups. CXCL8 and CXCL3 were upregulated in organoids compared to biopsies (regardless of the inflammatory status at the site of biopsy). mRNA levels of IL1β and IFNgamma were less expressed in organoids compared to primary biopsies, suggesting that removal from the inflammatory milieu leads to loss of the inflammatory phenotype.
Conclusions: Organoids derived from inflamed tissue have equivalent transcriptional profiles to those from non-inflamed tissue from the same patient, or from healthy controls. We identified that CXCL8 and CXCL3 transcripts are induced upon culture in all samples tested, suggesting that these markers might be less suitable for investigating a response in inflammation. Expression of other genes was not seen following culture and these may hence be more suitable for measuring inflammation in organoids. Most importantly, we could not detect significant differences in gene expression of a number of inflammatory genes in cultured organoids derived from inflamed and non-inflamed tissue of HC and IBD patients, indicating that removal from the in vivo inflamed environment results in loss of the inflammatory phenotype.
A. CEULEMANS (1), S. VERHULST (2), M. VAN HAELE (1), L. VAN GRUNSVEN (2), T. ROSKAMS (1) / [1] Katholieke Universiteit Leuven, Leuven, Belgium, Department of Imaging & Pathology, Translational Cell & Tissue Research Group, [2] Vrije Universiteit Brussel, Brussels, Belgium, Department of Basic Biomedical Sciences, Liver Cell Biology Lab
Introduction: Hepatic progenitor cells (HPCs) are small cells with a relative large oval nucleus and a scanty cytoplasm situated in the canals of Hering. Phenotypically, HPCs express both markers of (immature) hepatocytes (e.g. alpha-fetoprotein) and markers of cholangiocytes (e.g. cytokeratin K7 and K19). The mechanisms facilitating proliferation and differentiation of human HPCs are still poorly understood.
Aim: In this study, we aimed to characterise human HPCs through isolating and comparing, on both protein and RNA level, HPC-enriched cell populations from adult human liver tissue using different isolation methods.
Methods: Fresh human liver tissue was collected from alcoholic steatohepatitis explant livers and HPC-enriched cell populations were obtained via three different isolation methods: side population (SP) which is based on the cell’s efflux capacity of Hoechst-33342 and isolation based on the membrane markers EpCAM and TROP-2. FACS-sorted cells and whole liver extracts were evaluated at both protein level (immunohistochemical staining) and RNA level (RNA sequencing). Pathway analysis was performed using Ingenuity Pathway Analysis.
Results: Immunohistochemical evaluation of the isolated fractions indicated the enrichment of HPCs in the SP, EpCAM-positive and TROP-2-positive cell populations. Pathway analysis of the RNA sequencing data from the different isolated HPC fractions shows an enrichment and activation of known HPC pathways like the Wnt/beta-catenin pathway, but also a pathway thus far not linked to HPC activation: IL-17A signalling. Upregulation of downstream targets like IL-8, CXCL1 and CCL20 indicate activation of the IL-17A pathway in HPCs. Interestingly, chemoattractants like IL-23A and IL-1b are upregulated in HPCs, possibly to recruit and activate IL-17A producing cells in the liver. IL-17A has already been linked with fibrogenesis through activation of stellate cells and with inflammation through inducing the production of immune cell chemoattractants. Upregulation of TGFbeta, VEGF, IL6 and PDGFB in HPCs is correspondent with the known link of IL-17A with fibrogenesis.
Conclusions: Our analysis indicates an important role for IL-17A signalling during HPC activation, associated fibrogenesis and inflammation in human alcoholic liver disease.
M. STAKENBORG (1), E. MERONI (1), G. GOVERSE (1), M. DI MATTEO (2), M. MAZZONE (2), G. MATTEOLI (3) / [1] KU, Leuven, Belgium, TARGID, [2] KU, Leuven, Belgium, Department of Oncology, [3] KU Leuven, , Belgium, TARGID
Introduction: Neutrophils are essential to maintain intestinal homeostasis as they provide a first line of defense against invading pathogens. During inflammation, neutrophils also aid in the recruitment of other immune cells and facilitate the immune response in the gut. However, during chronic inflammatory conditions, such as Inflammatory Bowel Disease (IBD), excessive neutrophil accumulation can lead to tissue damage, delayed tissue repair and loss of homeostasis.
Aim: Thus, in the current study, we aim to identify the role and function of neutrophils recruited during intestinal inflammation and their contribution in the pathogenesis of colitis.
Methods: To block neutrophil chemoattraction and cytotoxicity in response to its ligand hepatocyte growth factor, we used the neutrophil-specific Mrp8-Cre line backcrossed with Metfl/fl. Acute colitis was induced in MRP8Cre/WT METfl/fl (KO) mice and MRP8WT/WT METfl/fl littermate controls (WT) by 2,5% dextran sodium sulfate (DSS) in drinking water for 5 days. Disease progression was assessed via a standardized disease activity index (DAI) including body weight loss, stool consistency and blood in the faeces. During chronic colitis, mice were subjected to 3 cycles of 2.5% DSS for 5 days followed by 2 weeks of drinking water. Colonic immune cells were assessed by flow cytometry. Data are expressed as mean ± SEM; t-test was performed; p<0.05 is considered significant.
Results: During the third cycle of chronic DSS colitis, KO mice displayed a decreased DAI (p<0.01) and body weight loss (p<0.05) compared to WT mice. Moreover, flow cytometric analysis revealed a reduced amount of ROS+ neutrophils (WT; 3.17 ± 0.84 x 105, KO; 0.69 ± 0.26 x 105, p<0,05), eosinophils (WT; 3.00 ± 0.62 x 105, KO; 0.80± 0.20 x 105, p<0,05), and macrophages (WT; 9.98 ± 0.59 x 105, KO; 6.46 ± 0.73 x 105, p<0,05), implying a protective effect of MET deletion in MRP8+ neutrophils during chronic intestinal inflammation. To further elucidate the observed phenotype, we performed acute DSS colitis. In line, KO mice subjected to acute DSS colitis showed an improved disease course with reduced body weight loss and DAI and a comparable decrease in the amount of neutrophils, eosinophils and macrophages. Moreover, the percentage of FoxP3+ T regulatory cells was increased in KO mice compared to their WT counterparts (WT; 29,27 ± 2,14%, KO; 41,05 ± 2,11%, p<0,05), pointing towards a return to homeostasis in the KO colon. Strikingly, analysis of CD3+ CD4+ T cells showed a predominant decrease of the percentage IL17A+ Th17 (WT; 26,87 ± 1,85%, KO; 14,19 ± 2,11%, p<0,01) and IL17A+ IFNg+ Th1-like Th17 (WT; 10,62 ± 0,50%, KO; 4,70 ± 1,51%, p<0,01) in KO mice compared to WT mice, while no differences were observed in the percentage of IFNg+ Th1 cells (WT; 12,03 ± 0,61%, KO; 12,03 ± 3,00%, ns).
Conclusions: In the present study, we showed that MET is required for neutrophil chemoattraction and cytotoxicity during colitis. MET deletion in neutrophils seems to be essential to limit inflammation in the lamina propria. In addition, MET deletion in neutrophils was associated with a specific reduction of Th17 cells. Further understanding the mechanisms underlying neutrophil function during colitis will aid in the development of novel therapeutic strategies to treat IBD patients.
I. MANNAERTS (1), L. THOEN (1), F. CUBERO (2), I. COLLE (3), C. TRAUTWEIN (2), I. COLDHAM (4), L. VAN GRUNSVEN (1) / [1] Vrije Universiteit Brussel, Jette, Belgium, BMWE-LIVR, [2] UH Aachen, , Belgium, Internal Medicine III, [3] UZ Gent, Gent, Belgium, Hepatology and Gastroenterology, [4] University of Sheffield, Sheffield, United Kingdom (the), Synthetic Organic Chemistry
Introduction: Liver fibrosis or scarring of the liver is the consequence of prolonged hepatocytic damage that results in persisting hepatic stellate cell (HSC) activation. This makes stellate cells the primary targets for anti-fibrotic therapy and emphasizes the need to understand how HSCs contribute to fibrosis development. The unfolded protein response (UPR) is a cellular response related to ER stress. Chemical induction of ER stress has been shown to affect HSC activation. The nonsense-mediated RNA decay (NMD) pathway functions in RNA quality control. Aberrant mRNAs are rapidly degraded, and a subset of normal mRNAs is regulated by NMD.
Aim: To evaluate whether the endogenous UPR is essential for the earliest phases of mouse HSC (mHSC) activation and how this UPR is regulated.
Methods: In vitro and in vivo activated HSCs were analyzed for ER stress by qPCR, western blot and immunohistochemistry, in WT and JNK-Knock-out mice. ER stress inducers and NMD inhibitors are used in HSC cultures.
Results: The ER stress markers, XBP1spliced, Bip and Chop, showed an early peak in mRNA and protein expression already 10h after seeding primary mouse HSCs on plastic culture dishes, followed by a decreased expression at 24h. This temporarily increased ER stress is also seen in freshly isolated HSCs from mice 10h after 1 CCl4 injection, suggesting that ER stress is an early event of HSC activation also in vivo. HSCs cultured as 3D spheroids showed prevention of early ER stress and inhibition of HSC activation compared to HSCs plated on plastic. In 3D HSC cultures, chemical induced ER-stress is not sufficient to induce HSC activation. Treatment of HSCs with JNK inhibitors prevents the initial ER stress and reduces culture-induced activation of primary mouse HSC. This role for JNK was confirmed using JNK1 KO mice where decreased ER stress and activation were observed when isolated HSCs were plated. NMD inhibitors induce UPR and enhance HSC activation in vitro suggesting an active role for NMD in the regulation of the UPR and HSC activation.
Conclusions: ER stress induction is an early event during HSC activation in vitro and in vivo. In vitro, this acute ER stress is JNK1 dependent, but is not sufficient to drive the activation process. Ongoing work strongly suggests a potential role for NMD in the regulation of the UPR termination and HSC activation.
L. VAN DEN BOSSCHE (1), P. HINDRYCKX (1), L. DEVISSCHER (1), S. DEVRIESE (1), S. VAN WELDEN (1), T. HOLVOET (1), R. VILCHEZ-VARGAS (2), M. VITAL (3), D. PIEPER (3), J. VANDEN BUSSCHE (4), L. VANHAECKE (4), T. VAN DE WIELE (2), M. DE VOS (1), D. LAUKENS (1) / [1] Ghent University, Ghent, Belgium, Department of Gastroenterology, [2] Ghent University, Ghent, Belgium, Center for Microbial Ecology and Technology, [3] Helmholtz Centre for Infection Research (HZI), , Germany, Department of Medical Microbiology, [4] Ghent University, Ghent, Belgium, Department of Veterinary Public Health and Food Safety
Introduction: The promising results with secondary bile acids in experimental colitis suggest that they may represent an attractive and safe class of drugs for the treatment of inflammatory bowel diseases (IBD). However, the exact mechanism by which bile acid therapy confers protection from colitogenesis is currently unknown.
Aim: Since the gut microbiota plays a crucial role in the pathogenesis of IBD, and exogenous bile acid administration may affect the community structure of the microbiota, we examined the impact of the secondary bile acid ursodeoxycholic acid (UDCA) and its taurine/glycine conjugates on the fecal microbial community structure during experimental colitis.
Methods: Acute colitis was induced in mice by administration of 4% dextran sodium sulfate to the drinking water for 7 days. Mice were treated with 500 mg/kg/d UDCA, tauroursodeoxycholic acid (TUDCA), glycoursodeoxycholic acid (GUDCA), or placebo by oral gavage. At day 9 of colitis, fecal microbiota profiles were determined through 16S rRNA Illumina MiSeq sequencing and mice were sacrificed at day 10 to assess the severity of inflammation. Ultra-high performance liquid chromatography and high resolution mass spectrometry were performed on fecal samples to analyze the extent of biotransformation of orally administered UDCA, TUDCA and GUDCA.
Results: Daily administration of UDCA, TUDCA and GUDCA equally lowered the severity of colitis, as evidenced by reduced body weight loss, colonic shortening and expression of inflammatory cytokines. Illumina sequencing demonstrated that bile acid therapy during colitis did not restore fecal bacterial richness and diversity but normalized the colitis-associated increased ratio of Firmicutes to Bacteroidetes. Interestingly, administration of bile acids prevented the loss of Clostridium cluster XIVa and increased the abundance of Akkermansia muciniphila, bacterial species known to be particularly decreased in IBD patients. Orally administered UDCA, TUDCA and GUDCA were extensively metabolized in vivo, resulting in a similar fecal bile acid composition.
Conclusions: We conclude that UDCA, which is an FDA-approved drug for cholestatic liver disorders, could be an attractive treatment option to reduce dysbiosis and improve inflammation in human IBD.
J. Schouten (AZ Nikolaas) and H. Orlent (AZ Sint-Jan Brugge) will challenge C. Van Steenkiste (AZ Maria Middelares) with difficult cases of portal hypertension and variceal bleeding, which will be discussed according to the Baveno guidelines.
Personal experiences and challenges as a young Hepatology researcher abroad.
P. SAMBON (1), S. VARMA (2), M. KOMUTA (3), P. CLAPUYT (4), E. SOKAL (1) / [1] Cliniques Universitaires St Luc, Brussels, Belgium, Pediatric Gastroenterology and Hepatology, [2] Cliniques universitaires St-Luc, Brussels, Belgium, Pediatric Gastroenterology and Hepatology, [3] Cliniques Universitaires St Luc, Brussels, Belgium, Service of anatomical pathology , [4] Cliniques Universitaires St Luc, Brussels, Belgium, Pediatric Radiology Unit
Introduction: Concomitant presence of autoimmune hepatitis (AIH) and primary sclerosing cholangitis (PSC) is labelled as AIH-PSC overlap syndrome or autoimmune sclerosing cholangitis (ASC). Treatment of AIH with corticosteroids and azathioprine; and of the PSC component with ursodeoxycholic acid (UDCA) is the standard practice. Antibiotics are increasingly being shown to have benefit in PSC but their role in paediatric ASC is not well evaluated.
Aim: We investigated the response to oral antibiotics as initial or subsequent therapy in children with ASC.
Methods: Patients diagnosed with ASC on basis of biochemical, liver biopsy and radiology findings were included. They received metronidazole or vancomycin for 14 days [10-220] either at diagnosis (i.e. initial therapy) or during their maintenance period. When antibiotics were administered as initial therapy, steroid free induction regime was adopted. In children during the maintenance phase antibiotics were administered if they had not achieved biochemical remission with their standard treatment of steroids, azathioprine and UDCA. The outcome parameters to assess the efficacy of antibiotics were achievement of biochemical remission and additionally steroid avoidance when given in the initial therapy.
Results: Ten children with ASC were included, of which 6 received oral antibiotics (4 metronidazole, 2 vancomycin) at diagnosis and 4 received metronidazole during the maintenance period. All patients showed a significant decrease in their AST (-55%, p=0,005), ALT (-84%, p=0,003) and GGT (-53%, p=0,003), without significant difference across the two groups. All six children in the initial therapy group did not need corticosteroids and continued to be in remission until last follow up duration of 400 days [216-888]. Among the four children administered antibiotics in the maintenance phase, two showed biochemical remission and steroids could be tapered; while two did not show any benefit. There was transient biochemical relapse after stopping antibiotics in one responder, for which they were restarted and continued until last follow up while continuing to be in remission.
Conclusions: We demonstrate the benefit of antibiotics in ASC by achieving steroid free treatment when given at diagnosis as induction regime. When given in the maintenance phase they assist in achieving long term biochemical remission in an otherwise uncontrolled ASC.
E. FIANI (1), F. GUISSET (1), Q. FONTANGES (2), J. DEVIERE (1), A. LEMMERS (1) / [1] Erasme Hospital, Brussels, Belgium, (1) Gastroenterology, Hepatopancreatology and Digestive Oncology department, [2] Erasme Hospital, Brussels, Belgium, (2) Pathology department
Introduction: Esophagitis dissecans superficialis (EDS) is a rare desquamative disorder of the esophagus, characterized by sloughing of the superficial mucosa. It is a benign entity of uncertain etiology. Most cases of EDS are idiopathic but can be caused by medications, hot beverages, chemical irritants, celiac disease and many skin conditions.
Aim: Knowing that few case series have described this entity, we decided to review all the cases diagnosed in our center to characterize them.
Methods: The pathological institutional database of Erasme University Hospital (Brussels, Belgium) was searched for the diagnosis of EDS. We reviewed retrospectively the clinical and endoscopic findings as well as histological features of all cases of EDS (table 1). During this period of time, 21500 upper gastrointestinal endoscopies have been performed in our institution.
Results: From 2010 to 2016, we identified 7 cases of EDS diagnosed in our institution in this time period. During the same period, 21500 upper gastrointestinal endoscopies were performed (incidence 0.03%). The median age of presentation was 73 years old, with a female predominance (85%). Associated symptoms were variable from weight loss and nausea to epigastric pain, dysphagia and atypical chest pain. The most common co-morbidity found was treated hypertension in 3 patients. There were no skin diseases in any of these patients. Only one patient in our series had an identified potential causal factor (clindamycin), because of the sudden onset of symptoms upon initiating clindamycin for septic arthritis. Endoscopic findings evoked in 2 patients a suspicion of an esophageal tumor; the first one was described as a raised detached lesion of the distal third of the esophagus with suspicion of squamous cell carcinoma (Figure 1) and the second as a suspected tumor of the proximal third of the esophagus (Figure 2). For other patients, EDS was misdiagnosed as unspecific esophagitis in 3, reflux or mycotic esophagitis in 2. Only one patient was suspected to have sloughing esophagitis. Histologic features present in all of those cases were characterized by the presence of a sloughing and necrosis of the superficial layer of the esophageal squamous epithelium with negative anti HSV and anti CMV antibodies, negative periodic acid Schiff stain for fungal infections as well as absence of signs of dysplasia or signs of malignancy. In 2 patients, there was a presence of multiple bacterial colonies on the superficial epithelium. Acute inflammation was reported in 4 of the patients with the presence of eosinophils in the superficial epithelium described in 2 of these patients and of polymorphonuclear leukocytes in 2 other patients (figure 3). An endoscopic follow up 2 months after PPI treatment (with pantoprazole 40 mg once daily) was performed in 2 patients who had an atypical endoscopic presentation with suspicion of a previous suspicion of esophageal neoplasia. A complete healing of the esophageal lesions was observed in these 2 patients (Figure:4).
Conclusions: EDS is a rare benign entity that endoscopists must be aware of in order not to mistake it for other entities such as esophagitis or squamous cell carcinoma. The diagnosis is based on biopsies. The prognosis is good after stopping the causative agent and with PPI treatment.
P. BOSSUYT (1), F. BAERT (2), J. COENEGRACHTS (3), M. DE VOS (4), O. DEWIT (5), M. FERRANTE (6), F. FONTAINE (7), F. MANA (8), J. VANDERVOORT (9), T. MOREELS (10) / [1] Imelda Hospital, Bonheiden, Belgium, Department of gastroenterology, [2] AZ Delta, Roeselare, Belgium, Department of gastroentrology, [3] Jessa Hospital, Hasselt, Belgium, Department of gastroenterology, [4] University Hospital Ghent, Ghent, Belgium, Department of gastroenterology, [5] Cliniques Universitaires Saint-Luc, , Belgium, Department of gastroenterology, [6] UZ Leuven Gasthuisberg, Leuven, Belgium, Department of gastroenterology, [7] CHC, Liège, Belgium, Department of gastroenterology, [8] UZ Brussel, Jette, Belgium, Department of gastroenterology, [9] Onze-Lieve-Vrouw Ziekenhuis, Aalst, Belgium, Department of gastroenterology, [10] Antwerp University Hospital, Edegem, Belgium, Department of gastroenterology
Introduction: The natural history of ulcerative colitis (UC) is unpredictable. The current approach is gradual step-up (SU) therapy in the majority of patients. Data on the need for and factors influencing SU therapy beyond 5ASA or steroids are understudied.
Aim: To describe the first year SU therapy in patients with early UC failing on 5-ASA or steroids.
Methods: In this prospective multicentre observational trial patients with UC failing on 5-ASA and/or steroids where followed for 12 months. Patient characteristics, demographics, medical therapy, biomarkers, therapy adherence and quality of life were evaluated at every out-patient visit.
Results: A total of 103 patients (54% male, median age 40 years, median disease duration 17 months) were included. Only 2% were active smokers, while 51% were ex-smokers. Of the 103 patients 34%, 24% and 42% were 5-ASA-refractory, cortico-dependent and cortico-refractory respectively. After 1 year of follow up 81% of patients had mild or inactive UC based on the Mayo score. Sixty percent of patients had been treated with immunomodulators and 30% with biological therapy. Eighteen percent used combination therapy, representing only 54% of patients on anti-TNF therapy. The median time to initiation of immunomodulators and anti-TNF was 1 day and 55 days respectively, with a quicker initiation of anti-TNF treatment in cortico-dependent (34 days; 95% CI: 0-148) and cortico-refractory (57 days; 95% CI: 2-181) patients as compared to 5-ASA-refractory patients (97 days; 95% CI: 17-262). In total, 24/43 (56%) cortico-refractory patients started anti-TNF treatment. This was a significantly higher number compared to 4/25 (16%) of the cortico-dependent group (p= 0.002) and 7/35 (20%) of the 5-ASA-refractory group (p = 0.002). Biomarkers (CRP and platelet count) and clinical scores were numerically higher at initiation of anti-TNF therapy compared to immunomodulators. Whereas the use of faecal calprotectin was negligible (7%) in therapeutic decision making. Two patients underwent colectomy. Based on the results of the MMAS-8 questionnaire, patients with severe disease at baseline presented a lower median MMAS-8 score throughout the study period and thus were less adherent to therapy.
Conclusions: In patients with early UC a step up approach leads to good clinical outcomes at 1 year. Immunomodulators are initiated very early in patient flaring on 5-ASA or steroids, and up to 30% will be on anti-TNF treatment within 1 year, with cortico-refractory patients having the highest risk. Surprisingly, combination therapy is not used very often in daily clinical practice. The gradual SU and the acceleration of the therapy are based on sanguine biomarkers and clinical scores, not on faecal calprotectin levels.
M. DE BRUYN (1), R. RINGOLD (2), M. FERRANTE (3), G. VAN ASSCHE (4), G. OPDENAKKER (5), A. DUKLER (2), S. VERMEIRE (4) / [1] Rega Institute for Medical Research, KU Leuven, , Belgium, Department of Microbiology and Immunology, Laboratory of Immunobiology & Department of Clinical and Experimental Medicine, Translational Research Center for Gastrointestinal Disorders (TARGID), [2] Kepler Diagnostics, Inc., , United States (the), Kepler Diagnostic, Inc., [3] University hospitals Leuven, KU Leuven, leuven, Belgium, Department of Gastroenterology and Hepatology & Department of Clinical and Experimental Medicine, Translational Research Center for Gastrointestinal Disorders (TARGID), [4] University Hospitals Leuven, KU Leuven, Leuven, Belgium, Department of Gastroenterology and Hepatology & Department of Clinical and Experimental Medicine, Translational Research Center for Gastrointestinal Disorders (TARGID), [5] Rega Institute for Medical Research, KU Leuven, , Belgium, Department of Microbiology and Immunology, Laboratory of Immunobiology
Introduction: The need for surrogate markers to detect endoscopic healing in inflammatory bowel disease (IBD) is imminent. Previously, neutrophil gelatinase B-associated lipocalin and matrix metalloproteinase-9 (NGAL-MMP-9) complex was found to be superior to CRP for detection of endoscopic healing with infliximab (IFX) in patients with ulcerative colitis (UC) (de Bruyn M, Inflamm Bowel Dis. 2014). The combination of NGAL-MMP-9 with CRP and neutrophils increased the sensitivity and specificity. Cathelicidin LL-37 is an antimicrobial protein found in lysosomes of neutrophils and plays a role in innate immune defense. Chitinase 3-like 1 (CHI3L1) is secreted by neutrophils and is a growth factor for vascular endothelial cells and fibroblasts. Both markers were previously associated with IBD (Koon HW, Gastroenterology 2011 and Buisson A, Aliment Pharmacol Ther 2016).
Aim: To study if LL-37 and/or CHI3L1 could improve detection of endoscopic healing with IFX in UC patients.
Methods: Serum samples were obtained from 145 UC patients (41% female, median [interquartile range, IQR] age 41.3 [30.8-51.9] years) who underwent endoscopy following IFX initiation and from 75 healthy individuals who served as controls (56% female, median [IQR] age 33.6 [29.2-51.8] years). Endoscopic healing with IFX was defined as a Mayo endoscopic subscore of 0 or 1. CRP, NGAL-MMP-9 and neutrophils were previously determined, and LL-37 and CHI3L1 were measured with ELISA (Hycult Biotech and R&D systems, respectively). For all markers, optimal cut-offs were determined with ROC analysis and binary variables were entered in a logistic regression model to generate the Ulcerative Colitis Response Index (UCRI). Non-parametric statistical tests were performed and p-values <0.05 were considered significant.
Results: Median (IQR) time to serum sampling after start of IFX was 8.2 (6.0-14.0) weeks. 83 patients (57%) had endoscopic healing, whereas 62 patients (43%) did not have endoscopic healing. Median [IQR] LL-37 levels (ng/ml) were significantly lower in healers (24.3 [16.1-41.4]) compared to non-healers (37.3 [24.0-53.8], p=0.002), but remained elevated compared to controls (16.7 [10.2-27.1]; p<0.001). Median [IQR] CHI3L1 levels (ng/ml) were significantly lower in healers (20.9 [14.3-34.4]) compared to both non-healers (30.0 [22.7-53.9], p<0.001) and controls (31.9 [19.6-48.6], p=0.003). UCRI consisted of a combination of CRP (Odds ratio [95% confidence interval] 3.3 [1.4-7.5]), CHI3L1 (3.1 [1.3-7.7]), neutrophils (4.9 [2.1-11.2]) and LL-37 (2.5 [1.0-6.4]). The area under the curve (AUC) of UCRI was 0.83 and quartile (Q)1 (0.0-2.6) was able to discriminate healing with 54% sensitivity, 92% specificity, 60% negative predictive value (NPV) and 90% positive predictive value (PPV), whereas Q4 (7.2-9.8) was able to discriminate non-healing with 37% sensitivity, 95% specificity, 67% NPV and 85% PPV. Finally, UCRI could detect endoscopic healing as early as 3 weeks after IFX initiation (Hazard ratio [95% CI] 4.1 [2.6-6.5]).
Conclusions: In the search for surrogate markers of endoscopic healing, UCRI (CRP, CHI3L1, neutrophils and LL-37) was shown to accurately identify UC patients who fail to achieve healing with IFX and may help in early decision making to switch treatment.
S. VAN HEES (1), P. MICHIELSEN (1), L. VONGHIA (1), S. FRANCQUE (1), T. VANWOLLEGHEM (1) / [1] Antwerp University Hospital, Edegem, Belgium, Department of Gastroenterology and Hepatology
Introduction: With the recent introduction of direct acting antivirals (DAA), eradication of the Hepatitis C Virus (HCV) has become possible in almost all patients. The effect of DAA’s on liver-related manifestations of HCV has been extensively studied, but their effect on extrahepatic manifestations remains unknown.
Aim: We studied the effect of DAA on extrahepatic manifestations of the Hepatitis C Virus.
Methods: This is a retrospective analysis of all DAA treated patients from the Antwerp University Hospital showing extrahepatic manifestations of HCV. Patients were included if they showed extrahepatic manifestations at the time of DAA treatment start. Wilcoxon-rank and chi-square tests were used to test for associations between treatment-related factors and clinical disease improvement of HCV extrahepatic manifestations.
Results: A total of 10 patients were included. They showed either non-Hodgkin B-cell lymphoma (NHL) (n=4), cryoglobulinemia/vasculitis/neuropathy (n=1), arthritis (n=2), or cryoglobuline-associated nephropathy (n=3). Patients were treated with sofosbuvir-daclatasvir±ribavirine (n=5), simeprevir-sofosbuvir (n=1), ledipasvir-sofosbuvir (n=1) or Peg-interferon-Ribavirine combined with either sofosbuvir (n=2) or simeprevir (n=1). All patients achieved an end-of-treatment response. Four patients had genotype 1a, two genotype 1b, one genotype 2, three genotype 3. Following end-of-treatment response, partial or complete clinical remission of NHL was observed, whereas no clinical improvement was observed in case of vasculitis/neuropathy or nephropathy. Cryoglobulins disappeared from the blood after HCV treatment. Signs of arthritis improved slightly in 1/2 patient. Wilcoxon signed ranks test confirmed that overall end-of-treatment response was not associated with clinical disease improvement (p=0.739). Chi-square test revealed that clinical disease improvement upon end-of-treatment response significantly depended on the type of extrahepatic manifestation (p=0.030). There was no association between HCV genotype and clinical improvement of extrahepatic manifestations after treatment stop (chi-square: p=0.267).
Conclusions: Clinical improvement of HCV-related extrahepatic manifestations after DAA treatment largely depends upon the type of manifestation. Remission was observed in case of HCV-associated NHL and cryoglobulinemia, but not in case of neuro- and nephropathy.