Histological Disease Activity Research Articles

Incremental Benefit of Achieving Endoscopic and Histologic Remission in Patients With Ulcerative Colitis: A Systematic Review and Meta-Analysis

Clinical remission, defined by a composite of patient reported outcomes and Mayo endoscopy subscore (MES) 0 or 1 is a recommended treatment target in patients with ulcerative colitis (UC). We estimated whether incorporating more rigorous remission definitions, of endoscopic remission (MES 0) and histologic remission, affects risk of relapse. Through a systematic review, we identified cohort studies in adults with UC in clinical remission that reported a minimum 12-month risk of clinical relapse, based on MES (0 vs 1) and/or histologic disease activity, in patients with endoscopic remission. Using random effects meta-analysis, we calculated relative and absolute risk of clinical relapse in patients with UC achieving different treatment targets. In a meta-analysis of 17 studies that included 2608 patients with UC in clinical remission, compared to patients achieving MES 1, patients achieving MES 0 had a 52% lower risk of clinical relapse (relative risk, 0.48; 95% CI, 0.37-0.62). The median 12-month risk of clinical relapse in patients with MES 1 was 28.7%; the estimated annual risk of clinical relapse in patients with MES 0 was 13.7% (95% CI, 10.6-17.9). In a meta-analysis of 10 studies in patients in endoscopic remission (MES 0), patients who achieved histologic remission had a 63% lower risk of clinical relapse vs patients with persistent histologic activity (relative risk, 0.37; 95% CI, 0.24-0.56). Estimated annual risk of clinical relapse in who achieved achieving histologic remission was 5.0% (95% CI, 3.3-7.7). In a systematic review and meta-analysis of patients with UC in clinical remission, we observed that patients achieving more rigorous treatment endpoints (endoscopic and histologic remission) have a substantially lower risk of clinical relapse compared with patients achieving clinical remission.

Incretin combination therapy for the treatment of non-alcoholic steatohepatitis.

To test specific mono-agonists to the glucagon-like peptide-1 receptor (GLP-1R), glucagon receptor (GCGR) and glucose-dependent insulinotropic peptide receptor (GIPR), individually and in combination, in a mouse model of diet-induced non-alcoholic steatohepatitis (NASH) and fibrosis in order to decipher the contribution of their activities and potential additive effects to improving systemic and hepatic metabolism. We induced NASH by pre-feeding C57BL/6J mice a diet rich in fat, fructose and cholesterol for 36 weeks. This was followed by 8 weeks of treatment with the receptor-specific agonists 1-GCG (20 μg/kg twice daily), 2-GLP1 (3 μg/kg twice daily) or 3-GIP (30 μg/kg twice daily), or the dual (1 + 2) or triple (1 + 2 + 3) combinations thereof. A dual GLP-1R/GCGR agonistic peptide, 4-dual-GLP1/GCGR (30 μg/kg twice daily), and liraglutide (100 μg/kg twice daily) were included as references. Whereas low-dose 1-GCG or 3-GIP alone did not influence body weight, liver lipids and histology, their combination with 2-GLP1 provided additional weight loss, reduction in liver triglycerides and improvement in histological disease activity score. Notably, 4-dual-GLP-1R/GCGR and the triple combination of selective mono-agonists led to a significantly stronger reduction in the histological non-alcoholic fatty liver disease activity score compared to high-dose liraglutide, at the same extent of body weight loss. GCGR and GIPR agonism provide additional, body weight-independent improvements on top of GLP-1R agonism in a murine model of manifest NASH with fibrosis.

Correlation of Patient-Reported Outcome (PRO-2) with Endoscopic and Histological Features in Ulcerative Colitis and Crohn's Disease Patients.

Methods A cross-sectional study was conducted in consecutive newly diagnosed patients with inflammatory bowel disease in a tertiary care referral center. The initial evaluation included patient-reported outcome for stool frequency subscore and rectal bleeding. Endoscopic activity was determined using the Mayo scoring system for ulcerative colitis and the Simple Endoscopic Score for Crohn's disease. Histopathological activity was assessed using a validated numeric scoring system. Results We included 159 patients (63 Crohn's disease with colonic involvement and 96 with ulcerative colitis). We found significant correlation between the Mayo endoscopic subscoring system and histology activity in ulcerative colitis, while no correlation was found in patients with Crohn's disease. Patient-reported outcome showed inverse correlation with endoscopic and histological activity in Crohn's disease (rs = −0.67; rs = −0.72), while positive correlation was found in ulcerative colitis (rs = 0.84; rs = 0.75). Interpretation and Conclusions. Patient-reported outcome is a practical and noninvasive tool for assessment of disease activity in ulcerative colitis patients but not in Crohn's disease.

High Protein Intake Is Associated With Histological Disease Activity in Patients With NAFLD.

Overconsumption of carbohydrates and lipids are well known to cause nonalcoholic fatty liver disease (NAFLD), while the role of nutritional protein intake is less clear. In Western diet, meat and other animal products are the main protein source, with varying concentrations of specific amino acids. Whether the amount or composition of protein intake is associated with a higher risk for disease severity has not yet been examined. In this study, we investigated associations of dietary components with histological disease activity by analyzing detailed 14‐day food records in a cohort of 61 patients with biopsy‐proven NAFLD. Furthermore, we used 16S ribosomal RNA gene sequencing to detect associations with different abundances of the gut microbiota with dietary patterns. Patients with definite nonalcoholic steatohepatitis (NAFLD activity score of 5‐8 on liver biopsy) had a significantly higher daily relative intake of protein compared with patients with a NAFLD activity score of 0‐4 (18.0% vs. 15.8% of daily protein‐based calories, P = 0.018). After adjustment for several potentially confounding factors, a higher protein intake (≥17.3% of daily protein‐based calories) remained associated with definite nonalcoholic steatohepatitis, with an odds ratio of 5.09 (95% confidence interval 1.22‐21.25, P = 0.026). This association was driven primarily by serine, glycine, arginine, proline, phenylalanine, and methionine. A higher protein intake correlated with a lower Bacteroides abundance and an altered abundance of several other bacterial taxa. Conclusion: A high protein intake was independently associated with more active and severe histological disease activity in patients with NAFLD. Further studies are needed to investigate the potential harmful role of dietary amino acids on NAFLD, with special attention to meat as their major source.

Endoscopic and Histological Assessment, Correlation, and Relapse in Clinically Quiescent Ulcerative Colitis (MARQUEE).

It is difficult to predict relapse in quiescent ulcerative colitis (UC), but newer endoscopic and histological indices could improve this. This study aimed to determine in UC patients in clinical remission (1) the prevalence of active endoscopic and histological disease; (2) the correlation between endoscopic and histological scores; and (3) the predictive power of these scores for clinical relapse. This multicenter prospective cohort study conducted by the Crohn's and Colitis Foundation Clinical Research Alliance included 100 adults with UC in clinical remission undergoing surveillance colonoscopy for dysplasia. Endoscopic activity was assessed using the Mayo endoscopic score (MES), ulcerative colitis endoscopic index of severity (UCEIS), and ulcerative colitis colonoscopic index of severity (UCCIS). Histology was assessed with the Riley index subcomponents, total Riley score, and basal plasmacytosis. Only 5% of patients had an MES of 0, whereas 38% had a score of 2 to 3; using the UCEIS, the majority of patients had at least mild activity, and 15% had more severe activity. Many patients also had evidence of histological disease activity. The correlations among endoscopic indices, histological subcomponents, and total score were low; the highest correlations occurred with the subcomponent architectural irregularity (ρ = 0.43-0.44), total Riley score (ρ = 0.35-0.37), and basal plasmacytosis (ρ = 0.35-0.36). Nineteen patients relapsed clinically over 1 year, with the subcomponent architectural irregularity being the most predictive factor (P = 0.0076). This multicenter prospective study found a high prevalence of both endoscopic and histological disease activity in clinically quiescent UC. The correlations between endoscopy and histology were low, and the power to predict clinical relapse was moderate.

P283 Patient-reported symptoms over a period of 14 days reliably predict endoscopic and histological disease activity in ulcerative colitis (UC)

Abstract Background Disease activity assessment in UC is most accurately evaluated by endoscopy and biopsy, which often correlates poorly with current symptoms. We investigated whether a machine learning classification algorithm using patient-reported Simple Clinical Colitis Activity Index (SCCAI) over 14 days and signature features could reliably distinguish endoscopic and histopathologic activity from remission. Methods The TrueColours ulcerative colitis (UC) monitoring platform was used to collect symptoms (SCCAI, daily) and endoscopic/histopathological activity, as per the UCEIS and Nancy indices (twice in 6 months) in 233 patients. The longitudinal data may be seen as a trajectory. The signature is a collection of statistics which efficiently summarises the trajectory and serves as a non-parametric hierarchical method for longitudinal data representation. Signature features were used as input to the extreme gradient boosting classification algorithm to categorise subjects into remission or active disease groups. Remission: UCEIS ≤1 AND Nancy ≤1; Active disease: UCEIS ≥4 AND Nancy ≥2. The advanced signature-based approach was compared with a baseline model with manually constructed features (mean SCCAI score over 14 days preceding endoscopy). Cross-validation was used to report metrics, Table 1. Results Disease activity classification with signature features significantly outperformed mean-SCCAI based approach (AUC 0.87 vs. AUC 0.79 correspondingly, p = 0.011, Figure 1). All other statistical metrics (including sensitivity (0.89), specificity (0.78), PPV (0.82) and NPV) demonstrated the same trend, favouring the signature method. The decision-tree-based ensemble gradient boosting classification algorithm (XGBoost) has the intrinsic ability to rank feature importance. The signature feature of the fourth item within the SCCAI (blood in stool) dominated, followed by cross-correlation between the first (daily bowel frequency) and fourth item. Conclusion Gradient boosting classification algorithm with signature features extracted from the longitudinal six-item symptoms of the SCCAI, reliably identified subjects with endoscopic and histopathological disease activity. The developed algorithm may be used with the TrueColours platform as a decision support tool.

P409 Endoscopic and histological activity is associated with non-response to biologic therapy in ulcerative colitis

Abstract Background Ulcerative colitis (UC) is a chronic inflammatory bowel disease, characterised by a relapsing and remitting course. Biologic therapy in UC has a 30% rate of primary non-response (PNR) and a further 5% loss of response per year. Prognostic indicators of non-response include younger age and longer disease duration. Emerging evidence suggests that endoscopic and histological activity is associated with adverse outcomes such as clinical relapse and colectomy. However, the role of endoscopic or histological disease activity in predicting non-response to biologic therapy is unclear. We aim to investigate the association of endoscopic and histological activity with non-response to biologic therapy in UC. Methods Patients with UC treated with biologic therapy between 2014 and 2019 were studied. Baseline endoscopic disease activity was assessed using the Mayo endoscopic subscore from endoscopy performed prior to commencing biologic therapy. Patients who underwent colonoscopy 3–9 months after commencing biologic therapy were selected for histopathological review. Sigmoid biopsies were evaluated by an expert gastrointestinal pathologist. Histological parameters were scored on a 4-point scale, with blinding to clinical and endoscopic outcomes. Results Sixty-six patients were followed for a median of 16 months (IQR 6–30), 12 (18%) of whom did not respond to biologic therapy initially and 3 (6%) of whom lost response secondarily. Baseline endoscopic activity was associated with non-response (OR 6.1, 95% confidence interval 1.3 – 29.4, for every 1-point increase in the 4-point Mayo endoscopic subscore). 23 patients were identified for histopathological review. Four (17%) patients exhibited PNR. Baseline crypt atrophy was associated with PNR (p = 0.014) and baseline bifid crypts and crypt atrophy were associated with 52-week clinical relapse (p = 0.482 and p = 0.0497). Conclusion For patients with UC, endoscopic disease activity assessed prior to commencing biologic therapy has the potential to predict non-response to biologic therapy. In addition, specific histological parameters relating to crypt inflammation and crypt architecture may also predict non-response. Further prospective studies should be conducted to investigate the clinical utility of assessing endoscopic and histological disease activity prior to commencing biologic therapy.

P418 Filgotinib decreases molecular markers of JAK1 signal transduction in Crohn’s disease: concordance with endoscopy and histopathology

Abstract Background Filgotinib (FIL) is a JAK1 inhibitor under phase 3 clinical evaluation for treatment of IBD. We conducted a post hoc analysis in a subset of patients with moderately to severely active CD from FITZROY (NCT02048618) to assess the effect of FIL (200 mg QD) on molecular markers of JAK1-related signalling (STAT1 and STAT3 phosphorylation) within epithelium (Ep) and non-Ep regions of intestinal mucosa and explore their correlation to histologic and endoscopic indices. Methods Biopsies were collected at baseline (BL) and week 10 (W10) from the most affected area of each predefined bowel segment (ileum, ascending, transverse, descending colon, and rectum). Within-subject matched biopsies for all segments from FIL (n = 42) and placebo (PBO; n = 18) treated patients were scored for histologic (GHAS) and endoscopic (SES-CD) disease activity totalling to 300 segments. Using specific antibodies and IHC, %pSTAT1 and %pSTAT3 positive nuclei within Ep and non-Ep regions of each biopsy were quantified using machine learning (Visiopharm v.2019.06). Basal pSTAT levels assessed from 182 nondiseased (SES-CD = 0 and GHAS = 0) segments were used to determine a threshold for categorising segments as either low or high molecular disease activity (MDA). Agreement between endoscopy or histology and MDA was evaluated by κ and % agreement by segment and all segments combined. Results Median basal pSTAT1 was similar between colonic Ep and non-Ep regions (1%–2%), but higher in ileal Ep (5%). Median basal pSTAT3 was higher in non-Ep (3%–5%) vs. Ep (1%–2%) regions across all segments. At BL, MDA was elevated in segments with ulceration (~10%, pSTAT1 and pSTAT3). In segments with GHAS activity subscore ≥2 at BL, both Ep MDA (10%–30%) and non-Ep MDA were elevated (25%–35%) and correlated to histologic activity. In segments with low BL MDA, significantly fewer segments with FIL showed MDA worsening at W10 compared with PBO (both pSTAT1 and pSTAT3). In segments with high BL MDA, FIL treatment improved significantly more segments than PBO; this was evident for pSTAT3 only (table). Concordance between MDA and endoscopy was mostly fair to moderate (κ 0.3–0.5), whereas MDA and histology showed moderate to good concordance (κ 0.4–0.8). Conclusion Filgotinib treatment improved JAK1-related MDA within the mucosa of CD patients. Concordance between MDA and clinical indices was highest with histology.

OP31 Meta–omics reveals microbiome-driven proteolysis as a contributing factor to the severity of ulcerative colitis disease activity

Abstract Background Ulcerative colitis (UC) is characterised by an aberrant immune response directed towards the gut microbiota. It remains poorly understood which mechanisms govern pathogenic roles of the microbiome in UC. Methods A prospective cohort of 40 UC patients were phenotyped with clinical (partial Mayo), biochemical (faecal calprotectin), endoscopic (Mayo endoscopic sub-score and ulcerative colitis Endoscopic Index of Severity) and histologic (Geboes) disease activity indices. Individual patient-matched faecal samples collected within 24 h prior to the lower endoscopy underwent 16S, shotgut metagenomic, metabolomic, metaproteomic, and metapeptidomic sequencing in addition to proteomic sequencing of paired serum samples. Sequencing datasets were integrated and analysed for microbial determinants of UC disease severity. Confirmatory in-vivo and in-vitro experiments were performed in Caco-2 epithelial monolayers and IL10 knockout mice. Results All meta-omics displayed large-scale shifts related to disease severity, with the metabolome and metaproteome best predicting disease severity and the metapeptidome demonstrating an increase of peptide fragments in UC patients with high severity. Broad-scale analyses and taxonomic inferences identified Bacteroides proteases as a distinguishing feature of disease severity, with the Bacteroides association being driven by changes in protease protein expression. Using a Caco-2 model we confirmed that epithelial disruption by Bacteroides vulgatus was prevented by protease inhibition, without influencing growth rates. Guided by the metagenome and metaproteome we transplanted a high-severity UC faecal sample with an increased abundance of proteases related to Bacteroides vulgatus into an IL10−/− gnotobiotic mouse model, and observed that colitis induced by transplant of faeces was attenuated by oral administration of protease inhibitors. Conclusion Through an integrated multi-omics data analysis we demonstrate that certain members of the microbiome, such as Bacteroides vulgatus, may contribute to exacerbating disease activity in UC through protease activity. In-vivo and in-vitro experiments provide evidence that bacterial protease inhibition may be a novel therapeutic approach in UC.

P688 Effect of golimumab induction therapy on histologic disease profile in patients with moderate to severe ulcerative colitis: Results from the randomised, placebo-controlled Phase 3 PURSUIT-SC trial

Abstract Background Histologic improvement is an emerging goal in ulcerative colitis (UC) management. We previously demonstrated in the 6-week, multicentre, randomised, placebo-controlled, double-blind Program of Ulcerative colitis Research Studies Utilising an Investigational Treatment [PURSUIT-SC] Phase 3 trial that SC induction therapy with the tumour necrosis factor-α inhibitor, golimumab (GLM), led to statistically significant and clinically meaningful improvements in the signs and symptoms of UC compared with placebo, including clinical remission, mucosal healing, and improved health-related QoL, in patients with moderate to severe active UC.1 In the present report, we explored the histologic effects of GLM induction therapy in PURSUIT-SC by examining data from a prespecified histology sub-study. Methods The Phase 3 PURSUIT-SC trial evaluated the safety and efficacy of induction therapy with GLM in 774 patients with moderate to severe UC disease activity (defined as Mayo scores of 6–12, including an endoscopic subscore of ≥ 2). Patients were randomised 1:1:1 to receive SC induction treatment with placebo at Weeks 0 and 2, GLM 200 mg at Week 0 followed by 100 mg at Week 2 (GLM 200–>100 mg), or GLM 400 mg at Week 0 followed by 200 mg at Week 2 (GLM 400–>200 mg). Rectal biopsies were collected during endoscopy at screening and Week 6 in those randomised patients who consented to participate in the histology sub-study (N=98). During the screening endoscopy, two adjacent biopsies were collected 15 to 20 cm beyond the anal verge in an area that was most representative of the inflammation observed. At Week 6, two additional biopsies were to be obtained from the same location biopsied at screening. Biopsies were graded using the Geboes Score (GS) by a single expert gastrointestinal histopathologist, who was blinded to subject, treatment and study visit. The GS comprises seven histologic grades assessing structural changes and inflammatory disease activity. Higher GS grade scores indicate more severe histologic disease. Results Evaluable biopsy data were available in 89 patients (placebo, n = 30; GLM 200–>100 mg, n = 30; GLM 400–>200 mg, n = 29). At Week 6, compared with placebo, treatment with GLM induction therapy led to a decrease in the proportion of biopsies with GS grade scores of > 1 and an increase in the proportion of biopsies with GS grade scores of ≤ 1, with the greatest improvements being observed in the GLM 400–>200 mg group (figure). Conclusion The findings of this exploratory histologic sub-study in the PURSUIT-SC trial suggest that GLM induction therapy led to an improvement in histologic disease activity compared with placebo in patients with moderate-to-severe UC. Reference

P497 Is mucosal healing sufficient paramater for anti-TNF discontinuation?

Abstract Background Long-term treatment with biologic therapy alongside with immunomodulators in patients with inflammatory bowel disease (IBD) can be associated with severe side effects. Nowadays, there is not a clear recommendation for de-escalation of therapy in patients on biologic treatment. The aim of the study was to evaluate relapse rate in 4 ili 2 years follow-up in patients with mucosal healing after two years of anti-TNF treatment, regardless achieved histological remission. Methods 52 patients with IBD after two years treatment with anti-TNF agents (Infliksimab (IFX) and Adalimumab (ADA)) and achieved mucosal healing, were enrolled in this retrospective study. Mayo endoscopic sub-score (0,1) and SES (<2) was used to define mucosal healing in UC and CD patients, respectively. For the assessment of histological activity Nancy and Global Histologic Disease Activity Score (GHAS) were used. Histologic remission was defined by Nancy’s score ≤ 1 and GHAS ≤ 4. Data were collected for 4 years follow-up period after discontinuation of anti-TNF agents. Statistical analysis was carried out using SPSS 20.0 (Chicago, IL). Results From 52 patients 40.38% (21/52) patients had UC, and 59.61 % (31/52) had CD with an average disease duration of 11.57 (SD ± 6.28) years. Gender distribution was relatively equal, 46% men (24/52), with an average age of 40.66 (SD ± 11.15) years. All of the patients were on anti-TNF therapy and had mucosal healing at the moment of discontinuation, 55% (29/52) were on IFX, 35% (23/52) were on ADA, respectively. 48.07% (25/52), were with pathohistological remission of disease. In 4 years follow-up after anti-TNF therapy discontinuation, 59.61% (31/52) relapsed, with an estimated period of relapse time of 15.4 (SD ± 9.83) months. Using Spearman`s correlation moderate relation was revealed between histological activity and relapse (rho 0.467, p < 0.001). Probability of relapse within 4 years in this group of patients with histologically active disease was higher (p < 0.01, OR 2.72, 95% Cl 1.24–5.92). Conclusion Achieving both, endoscopic and histological remission, may be a good parameter in making a decision when to de-escalate anti-TNF therapy.

Novel Inflammatory-Nutritional Biomarkers as Predictors of Histological Activity in Crohn's Disease.

There has increasingly been an interest in histological remission as a therapeutic endpoint in inflammatory bowel disease. The aim of this study was to evaluate the utility of a variety of inflammatory - nutritional markers for predicting histological disease activity in patients diagnosed with Crohn's disease. Patients with Crohn's disease that had requisite endoscopic, pathological, and laboratory data were retrospectively enrolled in the study. Relevant clinical, laboratory, endoscopic, and pathological data were abstracted. The neutrophil:lymphocyte ratio (NLR), lymphocyte:monocyte ratio (LMR), platelet:lymphocyte ratio (PLR), red blood cell distribution width (RDW), modified Glasgow Prognostic score (mGPS), Prognostic Nutritional Index (PNI), Geriatric Nutritional Risk index (GNRI), CRP/Albumin ratio (CAR), Iron:Ferritin ratio (Fe:F) and the Systemic immune inflammation index (SII) were calculated. The cohort was stratified by presence of histological disease on colonoscopy, and groups were compared with appropriate statistical methods. When comparing patients without histological disease to those with disease, there was a statistically significant difference in CAR (2.9 ± 1.5 vs. 4.2 ± 2, p = 0.001), RDW (13.4 ± 1.3 vs. 14.5 ± 1.8, p = 0.008), PNI (52.4 ± 6.2 vs. 47.4 ± 9.3, p = 0.03), and mGPS (0.2 ± 0.4 vs. 0.6 ± 0.7, p = 0.01). For predicting histological activity, ROC analyses indicated an optimal cutoff of 0.3 for CAR (AUC 0.8, PPV 90.5%), 13.5 for RDW (AUC 0.7, PPV 84.1), 86.1 for PNI (AUC 0.7, PPV 86.1) and > 0 for mGPS (AUC 0.6, PPV 85.2%). The NLR, LMR, PLR, GNRI, Fe: F, and SII did not meet statistical significance (p = 0.4, 0.08, 0.2, 0.5, 0.6, and 0.3, respectively). We report on ten biomarkers, many of them never studied in Crohn's disease, which can help in predicting the presence of active histological disease. Larger prospective studies are needed to investigate the utility of these biomarkers alone and in combination.

Leukocyte Apheresis Using a Fiber Filter Suppresses Colonic Injury Through Calcitonin Gene-Related Peptide Induction.

The aim of this study was to address whether the therapeutic effect of leukocytapheresis (LCAP) depends on calcitonin gene- related peptide (CGRP) induction. An HLA-B27 transgenic rat model was treated with an LCAP column. The effects of LCAP on clinical, endoscopic, and histologic disease activity, the colony-forming ability of colony-forming unit (CFU)-granulocyte macrophages (GMs), colonic blood flow, and tissue expression of tumor necrosis factor (TNF)-α and CGRP were examined. Changes in the effects of LCAP after pretreatment with the CGRP antagonist CGRP8-37 were also observed. A dextran sulfate sodium-induced colitis rat model included treatment with CGRP, and the effect was assessed based on clinical, endoscopic, and histologic disease activity, colonic blood flow, the colony-forming ability of CFU-GMs, and tissue expression of inflammatory cytokines and CGRP receptor families. LCAP improved disease activity, enhanced colonic blood flow, and induced the bone marrow colony-forming ability of CFU-GMs with an increase in CGRP mRNA levels. These effects were abolished by pretreatment with CGRP8-37. The administration of CGRP suppressed colitis, promoting colonic blood flow, inducing bone marrow-derived cells, downregulating inflammatory cytokines, and upregulating receptor activity-modifying protein-1. The mRNA and protein levels of inflammatory cytokines in lipopolysaccharide-stimulated mononuclear cells were also decreased after CGRP treatment. The therapeutic effects of LCAP depend on CGRP induction. CGRP can effectively suppress colitis through the downregulation of inflammatory events and upregulation of protective events.

The SGLT2 Inhibitor Canagliflozin Prevents Carcinogenesis in a Mouse Model of Diabetes and Non-Alcoholic Steatohepatitis-Related Hepatocarcinogenesis: Association with SGLT2 Expression in Hepatocellular Carcinoma.

The aim of the present study is to investigate the effects of canagliflozin, a selective sodium-glucose co-transporter 2 (SGLT2) inhibitor, on non-alcoholic steatohepatitis (NASH) and NASH-related hepatocellular carcinoma (HCC) in a mouse model of diabetes and NASH-HCC. First, mice aged five weeks were divided into two groups (vehicle group and canagliflozin group) and were treated for three weeks. Then, mice aged five weeks were divided into three groups of nine animals each: the vehicle group, early canagliflozin group (treated from five to nine weeks), and continuous canagliflozin group (treated from five to 16 weeks). Canagliflozin was administered at a dose of 30 mg/kg in these experiments. In addition, the in vitro effects of canagliflozin were investigated using HepG2 cells, a human HCC cell line. At the age of eight or 16 weeks, the histological non-alcoholic fatty liver disease activity score was lower in the canagliflozin-treated mice than in vehicle-treated mice. There were significantly fewer hepatic tumors in the continuous canagliflozin group than in the vehicle group. Immunohistochemistry showed significantly fewer glutamine synthetase-positive nodules in the continuous canagliflozin group than in the vehicle group. Expression of α-fetoprotein mRNA, a marker of HCC, was downregulated in the continuous canagliflozin group when compared with the vehicle group. At 16 weeks, there was diffuse SGLT1 expression in the hepatic lobules and strong expression by hepatocytes in the vehicle group, while SGLT2 expression was stronger in liver tumors than in the lobules. In the in vitro study, canagliflozin (10 μM) suppressed the proliferation of HepG2 cells. Flow cytometry showed that canagliflozin reduced the percentage of HepG2 cells in the G2/M phase due to arrest in the G1 phase along with decreased expression of cyclin D and Cdk4 proteins, while it increased the percentage of cells in the G0/1 phase. Canagliflozin also induced apoptosis of HepG2 cells via activation of caspase 3. In this mouse model of diabetes and NASH/HCC, canagliflozin showed anti-steatotic and anti-inflammatory effects that attenuated the development of NASH and prevented the progression of NASH to HCC, partly due to the induction of cell cycle arrest and/or apoptosis as well as the reduction of tumor growth through the direct inhibition of SGLT2 in tumor cells.

Association of Histologic Disease Activity With Progression of Nonalcoholic Fatty Liver Disease

The histologic evolution of the full spectrum of nonalcoholic fatty liver disease (NAFLD) and factors associated with progression or regression remain to be definitively established. To evaluate the histologic evolution of NAFLD and the factors associated with changes in disease severity over time. A prospective cohort substudy from the Nonalcoholic Steatohepatitis Clinical Research Network (NASH CRN) NAFLD Database study, a noninterventional registry, was performed at 8 university medical research centers. Masked assessment of liver histologic specimens was performed, using a prespecified protocol to score individual biopsies. Participants included 446 adults with NAFLD enrolled in the NASH CRN Database studies between October 27, 2004, and September 13, 2013, who underwent 2 liver biopsies 1 or more year apart. Data analysis was performed from October 2016 to October 2018. Progression and regression of fibrosis stage, using clinical, laboratory, and histologic findings, including the NAFLD activity score (NAS) (sum of scores for steatosis, lobular inflammation, and ballooning; range, 0-8, with 8 indicating more severe disease). A total of 446 adults (mean [SD] age, 47 [11] years; 294 [65.9%] women) with NAFLD (NAFL, 86 [19.3%]), borderline NASH (84 [18.8%]), and definite NASH (276 [61.9%]) were studied. Over a mean (SD) interval of 4.9 (2.8) years between biopsies, NAFL resolved in 11 patients (12.8%) and progressed to steatohepatitis in 36 patients (41.9%). Steatohepatitis resolved in 24 (28.6%) of the patients with borderline NASH and 61 (22.1%) of those with definite NASH. Fibrosis progression or regression by at least 1 stage occurred in 132 (30%) and 151 [34%] participants, respectively. Metabolic syndrome (20 [95%] vs 108 [72%]; P = .03), baseline NAS (mean [SD], 5.0 [1.4] vs 4.3 [1.6]; P = .005), and smaller reduction in NAS (-0.2 [2] vs -0.9 [2]; P < .001) were associated with progression to advanced (stage 3-4) fibrosis vs those without progression to stage 3 to 4 fibrosis. Fibrosis regression was associated with lower baseline insulin level (20 vs 33 μU/mL; P = .02) and decrease in all NAS components (steatosis grade -0.8 [0.1] vs -0.3 [0.9]; P < .001; lobular inflammation -0.5 [0.8] vs -0.2 [0.9]; P < .001; ballooning -0.7 [1.1] vs -0.1 [0.9]; P < .001). Only baseline aspartate aminotransferase (AST) levels were associated with fibrosis regression vs no change and progression vs no change on multivariable regression: baseline AST (regression: conditional odds ratio [cOR], 0.6 per 10 U/L AST; 95% CI, 0.4-0.7; P < .001; progression: cOR, 1.3; 95% CI, 1.1-1.5; P = .002). Changes in the AST level, alanine aminotransferase (ALT) level, and NAS were also associated with fibrosis regression and progression (ΔAST level: regression, cOR, 0.9; 95% CI, 0.6-1.2; P = .47; progression, cOR, 1.3; 95% CI, 1.0-1.6; P = .02; ΔALT level: regression, cOR, 0.7 per 10 U/L AST; 95% CI, 0.5-0.9; P = .002; progression, cOR, 1.0 per 10 U/L AST; 95% CI, 0.9-1.2; P = .93; ΔNAS: regression, cOR, 0.7; 95% CI, 0.6-0.9; P = .001; progression, cOR, 1.3; 95% CI, 1.1-1.5; P = .01). Improvement or worsening of disease activity may be associated with fibrosis regression or progression, respectively, in NAFLD.

757 Histologic Improvement With Vedolizumab vs. Adalimumab in Ulcerative Colitis: Results From VARSITY

INTRODUCTION: Histologic remission is associated with superior long-term clinical outcomes in ulcerative colitis (UC). VARSITY, the first head-to-head comparison of biologic agents in UC (NCT02497469; EudraCT 2015-000939-33), showed superior clinical remission and endoscopic improvement at Week 52 with vedolizumab (VDZ), a gut-selective, humanized, α4β7 integrin monoclonal antibody, vs adalimumab (ADA), a systemic, human, anti-tumor necrosis factor (anti-TNF) monoclonal antibody. 1 Both VDZ and ADA were generally safe and well-tolerated. 1 This analysis compared histologic improvements with VDZ vs ADA in VARSITY. METHODS: Patients with moderately to severely active UC were randomized 1:1 to active VDZ intravenous (IV) infusions (300 mg)/placebo subcutaneous (SC) injections or placebo IV/active ADA SC (160/80/40 mg). Prespecified histologic exploratory endpoints included histologic remission (Geboes score &lt; 2 or Robarts Histopathology Index [RHI] score &lt; 3) and minimal histologic disease activity (Geboes score &lt; 3.2 or RHI score &lt; 5) at Week 14 and Week 52. Histologic remission was also assessed based on previous anti-TNF use. RESULTS: A total of 769 patients received ≥1 dose of VDZ (n = 383) or ADA (n = 386). Median (range) duration of exposure was 477 (127, 630) days for VDZ and 420 (71, 454) days for ADA. Mean (standard deviation) baseline histologic disease activity was similar between groups (Geboes: VDZ, 15.0 [4.92]; ADA, 15.1 [5.03]; RHI: VDZ, 19.5 [8.74]; ADA, 19.6 [8.89]). Histologic remission induced by VDZ at Week 52 was greater than with ADA in the overall (Geboes or RHI), anti-TNF naïve (Geboes or RHI), and anti-TNF failure (RHI only) groups (Table 1). Histologic remission at Week 14 favored VDZ over ADA, with larger differentiation when using RHI (Table 1). VDZ also outperformed ADA in achieving minimal histologic disease activity at Weeks 14 and 52 (Table 1). CONCLUSION: VARSITY showed that use of VDZ, compared with ADA, achieved higher rates of histologic remission and minimal histologic disease activity at Weeks 14 and 52 in patients with moderately to severely active UC. These data support the use of VDZ over ADA in UC.

Urinary Angiotensinogen Predicts Renal Disease Activity in Lupus Nephritis.

Aims: A noninvasive indicator of renal histological lesions and disease activity in lupus nephritis (LN) is needed for timely and targeted treatment before overt renal injury. Here, we tested the utility of urinary angiotensinogen (UAGT) to predict renal disease activity in LN. Results: A prospective, three-stage study was performed in patients with LN. In stage I, UAGT was measured in 140 newly diagnosed LN patients. UAGT significantly increased in LN patients, correlating well with kidney angiotensinogen expression and histological activity. Patients with LN class IV exhibited the highest UAGT compared with other histopathological classes of LN. For identifying LN class IV, a particularly aggressive type of LN, UAGT outperformed the conventional clinical measures and improved their performance. In stage II, UAGT was monitored in 61 subjects from stage I for up to 12 months. UAGT decreased after induction therapy and remained low in patients with LN remission during follow-up. For predicting therapy success at month 12, the area under the receiver operating characteristics curve of UAGT reduction at month 4 was 0.83, outperforming that of 24-h proteinuria. In stage III, UAGT was monitored in 12 LN patients before, during, and after the onset of renal flares. An elevation in UAGT predicted recurrence of LN, and a decline in UAGT after a renal flare heralded the remission of disease before conventional clinical measures. Innovation and Conclusion: UAGT in LN is a promising indicator for dynamic surveillance of renal disease activity and prediction of renal flares. Antioxid. Redox Signal. 31, 1289-1301.

Endoscopic, Radiologic, and Histologic Healing With Vedolizumab in Patients With Active Crohn’s Disease

Vedolizumab is a gut-selective monoclonal antibody for the treatment of moderately to severely active Crohn's disease (CD). We performed a prospective study of endoscopic, radiologic, and histologic healing in patients with CD who received vedolizumab therapy. We performed a phase 3b, open-label, single-group study of 101 patients with at least 3 months of active CD (a CD Activity Index [CDAI] score of 220-450, a simple endoscopic score for CD [SES-CD] of 7 or more, 1 or more mucosal ulcerations [identified by endoscopy], and failure of conventional therapy) from March 2015 through December 2017. Among the patients enrolled, 54.5% had previous failure of 1 or more tumor necrosis factor (TNF) antagonists and 44.6% had severe endoscopic disease activity (SES-CD scores above 15) at baseline. Participants received vedolizumab (300 mg intravenously) at weeks 0, 2, and 6, and then every 8 weeks thereafter, for 26 weeks (primary study) or 52 weeks (substudy, 56 patients). The primary endpoint at week 26 was endoscopic remission (SES-CD score of 4 or less); other endpoints included endoscopic response (50% reduction in SES-CD), radiologic remission (magnetic resonance index of activity score below 7), and histologic response (modified global histologic disease activity score of 4 or less). At week 26, 11.9% of patients were in endoscopic remission (95% confidence interval [CI] 6.3-9.8); at week 52, 17.9% of the patients were in endoscopic remission (95% CI 8.9-30.4). Higher proportions of patients naïve to TNF antagonists achieved endoscopic remission than patients with TNF-antagonist-failure at weeks 26 and 52. Higher proportion of patients with moderate CD (SES-CD scores, 7-15) achieved endoscopic remission at weeks 26 and 52 than patients with severe CD (SES-CD scores above 15). The proportion of patients with complete mucosal healing increased over time, with greater rates of healing in the colon than in the ileum. Remission was detected by magnetic resonance enterography in 21.9% of patients at week 26 (95% CI 9.3-40.0) and in 38.1% at week 52 (95% CI 18.1-61.6). At week 26, 24.4% of patients had a histologic response in the colon (95% CI 15.3-35.4) and 28.3% of patients had a histologic response in theileum (95% CI 17.5-41.4). At week 52, 20.5% of patients had a histologic response in the colon (95% CI 9.8-35.3) and 34.3% of patients had a histologic response in the ileum (95% CI 19.1-52.2). There were no notable safety issues, including worsening of extraintestinal manifestations. In a phase 3b trial, we found that 26 and 52 weeks of treatment with vedolizumab (300 mg, at weeks 0, 2, and 6, and then every 8 weeks thereafter) induces endoscopic, radiologic, and histologic healing in patients with moderately to severely active CD. ClinicalTrials.gov no: NCT02425111.

SUN-039 URINARY ALPHA-M SUBUNIT OF INTEGRIN MAC-1 REFLECTS HISTOLOGICAL DISEASE ACTIVITY IN LUPUS NEPHRITIS

Immunological mechanisms are indispensable for the onset and progress of glmerulonephritis; in particular, neutrophils and macrophages play a central role in acute glomerular inflammation. Their detection in kidneys from patients with LN is associated with disease activation and poor renal prognosis. Integrin Mac-1 comprising a unique α subunit (αm; CD11b) complexed with a common β2 subunit (CD18), and CD16b recognizing pathogenic IgGs are released from specific leukocyte subsets upon cell activation under inflammatory conditions including glomerulonephritis. This study aimed to investigate the association of urinary CD11b (U-CD11b) and CD16b (U-CD16b) with the histopathological activity of various glomerular diseases, particularly lupus nephritis (LN). Biological samples from 272 patients with glomerular diseases including 118 LN between 2008 and 2014 in Nagoya University (N-KDR cohort), and 44 LN patients including 15 longitudinal samples from the independent another LN population (Gunma/Saitama cohort) were subjected to the analysis. Urinary concentrations of U-CD11b, U-CD16b, hemoglobin scavenger receptor CD163 (U-CD163), and MCP-1 (U-MCP-1) were measured by ELISA. On renal LN specimens obtained from N-KDR cohort, glomerular CD11b+cells for neutrophils and monocytes/macrophages, and histological disease activity according to the biopsy activity index (BAI) were histologically evaluated. Receiver operating characteristic (ROC) curves for U-CD11b and U-CD16b to predict ISN/RPS class III/IV LN were also generated. To further corroborate the clinical findings and identify the specific timing of U-CD11b elevation after disease initiation, a murine model of antibody-mediated glomerulonephritis was induced by rabbit anti-mouse nephrotoxic serum in male C57BL/6J mice (NTS-GN). Among various forms of glomerular diseases including IgA nephropathy, minimal change nephrotic syndrome, membranous nephropathy, diabetic nephropathy, ANCA-associated vasculitis (AAV) and LN, U-CD11b and U-CD16b levels were significantly increased in LN patients from N-KDR cohort (U-CD11b/U-Cr; 36.2±50.4 ng/mg, U-CD16b/U-Cr; 2159 ± 2483 ng/mg), particularly in International Society of Nephrology/Renal Pathology Society (ISN/RPS) class III and IV. The significant U-CD11b elevations in class III and IV LN were further validated in collected samples from Gunma/Saitama cohort, and were decreased in the 15 longitudinal samples after the immunosuppressive therapy. In LN patients, U-CD11b, but not U-CD16b, was markedly correlated with the number of glomerular leukocytes and histopathological activity. Receiver operating characteristic curve analysis predicting proliferative LN revealed a superior level of U-CD11b (the area under the curve of U-CD11b/U-Cr: 0.916; cut-off: 24.0 ng/mg; sensitivity: 93.3%; specificity:84.1%) to those of CD16b, the scavenger receptor CD163, and monocyte chemotactic protein-1. In anti-mouse nephrotoxic serum glomerulonephritis, U-CD11b elevation was significantly correlated with renal damage and was reduced by corticosteroid treatment, as in human patients. Our results collectively suggest that U-CD11b is a useful biomarker for estimating the histopathological activity, particularly glomerular leukocyte accumulation, in LN.

Mucosal delivery of Lactococcus lactis carrying an anti-TNF scFv expression vector ameliorates experimental colitis in mice

BackgroundAnti-Tumor Necrosis Factor-alpha therapy has become clinically important for treating inflammatory bowel disease. However, the use of conventional immunotherapy requires a systemic exposure of patients and collateral side effects. Lactic acid bacteria have been shown to be effective as mucosal delivering system for cytokine and single domain antibodies, and it is amenable to clinical purposes. Therefore, lactic acid bacteria may function as vehicles for delivery of therapeutic antibodies molecules to the gastrointestinal tract restricting the pharmacological effect towards the gut. Here, we use the mucosal delivery of Lactococcus lactis carrying an anti-TNFα scFv expression plasmid on a DSS-induced colitis model in mice.ResultsExperimental colitis was induced with DSS administered in drinking water. L. lactis carrying the scFv expression vector was introduced by gavage. After four days of treatment, animals showed a significant improvement in histological score and disease activity index compared to those of untreated animals. Moreover, treated mice display IL-6, IL17A, IL1β, IL10 and FOXP3 mRNA levels similar to health control mice. Therefore, morphological and molecular markers suggest amelioration of the experimentally induced colitis.ConclusionThese results provide evidence for the use of this alternative system for delivering therapeutic biopharmaceuticals in loco for treating inflammatory bowel disease, paving the way for a novel low-cost and site-specific biotechnological route for the treatment of inflammatory disorders.