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Tu1982 MMP9 Catalytic Activity Is Elevated in Colon Tissue of Patients with IBD
AGA Abstracts
Tu1979
Tu1981
Capsule Endoscopy Reveals Small Intestinal Mucosal Crohn's Disease Healing After Treatment With Adalimumab: Preliminary Results of the SIMCHA Study Ernest G. Seidman, Uri Kopylov, Che-Yung Chao, Marc Girardin, Michael Starr
The Oxido-reductases Enzymes (TDO2 and SOD2) in Colonic Mucosa are Markers Associated with Histological Activity and Clinical Course in Ulcerative Colitis Jesus K. Yamamoto-Furusho, Lucero A. Salazar-Salas, Gabriela Fonseca-Camarillo
Background Mucosal healing is increasingly recognized as an important treatment goal in Crohn's disease (CD). Most studies have relied on the results of ileocolonoscopy, without assessing the vast majority of the small bowel (SB). Videocapsule endoscopy (VCE) is a wellestablished non-invasive and accurate diagnostic modality for the evaluation of the extent and severity of SB CD. The aim of this study was to investigate SB mucosal healing of CD after 6 months of adalimumab therapy. Methods Prospective single center study in consecutive adult patients (> 18 y) with moderate to severe CD involving the SB, defined by a baseline VCE exam at diagnosis with a Lewis score > 790 (normal <135, mild disease 135790) in at least one tertile. Exclusion criteria included use of drugs known to induce SB lesions such as NSAIDs for the previous month. Patients were also excluded for a history suggestive of obstructive symptoms, known strictures or a failed patency capsule exam. Patients were all treated with adalimumab monotherapy for 24 weeks. Primary endpoint was the Lewis score on repeat VCE at 24 weeks. Mucosal healing was defined as a repeat Lewis score < 350, whereas partial response was defined as a > 50% decrease in repeat Lewis score. Secondary outcomes included clinical index of remission (Harvey Bradshaw Index < 5) and fecal calprotectin. Results Interim results are available for the first 10 consenting patients (5 M, 5 F) recruited (2012-15). Mean baseline Lewis score was 2305 (range 790- 6340; 95% CI 952-3108). Mean repeat Lewis score was 673 (range 112-2734; 95% CI 178-1096; p = 0.0047). Complete mucosal healing was observed in 4/10 cases, and partial response in 5 others. A persistent stricture resulted in a limited (38.3%) improvement in 1 other case. The mean decrease in Lewis score was 1632 (95% CI 643- 2622), representing a mean decrease of 74.4%. Baseline VCE demonstrated one or more ulcerated SB strictures in 3 cases; two had non-ulcerated strictures at week 24 that were traversed. No capsule retention or other adverse events were observed. The HBI was elevated (> 5) in 6 cases prior to therapy. Repeat HBI was consistent with clinical remission in all 6 at week 24. Mean fecal calprotectin decreased (300.3 to 169.7), but the difference did not achieve significance. Conclusions This is the first study using VCE demonstrating highly significant improvement of small intestinal mucosal Crohn's disease after treatment with adalimumab for 6 months. Our preliminary data suggest that VCE is a safe and effective method to reassess small bowel mucosal healing in CD.
Introduction: Ulcerative Colitis (UC) is a polygenic and multifactorial disease. In animal models has been observed that elevation of some of these enzymes in the pathway of tryptophan were found to be reduced in severe colitis conditions. The TDO2 gene encodes for an oxidoreductase enzyme participating in the pathway of tryptophan and having an immunosuppressive function. On the other hand, the SOD2 gene encodes an antioxidant enzyme (superoxide dismutase magnesium) located in the mitochondrial matrix that protects cells from oxidative stress by scavenging free radicals. No previous studies have evaluated the role of oxidoreductases enzymes (TDO2 and SOD2) expression in the colonic mucosa from patients with UC. The aim was to measure the gene expression of SOD2 and TDO2 in the colonic mucosa from patients with UC and normal controls as well as to determine its association with clinical characteristics of the disease. Methods: We evaluated a total of 40 patients with definitive diagnosis of UC and 20 normal controls without endoscopic and histologic evidence of colonic inflammation.The TDO2 and SOD2 relative gene expression was measured by real time polymerase chain reaction (PCR) using specific primers and GAPDH gene as a reference was analyzed for normalization purposes and quality control. Statistical analysis was performed using SPSS version 20. A value of P <0.05 was considered as significant. Results: We studied 40 patients with UC (20 active and 20 in remission, 50% female and 50% male (mean age at diagnosis of 40 years). The extent of disease 52.8% had pancolitis, 16.7% left colitis and 30.6% proctosigmoiditis. In the clinical course, 66.7% had an intermittent activity, 30.6% initial activity and then long-term remission and 2.8% continuous activity. The degree of UC activity was 16.7% severe, 27.8% moderate, 8.3% mild and 47.2% were in remission. The TDO2 and SOD2 gene expression were significantly higher in patients with active UC compared to the normal control group without inflammation (P= 0.001 and P= 0.001, respectively) and UC in remission and controls (P=0.04 and P= 0.001, respectively). The gene expression of SOD2 was associated with severe histological activity (OR = 26.4, 95%IC: 4.4-157.2, P = 0.00004). The over-expression of TDO2 was associated with clinical course characterized by the presence of initial activity and then prolonged remission more than 5 years (P=0.06, OR= 4.7, 95%CI: 0.85 to 26). Conclusions: The genes TDO2 and SOD2 were up-regulated in the colonic mucosa from patients with active UC. The gen TDO2 was associated with benign clinical course of UC and the gene expression of SOD2 was associated with histologial activity in the colonic mucosa of UC patients.
Tu1980 Correlation of Mucosal MMP9 Expression with Mayo Score and Endoscopic Sub-Score in Patients with Moderately to Severely Active Ulcerative Colitis Barrett Levesque, Bal R. Bhandari, Guang Chen, Erik G. Huntzicker, Jacky Woo, Bittoo Kanwar, Yuanyuan Xiao, Huan Wang, Dorothy M. French, Brian G. Feagan, William Sandborn
Tu1982 MMP9 Catalytic Activity Is Elevated in Colon Tissue of Patients with IBD Andrew E. Greenstein, Maile Velasquez, Ted Sullivan, Robert Brockett, Bittoo Kanwar, Dorothee Saddier Axe, Huan Wang, Guang Chen, Hyock-Joo Kwon, Erik Huntzicker, Igor Mikaelian, Jacky Woo
Background: Elevated expression levels of mucosal MMP-9 have been reported to be associated with ulcerative colitis severity by histological scores. In this study, we evaluated mucosal MMP-9 expression levels with standard clinical measurements of disease activity: Mayo Clinic score, endoscopic sub-score, frequency of bowel movements and bloody stools. We also compared the utility of mucosal MMP-9 expression with fecal inflammatory markers to assess the degree of disease activity. Methods: Immunohistochemistry staining was performed on baseline and post-dose (Day 36) colonic biopsy tissue obtained from a phase I study to evaluate the safety and therapeutic potential of GS-5745, a humanized antibody against MMP-9. "H-score" is a composite score that quantifies both the percentage and the intensity of MMP-9 staining. Wilcoxon or Kruskal-Wallis rank sum test was used to compare H-score values in patients with varying severity of disease; Spearman correlation coefficient was used to determine the level of correlation. Results: When stratified by baseline Mayo score, patients with a Mayo score of 9-11 (n=14, median H-score 89.71) had significantly higher baseline level of H-score (p < 0.05) compared to patients with a Mayo score of 6-8 (n=32, median H-score 64.86; Table 1). When patients were stratified by post-treatment Mayo score, an incremental increase in MMP9 H-score was observed amongst patients with overall Mayo score from 1-5, 6-8 and 9-11 (median H-score of 31.99, 61.68 and 82.97, respectively; p< 0.01). Baseline median H-Score were significantly higher (p<0.01) in patients with an endoscopic sub-score of 3 (n=17, median score at 89.38) compared to patients with a sub-score 2 (n =29, median score at 56.85). Patients who achieved mucosal healing (endoscopic sub-score 0-1) at Day 36 had significantly lower H-score (median score at 29.92) than patients who did not achieve mucosal healing (H-score at 43.99 and 90.36 for subjects with an endoscopic sub-score of 2 or 3, respectively). Patients who showed clinical response to GS-5745 had significantly larger reductions in H-score compared to nonresponding patients and patients receiving placebo (median % change from baseline -49.87, -2.55, and -9.86, respectively; p<0.01). Compared with fecal inflammatory markers (fecal calprotectin and fecal lactoferrin), mucosal MMP-9 expression exhibited the highest association with Mayo score, endoscopic score and stool frequency. Conclusion: The MMP-9 Hscore was highly associated with overall Mayo score and endoscopic sub-score in patients with moderately to severely active ulcerative colitis. These data further support MMP-9 as a relevant target for the treatment of ulcerative colitis.
Introduction: Matrix metalloproteinase 9 (MMP9) is highly expressed in ulcerative colitis (UC) and Crohn's disease (CD) with limited expression in the healthy colon. MMP9 is synthesized and secreted as an inactive pro-enzyme which can proteolyze a variety of tissueassociated substrates upon activation. Neutrophils, macrophages and epithelial cells in regions of active disease produce MMP9, but it is unclear if this MMP9 is active or if the level of active MMP9 correlates with disease severity. Methods: Frozen and formalin fixed colon tissue from patients with CD (n=17), UC (n=22), or non-IBD controls (n=17) were analyzed. A multiplexed ELISA-based assay was developed to quantify both total and active MMP9. To detect active MMP9, an antibody was generated by immunizing rabbits with a peptide corresponding to the non-native N-terminus of MMP9 after pro-domain removal (Phe107). Total and active MMP9 were assessed by western blot analysis using this reagent and a commercial antibody. Disease severity was determined using the Geboes UC scoring criteria, and MMP9 levels were detected by IHC in matched formalin fixed colon tissue. Results: Western blot analysis demonstrated the presence of pro-MMP9 at variable levels (major band at 92 kD) in all subjects while active MMP9 (bands at 70-84 kD) was detected in IBD subjects (Fig. 1B). The level of active MMP9 in these western blots was consistent with the quantitative ELISA based activity assays. Significantly higher levels of active MMP9 were detected in CD or UC versus non-IBD subjects ( p < 0.0001) (Fig. 1B). The levels of active MMP9 were not correlated with the levels of total MMP9. The level of active MMP9 correlated with the histologic severity of ulcerative colitis as assessed by Geboes score (Spearman correlation = 0.754). Conclusions: We developed an antibody that recognizes active MMP9 and a quantitative ELISA assay that enables accurate determination of active and pro-MMP9 in colon tissue. Our data demonstrate that the N-terminus of active MMP9 in tissue is phenylalanine 107. Given the limited correlation between total MMP9 or MMP9/TIMP1 ratio with active MMP9, this study also revealed caveats in inferring MMP9 activity from levels of total MMP9. The abundance of MMP9 catalytic activity in individual CD and UC colon samples provides further association of MMP9 function with IBD. These findings support the clinical assessment of Gilead's MMP9 inhibitory mAb, GS-5745, in IBD patients and could be used to investigate the local role of MMP9 proteolysis in other diseases.
Table1: MMP-9 H-score at baseline and at Day 36 in UC subjects stratified by overall Mayo Score and Endoscopic Sub-score
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Tu1983 The Investigation of Medium- to Long-Term Prognosis of Ulcerative Colitis Patients Using Ulcerative Colitis Endoscopic Index of Severity (UCEIS) Mari Arai, Makoto Naganuma, Shinya Sugimoto, Hiroki Kiyohara, Keiko Ohno, Kiyoto Mori, Kosaku Nanki, Makoto Mutaguchi, Shinta Mizuno, Rieko Bessho, Yoshihiro Nakazato, Nagamu Inoue, Haruhiko Ogata, Yasushi Iwao, Takanori Kanai
Tu1985 Correlation of Fecal Lactoferrin Concentration and Extent of Colonic Involvement in Patients With Inflammatory Bowel Disease Marrieth Rubio, Joshua G. Gazo, Vu Q. Nguyen, James H. Boone, Dario Sorrentino
Background Mucosal healing is an important goal in the management of ulcerative colitis (UC). Various scoring systems have been used to assess mucosal healing in UC. Recently, the Ulcerative Colitis Endoscopic Index of Severity (UCEIS) has been reported. The UCEIS assesses the vascular pattern, bleeding, and erosions and ulcers, on a scale of 0 (remission) to 8 (severe). Although the UCEIS is a validated scoring system, there has been no study to evaluate the usefulness of the UCEIS in clinical settings. Aim In this study, we aimed to assess the correlation between the UCEIS and existing scores and to predict clinical outcome of UC patients with clinical remission (CR) using the UCEIS. Methods We enrolled UC patients undergoing colonoscopy at Keio University Hospital between April 2012 and March 2013. We reviewed clinical characteristics and endoscopic scores at the time of the colonoscopy and then checked the clinical remission rate in the patients with CR until September 2015 retrospectively. UCEIS and Mayo endoscopic scores were assessed by the single operator at the procedure of colonoscopy. CR and relapse were defined as a Partial Mayo (pMayo) score £ 1 and pMayo score ‡ 3, Lichtiger score ‡ 5, alteration or addition of medical treatments, respectively. Results A colonoscopy was performed on a total of 215 UC patients within the aforementioned period and 117 of those maintained CR. The correlation between the UCEIS score and the Mayo endoscopic score was strong (r=0.940), whereas the Lichtiger score (r= 0.611) and CRP (r=0.397) had moderate to poor correlation with the UCEIS score. In patients with CR, 87.6% maintained CR at 1 year and 75.2% at 2 years. The follow-up period duration had a range of 0 - 41 months, with a mean duration of 22.7 months. During the follow-up period, the lower UCEIS score showed a higher rate of CR and patients with the UCEIS score of 0-1 had a significantly higher rate of CR than those with the UCEIS score of 2 or more (p<0.001). Absence of bleeding and mucosal damage on colonoscopy was associated with maintaining CR. Among patients with Mayo endoscopic score of 0, patients with the UCEIS score of 0 had a higher rate (95%) of CR compared with patients with the UCEIS score of 1 (78%) or 2 (75%). Conclusion The UCEIS is useful to predict a medium- to long-term prognosis of UC patients with CR. Mucosal healing may be defined with a UCEIS score of £ 1. Absence of bleeding and mucosal damage on colonoscopy are more associated with maintaining CR.
Introduction: Endoscopy is increasingly used as an objective parameter for mucosal inflammation and disease activity in inflammatory bowel disease [IBD]. However, endoscopy is both expensive and invasive which limits its role during patient management. Several studies have demonstrated the potential utility of noninvasive fecal biomarkers, such as calprotectin and lactoferrin, for detecting and monitoring intestinal inflammation in IBD. However, it is unclear whether their values correlate with disease extent. Aim: In this study, we evaluated the correlation between lactoferrin concentrations and the extent of colonic disease as determined by endoscopy in Crohn's Disease (CD) and Ulcerative Colitis (UC) patients. Methods: A retrospective study was performed on IBD patients at Carilion Clinic, a tertiary referral medical center, examining extent of colonic disease as determined by endoscopy. Patients with any evidence of small bowel involvement were excluded. Fecal lactoferrin levels within 30 (mean 13±11) days of endoscopy were collected. The extent of colonic disease was scored 0 to 7 from no disease (0), Ileocecal valve only (1), 1 segment (2), 2 segments (3), 3 segments (4), 4 segments (5), 5 segments (6) and pancolitis (7). The cecum, ascending, transverse, descending, rectum, and sigmoid colon are each considered one segment. Quantitative lactoferrin was determined by enzyme-linked immune assay and reported as µg/g. Spearman's correlation coefficient was used to assess the strength of correlation between lactoferrin levels and colonic disease extent. Results: We identified 82 IBD patients; 38(46%) with CD and 44(54%) with UC. 52 patients were women. The average age was 37±18 years. Average disease duration was 5.4±7.5 years. Lactoferrin levels for groupings of colonic extent were Group 0-1 (N=14) median 7.1 interquartile range 25%75% (IQR) [0 - 152.3], Group 2-3 (N=29) median=92.7 IQR [24.9 - 1133.7], Group 4-5 (N=14) median=589.0 IQR [144.7 - 2269.5], and Group 6-7 (N=25) median=2168.1 IQR [482.2 - 3662.8]. Fecal lactoferrin had a moderate to strong positive correlation with the extent of colonic involvement (R=0.577, p<0.01). The correlation was similar when stratified across patients with CD (R=0.556, p<0.01) or UC (R=0.562, p<0.01). Conclusion: This study confirms the clinical utility of fecal biomarkers to evaluate inflammation in IBD. Fecal lactoferrin showed a moderate to strong positive correlation with the extent of colonic involvement in patients with active CD and UC. A higher lactoferrin level was predictive of a greater extent of colonic disease, regardless of whether the patients had CD or UC. This finding may bear clinical significance in monitoring evolution of IBD of the colon over time.
Tu1984 Utility of Capsule Endoscopy for Monitoring Crohn's Disease Activity Gil Y. Melmed, Marla Dubinsky, David Rubin, Mark Fleisher, Shabana F. Pasha, Ignacio Fernandez-Urien, Bruno Rosa, Neofytos Papageorgiou, Ari J. Bergwerk, Michal KedarDatel, Jonathan A. Leighton
Tu1986 TNF-Alpha Activity in Plasma From Patients With Inflammatory Bowel Disease Under Adalimumab and Infliximab Treatment Evaluated in a CellBased Assay Pablo M. Linares, H de la Fuente, María Chaparro, Iván Guerra, Pedro L. Majano, Marisa Iborra, Jose Luis Cabriada Nuño, Luis Bujanda, M Barreiro-de Acosta, Valle GarcíaSánchez, Ignacio Marin-Jimenez, David Bernardo, Javier P. Gisbert
Introduction: Treatment paradigms for Crohn's disease (CD) have increasingly emphasized a ‘treat-to-target' approach that suggests the need for serial monitoring of mucosal disease activity. Capsule endoscopy (CE) offers a safe, less invasive alternative to ileocolonoscopy for the evaluation of the small bowel (SB) in patients with CD. The utility of CE for longitudinal monitoring of CD activity has not been established. Methods: We conducted a prospective, longitudinal, multicenter trial of patients with CD using the small bowel capsule endoscopy system, with a planned sample size of n=75. After SB patency was confirmed, subjects underwent colonoscopy and CE at baseline and at 6 months. Physician global assessment (PGA), Crohn's disease activity index (CDAI), C-reactive protein (CRP), and fecal calprotectin (FC) were assessed at both timepoints. The Simple Endoscopic Score (SES-CD) was used to evaluate endoscopic disease activity of the terminal ileum, and the Lewis score and the Capsule Endoscopy Crohn's Disease Endoscopic Index of Severity (CECDEIS) were used for CE; all endoscopic evaluations were performed by blinded central readers. CE scoring indices were analyzed separately for the entire SB and for the terminal ileum (Lewis-TI, CECDEIS-TI). The primary measure was correlation between changes in CE and PGA over 6 months. Secondary measures assessed correlations between changes in CE with changes in colonoscopy and clinical parameters. We report the results of an a priori planned interim analysis to assess feasibility. Results: Twenty subjects completed evaluations at both timepoints. Subjects had a mean age of 32.7 years, and 60% were female. At the 6 month assessments, numeric changes consistent with clinical improvement in endoscopic and clinical parameters were noted (Table). Changes in the Lewis score correlated with changes in SES-CD (p=0.008). When CE scores were limited to the terminal ileum, both
Background: L929 murine fibroblast is a TNF-alpha sensitive cell line. The usefulness of co-culturing those cells with plasma to assess its in vitro activity from patients with Crohn's disease (CD) before and during the induction doses of anti-TNF-alpha drug has not been studied so far. Aims: 1) To evaluate the correlation between TNF-alpha plasma in vitro activity and clinical activity in CD patients. 2) To assess the usefulness of measuring TNFalpha plasma in vitro activity to predict short-term remission with anti-TNF-alpha treatment. Methods: CD patients naïve to anti-TNF-alpha treatment were prospectively included in this multicenter study. Patients received 160/80 mg adalimumab at weeks 0 and 2, and 40 mg every-other-week thereafter, or infliximab 5 mg/kg at weeks 0, 2, 6 and every-twomonths thereafter. Remission was defined as a CDAI score <150, and response as a decrease of >70 points, after 14 weeks of treatment. Clinical evaluation was assessed and blood samples were obtained at baseline and at weeks 4 and 14. Receiver operating characteristic (ROC) curves were constructed and the area-under the ROC curves (AUCs) were calculated. TNF-alpha-sensitive, actinomycin D-treated murine L929 fibroblasts were employed to quantify TNF-alpha activity in heat-inactivated plasma samples from patients with CD. Human
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Lewis-TI and CECDEIS-TI changes correlated with SES-CD changes (p=0.01, p=0.005). However, changes in the Lewis and CECDEIS scores did not correlate with changes in PGA, CDAI, CRP, or FC. There were no serious adverse events. Conclusion: The use of CE for the assessment of mucosal disease activity over time correlates with ileocolonoscopy. This correlation is most consistent when CE activity scores are limited to TI evaluation, suggesting that CE may identify more proximal inflammation. This analysis demonstrates that CE is feasible and safe in patients with CD. Correlations with clinical parameters were not seen, although all clinical parameters and endoscopic scores improved over time. Continued evaluation of CE for longitudinal disease monitoring of patients with CD is warranted. Endoscopic and clinical assessments from baseline to 6 months
Tu1979
Tu1981
Capsule Endoscopy Reveals Small Intestinal Mucosal Crohn's Disease Healing After Treatment With Adalimumab: Preliminary Results of the SIMCHA Study Ernest G. Seidman, Uri Kopylov, Che-Yung Chao, Marc Girardin, Michael Starr
The Oxido-reductases Enzymes (TDO2 and SOD2) in Colonic Mucosa are Markers Associated with Histological Activity and Clinical Course in Ulcerative Colitis Jesus K. Yamamoto-Furusho, Lucero A. Salazar-Salas, Gabriela Fonseca-Camarillo
Background Mucosal healing is increasingly recognized as an important treatment goal in Crohn's disease (CD). Most studies have relied on the results of ileocolonoscopy, without assessing the vast majority of the small bowel (SB). Videocapsule endoscopy (VCE) is a wellestablished non-invasive and accurate diagnostic modality for the evaluation of the extent and severity of SB CD. The aim of this study was to investigate SB mucosal healing of CD after 6 months of adalimumab therapy. Methods Prospective single center study in consecutive adult patients (> 18 y) with moderate to severe CD involving the SB, defined by a baseline VCE exam at diagnosis with a Lewis score > 790 (normal <135, mild disease 135790) in at least one tertile. Exclusion criteria included use of drugs known to induce SB lesions such as NSAIDs for the previous month. Patients were also excluded for a history suggestive of obstructive symptoms, known strictures or a failed patency capsule exam. Patients were all treated with adalimumab monotherapy for 24 weeks. Primary endpoint was the Lewis score on repeat VCE at 24 weeks. Mucosal healing was defined as a repeat Lewis score < 350, whereas partial response was defined as a > 50% decrease in repeat Lewis score. Secondary outcomes included clinical index of remission (Harvey Bradshaw Index < 5) and fecal calprotectin. Results Interim results are available for the first 10 consenting patients (5 M, 5 F) recruited (2012-15). Mean baseline Lewis score was 2305 (range 790- 6340; 95% CI 952-3108). Mean repeat Lewis score was 673 (range 112-2734; 95% CI 178-1096; p = 0.0047). Complete mucosal healing was observed in 4/10 cases, and partial response in 5 others. A persistent stricture resulted in a limited (38.3%) improvement in 1 other case. The mean decrease in Lewis score was 1632 (95% CI 643- 2622), representing a mean decrease of 74.4%. Baseline VCE demonstrated one or more ulcerated SB strictures in 3 cases; two had non-ulcerated strictures at week 24 that were traversed. No capsule retention or other adverse events were observed. The HBI was elevated (> 5) in 6 cases prior to therapy. Repeat HBI was consistent with clinical remission in all 6 at week 24. Mean fecal calprotectin decreased (300.3 to 169.7), but the difference did not achieve significance. Conclusions This is the first study using VCE demonstrating highly significant improvement of small intestinal mucosal Crohn's disease after treatment with adalimumab for 6 months. Our preliminary data suggest that VCE is a safe and effective method to reassess small bowel mucosal healing in CD.
Introduction: Ulcerative Colitis (UC) is a polygenic and multifactorial disease. In animal models has been observed that elevation of some of these enzymes in the pathway of tryptophan were found to be reduced in severe colitis conditions. The TDO2 gene encodes for an oxidoreductase enzyme participating in the pathway of tryptophan and having an immunosuppressive function. On the other hand, the SOD2 gene encodes an antioxidant enzyme (superoxide dismutase magnesium) located in the mitochondrial matrix that protects cells from oxidative stress by scavenging free radicals. No previous studies have evaluated the role of oxidoreductases enzymes (TDO2 and SOD2) expression in the colonic mucosa from patients with UC. The aim was to measure the gene expression of SOD2 and TDO2 in the colonic mucosa from patients with UC and normal controls as well as to determine its association with clinical characteristics of the disease. Methods: We evaluated a total of 40 patients with definitive diagnosis of UC and 20 normal controls without endoscopic and histologic evidence of colonic inflammation.The TDO2 and SOD2 relative gene expression was measured by real time polymerase chain reaction (PCR) using specific primers and GAPDH gene as a reference was analyzed for normalization purposes and quality control. Statistical analysis was performed using SPSS version 20. A value of P <0.05 was considered as significant. Results: We studied 40 patients with UC (20 active and 20 in remission, 50% female and 50% male (mean age at diagnosis of 40 years). The extent of disease 52.8% had pancolitis, 16.7% left colitis and 30.6% proctosigmoiditis. In the clinical course, 66.7% had an intermittent activity, 30.6% initial activity and then long-term remission and 2.8% continuous activity. The degree of UC activity was 16.7% severe, 27.8% moderate, 8.3% mild and 47.2% were in remission. The TDO2 and SOD2 gene expression were significantly higher in patients with active UC compared to the normal control group without inflammation (P= 0.001 and P= 0.001, respectively) and UC in remission and controls (P=0.04 and P= 0.001, respectively). The gene expression of SOD2 was associated with severe histological activity (OR = 26.4, 95%IC: 4.4-157.2, P = 0.00004). The over-expression of TDO2 was associated with clinical course characterized by the presence of initial activity and then prolonged remission more than 5 years (P=0.06, OR= 4.7, 95%CI: 0.85 to 26). Conclusions: The genes TDO2 and SOD2 were up-regulated in the colonic mucosa from patients with active UC. The gen TDO2 was associated with benign clinical course of UC and the gene expression of SOD2 was associated with histologial activity in the colonic mucosa of UC patients.
Tu1980 Correlation of Mucosal MMP9 Expression with Mayo Score and Endoscopic Sub-Score in Patients with Moderately to Severely Active Ulcerative Colitis Barrett Levesque, Bal R. Bhandari, Guang Chen, Erik G. Huntzicker, Jacky Woo, Bittoo Kanwar, Yuanyuan Xiao, Huan Wang, Dorothy M. French, Brian G. Feagan, William Sandborn
Tu1982 MMP9 Catalytic Activity Is Elevated in Colon Tissue of Patients with IBD Andrew E. Greenstein, Maile Velasquez, Ted Sullivan, Robert Brockett, Bittoo Kanwar, Dorothee Saddier Axe, Huan Wang, Guang Chen, Hyock-Joo Kwon, Erik Huntzicker, Igor Mikaelian, Jacky Woo
Background: Elevated expression levels of mucosal MMP-9 have been reported to be associated with ulcerative colitis severity by histological scores. In this study, we evaluated mucosal MMP-9 expression levels with standard clinical measurements of disease activity: Mayo Clinic score, endoscopic sub-score, frequency of bowel movements and bloody stools. We also compared the utility of mucosal MMP-9 expression with fecal inflammatory markers to assess the degree of disease activity. Methods: Immunohistochemistry staining was performed on baseline and post-dose (Day 36) colonic biopsy tissue obtained from a phase I study to evaluate the safety and therapeutic potential of GS-5745, a humanized antibody against MMP-9. "H-score" is a composite score that quantifies both the percentage and the intensity of MMP-9 staining. Wilcoxon or Kruskal-Wallis rank sum test was used to compare H-score values in patients with varying severity of disease; Spearman correlation coefficient was used to determine the level of correlation. Results: When stratified by baseline Mayo score, patients with a Mayo score of 9-11 (n=14, median H-score 89.71) had significantly higher baseline level of H-score (p < 0.05) compared to patients with a Mayo score of 6-8 (n=32, median H-score 64.86; Table 1). When patients were stratified by post-treatment Mayo score, an incremental increase in MMP9 H-score was observed amongst patients with overall Mayo score from 1-5, 6-8 and 9-11 (median H-score of 31.99, 61.68 and 82.97, respectively; p< 0.01). Baseline median H-Score were significantly higher (p<0.01) in patients with an endoscopic sub-score of 3 (n=17, median score at 89.38) compared to patients with a sub-score 2 (n =29, median score at 56.85). Patients who achieved mucosal healing (endoscopic sub-score 0-1) at Day 36 had significantly lower H-score (median score at 29.92) than patients who did not achieve mucosal healing (H-score at 43.99 and 90.36 for subjects with an endoscopic sub-score of 2 or 3, respectively). Patients who showed clinical response to GS-5745 had significantly larger reductions in H-score compared to nonresponding patients and patients receiving placebo (median % change from baseline -49.87, -2.55, and -9.86, respectively; p<0.01). Compared with fecal inflammatory markers (fecal calprotectin and fecal lactoferrin), mucosal MMP-9 expression exhibited the highest association with Mayo score, endoscopic score and stool frequency. Conclusion: The MMP-9 Hscore was highly associated with overall Mayo score and endoscopic sub-score in patients with moderately to severely active ulcerative colitis. These data further support MMP-9 as a relevant target for the treatment of ulcerative colitis.
Introduction: Matrix metalloproteinase 9 (MMP9) is highly expressed in ulcerative colitis (UC) and Crohn's disease (CD) with limited expression in the healthy colon. MMP9 is synthesized and secreted as an inactive pro-enzyme which can proteolyze a variety of tissueassociated substrates upon activation. Neutrophils, macrophages and epithelial cells in regions of active disease produce MMP9, but it is unclear if this MMP9 is active or if the level of active MMP9 correlates with disease severity. Methods: Frozen and formalin fixed colon tissue from patients with CD (n=17), UC (n=22), or non-IBD controls (n=17) were analyzed. A multiplexed ELISA-based assay was developed to quantify both total and active MMP9. To detect active MMP9, an antibody was generated by immunizing rabbits with a peptide corresponding to the non-native N-terminus of MMP9 after pro-domain removal (Phe107). Total and active MMP9 were assessed by western blot analysis using this reagent and a commercial antibody. Disease severity was determined using the Geboes UC scoring criteria, and MMP9 levels were detected by IHC in matched formalin fixed colon tissue. Results: Western blot analysis demonstrated the presence of pro-MMP9 at variable levels (major band at 92 kD) in all subjects while active MMP9 (bands at 70-84 kD) was detected in IBD subjects (Fig. 1B). The level of active MMP9 in these western blots was consistent with the quantitative ELISA based activity assays. Significantly higher levels of active MMP9 were detected in CD or UC versus non-IBD subjects ( p < 0.0001) (Fig. 1B). The levels of active MMP9 were not correlated with the levels of total MMP9. The level of active MMP9 correlated with the histologic severity of ulcerative colitis as assessed by Geboes score (Spearman correlation = 0.754). Conclusions: We developed an antibody that recognizes active MMP9 and a quantitative ELISA assay that enables accurate determination of active and pro-MMP9 in colon tissue. Our data demonstrate that the N-terminus of active MMP9 in tissue is phenylalanine 107. Given the limited correlation between total MMP9 or MMP9/TIMP1 ratio with active MMP9, this study also revealed caveats in inferring MMP9 activity from levels of total MMP9. The abundance of MMP9 catalytic activity in individual CD and UC colon samples provides further association of MMP9 function with IBD. These findings support the clinical assessment of Gilead's MMP9 inhibitory mAb, GS-5745, in IBD patients and could be used to investigate the local role of MMP9 proteolysis in other diseases.
Table1: MMP-9 H-score at baseline and at Day 36 in UC subjects stratified by overall Mayo Score and Endoscopic Sub-score
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Tu1983 The Investigation of Medium- to Long-Term Prognosis of Ulcerative Colitis Patients Using Ulcerative Colitis Endoscopic Index of Severity (UCEIS) Mari Arai, Makoto Naganuma, Shinya Sugimoto, Hiroki Kiyohara, Keiko Ohno, Kiyoto Mori, Kosaku Nanki, Makoto Mutaguchi, Shinta Mizuno, Rieko Bessho, Yoshihiro Nakazato, Nagamu Inoue, Haruhiko Ogata, Yasushi Iwao, Takanori Kanai
Tu1985 Correlation of Fecal Lactoferrin Concentration and Extent of Colonic Involvement in Patients With Inflammatory Bowel Disease Marrieth Rubio, Joshua G. Gazo, Vu Q. Nguyen, James H. Boone, Dario Sorrentino
Background Mucosal healing is an important goal in the management of ulcerative colitis (UC). Various scoring systems have been used to assess mucosal healing in UC. Recently, the Ulcerative Colitis Endoscopic Index of Severity (UCEIS) has been reported. The UCEIS assesses the vascular pattern, bleeding, and erosions and ulcers, on a scale of 0 (remission) to 8 (severe). Although the UCEIS is a validated scoring system, there has been no study to evaluate the usefulness of the UCEIS in clinical settings. Aim In this study, we aimed to assess the correlation between the UCEIS and existing scores and to predict clinical outcome of UC patients with clinical remission (CR) using the UCEIS. Methods We enrolled UC patients undergoing colonoscopy at Keio University Hospital between April 2012 and March 2013. We reviewed clinical characteristics and endoscopic scores at the time of the colonoscopy and then checked the clinical remission rate in the patients with CR until September 2015 retrospectively. UCEIS and Mayo endoscopic scores were assessed by the single operator at the procedure of colonoscopy. CR and relapse were defined as a Partial Mayo (pMayo) score £ 1 and pMayo score ‡ 3, Lichtiger score ‡ 5, alteration or addition of medical treatments, respectively. Results A colonoscopy was performed on a total of 215 UC patients within the aforementioned period and 117 of those maintained CR. The correlation between the UCEIS score and the Mayo endoscopic score was strong (r=0.940), whereas the Lichtiger score (r= 0.611) and CRP (r=0.397) had moderate to poor correlation with the UCEIS score. In patients with CR, 87.6% maintained CR at 1 year and 75.2% at 2 years. The follow-up period duration had a range of 0 - 41 months, with a mean duration of 22.7 months. During the follow-up period, the lower UCEIS score showed a higher rate of CR and patients with the UCEIS score of 0-1 had a significantly higher rate of CR than those with the UCEIS score of 2 or more (p<0.001). Absence of bleeding and mucosal damage on colonoscopy was associated with maintaining CR. Among patients with Mayo endoscopic score of 0, patients with the UCEIS score of 0 had a higher rate (95%) of CR compared with patients with the UCEIS score of 1 (78%) or 2 (75%). Conclusion The UCEIS is useful to predict a medium- to long-term prognosis of UC patients with CR. Mucosal healing may be defined with a UCEIS score of £ 1. Absence of bleeding and mucosal damage on colonoscopy are more associated with maintaining CR.
Introduction: Endoscopy is increasingly used as an objective parameter for mucosal inflammation and disease activity in inflammatory bowel disease [IBD]. However, endoscopy is both expensive and invasive which limits its role during patient management. Several studies have demonstrated the potential utility of noninvasive fecal biomarkers, such as calprotectin and lactoferrin, for detecting and monitoring intestinal inflammation in IBD. However, it is unclear whether their values correlate with disease extent. Aim: In this study, we evaluated the correlation between lactoferrin concentrations and the extent of colonic disease as determined by endoscopy in Crohn's Disease (CD) and Ulcerative Colitis (UC) patients. Methods: A retrospective study was performed on IBD patients at Carilion Clinic, a tertiary referral medical center, examining extent of colonic disease as determined by endoscopy. Patients with any evidence of small bowel involvement were excluded. Fecal lactoferrin levels within 30 (mean 13±11) days of endoscopy were collected. The extent of colonic disease was scored 0 to 7 from no disease (0), Ileocecal valve only (1), 1 segment (2), 2 segments (3), 3 segments (4), 4 segments (5), 5 segments (6) and pancolitis (7). The cecum, ascending, transverse, descending, rectum, and sigmoid colon are each considered one segment. Quantitative lactoferrin was determined by enzyme-linked immune assay and reported as µg/g. Spearman's correlation coefficient was used to assess the strength of correlation between lactoferrin levels and colonic disease extent. Results: We identified 82 IBD patients; 38(46%) with CD and 44(54%) with UC. 52 patients were women. The average age was 37±18 years. Average disease duration was 5.4±7.5 years. Lactoferrin levels for groupings of colonic extent were Group 0-1 (N=14) median 7.1 interquartile range 25%75% (IQR) [0 - 152.3], Group 2-3 (N=29) median=92.7 IQR [24.9 - 1133.7], Group 4-5 (N=14) median=589.0 IQR [144.7 - 2269.5], and Group 6-7 (N=25) median=2168.1 IQR [482.2 - 3662.8]. Fecal lactoferrin had a moderate to strong positive correlation with the extent of colonic involvement (R=0.577, p<0.01). The correlation was similar when stratified across patients with CD (R=0.556, p<0.01) or UC (R=0.562, p<0.01). Conclusion: This study confirms the clinical utility of fecal biomarkers to evaluate inflammation in IBD. Fecal lactoferrin showed a moderate to strong positive correlation with the extent of colonic involvement in patients with active CD and UC. A higher lactoferrin level was predictive of a greater extent of colonic disease, regardless of whether the patients had CD or UC. This finding may bear clinical significance in monitoring evolution of IBD of the colon over time.
Tu1984 Utility of Capsule Endoscopy for Monitoring Crohn's Disease Activity Gil Y. Melmed, Marla Dubinsky, David Rubin, Mark Fleisher, Shabana F. Pasha, Ignacio Fernandez-Urien, Bruno Rosa, Neofytos Papageorgiou, Ari J. Bergwerk, Michal KedarDatel, Jonathan A. Leighton
Tu1986 TNF-Alpha Activity in Plasma From Patients With Inflammatory Bowel Disease Under Adalimumab and Infliximab Treatment Evaluated in a CellBased Assay Pablo M. Linares, H de la Fuente, María Chaparro, Iván Guerra, Pedro L. Majano, Marisa Iborra, Jose Luis Cabriada Nuño, Luis Bujanda, M Barreiro-de Acosta, Valle GarcíaSánchez, Ignacio Marin-Jimenez, David Bernardo, Javier P. Gisbert
Introduction: Treatment paradigms for Crohn's disease (CD) have increasingly emphasized a ‘treat-to-target' approach that suggests the need for serial monitoring of mucosal disease activity. Capsule endoscopy (CE) offers a safe, less invasive alternative to ileocolonoscopy for the evaluation of the small bowel (SB) in patients with CD. The utility of CE for longitudinal monitoring of CD activity has not been established. Methods: We conducted a prospective, longitudinal, multicenter trial of patients with CD using the small bowel capsule endoscopy system, with a planned sample size of n=75. After SB patency was confirmed, subjects underwent colonoscopy and CE at baseline and at 6 months. Physician global assessment (PGA), Crohn's disease activity index (CDAI), C-reactive protein (CRP), and fecal calprotectin (FC) were assessed at both timepoints. The Simple Endoscopic Score (SES-CD) was used to evaluate endoscopic disease activity of the terminal ileum, and the Lewis score and the Capsule Endoscopy Crohn's Disease Endoscopic Index of Severity (CECDEIS) were used for CE; all endoscopic evaluations were performed by blinded central readers. CE scoring indices were analyzed separately for the entire SB and for the terminal ileum (Lewis-TI, CECDEIS-TI). The primary measure was correlation between changes in CE and PGA over 6 months. Secondary measures assessed correlations between changes in CE with changes in colonoscopy and clinical parameters. We report the results of an a priori planned interim analysis to assess feasibility. Results: Twenty subjects completed evaluations at both timepoints. Subjects had a mean age of 32.7 years, and 60% were female. At the 6 month assessments, numeric changes consistent with clinical improvement in endoscopic and clinical parameters were noted (Table). Changes in the Lewis score correlated with changes in SES-CD (p=0.008). When CE scores were limited to the terminal ileum, both
Background: L929 murine fibroblast is a TNF-alpha sensitive cell line. The usefulness of co-culturing those cells with plasma to assess its in vitro activity from patients with Crohn's disease (CD) before and during the induction doses of anti-TNF-alpha drug has not been studied so far. Aims: 1) To evaluate the correlation between TNF-alpha plasma in vitro activity and clinical activity in CD patients. 2) To assess the usefulness of measuring TNFalpha plasma in vitro activity to predict short-term remission with anti-TNF-alpha treatment. Methods: CD patients naïve to anti-TNF-alpha treatment were prospectively included in this multicenter study. Patients received 160/80 mg adalimumab at weeks 0 and 2, and 40 mg every-other-week thereafter, or infliximab 5 mg/kg at weeks 0, 2, 6 and every-twomonths thereafter. Remission was defined as a CDAI score <150, and response as a decrease of >70 points, after 14 weeks of treatment. Clinical evaluation was assessed and blood samples were obtained at baseline and at weeks 4 and 14. Receiver operating characteristic (ROC) curves were constructed and the area-under the ROC curves (AUCs) were calculated. TNF-alpha-sensitive, actinomycin D-treated murine L929 fibroblasts were employed to quantify TNF-alpha activity in heat-inactivated plasma samples from patients with CD. Human
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Lewis-TI and CECDEIS-TI changes correlated with SES-CD changes (p=0.01, p=0.005). However, changes in the Lewis and CECDEIS scores did not correlate with changes in PGA, CDAI, CRP, or FC. There were no serious adverse events. Conclusion: The use of CE for the assessment of mucosal disease activity over time correlates with ileocolonoscopy. This correlation is most consistent when CE activity scores are limited to TI evaluation, suggesting that CE may identify more proximal inflammation. This analysis demonstrates that CE is feasible and safe in patients with CD. Correlations with clinical parameters were not seen, although all clinical parameters and endoscopic scores improved over time. Continued evaluation of CE for longitudinal disease monitoring of patients with CD is warranted. Endoscopic and clinical assessments from baseline to 6 months