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15 results on '"M. Wildenberg"'

1. P110 activation protein is strongly expressed in intestinal fibrosis in inflammatory bowel disease patients

Author

D Lartey, M van Roest, J Verhoeff, J Grootjans, C Buskens, J van der Bilt, M Wildenberg, G D'Haens, and M Löwenberg

Subjects
Gastroenterology, General Medicine
Abstract

Background Intestinal fibrosis is a common complication of inflammatory bowel disease (IBD) with limited diagnostic modalities to detect and determine the degree of fibrosis. Fibroblast activation protein alpha (FAP) is a protein with both dipeptidyl peptidase and endopeptidase properties which is virtually absent in healthy tissues and has increased expression in chronic inflammatory and fibrotic diseases. In this study, we investigated the presence of FAP in intestinal fibrosis caused by Crohn’s disease (CD) and ulcerative colitis (UC). Methods For this study, transmural surgical resection specimens were retrieved from the Tytgat institute IBD biobank located at Amsterdam University Medical Center. IBD patients undergoing an ileocecal resection for stricturing CD or a subtotal colectomy for therapy-refractory disease, and patients undergoing a right-sided hemicolectomy or ileocecal resection for dysplasia or non-metastasized colorectal cancer (CRC), were included. All patients had an established diagnosis of (CD, UC, dysplasia or CRC) by previous endoscopy and biopsies and had provided informed consent prior to surgery. FAP protein and gene expression levels were determined using immunohistochemistry staining and quantitative polymerase chain reaction (qPCR). Mass cytometry was used to identify FAP–expressing cells. Results In total, 59 ileal and 35 colonic samples were stained and quantified to asses FAP expression (figure 1). A 1.7-fold and 1.9-fold increase of FAP protein expression was seen in inflamed (mean 22.3, p=0.0069); and stenotic (mean 25.5 p=0.0002) CD ileum compared to healthy non-IBD tissue (mean 13.1). Inflamed UC colon (mean 26.8, p = 0.0406) showed a 1.4-fold increase compared to healthy colon (mean 19.2). For the gene expression, 66 ileal and 42 colonic samples were processed and used for qPCR analysis. A 5-fold and 6.4-fold increase of FAP was found in inflamed (mean 0.011, p=0.0475) and stenotic (mean 0.014, p=0.0042) CD ileum, and a 1.7-fold increase was seen in inflamed UC colon (mean 0.278, p=0.0195) compared to controls (means: ileum 0.002; colon 0.165). Conclusion This is the first study reporting the potential of FAP as a fibrotic biomarker not only in stricturing CD, but also in therapy-refractory UC. Significantly increased FAP protein and gene expression is found in inflamed and stenotic ileum in CD and inflamed colon in UC. Additionally, CD90+GP38+ cells have been identified as the most present FAP expressing cells.

Published
2023
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2. P107 Decreased epithelial keratinization contributes to defective wound healing in Crohn's disease fistula

Author

M Becker drs., P Koelink, W Bemelman, G D'Haens, C Buskens, and M Wildenberg

Subjects
Gastroenterology, General Medicine
Abstract

Background Up to 30% of Crohn's Disease (CD) patients suffer from perianal fistula creating a high disease burden. Treating CD fistula is challenging compared to cryptoglandular fistula, which have a significantly better wound healing response. Previous studies suggested epithelial to mesenchymal transition as an inducer of fistula formation, as markers were observed in fistula tracts. However, most studies compared CD fistula to normal mucosa, thus disregarding whether any observed difference was in fact a sign of healing (as CD fistula will have more active healing than uninjured mucosa) or a sign of pathology (as healing is not occurring effectively). Therefore, in this study we evaluated CD fistula, as well as both healthy mucosa (no active healing) and cryptoglandular fistula (effective healing) in order to determine the role of differences found. Methods Biopsies from the internal opening (n=20) of perianal fistula of CD patients were collected and compared to two publicly available datasets obtained from un-inflamed rectum (RISK cohort and GSE83245). In addition, curettage material of CD (n=54) and cryptoglandular fistula (n=15) was obtained. Expression profiling was performed using RNASeq. Results Comparing the mucosa at the internal opening of CD fistula to publicly available datasets of healthy rectum, 770 genes were differentially expressed. Pathway analysis revealed upregulation of proliferative and inflammatory processes, consistent with expected immune and wound healing responses. Interestingly, strongest upregulation was seen for processes of cornification and keratinization, including expression of IVL (involucrin) and FLGR2 (fillagrin2), involved in keratinocyte differentiation. To evaluate whether this was contributing to active wound healing or rather to persistent disease, we analyzed the actual tract in CD and cryptoglandular fistula. Strikingly, cornification and keratinization processes were more pronounced in cryptoglandular fistula, suggesting a role in effective healing rather than the persistent pathology seen in CD. Similar data was observed for TGFb, previously detected in CD fistula and thus considered a pathological contributor. While we confirm increased TGFb activity in CD fistula compared to normal mucosa, activity in cryptoglandular fistula tracts was higher than in CD fistula tracts. Again, this would suggest TGFb signaling as part of the wound healing response rather than part of the pathogenic mechanism. Conclusion Perianal fistulas show upregulation of keratinization and cornification pathways compared to healthy mucosa. Comparing CD and cryptoglandular fistula, we saw more cornification in the better healing cryptoglandular fistula, suggesting involvement in effective healing rather than pathology.

Published
2023
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3. P166 Calprotectin in patients with perianal fistula: the CANALS study

Author

M Becker drs., T Stevens, F de Voogd, M Wildenberg, K Gecse, and C Buskens

Subjects
Gastroenterology, General Medicine
Abstract

Background Previous studies suggest that faecal calprotectin can be used to discriminate Crohn’s disease (CD) perianal fistulas from cryptoglandular (CG) perianal fistulas. The aim of this research was to prospectively validate these findings and further investigate local calprotectin production inside fistula tracts. The aim of the current study was to prospectively analyse the clinical value of calprotectin levels in both the faeces and the fistula tract. Methods In this prospective study, all consecutive patients with an active perianal fistula undergoing examination under anesthetics were included. Faecal and scraping calprotectin levels collected during surgery were measured. The primary objective was to differentiate between active CD perianal fistulas and CG perianal fistulas based on faecal calprotectin. Secondary outcome parameters were the levels of local calprotectin in fistula tracts and the correlation with fistula characteristics and clinical prognostic parameters like the Geldof Classification system. Results Sixty-three fistula patients were included in this study (CD n=45; CG n=18). Faecal calprotectin values were significantly higher in CD patients compared to CG patients (396.8 [61.9-1136.8] vs. 47.3 [14.6-233.6] ug/g, p=0.004). The Area Under the Curve (AUC) was 0.733 (95% CI [0.644 to 0.903] with an optimal FC cut-off value of 344ug/g. Faecal calprotectin was not related to calprotectin levels in fistula scrapings which was not significantly different in CD and cryptoglandular fistulas (p=0.893). Calprotectin in fistula scrapings was also not related to fistula characteristics like number of tracts, duration of disease or (previous) seton drainage. However, a significant correlation was found between scraping calprotectin and fistula disease severity and prognosis as defined by a new Geldof classification system: Patients in group 2a (amenable for fistula repair) had significantly lower calprotectin levels (140 [31.0-149.0])ug/g) when compared to class 2b (symptom control) (706 [198.5 – 1936]ug/g, p=0.054) and class 2c (gradually debilitating disease) (3464.0 [328.5-5692.0]ug/g, p=0.008). Patients in class 3 (exhausted perineum) had the highest calprotectin levels highest (5928.0 [2605.0-6000]ug/g). Conclusion CD and CG perianal fistulas can be distinguished by measuring faecal calprotectin levels. The difference can not be explained by local fistula calprotectin production as calprotectin in fistula scrapings is not predictive of CD. However, the results of this study suggest that scraping calprotectin levels may be used to classify fistula patients and predict outcome of fistula treatment.

Published
2023
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4. P096 Characterization of suppressive immune cell subsets in mouse models of colitis-associated colorectal cancer

Author

S Frigerio, J Saris, R Franco Fernandez, P Koelink, M van Roest, M Wildenberg, G D'Haens, and J Grootjans

Subjects
Gastroenterology, General Medicine
Abstract

Background Chronic colonic inflammation in inflammatory bowel disease (IBD) patients increases the risk of colitis-associated cancer (CAC). Cancer development in CAC is different from that observed in sporadic colorectal cancer (CRC). Most studies have focused on the role of pro-inflammatory immune cell subsets in CAC, yet it is becoming increasingly clear that immunosuppression may also drive cancer progression. We hypothesize that emergence of immunosuppressive cell subsets as a result of chronic intestinal inflammation, dampens anti-tumor immune responses and accelerates the development of CAC. Methods To simulate cancer formation in the background of chronic inflammation, mice were treated with azoxymethane (AOM) followed by three cycles of dextran sodium sulphate (DSS). Apc Min/+ mice treated with AOM were used to model sporadic CRC. After 10 weeks, immune cells from the proximal and distal colonic lamina propria, mesenteric lymph nodes (MLNs) and tumors were obtained for microscopy analyses and processed to obtain single cell suspensions for flow cytometry. Immune cell populations were studied by conventional flow cytometry and immunofluorescence. Results We observed a significant increase in absolute numbers of PD1+CTLA4+TIM3+ ‘exhausted’ CD4+ T cells in the lamina propria from proximal colons in CAC mice as compared to sporadic CRC mice. Interestingly, these numbers showed a positive correlation with the inflammation score in AOM/DSS, demonstrating that chronic inflammation is associated with the emergence of ‘exhausted’ T cells. In addition, we observed a trend to higher numbers of M2-like (MHCII-CD206+) suppressive macrophages and CD4+FoxP3+ regulatory T cells (Tregs) in tumors from CAC mice compared to sporadic CRC. When analysing T cell immune responses in the MLNs, we found a significant decrease in the ratio IFNγ/IL10-producing CD8+ T cells in CAC mice as compared to CRC mice, suggesting a shift towards an anti-inflammatory T cell response in CAC. Conclusion In summary, suppressive immune cell subsets were increased in colonic mucosa and tumors from CAC mice as a result of chronic inflammation, which may be associated with the development of dysplasia. Further functional studies are necessary to prove the role of suppressive immune subsets in IBD-associated dysplasia- and carcinoma progression.

Published
2023
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5. P084 Response to appendectomy in refractory Ulcerative Colitis appears based on two distinct inflammatory phenotypes

Author

M Becker drs., L Heuthorst, M van Roest, J van der Bilt, C Buskens, and M Wildenberg

Subjects
Gastroenterology, General Medicine
Abstract

Background Appendectomy has been suggested to decrease the risk of disease flares in UC, but we recently showed that even in therapy refractory patients, appendectomy may improve symptoms in a subset of patients. Despite several hypotheses, the effector mechanism of this phenomenon remains unclear. In this study we characterized gene expression profiles of appendices from therapy refractory UC patients either responding or non-responding to appendectomy, with the aim to identify pathways involved in response as well as guide selection of patients who are optimal candidates for appendectomy. Methods Appendix specimen from responders (N=10) and non-responders (N=10) to appendectomy for therapy refractory UC were included (COSTA-study NCT03912714 or off-label appendectomy). Endoscopic response was defined as steroid-free remission at 12 months without intensified medical treatment. The control group consisted of patients with acute appendicitis(N=2). Post-operative biopsies were taken from the appendix and the coecal base and snap frozen. RNA was isolated and expression profiling was performed using RNAseq. Results Unsupervised clustering indicated two separate clusters, one containing a more stromal phenotype expressing ADH1B, IGFBP5, Desmin and FLNA and the other expressing a more intestinal/B cell phenotype (OLMF4, PIGR, CEACAM5 and IHGA2). Stainings for the presence of collagen (stromal) versus pancytokeratin (epithelial) confirmed this analysis on protein level. Both clusters contained subclusters of responders and non-responders. In both the ‘responder’ subclusters, markers of immune activation were increased, but the type of response differed between ‘stromal responders’ and ‘epithelial responders’. In ‘stromal responders’ increased expression of acute phase genes was present (FABP4, ADIPOQ, SAA1 and CXCL8). In the ‘epithelial responders’, immune activity was more characterized by increased mitochondrial activation/oxidative phosphorylation. In both subclusters, non-responders lacked these immune activation markers. We also evaluated the expression of the differentiating genes in biopsies of the coecal bottom. Indeed, the coecal samples also largerly clustered according to response based on the gene expression profile of the matching appendices. Conclusion Patients responding to appendectomy showed two distinct gene expression patterns, suggesting two separate mechanisms of action. Although in both cases it clearly identified the requirement of active immune responses for response, the identifying markers differed between the two groups. This finding has consequences for both further functional studies into the effector mechanism of appendectomy in UC, but also in future efforts to identify patients suitable for this surgery.

Published
2023
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6. DOP29 Single cell analysis of Crohn’s disease fistula; comparison of different locations

Author

M Becker drs., P Koelink, W Bemelman, C Buskens, and M Wildenberg

Subjects
Gastroenterology, General Medicine
Abstract

Background A common complication of Crohn’s disease(CD) are perianal fistulae, highly impacting the quality of life in these patients. The pathogenesis of these fistulas is incompletely understood. Treatment is often given based on knowledge of disease processes in the inflamed intestine itself, but whether the immune activity in the fistula tract is similar remains unknown. Full characterization of the intestinal mucosa versus the fistula tract itself may reveal immunological differences between the two, and generate leads for both development of new interventional options as well as allow better stratification of patients. Methods Matched biopsies were obtained from the rectal mucosa, the internal opening of the fistula and the tract itself. A total of 30 patients were included (20 CD; 10 non-IBD patients). Tissues were processed to cell suspensions and selected for live CD66b- cells. Single cell RNASeq analysis was performed using 10x Chromium. In total, 111.955 immune cells were annotated and used for analysis. Results T cells comprised the largest cluster of immune cells, which was subclustered into 19 clusters. Both the internal opening and the fistula tracts contained elevated proportions of effector memory and cycling T cells. Conversely, the rectum contained more central and tissue resident memory cells. T cells expressing IL-22, which were previously described in fistula were detected in considerable number in the rectum and internal opening, but were scarce in the fistula tract itself. These cells were clearly distinct from the IL17A producing subset, and IL17F expressing cells and mainly found in the fistula tracts. Only the tract contained a clear subset of CXCL13 expressing T cells, suggesting follicular helper T cells. Presence of these cells was strongly correlated to the presence of lymphoid inducer cells as well as IgG producing B cells rather than the IgA subset found in the rectum. Clusters of myeloid cells were largely derived from the fistula tracts. Subclustering into 10 clusters indicated a shift from anti-inflammatory to pro-inflammatory macrophages in the tracts compared to the rectal mucosa. Finally, a striking difference in NK cells subsets was observed, with significantly more CD56bright NK cells present in the rectal samples while the internal opening and the tracts were comparable. Conclusion Cellular composition and activity of the immune cell compartment differs considerably between the rectum, the internal opening of the fistula and the tract. Subsequently, therapy targeted at processes involved in mucosal inflammation may not actually target the fistula itself, which may explain the low success rate in fistula resolution. Careful analysis of the fistula may help to identify targets useful in the treatment.

Published
2023
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7. Guidelines for the use and interpretation of assays for monitoring autophagy (4th edition)

Author

D. KLIONSKY, A. ABDEL-AZIZ, S. ABDELFATAH, M. ABDELLATIF, A. ABDOLI, S. ABEL, H. ABELIOVICH, M. ABILDGAARD, Y. ABUDU, A. ACEVEDO-AROZENA, I. ADAMOPOULOS, K. ADELI, T. ADOLPH, A. ADORNETTO, E. AFLAKI, G. AGAM, A. AGARWAL, B. AGGARWAL, M. AGNELLO, P. AGOSTINIS, J. AGREWALA, A. AGROTIS, P. AGUILAR, S. AHMAD, Z. AHMED, U. AHUMADA-CASTRO, S. AITS, S. AIZAWA, Y. AKKOC, T. AKOUMIANAKI, H. AKPINAR, A. AL-ABD, L. AL-AKRA, A. AL-GHARAIBEH, M. ALAOUI-JAMALI, S. ALBERTI, E. ALCOCER-GOMEZ, C. ALESSANDRI, M. ALI, M. AL-BARI, S. ALIWAINI, J. ALIZADEH, E. ALMACELLAS, A. ALMASAN, A. ALONSO, G. ALONSO, N. ALTAN-BONNET, D. ALTIERI, S. ALVES, C. DA COSTA, M. ALZAHARNA, M. AMADIO, C. AMANTINI, C. AMARAL, S. AMBROSIO, A. AMER, V. AMMANATHAN, Z. AN, S. ANDERSEN, S. ANDRABI, M. ANDRADE-SILVA, A. ANDRES, S. ANGELINI, D. ANN, U. ANOZIE, M. ANSARI, P. ANTAS, A. ANTEBI, Z. ANTON, T. ANWAR, L. APETOH, N. APOSTOLOVA, T. ARAKI, Y. ARAKI, K. ARASAKI, W. ARAUJO, J. ARAYA, C. ARDEN, M. AREVALO, S. ARGUELLES, E. ARIAS, J. ARIKKATH, H. ARIMOTO, A. ARIOSA, D. ARMSTRONG-JAMES, L. ARNAUNE-PELLOQUIN, A. AROCA, D. ARROYO, I. ARSOV, R. ARTERO, D. ASARO, M. ASCHNER, M. ASHRAFIZADEH, O. ASHUR-FABIAN, A. ATANASOV, A. AU, P. AUBERGER, H. AUNER, L. AURELIAN, R. AUTELLI, L. AVAGLIANO, Y. AVALOS, S. AVEIC, C. AVELEIRA, T. AVINWITTENBERG, Y. AYDIN, S. AYTON, S. AYYADEVARA, M. AZZOPARDI, M. BABA, J. BACKER, S. BACKUES, D. BAE, O. BAE, S. BAE, E. BAEHRECKE, A. BAEK, S. BAEK, G. BAGETTA, A. BAGNIEWSKA-ZADWORNA, H. BAI, J. BAI, X. BAI, Y. BAI, N. BAIRAGI, S. BAKSI, T. BALBI, C. BALDARI, W. BALDUINI, A. BALLABIO, M. BALLESTER, S. BALAZADEH, R. BALZAN, R. BANDOPADHYAY, S. BANERJEE, Y. BAO, M. BAPTISTA, A. BARACCA, C. BARBATI, A. BARGIELA, D. BARILA, P. BARLOW, S. BARMADA, E. BARREIRO, G. BARRETO, J. BARTEK, B. BARTEL, A. BARTOLOME, G. BARVE, S. BASAGOUDANAVAR, D. BASSHAM, R. JR, A. BASU, H. BATOKO, I. BATTEN, E. BAULIEU, B. BAUMGARNER, J. BAYRY, R. BEALE, I. BEAU, F. BEAUMATIN, L. BECHARA, G. BECK, M. BEERS, J. BEGUN, C. BEHRENDS, G. BEHRENS, R. BEI, E. BEJARANO, S. BEL, C. BEHL, A. BELAID, N. BELGAREH-TOUZE, C. BELLAROSA, F. BELLEUDI, M. PEREZ, R. BELLO-MORALES, J. BELTRAN, S. BELTRAN, D. BENBROOK, M. BENDORIUS, B. BENITEZ, I. BENITO-CUESTA, J. BENSALEM, M. BERCHTOLD, S. BEREZOWSKA, D. BERGAMASCHI, M. BERGAMI, A. BERGMANN, L. BERLIOCCHI, C. BERLIOZ-TORRENT, A. BERNARD, L. BERTHOUX, C. BESIRLI, S. BESTEIRO, V. BETIN, R. BEYAERT, J. BEZBRADICA, K. BHASKAR, I. BHATIA-KISSOVA, R. BHATTACHARYA, S. BHATTACHARYA, S. BHATTACHARYYA, M. BHUIYAN, S. BHUTIA, L. BI, X. BI, T. BIDEN, K. BIJIAN, V. BILLES, N. BINART, C. BINCOLETTO, A. BIRGISDOTTIR, G. BJORKOY, G. BLANCO, A. BLAS-GARCIA, J. BLASIAK, R. BLOMGRAN, K. BLOMGREN, J. BLUM, E. BOADA-ROMERO, M. BOBAN, K. BOESZEBATTAGLIA, P. BOEUF, B. BOLAND, P. BOMONT, P. BONALDO, S. BONAM, L. BONFILI, J. BONIFACINO, B. BOONE, M. BOOTMAN, M. BORDI, C. BORNER, B. BORNHAUSER, G. BORTHAKUR, J. BOSCH, S. BOSE, L. BOTANA, J. BOTAS, C. BOULANGER, M. BOULTON, M. BOURDENX, B. BOURGEOIS, N. BOURKE, G. BOUSQUET, P. BOYA, P. BOZHKOV, L. BOZI, T. BOZKURT, D. BRACKNEY, C. BRANDTS, R. BRAUN, G. BRAUS, R. BRAVO-SAGUA, J. BRAVO-SAN PEDRO, P. BREST, M. BRINGER, A. BRIONES-HERRERA, V. BROADDUS, P. BRODERSEN, E. ALVAREZ, J. BRODSKY, S. BRODY, P. BRONSON, J. BRONSTEIN, C. BROWN, R. BROWN, P. BRUM, J. BRUMELL, N. BRUNETTI-PIERRI, D. BRUNO, R. BRYSON-RICHARDSON, C. BUCCI, C. BUCHRIESER, M. BUENO, L. BUITRAGO-MOLINA, S. BURASCHI, S. BUCH, J. BUCHAN, E. BUCKINGHAM, H. BUDAK, M. BUDINI, G. BULTYNCK, F. BURADA, J. BURGOYNE, M. BURON, V. BUSTOS, S. BUTTNER, E. BUTTURINI, A. BYRD, I. CABAS, S. CABRERA-BENITEZ, K. CADWELL, J. CAI, L. CAI, Q. CAI, M. CAIRO, J. CALBET, G. CALDWELL, K. CALDWELL, J. CALL, R. CALVANI, A. CALVO, M. BARRERA, N. CAMARA, J. CAMONIS, N. CAMOUGRAND, M. CAMPANELLA, E. CAMPBELL, F. CAMPBELL-VALOIS, S. CAMPELLO, I. CAMPESI, J. CAMPOS, O. CAMUZARD, J. CANCINO, D. DE ALMEIDA, L. CANESI, I. CANIGGIA, B. CANONICO, C. CANTI, B. CAO, M. CARAGLIA, B. CARAMES, E. CARCHMAN, E. CARDENAL-MUNOZ, C. CARDENAS, L. CARDENAS, S. CARDOSO, J. CAREW, G. CARLE, G. CARLETON, S. CARLONI, D. CARMONA-GUTIERREZ, L. CARNEIRO, O. CARNEVALI, J. CAROSI, S. CARRA, A. CARRIER, L. CARRIER, B. CARROLL, A. CARTER, A. CARVALHO, M. CASANOVA, C. CASAS, J. CASAS, C. CASSIOLI, E. CASTILLO, K. CASTILLO, S. CASTILLO-LLUVA, F. CASTOLDI, M. CASTORI, A. CASTRO, M. CASTRO-CALDAS, J. CASTRO-HERNANDEZ, S. CASTRO-OBREGON, S. CATZ, C. CAVADAS, F. CAVALIERE, G. CAVALLINI, M. CAVINATO, M. CAYUELA, P. RICA, V. CECARINI, F. CECCONI, M. CECHOWSKA-PASKO, S. CENCI, V. CEPERUELO-MALLAFRE, J. CERQUEIRA, J. CERUTTI, D. CERVIA, V. CETINTAS, S. CETRULLO, H. CHAE, A. CHAGIN, C. CHAI, G. CHAKRABARTI, O. CHAKRABARTI, T. CHAKRABORTY, M. CHAMI, G. CHAMILOS, D. CHAN, E. CHAN, H. CHAN, M. CHAN, Y. CHAN, P. CHANDRA, C. CHANG, H. CHANG, K. CHANG, J. CHAO, T. CHAPMAN, N. CHARLET-BERGUERAND, S. CHATTERJEE, S. CHAUBE, A. CHAUDHARY, S. CHAUHAN, E. CHAUM, F. CHECLER, M. CHEETHAM, C. CHEN, G. 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CRESPO, A. CRIOLLO, V. CRIPPA, R. CRISTOFANI, T. CSIZMADIA, A. CUADRADO, B. CUI, J. CUI, Y. CUI, E. CULETTO, A. CUMINO, A. CYBULSKY, M. CZAJA, S. CZUCZWAR, S. D'ADAMO, M. D'AMELIO, D. D'ARCANGELO, A. D'LUGOS, G. D'ORAZI, J. DA SILVA, H. DAFSARI, R. DAGDA, Y. DAGDAS, M. DAGLIA, X. DAI, Y. DAI, J. DAL COL, P. DALHAIMER, L. DALLA VALLE, T. DALLENGA, G. DALMASSO, M. DAMME, I. DANDO, N. DANTUMA, A. DARLING, H. DAS, S. DASARATHY, S. DASARI, S. DASH, O. DAUMKE, A. DAUPHINEE, J. DAVIES, V. DAVILA, R. DAVIS, T. DAVIS, S. NAIDU, F. DE AMICIS, K. DE BOSSCHER, F. DE FELICE, L. DE FRANCESCHI, C. DE LEONIBUS, M. BARBOSA, G. DE MEYER, A. DE MILITO, C. DE NUNZIO, C. DE PALMA, M. DE SANTI, C. DE VIRGILIO, D. DE ZIO, J. DEBNATH, B. DEBOSCH, J. DECUYPERE, M. DEEHAN, G. DEFLORIAN, J. DEGREGORI, B. DEHAY, G. DEL RIO, J. DELANEY, L. DELBRIDGE, E. DELORME-AXFORD, M. DELPINO, F. DEMARCHI, V. DEMBITZ, N. DEMERS, H. DENG, Z. DENG, J. DENGJEL, P. DENT, D. DENTON, M. DEPAMPHILIS, C. DER, V. DERETIC, A. 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PALMIERI, G. PALMISANO, C. PALUMBO, F. PAMPALONI, L. PAN, Q. PAN, W. PAN, X. PAN, G. PANASYUK, R. PANDEY, U. PANDEY, V. PANDYA, F. PANENI, S. PANG, E. PANZARINI, D. PAPADEMETRIO, E. PAPALEO, D. PAPINSKI, D. PAPP, E. PARK, H. PARK, J. PARK, S. PARK, A. PAROLA, J. PARYS, A. PASQUIER, B. PASQUIER, J. PASSOS, N. PASTORE, H. PATEL, D. PATSCHAN, S. PATTINGRE, G. PEDRAZA-ALVA, J. PEDRAZA-CHAVERRI, Z. PEDROZO, G. PEI, J. PEI, H. PELED-ZEHAVI, J. PELLEGRINI, J. PELLETIER, M. PENALVA, D. PENG, Y. PENG, F. PENNA, M. PENNUTO, F. PENTIMALLI, C. PEREIRA, G. PEREIRA, L. PEREIRA, L. DE ALMEIDA, N. PERERA, A. PEREZOLIVA, M. PEREZ-PEREZ, P. PERIYASAMY, A. PERL, C. PERROTTA, I. PERROTTA, R. PESTELL, M. PETERSEN, I. PETRACHE, G. PETROVSKI, T. PFIRRMANN, A. PFISTER, J. PHILIPS, H. PI, A. PICCA, A. PICKRELL, S. PICOT, G. PIERANTONI, M. PIERDOMINICI, P. PIERRE, V. PIERREFITE-CARLE, K. PIERZYNOWSKA, F. PIETROCOLA, M. PIETRUCZUK, C. PIGNATA, F. PIMENTELMUINOS, M. PINAR, R. PINHEIRO, R. PINKAS-KRAMARSKI, P. 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TUXWORTH, J. TYLER, E. TYUTEREVA, Y. UCHIYAMA, A. UGUNKLUSEK, H. UHLIG, I. ULASOV, M. UMEKAWA, C. UNGERMANN, R. UNNO, S. URBE, E. URIBE-CARRETERO, S. USTUN, V. UVERSKY, T. VACCARI, M. VACCARO, B. VAHSEN, H. VAKIFAHMETOGLU-NORBERG, R. VALDOR, M. VALENTE, A. VALKO, R. VALLEE, A. VALVERDE, G. VAN DEN BERGHE, S. VAN DER VEEN, L. VAN KAER, J. VAN LOOSDREGT, S. VAN WIJK, W. VANDENBERGHE, I. VANHOREBEEK, M. VANNIER-SANTOS, N. VANNINI, M. VANRELL, C. VANTAGGIATO, G. VARANO, I. VARELA-NIETO, M. VARGA, M. VASCONCELOS, S. VATS, D. VAVVAS, I. VEGANAREDO, S. VEGA-RUBIN-DE-CELIS, G. VELASCO, A. VELAZQUEZ, T. VELLAI, E. VELLENGA, F. VELOTTI, M. VERDIER, P. VERGINIS, I. VERGNE, P. VERKADE, M. VERMA, P. VERSTREKEN, T. VERVLIET, J. VERVOORTS, A. VESSONI, V. VICTOR, M. VIDAL, C. VIDONI, O. VIEIRA, R. VIERSTRA, S. VIGANO, H. VIHINEN, V. VIJAYAN, M. VILA, M. VILAR, J. VILLALBA, A. VILLALOBO, B. VILLAREJO-ZORI, F. VILLARROYA, J. VILLARROYA, O. VINCENT, C. VINDIS, C. VIRET, M. VISCOMI, D. VISNJIC, I. VITALE, D. VOCADLO, O. VOITSEKHOVSKAJA, C. VOLONTE, M. VOLTA, M. VOMERO, C. VON HAEFEN, M. VOOIJS, W. VOOS, L. VUCICEVIC, R. WADE-MARTINS, S. WAGURI, K. WAITE, S. WAKATSUKI, D. WALKER, M. WALKER, S. WALKER, J. WALTER, F. WANDOSELL, B. WANG, C. WANG, D. WANG, F. WANG, G. WANG, H. WANG, J. WANG, K. WANG, L. WANG, M. WANG, N. WANG, P. WANG, Q. WANG, W. WANG, X. WANG, Y. WANG, Z. WANG, G. WARNES, V. WARNSMANN, H. WATADA, E. WATANABE, M. WATCHON, T. WEAVER, G. WEGRZYN, A. WEHMAN, H. WEI, L. WEI, T. WEI, Y. WEI, O. WEIERGRABER, C. WEIHL, G. WEINDL, R. WEISKIRCHEN, A. WELLS, R. WEN, X. WEN, A. WERNER, B. WEYKOPF, S. WHEATLEY, J. WHITTON, A. WHITWORTH, K. WIKTORSKA, M. WILDENBERG, T. WILEMAN, S. WILKINSON, D. WILLBOLD, B. WILLIAMS, R. WILLIAMS, P. WILLIAMSON, R. WILSON, B. WINNER, N. WINSOR, S. WITKIN, H. WODRICH, U. WOEHLBIER, T. WOLLERT, E. WONG, J. WONG, R. WONG, V. WONG, W. WONG, A. WU, C. WU, J. WU, K. WU, M. WU, S. WU, W. WU, X. WU, Y. WU, R. XAVIER, H. XIA, L. XIA, Z. XIA, G. XIANG, J. XIANG, M. XIANG, W. XIANG, B. XIAO, G. XIAO, H. XIAO, J. XIAO, L. XIAO, S. XIAO, Y. XIAO, B. XIE, C. XIE, M. XIE, Y. XIE, Z. XIE, M. XILOURI, C. XU, E. XU, H. XU, J. XU, L. XU, W. XU, X. XU, Y. XUE, S. YAKHINE-DIOP, M. YAMAGUCHI, O. YAMAGUCHI, A. YAMAMOTO, S. YAMASHINA, S. YAN, Z. YAN, Y. YANAGI, C. YANG, D. YANG, H. YANG, J. YANG, L. YANG, M. YANG, P. YANG, Q. YANG, S. YANG, W. YANG, X. YANG, Y. YANG, H. YAO, S. YAO, X. YAO, Y. YAO, T. YASUI, M. YAZDANKHAH, P. YEN, C. YI, X. YIN, Y. YIN, Z. YIN, M. YING, Z. YING, C. YIP, S. YIU, Y. YOO, K. YOSHIDA, S. YOSHII, T. YOSHIMORI, B. YOUSEFI, B. YU, H. YU, J. YU, L. YU, M. YU, S. YU, V. YU, W. YU, Z. YU, J. YUAN, L. YUAN, S. YUAN, Y. YUAN, Z. YUAN, J. YUE, Z. YUE, J. YUN, R. YUNG, D. ZACKS, G. ZAFFAGNINI, V. ZAMBELLI, I. ZANELLA, Q. ZANG, S. ZANIVAN, S. ZAPPAVIGNA, P. ZARAGOZA, K. ZARBALIS, A. ZAREBKOHAN, A. ZARROUK, S. ZEITLIN, J. ZENG, E. ZEROVNIK, L. ZHAN, B. ZHANG, D. ZHANG, H. ZHANG, J. ZHANG, K. ZHANG, L. ZHANG, M. ZHANG, P. ZHANG, S. ZHANG, W. ZHANG, X. ZHANG, Y. ZHANG, Z. ZHANG, H. ZHAO, L. ZHAO, S. ZHAO, T. ZHAO, X. ZHAO, Y. ZHAO, G. ZHENG, K. ZHENG, L. ZHENG, S. ZHENG, X. ZHENG, Y. ZHENG, Z. ZHENG, B. ZHIVOTOVSKY, Q. ZHONG, A. ZHOU, B. ZHOU, C. ZHOU, G. ZHOU, H. ZHOU, J. ZHOU, K. ZHOU, R. ZHOU, X. ZHOU, Y. ZHOU, Z. ZHOU, B. ZHU, C. ZHU, G. ZHU, H. ZHU, W. ZHU, Y. ZHU, H. ZHUANG, X. ZHUANG, K. ZIENTARA-RYTTER, C. ZIMMERMANN, E. ZIVIANI, T. ZOLADEK, W. ZONG, D. ZOROV, A. ZORZANO, W. ZOU, Z. ZOU, S. ZURYN, W. ZWERSCHKE, B. BRAND-SABERI, C. KENCHAPPA, S. OSHIMA, Y. RONG, J. SLUIMER, and C. STALLINGS

Subjects
flux, macroautophagy, phagophore, stress, vacuole, Autophagosome, LC3, lysosome, neurodegeneration, cancer
Abstract

In 2008, we published the first set of guidelines for standardizing research in autophagy. Since then, this topic has received increasing attention, and many scientists have entered the field. Our knowledge base and relevant new technologies have also been expanding. Thus, it is important to formulate on a regular basis updated guidelines for monitoring autophagy in different organisms. Despite numerous reviews, there continues to be confusion regarding acceptable methods to evaluate autophagy, especially in multicellular eukaryotes. Here, we present a set of guidelines for investigators to select and interpret methods to examine autophagy and related processes, and for reviewers to provide realistic and reasonable critiques of reports that are focused on these processes. These guidelines are not meant to be a dogmatic set of rules, because the appropriateness of any assay largely depends on the question being asked and the system being used. Moreover, no individual assay is perfect for every situation, calling for the use of multiple techniques to properly monitor autophagy in each experimental setting. Finally, several core components of the autophagy machinery have been implicated in distinct autophagic processes (canonical and noncanonical autophagy), implying that genetic approaches to block autophagy should rely on targeting two or more autophagy-related genes that ideally participate in distinct steps of the pathway. Along similar lines, because multiple proteins involved in autophagy also regulate other cellular pathways including apoptosis, not all of them can be used as a specific marker for bona fide autophagic responses. Here, we critically discuss current methods of assessing autophagy and the information they can, or cannot, provide. Our ultimate goal is to encourage intellectual and technical innovation in the field.

Published
2021

8. P103 Identifying CES-1 positive myeloid cells with mass cytometry as a potential therapeutic target for Crohn’s disease

Author

I Hageman MD, A Elfiky, M Becker, V Joustra, C Buskens, A Li Yim, T Hakvoort, J Verhoeff, J Garcia Vallejo, G D’Haens, M Wildenberg, and W De Jonge

Subjects
Gastroenterology, General Medicine
Abstract

Background Myeloid cells are key players in the sustained inflammatory process in Crohn’s disease (CD), and are therefore therapeutic targets. Human mononuclear myeloid cells express the cell selective expression of carboxylesterase 1 (CES-1). Esterase-sensitive motif (ESM) technology has been developed to specifically target ESM tagged small molecules on human mononuclear myeloid cells, based on their CES-1 expression, creating a larger therapeutic window with less side effects. This study investigates the presence of CES-1 expressing cells in PBMCs, intestinal biopsies and fistula cells of CD patients, highlighting the potential of ESM-technology in CD. This could benefit CD treatment in non-responsive patients or patients with a fistulizing phenotype. Methods Intestinal biopsies were collected from CD patients who underwent endoscopy. PBMCs were collected from CD patients who were receiving Vedolizumab (VDZ) treatment and categorized as responders or non-responders. Criteria for response was based on clinical (HBI ≤4) and biochemical (fecal calprotectine ≤250 µg/g) and/or endoscopic assessment (≥50% reduction in SES-CD score). Fistula tissue was of CD patients was obtained during surgery. CES1-positive cells were phenotyped by mass cytometry with a panel of 32-markers, including the myeloid markers CES-1, CD14, CD16, HLA-DR, CD11c, CD206. Results We demonstrated the presence of CD14+HLADR+CD11a+CD44+CD11b+ myeloid cells exclusively expressing CES-1 in intestinal biopsies of CD patients (n=7). CES-1 expression did not differ between inflamed and uninflamed tissue. In PBMCs of VDZ responders (n=5) and non-responders (n=6), we were able to identify different subsets of myeloid cells such as classical (CD14++CD16-), intermediate(CD14+CD16+) and non-classical (CD14-CD16++) monocytes, DCs and CD2+ DC precursors all demonstrating CES-1 expression. Percentage of CES-1 positive cells in inflamed PBMCs (or non-responders) was higher than in uninflamed PBMCs (responders) in all of the above-mentioned subsets. Furthermore, we demonstrated different myeloid populations within fistula cells (n=13) such as pDCs (CD123+CD68+HLA-DR+), DCs (CD11c+CD11b+HLA-DR+ CD44+), CD141+DCs (CD141+CD49d+HLA-DR+),Macrophage (CD14+CD11c+CD11b+HLA-DR+CD44+) and M2 Macrophages (CD14+CD163+CD206+CD11c+CD11b+HLA-DR+CD44+), all expressing CES-1. Conclusion We demonstrated specific CES-1 positive myeloid cells within intestinal biopsies, PBMCs of non-responsive patients, and fistulae samples of CD patients. Targeting CES-1+ with ESM-technology could provide new therapeutic targets for CD.

Published
2022
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9. P077 Different granulocyte subsets are involved in the pathogenesis in Crohn’s disease fistula

Author

M Becker drs, P Koelink, M de Krijger, W Bemelman, W de Jonge, C Buskens, and M Wildenberg

Subjects
Gastroenterology, General Medicine
Abstract

Background Early in inflammation granulocytes migrate from the circulation into inflamed areas and rapidly accumulate. They form an important role in first defense as well as the orchestration of downstream immune responses. In the circulation, several different subsets of granulocytes have been described, for example immature, intermediate and mature granulocytes. Surprisingly, despite their abundance and prominent function, this cell type is rarely studied from a tissue perspective, most likely due to their short lifespan ex vivo. One example of gastrointestinal disease with an extremely high number of granulocytes is the occurrence of perianal fistula which contain a considerable amount of granulation tissue. As curettage of this tissue is part of the treatment, the fistula tract is good source to study tissue derived granulocyte subsets. The aim of this study was to characterize granulocyte subsets in the fistula tract. Methods Curettage material of perianal fistula tracts was obtained during surgical intervention from patients with CD (n=15) and cryptoglandular fistulas (n=5). From 2 CD patients venous blood was taken and a ficoll step was performed. Single-cell suspensions were stained with a 35-antibody panel, including CD66, CD14, CD33, CD16 and CD11b. Samples were analyzed using mass cytometry (CyTOF). Results Granulocytes, characterized as CD45+CD66a+ cells, were the main cellular component of the fistula tract (64%±24) compared to other immune cells. This was more pronounced in cryptoglandular fistula compared to CD fistula (81% vs. 67%; p=0.019). Within these cells we could differentiate between 4 subsets, mature granulocytes (CD33+CD11b+CD16+), intermediate (CD33intCD11b+CD16int), immature granulocytes (CD33intCD11b-CD16int) and eosinophils (CD33-CD11b-CD16-). Immature granulocytes were the most abundant in de fistula tract (31%±15), closely followed by the mature (30%±23) and intermediate (23±9). No significant difference was seen in the percentages of granulocyte subsets between CD and cryptoglandular fistula. Conclusion Here, we show the presence of different subtypes of granulocytes in fistula derived granulation tissue. We show a heterogeneous group of cells ranging from immature neutrophils to mature neutrophils. Due to their high number in active ulcerating tissue, further evaluation of granulocyte subsets and function could be an interesting target for medical treatment options.

Published
2022
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10. LB01 SIX WEEKS OF SOFOSBUVIR/LEDIPASVIR TREATMENT OF ACUTE HEPATITIS C VIRUS GENOTYPE 1 MONOINFECTION: FINAL RESULTS OF THE THE GERMAN HEPNET ACUTE HCV IV STUDY

Author

K. Deterding, C.D. Spinner, E. Schott, T.M. Welzel, G. Gerken, H. Klinker, U. Spengler, J. Wiegand, J. Schulze zur Wiesch, A. Pathil, M. Cornberg, A. Umgelter, C. Zöllner, S. Zeuzem, A. Papkalla, K. Weber, S. Hardtke, H. Leyen, A. Koch, D. Witzendorff, M. Manns, H. Wedemeyer, C.M. Preda, C.P. Popescu, C. Baicus, M. Manuc, R. Voiosu, E. Ceausu, L. Fulger, A. Nisanian, C.S. Pop, A. Oproiu, A. Arezzo, R. Passera, A. Bullano, Y. Mintz, A. KEDAR, L. Boni, E. Cassinotti, R. Rosati, U. Fumagalli, M. Sorrentino, M. Brizzolari, N. Di Lorenzo, A.L. Gaspari, D. Andreone, E. De Stefani, G. Navarra, S. Lazzara, M. Degiuli, K. Shishin, I. Khatkov, I. Kazakov, R. Schrittwieser, T. Carus, A. Corradi, G. Sitzman, A. Lacy, S. Uranues, A. Szold, M.A. Bonino, M. Morino, J. Strömberg, G. Sandblom, R. Coelen, M. Gaspersz, T. Labeur, J. Vugt, S. Dieren, F. Willemssen, C.Y. Nio, J. IJzermans, H.‐J. Klümpen, B. Groot Koerkamp, T. Gulik, R. Sturgess, D. Palmer, J. Trojan, A. Hoffmeister, B. Neu, S. Kasper, A. Dechêne, C. Jürgensen, J. Schirra, R. Jakobs, A. Høgset, L. Finnesand, A.E. Abd Elrazek, S. Saab, T. Salem, M. Abdel‐Aty, B. Hawary, A. Ismail, M. Zayied, M. Alboraie, R. Orenstein, E. Dubberke, C.H. Lee, S. Khanna, G. Hecht, S. Wong, T. Kwong, X. Wang, R.S.Y. Tang, S.C. Ng, J.J.Y. Sung, J. Yu, S. Ott, G.H. Waetzig, A. Rehmann, J. Moltzau‐Anderson, R. Bharti, J.A. Grasis, L. Cassidy, A. Tholey, H. Fickenscher, D. Seegert, P. Rosenstiel, S. Schreiber, T. Mazzawi, G.A. Lied, M. El‐Salhy, O.H. Gilja, J.G. Hatlebakk, T. Hausken, S.T. Witt, O. Bednarska, A. Icenhour, S. Elsenbruch, M. Ström, J.D. Söderholm, M. Engström, E.A. Mayer, Å. Keita, S. Walter, P.K. Kump, P. Wurm, H.P. Gröchenig, H.H. Wenzl, W. Petritsch, B. Halwachs, M. Wagner, V. Stadlbauer‐Köllner, A.J. Eherer, K.M. Hoffmann, A. Deutschmann, G. Reicht, L. Reiter, P. Slawitsch, G. Gorkiewicz, C. Hoegenauer, Y. Zhou, R. Kakuturu, D. Jung, K.K. Jørgensen, I.C. Olsen, G.L. Goll, M. Lorentzen, N. Bolstad, E.A. Haavardsholm, K.E. Lundin, C. Mørk, J. Jahnsen, T.K. Kvien, B.G. Feagan, B.E. Sands, G. Rossiter, X. Li, K. Usiskin, X. Zhan, J.‐F. Colombel, W.J. Sandborn, J. Panés, M. Ferrante, E. Louis, G. D'Haens, D. Franchimont, A. Kaser, O. Dewit, U. Seidler, K.‐J. Kim, M.F. Neurath, P. Scholl, S. Visvanathan, S.J. Padula, I. Herichova, N. Sha, D. Hall, W.O. Böcher, F. Bloemendaal, A. Levin, M. Wildenberg, P. Koelink, S. Verbeek, J. Claassens, B. Mcrae, G. Vidarsson, G.R. Brink, M. Badke, S. Rose‐John, M.E. Spehlmann, L. Peyrin‐Biroulet, J. Gatlin, M. Soloman, D. Unett, H. Al‐Shamma, D. Behan, J. Langhorst, J. Boone, A. Rueffer, G. Dobos, K. Beiderwellen, T. Lauenstein, W.S. Ngu, R. Bevan, Z.P. Tsiamoulos, P. Bassett, Z. Hoare, M. Rutter, N. Totton, T.J. Lee, A.V. Ramadas, J. Silcock, J. Painter, L.J. Neilson, B.P. Saunders, C.J. Rees, A. Schmidt, S. Goelder, H. Messmann, M. Goetz, T. Kratt, A. Meining, M. Birk, J. Delius, M. Albert, J.Y.W. Escher, A. Lau, R. Hoffman, K. Wiest, null Caca, A. Siddiqui, D. Wilson, M. Cangelosi, R. Rameshshanker, P. Wall, K. Cocks, T. Doulton, A. Yusuf, C. Hancock, R. Valori, A. Aravani, J. Rashbass, S. Vernon, E.J.A. Morris, J.H. ‐Choi, D.‐W. Seo, T.J. Song, D.H. Park, S.S. Lee, S.K. Lee, ‐H. Kim, P. Somani, and M. Sharma

Subjects
0301 basic medicine, Ledipasvir, Sofosbuvir, business.industry, Gastroenterology, Virology, Virus, 03 medical and health sciences, chemistry.chemical_compound, 030104 developmental biology, 0302 clinical medicine, Oncology, chemistry, Genotype, Medicine, 030211 gastroenterology & hepatology, Acute hepatitis C, business, medicine.drug
Published
2016
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12. Potential to reduce pesticides in intensive apple production through management practices could be challenged by climatic extremes.

Author

Zaller JG, Oswald A, Wildenberg M, Burtscher-Schaden H, Nadeem I, Formayer H, and Paredes D

Subjects
Malus, Pesticides, Climate, Agriculture
Abstract

Apples are the third most produced fruit in the world, but their production is often pesticide-intensive. Our objective was to identify options for pesticide reduction using farmer records from 2549 commercial apple fields in Austria during five years between 2010 and 2016. Using generalized additive mixed modeling, we examined how pesticide use was related to farm management, apple varieties, and meteorological parameters, and how it affected yields and toxicity to honeybees. Apple fields received 29.5 ± 8.6 (mean ± SD) pesticide applications per season at a rate of 56.7 ± 22.7 kg ha -1 , which included a total of 228 pesticide products with 80 active ingredients. Over the years, fungicides accounted for 71 % of the pesticide amounts applied, insecticides for 15 %, and herbicides for 8 %. The most frequently used fungicides were sulfur (52 %), followed by captan (16 %) and dithianon (11 %). Of insecticides, paraffin oil (75 %) and chlorpyrifos/chlorpyrifos-methyl (6 % combined) were most frequently used. Among herbicides, glyphosate (54 %), CPA (20 %) and pendimethalin (12 %) were most often used. Pesticide use increased with increasing frequency of tillage and fertilization, increasing field size, increasing spring temperatures, and drier summer conditions. Pesticide use decreased with increasing number of summer days with maximum temperatures >30 °C and number of warm, humid days. Apple yields were significantly positively related to the number of heat days, warm humid nights, and pesticide treatment frequency, but were not affected by frequency of fertilization and tillage. Honeybee toxicity was not related to insecticide use. Pesticide use and yield were significantly related to apple varieties. Our analysis shows that pesticide use in the apple farms studied can be reduced by less fertilization and tillage, partly because yields were >50 % higher than the European average. However, weather extremes related to climate change, such as drier summers, could challenge plans to reduce pesticide use., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 The Authors. Published by Elsevier B.V. All rights reserved.)

Published
2023
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13. Peripheral Blood DNA Methylation Profiles Do Not Predict Endoscopic Post-Operative Recurrence in Crohn's Disease Patients.

Author

Joustra VW, Li Yim AYF, de Bruyn JR, Duijvestein M, Hageman IL, de Jonge WJ, Henneman P, Wildenberg M, and D'Haens G

Subjects
DNA Methylation, Endoscopy, Epigenome, Humans, Postoperative Period, Recurrence, Crohn Disease genetics, Crohn Disease surgery
Abstract

Prediction of endoscopic post-operative recurrence (POR) in Crohn's disease (CD) patients following ileocolonic resection (ICR) using clinical risk factors alone has thus far been inadequate. While peripheral blood leukocyte (PBL) DNA methylation has shown promise as a tool for predicting recurrence in cancer, no data in CD patients exists. Therefore, this study explored the association and predictive value of PBL DNA methylation in CD patients following ICR. From a cohort of 117 CD patients undergoing ICR, epigenome-wide PBL methylation profiles from 25 carefully selected patients presenting either clear endoscopic remission ( n = 12) or severe recurrence ( n = 13) were assessed using the Illumina MethylationEPIC (850K) array. No statistically significant differentially methylated positions (DMPs) or regions (DMRs) associated with endoscopic POR were identified (FDR p ≤ 0.05), further evidenced by the low accuracy (0.625) following elastic net classification analysis. Nonetheless, interrogating the most significant differences in methylation suggested POR-associated hypermethylation in the MBNL1 , RAB29 and LEPR genes, respectively, which are involved in intestinal fibrosis, inflammation and wound healing. Notably, we observed a higher estimated proportion of monocytes in endoscopic POR compared to remission. Altogether, we observed limited differences in the genome-wide DNA methylome among CD patients with and without endoscopic POR. We therefore conclude that PBL DNA methylation is not a feasible predictive tool in post-operative CD.

Published
2022
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14. High-Dose Vitamin D Does Not Prevent Postoperative Recurrence of Crohn's Disease in a Randomized Placebo-Controlled Trial.

Author

de Bruyn JR, Bossuyt P, Ferrante M, West RL, Dijkstra G, Witteman BJ, Wildenberg M, Hoentjen F, Franchimont D, Clasquin E, van der Bilt JD, Tollens T, Bemelman WA, D'Hoore A, Duijvestein M, and D'Haens GR

Subjects
Humans, Neoplasm Recurrence, Local, Quality of Life, Vitamin D, Vitamins, Crohn Disease prevention & control, Crohn Disease surgery
Abstract

Background & Aims: Vitamin D deficiency is common in Crohn's disease (CD). High-dose vitamin D had anti-inflammatory effects in preclinical studies and trials of patients with CD. We performed a randomized trial to determine whether high-dose vitamin D prevents postoperative recurrence of CD after ileocolonic resection., Methods: Patients with CD after ileocolonic resection with ileocolonic anastomosis were assigned randomly to groups given weekly 25,000 IU oral vitamin D (n = 72) or placebo (n = 71) for 26 weeks, at 17 hospitals in The Netherlands and Belgium, from February 2014 through June 2017. Patients were assessed at baseline and at weeks 2, 6, 12, and 26 for laboratory and clinical parameters, and underwent ileocolonoscopy at 26 weeks. The primary end point was endoscopic recurrence (modified Rutgeerts score, ≥i2b, as assessed by blinded readers) at 26 weeks. Secondary end points included clinical recurrence (Crohn's disease activity index, ≥220), quality of life (measured by the 36-Item Short Form Health Survey, Inflammatory Bowel Disease Questionnaire, and EuroQol, a 5-dimension questionnaire), and outcomes associated with the baseline serum concentration of vitamin D., Results: In the vitamin D group, serum levels of 25-hydroxy vitamin D increased from a median of 42 nmol/L at baseline to 81 nmol/L at week 26 (P < .00001), whereas levels did not change significantly in the placebo group and remained unchanged at 43 nmol/L. In the intention-to-treat analysis, the proportion of patients with endoscopic recurrence at 26 weeks did not differ significantly between the vitamin D vs the placebo group (58% vs 66%; P = .37). The cumulative rate of clinical recurrence did not differ significantly between the groups (18.1% in the vitamin D group vs 18.3% in the placebo group; P = .91). Quality of life improved slightly over time in both groups, but did not differ significantly between groups (P = .07). There were few adverse events in either group., Conclusions: High-dose vitamin D, compared with placebo, did not reduce the incidence of postoperative endoscopic or clinical recurrence of CD in patients who underwent ileocolonic resection with ileocolonic anastomosis. ClinicalTrials.gov no: NCT02010762., (Copyright © 2021 The Authors. Published by Elsevier Inc. All rights reserved.)

Published
2021
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15. Hepatic uptake of conjugated bile acids is mediated by both sodium taurocholate cotransporting polypeptide and organic anion transporting polypeptides and modulated by intestinal sensing of plasma bile acid levels in mice.

Author

Slijepcevic D, Roscam Abbing RLP, Katafuchi T, Blank A, Donkers JM, van Hoppe S, de Waart DR, Tolenaars D, van der Meer JHM, Wildenberg M, Beuers U, Oude Elferink RPJ, Schinkel AH, and van de Graaf SFJ

Subjects
Animals, Cell Line, Cholesterol 7-alpha-Hydroxylase metabolism, Down-Regulation, Female, Humans, Ileum metabolism, Lipopeptides, Male, Mice, Inbred C57BL, Mice, Knockout, Protein Isoforms metabolism, Rats, Bile Acids and Salts metabolism, Enterocytes physiology, Fibroblast Growth Factors metabolism, Liver metabolism, Organic Anion Transporters, Sodium-Dependent metabolism, Symporters metabolism
Abstract

The Na + -taurocholate cotransporting polypeptide (NTCP/SLC10A1) is believed to be pivotal for hepatic uptake of conjugated bile acids. However, plasma bile acid levels are normal in a subset of NTCP knockout mice and in mice treated with myrcludex B, a specific NTCP inhibitor. Here, we elucidated which transport proteins mediate the hepatic uptake of conjugated bile acids and demonstrated intestinal sensing of elevated bile acid levels in plasma in mice. Mice or healthy volunteers were treated with myrcludex B. Hepatic bile acid uptake kinetics were determined in wild-type (WT), organic anion transporting polypeptide (OATP) knockout mice (lacking Slco1a/1b isoforms), and human OATP1B1-transgenic mice. Effects of fibroblast growth factor 19 (FGF19) on hepatic transporter mRNA levels were assessed in rat hepatoma cells and in mice by peptide injection or adeno-associated virus-mediated overexpression. NTCP inhibition using myrcludex B had only moderate effects on bile acid kinetics in WT mice, but completely inhibited active transport of conjugated bile acid species in OATP knockout mice. Cholesterol 7α-hydroxylase Cyp7a1 expression was strongly down-regulated upon prolonged inhibition of hepatic uptake of conjugated bile acids. Fgf15 (mouse counterpart of FGF19) expression was induced in hypercholanemic OATP and NTCP knockout mice, as well as in myrcludex B-treated cholestatic mice, whereas plasma FGF19 was not induced in humans treated with myrcludex B. Fgf15/FGF19 expression was induced in polarized human enterocyte-models and mouse organoids by basolateral incubation with a high concentration (1 mM) of conjugated bile acids., Conclusion: NTCP and OATPs contribute to hepatic uptake of conjugated bile acids in mice, whereas the predominant uptake in humans is NTCP mediated. Enterocytes sense highly elevated levels of (conjugated) bile acids in the systemic circulation to induce FGF15/19, which modulates hepatic bile acid synthesis and uptake. (Hepatology 2017;66:1631-1643)., (© 2017 The Authors. Hepatology published by Wiley Periodicals, Inc., on behalf of the American Association for the Study of Liver Diseases.)

Published
2017
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