TY - JOUR AU - Hård Af Segerstad, E.M.* AU - Mramba, L.K.* AU - Aronsson, C.A.* AU - Uusitalo, U.* AU - Virtanen, S.M.* AU - Agardh, D.* AU - TEDDY Study Group (Ziegler, A.-G.) C1 - 74256 C2 - 57256 TI - Early dietary fiber intake reduces celiac disease risk in genetically prone children: Insights from the TEDDY study. JO - Gastroenterology PY - 2025 SN - 0016-5085 ER - TY - JOUR AU - Kotlarz, D.M. C1 - 74206 C2 - 57368 TI - The paradox of type III interferons: A delicate balance between protection and pathology in colitis development and gut tissue repair. JO - Gastroenterology VL - 169 IS - 4 PY - 2025 SN - 0016-5085 ER - TY - JOUR AU - Kotlarz, D.M. C1 - 75185 C2 - 57831 TI - Unraveling the genetic mosaic of congenital enteropathies. JO - Gastroenterology VL - 169 IS - 6 PY - 2025 SN - 0016-5085 ER - TY - JOUR AU - Kotlarz, D.M. C1 - 68888 C2 - 55007 SP - 711-712 TI - Unveiling the protective role of BTNL-selected γδ T cells in inflammatory bowel disease. JO - Gastroenterology VL - 166 IS - 4 PY - 2024 SN - 0016-5085 ER - TY - JOUR AU - Kotlarz, D.M. C1 - 71655 C2 - 56123 SP - 1048-1049 TI - Defining the role of immune microniches in intestinal effector regulatory T-cell functionality. JO - Gastroenterology VL - 167 IS - 5 PY - 2024 SN - 0016-5085 ER - TY - JOUR AU - Kotlarz, D.M. C1 - 72038 C2 - 56353 TI - Unveiling a critical role of IL-7 in Celiac Disease - Insights from a novel human autoimmune organoid model. JO - Gastroenterology VL - 168 IS - 4 PY - 2024 SN - 0016-5085 ER - TY - JOUR AB - BACKGROUND & AIMS: Ulcerative colitis (UC) is characterized by severe inflammation and destruction of the intestinal epithelium, and is associated with specific risk single nucleotide polymorphisms in HLA class II. Given the recently discovered interactions between subsets of HLA-DP molecules and the activating natural killer (NK) cell receptor NKp44, genetic associations of UC and HLA-DP haplotypes and their functional implications were investigated. METHODS: HLA-DP haplotype and UC risk association analyses were performed (UC: n = 13,927; control: n = 26,764). Expression levels of HLA-DP on intestinal epithelial cells (IECs) in individuals with and without UC were quantified. Human intestinal 3-dimensional (3D) organoid cocultures with human NK cells were used to determine functional consequences of interactions between HLA-DP and NKp44. RESULTS: These studies identified HLA-DPA1∗01:03-DPB1∗04:01 (HLA-DP401) as a risk haplotype and HLA-DPA1∗01:03-DPB1∗03:01 (HLA-DP301) as a protective haplotype for UC in European populations. HLA-DP expression was significantly higher on IECs of individuals with UC compared with controls. IECs in human intestinal 3D organoids derived from HLA-DP401pos individuals showed significantly stronger binding of NKp44 compared with HLA-DP301pos IECs. HLA-DP401pos IECs in organoids triggered increased degranulation and tumor necrosis factor production by NKp44+ NK cells in cocultures, resulting in enhanced epithelial cell death compared with HLA-DP301pos organoids. Blocking of HLA-DP401-NKp44 interactions (anti-NKp44) abrogated NK cell activity in cocultures. CONCLUSIONS: We identified an UC risk HLA-DP haplotype that engages NKp44 and activates NKp44+ NK cells, mediating damage to intestinal epithelial cells in an HLA-DP haplotype-dependent manner. The molecular interaction between NKp44 and HLA-DP401 in UC can be targeted by therapeutic interventions to reduce NKp44+ NK cell-mediated destruction of the intestinal epithelium in UC. AU - Baumdick, M.E.* AU - Niehrs, A.* AU - Degenhardt, F.* AU - Schwerk, M.* AU - Hinrichs, O.* AU - Jordan-Paiz, A.* AU - Padoan, B.* AU - Wegner, L.H.M.* AU - Schloer, S.* AU - Zecher, B.F.* AU - Malsy, J.* AU - Joshi, V.R.* AU - Illig, C.* AU - Schröder-Schwarz, J.* AU - Möller, K.J.* AU - Martin, M.P.* AU - Yuki, Y.* AU - Ozawa, M.G.* AU - Sauter, J.* AU - Schmidt, A.H.* AU - Perez, D.* AU - Giannou, A.D.* AU - Carrington, M.N.* AU - Davis, R.S.* AU - Schumacher, U.* AU - Sauter, G.* AU - Huber, S.* AU - Puelles, V.G.* AU - Melling, N.* AU - Franke, A.* AU - International Inflammatory Bowel Disease Genetics Consortium (Gieger, C.) AU - Bunders, M.J.* C1 - 70155 C2 - 55027 SP - 946-962.e13 TI - HLA-DP on epithelial cells enables tissue damage by NKp44+ natural killer cells in ulcerative colitis. JO - Gastroenterology VL - 165 IS - 4 PY - 2023 SN - 0016-5085 ER - TY - JOUR AB - Background & Aims: Excess copper causes hepatocyte death in hereditary Wilson's disease (WD). Current WD treatments by copper-binding chelators may gradually reduce copper overload; they fail, however, to bring hepatic copper close to normal physiological levels. Consequently, lifelong daily dose regimens are required to hinder disease progression. This may result in severe issues due to nonadherence or unwanted adverse drug reactions and also due to drug switching and ultimate treatment failures. This study comparatively tested bacteria-derived copper binding agents—methanobactins (MBs)—for efficient liver copper depletion in WD rats as well as their safety and effect duration. Methods: Copper chelators were tested in vitro and in vivo in WD rats. Metabolic cage housing allowed the accurate assessment of animal copper balances and long-term experiments related to the determination of minimal treatment phases. Results: We found that copper-binding ARBM101 (previously known as MB-SB2) depletes WD rat liver copper dose dependently via fecal excretion down to normal physiological levels within 8 days, superseding the need for continuous treatment. Consequently, we developed a new treatment consisting of repetitive cycles, each of ∼1 week of ARBM101 applications, followed by months of in-between treatment pauses to ensure a healthy long-term survival in WD rats. Conclusions: ARBM101 safely and efficiently depletes excess liver copper from WD rats, thus allowing for short treatment periods as well as prolonged in-between rest periods. AU - Einer, C. AU - Munk, D.E.* AU - Park, E.* AU - Akdogan, B. AU - Nagel, J.* AU - Lichtmannegger, J. AU - Eberhagen, C. AU - Rieder, T.* AU - Vendelbo, M.H.* AU - Michalke, B. AU - Wimmer, R.* AU - Blutke, A. AU - Feuchtinger, A. AU - Dershwitz, P.* AU - DiSpirito, A.M.* AU - Islam, T.* AU - Castro, R.E.* AU - Min, B.K.* AU - Kim, T.W.* AU - Choi, S.* AU - Kim, D.* AU - Jung, C.* AU - Lee, H.* AU - Park, D.* AU - Im, W.* AU - Eun, S.Y.* AU - Cho, Y.H.* AU - Semrau, J.D.* AU - Rodrigues, C.M.P.* AU - Hohenester, S.* AU - Damgaard Sandahl, T.* AU - DiSpirito, A.A.* AU - Zischka, H. C1 - 68534 C2 - 53666 CY - 1600 John F Kennedy Boulevard, Ste 1800, Philadelphia, Pa 19103-2899 Usa SP - 187-200.e7 TI - ARBM101 (Methanobactin SB2) drains excess liver copper via biliary excretion in Wilson's disease rats. JO - Gastroenterology VL - 165 IS - 1 PB - W B Saunders Co-elsevier Inc PY - 2023 SN - 0016-5085 ER - TY - JOUR AU - Kotlarz, D.M. C1 - 67258 C2 - 54197 SP - 1337-1338 TI - Identification of the γδ IEL-secreted Paneth cell protective factor API5 as a promising therapeutic target for ATG16L1-associated Crohn's disease. JO - Gastroenterology VL - 164 IS - 7 PY - 2023 SN - 0016-5085 ER - TY - JOUR AU - Riemann, J.F.* AU - Protzer, U. C1 - 69372 C2 - 53885 CY - Tiergartenstrasse 17, D-69121 Heidelberg, Germany SP - 50-52 TI - SARS-CoV-2-the Pandemic and its Consequences Interview with Prof. Dr. Ulrike Protzer, Director of the Institute of Virology, Technical University of Munich/Helmholtz Center Munich - German Center for Environmental Health. JO - Gastroenterology VL - 18 PB - Springer Heidelberg PY - 2023 SN - 0016-5085 ER - TY - JOUR AU - Kotlarz, D.M. C1 - 65465 C2 - 52458 SP - 527-528 TI - Mucus sialylation maintains the peace in intestinal host microbe relations. JO - Gastroenterology VL - 163 IS - 2 PY - 2022 SN - 0016-5085 ER - TY - JOUR AB - OBJECTIVE: Lipidomic changes were causally linked to metabolic diseases, but the scenario for colorectal cancer (CRC) is less clear. We investigated the CRC lipidome for putative tumour-specific alterations through analysis of three independent retrospective patient cohorts from two clinical centers, to derive a clinically useful signature. DESIGN: Quantitative comprehensive lipidomic analysis was performed by direct infusion electrospray ionization coupled to tandem mass spectrometry (ESI-MS/MS) and high-resolution mass spectrometry (HR-MS) on matched non-diseased mucosa and tumor tissue in a discovery cohort (n=106). Results were validated in two independent cohorts (n=28, and n=20), associated with genomic and clinical data, and lipidomic data from a genetic mouse tumor model (Apc1638N). RESULTS: Significant differences were found between tumor and normal tissue for glycero-, glycerophospho- and sphingolipids in the discovery cohort. Comparison to the validation collectives unveiled that glycerophospholipids showed high interpatient variation and were strongly affected by preanalytical conditions, whereas glycero- and sphingolipids appeared more robust. Signatures of sphingomyelin (SM) and triacylglycerol (TG) species significantly differentiated cancerous from non-diseased tissue in both validation studies. Moreover, lipogenic enzymes were significantly upregulated in CRC, and FASN gene expression was prognostically detrimental. The TG profile was significantly associated with post-operative disease-free survival and lymphovascular invasion, and was essentially conserved in murine digestive cancer, but not associated with microsatellite status, KRAS or BRAF mutations, or T-cell infiltration. CONCLUSION: Analysis of the CRC lipidome revealed a robust TG-species signature with prognostic potential. A better understanding of the cancer-associated glycerolipid and sphingolipid metabolism may lead to novel therapeutic strategies. AU - Ecker, J.* AU - Benedetti, E.* AU - Kindt, A. AU - Höring, M.* AU - Perl, M.* AU - Machmüller, A.C. AU - Sichler, A.* AU - Plagge, J.* AU - Wang, Y.* AU - Zeissig, S.* AU - Shevchenko, A.* AU - Burkhardt, R.* AU - Krumsiek, J.* AU - Liebisch, G.* AU - Janssen, K.P.* C1 - 62103 C2 - 50453 CY - 1600 John F Kennedy Boulevard, Ste 1800, Philadelphia, Pa 19103-2899 Usa SP - 910-923.e19 TI - The colorectal cancer lipidome - identification of a robust tumor-specific lipid species signature. JO - Gastroenterology VL - 161 IS - 3 PB - W B Saunders Co-elsevier Inc PY - 2021 SN - 0016-5085 ER - TY - JOUR AB - Background & Aims: Pancreatic ductal adenocarcinoma (PDAC) is characterized by a fibroblast-rich desmoplastic stroma. Cancer-associated fibroblasts (CAFs) have been shown to display a high degree of interconvertible states including quiescent, inflammatory, and myofibroblastic phenotypes; however, the mechanisms by which this plasticity is achieved are poorly understood. Here, we aim to elucidate the role of CAF plasticity and its impact on PDAC biology. Methods: To investigate the role of mesenchymal plasticity in PDAC progression, we generated a PDAC mouse model in which CAF plasticity is modulated by genetic depletion of the transcription factor Prrx1. Primary pancreatic fibroblasts from this mouse model were further characterized by functional in vitro assays. To characterize the impact of CAFs on tumor differentiation and response to chemotherapy, various coculture experiments were performed. In vivo, tumors were characterized by morphology, extracellular matrix composition, and tumor dissemination and metastasis. Results: Our in vivo findings showed that Prrx1-deficient CAFs remain constitutively activated. Importantly, this CAF phenotype determines tumor differentiation and disrupts systemic tumor dissemination. Mechanistically, coculture experiments of tumor organoids and CAFs showed that CAFs shape the epithelial-to-mesenchymal phenotype and confer gemcitabine resistance of PDAC cells induced by CAF-derived hepatocyte growth factor. Furthermore, gene expression analysis showed that patients with pancreatic cancer with high stromal expression of Prrx1 display the squamous, most aggressive, subtype of PDAC. Conclusions: Here, we define that the Prrx1 transcription factor is critical for tuning CAF activation, allowing a dynamic switch between a dormant and an activated state. This work shows that Prrx1-mediated CAF plasticity has significant impact on PDAC biology and therapeutic resistance. AU - Feldmann, K.* AU - Maurer, C.* AU - Peschke, K.* AU - Teller, S.* AU - Schuck, K.* AU - Steiger, K.* AU - Engleitner, T.* AU - Öllinger, R.* AU - Nomura, A.* AU - Wirges, N.* AU - Papargyriou, A. AU - Jahan Sarker, R.S.* AU - Ranjan, R.A.* AU - Dantes, Z.* AU - Weichert, W.* AU - Rustgi, A.K.* AU - Schmid, R.M.* AU - Rad, R.* AU - Schneider, G.* AU - Saur, D.* AU - Reichert, M.* C1 - 60826 C2 - 49667 CY - 1600 John F Kennedy Boulevard, Ste 1800, Philadelphia, Pa 19103-2899 Usa SP - 346-361.e24 TI - Mesenchymal plasticity regulated by Prrx1 drives aggressive pancreatic cancer biology. JO - Gastroenterology VL - 160 IS - 1 PB - W B Saunders Co-elsevier Inc PY - 2021 SN - 0016-5085 ER - TY - JOUR AB - BACKGROUND AIMS: Throughout life, the intestinal epithelium undergoes constant self-renewal from intestinal stem cells. Together with genotoxic stressors and failing DNA repair, this self-renewal causes susceptibility towards malignant transformation. X-box binding protein 1 (XBP1) is a stress sensor involved in the unfolded protein response (UPR). We hypothesized that XBP1 acts as a signaling hub to regulate epithelial DNA damage responses. METHODS: Data from the TCGA were analyzed for association of XBP1 with CRC survival and molecular interactions between XBP1 andp53 pathway activity. The role of XBP1 in orchestrating p53-driven DNA damage response was tested in-vitro, in mouse models of chronic intestinal epithelial DNA damage (Xbp1/H2bfl/fl, Xbp1ΔIEC, H2bΔIEC, H2b/Xbp1ΔIEC) and via orthotopic tumor organoid transplantation. Transcriptome analysis of intestinal organoids was performed to identify molecular targets of Xbp1-mediated DNA damage response. RESULTS: In the TCGA dataset of CRC, low XBP1 expression was significantly associated with poor overall survival (OS) and reduced p53 pathway activity. In-vivo, H2b/Xbp1ΔIEC mice developed spontaneous intestinal carcinomas. Orthotopic tumor organoid transplantation revealed a metastatic potential of H2b/Xbp1ΔIEC-derived tumors. RNA sequencing of intestinal organoids (H2b/Xbp1fl/fl, H2bΔIEC, H2b/Xbp1ΔIEC, H2b/p53ΔIEC) identified a transcriptional program downstream of p53, in which XBP1 directs DNA damage-induced Ddit4l expression. DDIT4L inhibits mTOR-mediated phosphorylation of 4E-BP1. Pharmacological mTOR inhibition suppressed epithelial hyperproliferation via 4E-BP1. CONCLUSIONS: Our data suggest a crucial role for XBP1 in coordinating epithelial DNA damage responses and stem cell function via a p53-DDIT4L-dependent feedback mechanism. AU - Welz, L.* AU - Kakavand, N.* AU - Hang, X.* AU - Laue, G.* AU - Ito, G.* AU - Silva, M.G.* AU - Plattner, C.* AU - Mishra, N.* AU - Tengen, F. AU - Ogris, C. AU - Jesinghaus, M.* AU - Wottawa, F.* AU - Arnold, P.* AU - Kaikkonen, L.* AU - Stengel, S.* AU - Tran, F.* AU - Das, S.* AU - Kaser, A.* AU - Trajanoski, Z.* AU - Blumberg, R.* AU - Roecken, C.* AU - Saur, D.* AU - Tschurtschenthaler, M.* AU - Schreiber, S.* AU - Rosenstiel, P.* AU - Aden, K.* C1 - 63180 C2 - 51371 CY - 1600 John F Kennedy Boulevard, Ste 1800, Philadelphia, Pa 19103-2899 Usa SP - 223-237.e11 TI - Epithelial XBP1 coordinates TP53-driven DNA damage responses and suppression of intestinal carcinogenesis. JO - Gastroenterology VL - 162 IS - 1 PB - W B Saunders Co-elsevier Inc PY - 2021 SN - 0016-5085 ER - TY - JOUR AB - BACKGROUND & AIMS: Intestinal epithelial homeostasis depends on a tightly regulated balance between intestinal epithelial cell (IEC) death and proliferation. While the disruption of several IEC death regulating factors result in intestinal inflammation, the loss of the anti-apoptotic BCL2 family members BCL2 and BCL2L1 has no effect on intestinal homeostasis in mice. We investigated the functions of the antiapoptotic protein MCL1, another member of the BCL2 family, in intestinal homeostasis in mice. METHODS: We generated mice with IECspecific disruption of Mcl1 (Mcl1(Delta IEC) mice) or tamoxifeninducible IEC-specific disruption of Mcl1 (i-Mcl1(Delta IEC) mice); these mice and mice with full-length Mcl1 (controls) were raised under normal or germ-free conditions. Mice were analyzed by endoscopy and for intestinal epithelial barrier permeability. Intestinal tissues were analyzed by histology, in situ hybridization, proliferation assays, and immunoblots. Levels of calprotectin, a marker of intestinal inflammation, were measured in intestinal tissues and feces. RESULTS: Mcl1(Delta IEC) mice spontaneously developed apoptotic enterocolopathy, characterized by increased IEC apoptosis, hyperproliferative crypts, epithelial barrier dysfunction, and chronic inflammation. Loss of MCL1 retained intestinal crypts in a hyperproliferated state and prevented the differentiation of intestinal stem cells. Proliferation of intestinal stem cells in MCL1-deficient mice required WNT signaling and was associated with DNA damage accumulation. By 1 year of age, Mcl1(Delta IEC) mice developed intestinal tumors with morphologic and genetic features of human adenomas and carcinomas. Germ-free housing of Mcl1(Delta IEC) mice reduced markers of microbiotainduced intestinal inflammation but not tumor development. CONCLUSION: The antiapoptotic protein MCL1, a member of the BCL2 family, is required for maintenance of intestinal homeostasis and prevention of carcinogenesis in mice. Loss of MCL1 results in development of intestinal carcinomas, even under germ-free conditions, and therefore does not involve microbe-induced chronic inflammation. Mcl1(Delta IEC) mice might be used to study apoptotic enterocolopathy and inflammatory bowel diseases. AU - Healy, M.E.* AU - Boege, Y.* AU - Hodder, M.C.* AU - Boehm, F.* AU - Malehmir, M.* AU - Scherr, A.L.* AU - Jetzer, J.* AU - Chan, L.K.* AU - Parrotta, R.* AU - Jacobs, K.* AU - Clerbaux, L.* AU - Kreutzer, S.* AU - Campbell, A.* AU - Gilchrist, E.* AU - Gilroy, K.* AU - Rodewald, A.* AU - Honcharova-Biletska, H.* AU - Schimmer, R.* AU - Velez, K.* AU - Bueeler, S.* AU - Cammareri, P.* AU - Kalna, G.* AU - Wenning, A.S.* AU - McCoy, K.D.* AU - Gomez de Agueero, M.* AU - Schulze-Bergkamen, H.* AU - Klose, C.S.N.* AU - Unger, K. AU - Macpherson, A.J.* AU - Moor, A.E.* AU - Koehler, B.* AU - Sansom, O.J.* AU - Heikenwaelder, M.* AU - Weber, A.* C1 - 59865 C2 - 49080 CY - 1600 John F Kennedy Boulevard, Ste 1800, Philadelphia, Pa 19103-2899 Usa SP - 183-199 TI - MCL1 is required for maintenance of intestinal homeostasis and prevention of carcinogenesis in mice. JO - Gastroenterology VL - 159 IS - 1 PB - W B Saunders Co-elsevier Inc PY - 2020 SN - 0016-5085 ER - TY - JOUR AB - Background & Aims: Hepatitis B virus (HBV) infection persists because the virus-specific immune response is dysfunctional. Therapeutic vaccines might be used to end immune tolerance to the virus in patients with chronic infection, but these have not been effective in patients so far. In patients with chronic HBV infection, high levels of virus antigens might prevent induction of HBV-specific immune responses. We investigated whether knocking down expression levels of HBV antigens in liver might increase the efficacy of HBV vaccines in mice. Methods: We performed studies with male C57BL/6 mice that persistently replicate HBV (genotype D, serotype ayw)—either from a transgene or after infection with an adeno-associated virus that transferred an overlength HBV genome—and expressed HB surface antigen at levels relevant to patients. Small hairpin or small interfering (si)RNAs against the common 3′-end of all HBV transcripts were used to knock down antigen expression in mouse hepatocytes. siRNAs were chemically stabilized and conjugated to N-acetylgalactosamine to increase liver uptake. Control mice were given either entecavir or non-HBV–specific siRNAs and vaccine components. Eight to 12 weeks later, mice were immunized twice with a mixture of adjuvanted HBV S and core antigen, followed by a modified Vaccinia virus Ankara vector to induce HBV-specific B- and T-cell responses. Serum and liver samples were collected and analyzed for HBV-specific immune responses, liver damage, and viral parameters. Results: In both models of HBV infection, mice that express hepatocyte-specific small hairpin RNAs or that were given subcutaneous injections of siRNAs had reduced levels of HBV antigens, HBV replication, and viremia (1–3 log10 reduction) compared to mice given control RNAs. Vaccination induced production of HBV-neutralizing antibodies and increased numbers and functionality of HBV-specific, CD8+ T cells in mice with low, but not in mice with high, levels of HBV antigen. Mice with initially high titers of HBV and knockdown of HBV antigen expression, but not mice with reduced viremia after administration of entecavir, developed polyfunctional, HBV-specific CD8+ T cells, and HBV was eliminated. Conclusions: In mice with high levels of HBV replication, knockdown of HBV antigen expression along with a therapeutic vaccination strategy, but not knockdown alone, increased numbers of effector T cells and eliminated the virus. These findings indicate that high titers of virus antigens reduce the efficacy of therapeutic vaccination. Anti-HBV siRNAs and therapeutic vaccines are each being tested in clinical trials—their combination might cure chronic HBV infection. AU - Michler, T. AU - Kosinska, A. AU - Festag, J. AU - Bunse, T. AU - Su, J. AU - Ringelhan, M. AU - Imhof, H. AU - Grimm, D.* AU - Steiger, K.* AU - Mogler, C.* AU - Heikenwalder, M.* AU - Michel, M.L.* AU - Guzman, C.A.* AU - Milstein, S.* AU - Sepp-Lorenzino, L.* AU - Knolle, P.* AU - Protzer, U. C1 - 58800 C2 - 48336 SP - 1762-1775 TI - Knockdown of virus antigen expression increases therapeutic vaccine efficacy in high-titer hepatitis B virus carrier mice. JO - Gastroenterology VL - 158 IS - 6 PY - 2020 SN - 0016-5085 ER - TY - JOUR AB - BACKGROUND & AIMS: Altered interactions between the mucosal immune system and intestinal microbiota contribute to pathogenesis of inflammatory bowel diseases (IBD). It is not clear how inhibitors of cytokines, such as antagonists of tumor necrosis factor (anti-TNF), affect the intestinal microbiome. We investigated the effects of anti-TNF agents on gut microbe community structure and function in a longitudinal 2-step study of patients with IBD. We correlated our findings with outcomes of treatment and investigated patterns of metabolites in fecal samples before and after anti-TNF therapy. METHODS: We performed a prospective study of 2 cohorts of patients in Germany; the discovery cohort comprised 12 patients with IBD, 17 patients with rheumatic disease, and 19 healthy individuals (controls); fecal samples were collected at baseline and 2, 6, and 30 weeks after induction of anti-TNF therapy. The validation cohort comprised 23 patients with IBD treated with anti-TNF or vedolizumab (anti-alpha 4 beta 7 integrin) and 99 healthy controls; fecal samples were collected at baseline and at weeks 2, 6, and 14. Fecal microbiota were analyzed by V3-V4 16S ribosomal RNA gene amplicon sequencing. Clinical response and remission were determined by clinical disease activity scores. Metabolic network reconstruction and associated fecal metabolite level inference was performed in silico using the AGORA (Assembly of Gut Organisms through Reconstruction and Analysis) resource. Metabolomic analyses of fecal samples from a subset of patients were performed to validate metabolites associated with treatment outcomes. RESULTS: Anti-TNF therapy shifted the diversity of fecal microbiota in patients with IBD, but not with rheumatic disease, toward that of controls. Across timepoints, diversity indices did not vary significantly between patients with IBD who did or did not achieve clinical remission after therapy. In contrast, in silico modeling of metabolic interactions between gut microbes found metabolite exchange to be significantly reduced at baseline in fecal samples from patients with IBD and to be associated with later clinical remission. Predicted levels of butyrate and substrates involved in butyrate synthesis (ethanol or acetaldehyde) were significantly associated with clinical remission following anti-TNF therapy, verified by fecal metabolomic analyses. CONCLUSIONS: Metabolic network reconstruction and assessment of metabolic profiles of fecal samples might be used to identify patients with IBD likely to achieve clinical remission following anti-TNF therapy and increase our understanding of the heterogeneity of IBD. AU - Aden, K.* AU - Rehman, A.* AU - Waschina, S.* AU - Pan, W.H.* AU - Walker, A. AU - Lucio, M. AU - Nunez, A.M.* AU - Bharti, R.* AU - Zimmerman, J.* AU - Bethge, J.* AU - Schulte, B.* AU - Schulte, D.* AU - Franke, A.* AU - Nikolaus, S.* AU - Schroeder, J.O.* AU - Vandeputte, D.* AU - Raes, J.* AU - Szymczak, S.* AU - Waetzig, G.H.* AU - Zeuner, R.* AU - Schmitt-Kopplin, P. AU - Kaleta, C.* AU - Schreiber, S.* AU - Rosenstiel, P.* C1 - 56605 C2 - 47172 CY - 1600 John F Kennedy Boulevard, Ste 1800, Philadelphia, Pa 19103-2899 Usa SP - 1279-1292.e11 TI - Metabolic functions of gut microbes associate with efficacy of tumor necrosis factor antagonists in patients with inflammatory bowel diseases. JO - Gastroenterology VL - 157 IS - 5 PB - W B Saunders Co-elsevier Inc PY - 2019 SN - 0016-5085 ER - TY - JOUR AB - BACKGROUND AND AIMS: Cells in pancreatic ductal adenocarcinoma (PDAC) undergo autophagy, but its effects vary with tumor stage and genetic factors. We investigated the consequences of varying levels of the autophagy related 5 (Atg5) protein on pancreatic tumor formation and progression. METHODS: We generated mice that express oncogenic Kras in primary pancreatic cancer cells and have homozygous disruption of Atg5 (A5; Kras) or heterozygous disruption of Atg5 (A5(+/-); Kras), and compared them with mice with only oncogenic Kras (controls). Pancreata were analyzed by histology and immunohistochemistry. Primary tumor cells were isolated and used to perform transcriptome, metabolome, intracellular calcium, extracellular cathepsin activity, and cell migration and invasion analyses. The cells were injected into wild-type littermates, and orthotopic tumor growth and metastasis were monitored. Atg5 was knocked down in pancreatic cancer cell lines using small hairpin RNAs; cell migration and invasion were measured, and cells were injected into wild-type littermates. PDAC samples were obtained from independent cohorts of patients and protein levels were measured on immunoblot and immunohistochemistry; we tested the correlation of protein levels with metastasis and patient survival times. RESULTS: A5(+/-); Kras mice, with reduced Atg5 levels, developed more tumors and metastases, than control mice, whereas A5; Kras mice did not develop any tumors. Cultured A5(+/-); Kras primary tumor cells were resistant to induction and inhibition of autophagy, had altered mitochondrial morphology, compromised mitochondrial function, changes in intracellular Ca2thorn oscillations, and increased activity of extracellular cathepsin L and D. The tumors that formed in A5(+/-); Kras mice contained greater numbers of type 2 macrophages than control mice, and primary A5(+/-); Kras tumor cells had up-regulated expression of cytokines that regulate macrophage chemoattraction and differentiation into M2 macrophage. Knockdown of Atg5 in pancreatic cancer cell lines increased their migratory and invasive capabilities, and formation of metastases following injection into mice. In human PDAC samples, lower levels of ATG5 associated with tumor metastasis and shorter survival time. CONCLUSIONS: In mice that express oncogenic Kras in pancreatic cells, heterozygous disruption of Atg5 and reduced protein levels promotes tumor development, whereas homozygous disruption of Atg5 blocks tumorigenesis. Therapeutic strategies to alter autophagy in PDAC should consider the effects of ATG5 levels to avoid the expansion of resistant and highly aggressive cells. AU - Görgülü, K.* AU - Diakopoulos, K.N.* AU - Ai, J.* AU - Schoeps, B.* AU - Kabacaoglu, D.* AU - Karpathaki, A.F.* AU - Ciecielski, K.J.* AU - Kaya-Aksoy, E.* AU - Ruess, D.A.* AU - Berninger, A.* AU - Kowalski, M.* AU - Stevanovic, M.* AU - Wörmann, S.M.* AU - Wartmann, T.* AU - Zhao, Y.* AU - Halangk, W.* AU - Voronina, S.* AU - Tepikin, A.* AU - Schlitter, A.M.* AU - Steiger, K.* AU - Artati, A. AU - Adamski, J. AU - Aichler, M. AU - Walch, A.K. AU - Jastroch, M. AU - Hartleben, G. AU - Mantzoros, C.S.* AU - Weichert, W.* AU - Schmid, R.M.* AU - Herzig, S.* AU - Krüger, A.* AU - Sainz, B.* AU - Lesina, M.* AU - Algül, H.* C1 - 54480 C2 - 45622 CY - 1600 John F Kennedy Boulevard, Ste 1800, Philadelphia, Pa 19103-2899 Usa SP - 203-217.e20 TI - Levels of the autophagy-related 5 protein affect progression and metastasis of pancreatic tumors in mice. JO - Gastroenterology VL - 156 IS - 1 PB - W B Saunders Co-elsevier Inc PY - 2019 SN - 0016-5085 ER - TY - JOUR AB - BACKGROUND & AIMS: Hepatitis D virus (HDV) superinfection in patients with hepatitis B virus (HBV) is associated with rapid progression to liver cirrhosis and hepatocellular carcinoma. Treatment options are limited, and no vaccine is available. Although HDV-specific CD8(+) T cells are thought to control the virus, little is known about which HDV epitopes are targeted by virus-specific CD8(+)T cells or why these cells ultimately fail to control the infection. We aimed to define how HDV escapes the CD8(+) T-cell-mediated response. METHODS: We collected plasma and DNA samples from 104 patients with chronic HDV and HBV infection at medical centers in Europe and the Middle East, sequenced HDV, typed human leukocyte antigen (HLA) class I alleles from patients, and searched for polymorphisms in HDV RNA associated with specific HLA class I alleles. We predicted epitopes in HDV that would be recognized by CD8(+) T cells and corresponded with the identified virus polymorphisms in patients with resolved (n = 12) or chronic (n = 13) HDV infection. RESULTS: We identified 21 polymorphisms in HDV that were significantly associated with specific HLA class I alleles (P < .005). Five of these polymorphisms were found to correspond to epitopes in HDV that are recognized by CD8(+) T cells; we confirmed that CD8(+) T cells in culture targeted these HDV epitopes. HDV variant peptides were only partially cross-recognized by CD8(+) T cells isolated from patients, indicating that the virus had escaped detection by these cells. These newly identified HDV epitopes were restricted by relatively infrequent HLA class I alleles, and they bound most frequently to HLA-B. In contrast, frequent HLA class I alleles were not associated with HDV sequence polymorphisms. CONCLUSIONS: We analyzed sequences of HDV RNA and HLA class I alleles that present epitope peptides to CD8(+) T cells in patients with persistent HDV infection. We identified polymorphisms in the HDV proteome that associate with HLA class I alleles. Some variant peptides in epitopes from HDV were only partially recognized by CD8(+) T cells isolated from patients; these could be mutations that allow HDV to escape the immune response, resulting in persistent infection. HDV escape from the immune response was associated with uncommon HLA class I alleles, indicating that HDV evolves, at the population level, to evade recognition by common HLA class I alleles. AU - Karimzadeh, H. AU - Kiraithe, M.M.* AU - Oberhardt, V.* AU - Alizei, E.S.* AU - Bockmann, J.* AU - Zur Wiesch, J.S.* AU - Budeus, B.* AU - Hoffmann, D.* AU - Wedemeyer, H.* AU - Cornberg, M.* AU - Krawczyk, A.* AU - Rashidi-Alavijeh, J.* AU - Rodríguez-Frías, F.* AU - Casillas, R.* AU - Buti, M.* AU - Smedile, A.* AU - Alavian, S.M.* AU - Heinold, A.* AU - Emmerich, F.* AU - Panning, M.* AU - Gostick, E.* AU - Price, D.A.* AU - Timm, J.* AU - Hofmann, M.* AU - Raziorrouh, B.* AU - Thimme, R.* AU - Protzer, U. AU - Roggendorf, M. AU - Neumann-Haefelin, C.* C1 - 55520 C2 - 46400 CY - 1600 John F Kennedy Boulevard, Ste 1800, Philadelphia, Pa 19103-2899 Usa SP - 1820-1833 TI - Mutations in hepatitis D virus allow it to escape detection by CD8+ T cells and evolve at the population level. JO - Gastroenterology VL - 156 IS - 6 PB - W B Saunders Co-elsevier Inc PY - 2019 SN - 0016-5085 ER - TY - JOUR AB - BACKGROUND & AIMS: Wilson disease (WD) is an inherited disorder of copper metabolism that leads to copper accumulation and toxicity in the liver and brain. It is caused by mutations in the adenosine triphosphatase copper transporting b gene (ATP7B), which encodes a protein that transports copper from hepatocytes into the bile. We studied ATP7B-deficient cells and animals to identify strategies to decrease copper toxicity in patients with WD. METHODS: We used RNA-seq to compare gene expression patterns between wild-type and ATP7B-knockout HepG2 cells exposed to copper. We collected blood and liver tissues from Atp7b(-/-) and Atp7b(+/-) (control) rats (LPP) and mice; some mice were given 5 daily injections of an autophagy inhibitor (spautin-1) or vehicle. We obtained liver biopsies from 2 patients with WD in Italy and liver tissues from patients without WD (control). Liver tissues were analyzed by immunohistochemistry, immunofluorescence, cell viability, apoptosis assays, and electron and confocal microscopy. Proteins were knocked down in cell lines using small interfering RNAs. Levels of copper were measured in cell lysates, blood samples, liver homogenates, and subcellular fractions by spectroscopy. RESULTS: After exposure to copper, ATP7B-knockout cells had significant increases in the expression of 103 genes that regulate autophagy (including MAP1LC3A, known as LC3) compared with wild-type cells. Electron and confocal microscopy visualized more autophagic structures in the cytoplasm of ATP7B-knockout cells than wild-type cells after copper exposure. Hepatocytes in liver tissues from patients with WD and from Atp7b(-/-) mice and rats (but not controls) had multiple autophagosomes. In ATP7B-knockout cells, mammalian target of rapamycin (mTOR) had decreased activity and was dissociated from lysosomes; this resulted in translocation of the mTOR substrate transcription factor EB to the nucleus and activation of autophagy-related genes. In wild-type HepG2 cells (but not ATP7B-knockout cells), exposure to copper and amino acids induced recruitment of mTOR to lysosomes. Pharmacologic inhibitors of autophagy or knockdown of autophagy proteins ATG7 and ATG13 induced and accelerated the death of ATP7B-knockout HepG2 cells compared with wild-type cells. Autophagy protected ATP7B-knockout cells from copperinduced death. CONCLUSION: ATP7B-deficient hepatocytes, such as in those in patients with WD, activate autophagy in response to copper overload to prevent copper-induced apoptosis. Agents designed to activate this autophagic pathway might decrease copper toxicity in patients with WD. AU - Polishchuk, E.V.* AU - Merolla, A.* AU - Lichtmannegger, J. AU - Romano, A.* AU - Indrieri, A.* AU - Ilyechova, E.Y.* AU - Concilli, M.* AU - De Cegli, R.* AU - Crispino, R.* AU - Mariniello, M.* AU - Petruzzelli, R.* AU - Ranucci, G.* AU - Iorio, R.* AU - Pietrocola, F.* AU - Einer, C. AU - Borchard, S. AU - Zibert, A.* AU - Schmidt, H.H.* AU - Di Schiavi, E.* AU - Puchkova, L.V.* AU - Franco, B.* AU - Kroemer, G.* AU - Zischka, H. AU - Polishchuk, R.S.* C1 - 54776 C2 - 45886 CY - 1600 John F Kennedy Boulevard, Ste 1800, Philadelphia, Pa 19103-2899 Usa SP - 1173-1189.e5 TI - Activation of autophagy, observed in liver tissues from patients with Wilson disease and from Atp7b-deficient animals, protects hepatocytes from copper-induced apoptosis. JO - Gastroenterology VL - 156 IS - 4 PB - W B Saunders Co-elsevier Inc PY - 2019 SN - 0016-5085 ER - TY - JOUR AB - BACKGROUND & AIMS: The CYLD lysine 63 deubiquitinase gene (CYLD) encodes tumor suppressor protein that is mutated in familial cylindromatosus, and variants have been associated with Crohn disease (CD). Splice forms of CYLD that lack exons 7 and 8 regulate transcription factors and functions of immune cells. We examined the expression of splice forms of CYLD in colon tissues from patients with CD and their effects in mice. METHODS: We performed immunohistochemical analyses of colon tissues from patients with untreated CD and patients without inflammatory bowel diseases (controls). We obtained mice that expressed splice forms of CYLD (sCYLD mice) without or with SMAD7 (sCYLD/SMAD7 mice) from transgenes and CYLD-knockout mice (with or without transgenic expression of SMAD7) and performed endoscopic analyses. Colitis was induced in Rag1(-/-) mice by transfer of CD4(+) CD62L(+) T cells from C57/Bl6 or transgenic mice. T cells were isolated from mice and analyzed by flow cytometry and quantitative realtime polymerase chain reaction and intestinal tissues were analyzed by histology and immunohistochemistry. CYLD forms were expressed in mouse embryonic fibroblasts, primary T cells, and HEK293T cells, which were analyzed by immunoblot, mobility shift, and immunoprecipitation assays. RESULTS: The colonic lamina propria from patients with CD was infiltrated by T cells and had higher levels of sCYLD (but not full-length CYLD) and SMAD7 than tissues from controls. Incubation of mouse embryonic fibroblasts and T cells with transforming growth factor b increased their production of sCYLD and decreased full-length CYLD. Transgenic expression of sCYLD and SMAD7 in T cells prevented the differentiation of regulatory T cells and T-helper type 17 cells and increased the differentiation of T-helper type 1 cells. The same effects were observed in colon tissues from sCYLD/SMAD7 mice but not in those from CYLD-knockout SMAD7 mice. The sCYLD mice had significant increases in the numbers of T-helper type 1 cells and CD44high CD62Llow memory-effector CD4(+) T cells in the spleen and mesenteric lymph nodes compared with wild-type mice; sCYLD/SMAD7 mice had even larger increases. The sCYLD/ SMAD7 mice spontaneously developed severe colitis, with infiltration of the colon by dendritic cells, neutrophils, macrophages, and CD4(+) T cells and increased levels of Ifng, Il6, Il12a, Il23a, and Tnf mRNAs. Co-transfer of regulatory T cells from wild-type, but not from sCYLD/SMAD7, mice prevented the induction of colitis in Rag1(-/-) mice by CD4(+) T cells. We found increased levels of poly-ubiquitinated SMAD7 in sCYLD CD4(+)T cells. CYLD formed a nuclear complex with SMAD3, whereas sCYLD recruited SMAD7 to the nucleus, which inhibited the expression of genes regulated by SMAD3 and SMAD4. We found that sCYLD mediated lysine 63-linked ubiquitination of SMAD7. The sCYLD-SMAD7 complex inhibited transforming growth factor beta signaling in CD4(+)T cells. CONCLUSIONS: Levels of the spliced form of CYLD are increased in colon tissues from patients with CD. sCYLD mediates ubiquitination and nuclear translocation of SMAD7 and thereby decreases transforming growth factor beta signaling in T cells. This prevents immune regulatory mechanisms and leads to colitis in mice. AU - Tang, Y.* AU - Reissig, S.* AU - Glasmacher, E. AU - Regen, T.* AU - Wanke, F.* AU - Nikolaev, A.* AU - Gerlach, K.* AU - Popp, V.* AU - Karram, K.* AU - Fantini, M.C.* AU - Schattenberg, J.M.* AU - Galle, P.R.* AU - Neurath, M.F.* AU - Weigmann, B.* AU - Kurschus, F.C.* AU - Hövelmeyer, N.* AU - Waisman, A.* C1 - 54518 C2 - 45628 CY - 1600 John F Kennedy Boulevard, Ste 1800, Philadelphia, Pa 19103-2899 Usa SP - 692-707.e7 TI - Alternative splice forms of CYLD mediate ubiquitination of SMAD7 to prevent TGFB signaling and promote colitis. JO - Gastroenterology VL - 156 IS - 3 PB - W B Saunders Co-elsevier Inc PY - 2019 SN - 0016-5085 ER - TY - JOUR AB - Fewer than 1% of chronic hepatitis B virus infections per year are cured with antiviral treatment. This creates a need for long-term treatment, which poses challenges for patients and health systems. Because cure is accompanied by recovery of antiviral immunity, a combination of direct-acting antiviral agents and immunotherapy are likely to be required. Extensive efforts have been made to identify determinants of the failed immune response to hepatitis B virus in patients with chronic infection. We review mechanisms of immune dysfunction in patients with chronic hepatitis B virus infection, immunotherapy strategies in development, and the challenges associated with successful implementation of immunotherapy. AU - Gehring, A.* AU - Protzer, U. C1 - 54601 C2 - 45702 CY - 1600 John F Kennedy Boulevard, Ste 1800, Philadelphia, Pa 19103-2899 Usa SP - 325-337 TI - Targeting innate and adaptive immune responses to cure chronic HBV infection. JO - Gastroenterology VL - 156 IS - 2 PB - W B Saunders Co-elsevier Inc PY - 2018 SN - 0016-5085 ER - TY - JOUR AB - In vivo optical imaging modalities are mostly limited to cell cultures, superficial tissues, and intravital imaging since they lack either resolution or penetration depth.1 In contrast, optoacoustic (OA) imaging—combining features of optical and ultrasound imaging—has been used to visualize hemoglobin in depths of approximately 3 cm in patients with Crohn’s disease.2,3 Realizing an even higher resolution, raster-scanning OA mesoscopy (RSOM) provides intrinsic optical tissue contrast down to 10-20 μm resolution at still high penetration depths of several millimeters. AU - Knieling, F.* AU - Gonzales-Menezes, J.* AU - Claussen, J.* AU - Schwarz, M.A.* AU - Neufert, C.* AU - Fahlbusch, F.B.* AU - Rath, T.* AU - Thoma, O.M.* AU - Kramer, V.* AU - Menchicchi, B.* AU - Kersten, C.* AU - Scheibe, K.* AU - Schürmann, S.* AU - Carlé, B.* AU - Rascher, W.* AU - Neurath, M.F.* AU - Ntziachristos, V. AU - Waldner, M.J.* C1 - 52689 C2 - 44107 CY - Philadelphia SP - 807-809.e3 TI - Raster-scanning optoacoustic mesoscopy for gastrointestinal imaging at high resolution. JO - Gastroenterology VL - 154 IS - 4 PB - W B Saunders Co-elsevier Inc PY - 2018 SN - 0016-5085 ER - TY - JOUR AU - Weiss, K.H.* AU - Zischka, H. C1 - 52421 C2 - 43958 CY - Philadelphia SP - 15-17 TI - Copper directly affects intestinal lipid turnover. JO - Gastroenterology VL - 154 IS - 1 PB - W B Saunders Co-elsevier Inc PY - 2018 SN - 0016-5085 ER - TY - JOUR AB - BACKGROUND & AIMS: Non-alcoholic fatty liver disease (NAFLD) is associated with increased risk of hepatic, cardiovascular, and metabolic diseases. High-protein diets, rich in methionine and branched chain amino acids (BCAAs), apparently reduce liver fat but may induce insulin resistance. We investigated the effects of diets high in animal protein vs plant protein, which differ in levels of methionine and BCAA, in subjects with type 2 diabetes and NAFLD. We examined levels of liver fat, lipogenic indices, markers of inflammation, serum levels of fibroblast growth factor 21 (FGF21), and activation of signaling pathways in adipose tissue. METHODS: We performed a prospective study of individuals with type 2 diabetes and NAFLD at a tertiary medical center in Germany, from June 2013 through March 2015. We analyzed data from 37 subjects placed on a diet high in animal protein (AP, rich in meat and dairy foods; n=18) or plant protein (PP, mainly legume protein; n=19) without calorie restriction for 6 weeks. The diets were isocaloric with the same macronutrient composition (30% protein, 40% carbohydrates, and 30% fat). Participants were examined at the start of the study and after the 6-week diet period for body mass index, body composition, hip circumference, resting energy expenditure, and respiratory quotient. Body fat and intrahepatic fat were detected by magnetic resonance imaging and spectroscopy, respectively. Levels of glucose, insulin, liver enzymes, and inflammation markers as well as individual free fatty acids and free amino acids were measured in collected blood samples. Hyperinsulinemic euglycemic clamps were performed to determine whole-body insulin sensitivity. Subcutaneous adipose tissue samples were collected and analyzed for gene expression patterns and phosphorylation of signaling proteins. RESULTS: Post-prandial levels of BCAAs and methionine were significantly higher in subjects on the AP vs the PP diet. The AP and PP diets each reduced liver fat by 36%-48% within 6 weeks (P for AP diet=.0002, P for PP diet=.001). These reductions were unrelated to change in body weight but correlated with downregulation of lipolysis and lipogenic indices. Serum level of FGF21 decreased by 50% in each group (P for AP diet<.0002, P for PP diet<.0002); decrease in FGF21 correlated with loss of hepatic fat. In gene expression analyses of adipose tissue, expression of the FGF21 receptor cofactor klotho beta was associated with reduced expression of genes encoding lipolytic and lipogenic proteins. In subjects on each diet, levels of hepatic enzymes and markers of inflammation decreased, insulin sensitivity increased, and serum level of keratin 18 decreased. CONCLUSIONS: In a prospective study of patients with type 2 diabetes, we found diets high in protein (either animal or plant) to significantly reduce liver fat, independently of body weight, and reduce markers of insulin resistance and hepatic necroinflammation. The diets appear to mediate these changes via lipolytic and lipogenic pathways in adipose tissue. Negative effects of BCAA or methionine were not detectable. FGF21 level appears to be a marker of metabolic improvement. ClincialTrials.gov no: NCT02402985. AU - Markova, M.* AU - Pivovarova, O.* AU - Hornemann, S.* AU - Sucher, S.* AU - Frahnow, T.* AU - Wegner, K.* AU - Machann, J. AU - Petzke, K.J.* AU - Hierholzer, J.* AU - Lichtinghagen, R.* AU - Herder, C.* AU - Carstensen-Kirberg, M.* AU - Roden, M.* AU - Rudovich, N.N.* AU - Klaus, S.* AU - Thomann, R.* AU - Schneeweiss, R.* AU - Rohn, S.* AU - Pfeiffer, A.F.* C1 - 49803 C2 - 40958 CY - Philadelphia SP - 571-585.e8 TI - Isocaloric diets high in animal or plant protein reduce liver fat and inflammation in individuals with Type 2 Diabetes. JO - Gastroenterology VL - 152 IS - 3 PB - W B Saunders Co-elsevier Inc PY - 2017 SN - 0016-5085 ER - TY - JOUR AU - Pikarsky, E.* AU - Heikenwälder, M. C1 - 50156 C2 - 42038 CY - Philadelphia SP - 780-783 TI - Focal and Local: Ectopic lymphoid structures and aggregates of myeloid and other immune cells in liver. JO - Gastroenterology VL - 151 IS - 5 PB - W B Saunders Co-elsevier Inc PY - 2016 SN - 0016-5085 ER - TY - JOUR AU - Protzer, U. AU - Knolle, P.A.* C1 - 49681 C2 - 40813 CY - Philadelphia SP - 805-806 TI - "To be or not to be": Immune tolerance in chronic hepatitis B. JO - Gastroenterology VL - 151 IS - 5 PB - W B Saunders Co-elsevier Inc PY - 2016 SN - 0016-5085 ER - TY - JOUR AB - BACKGROUND & AIM: Anemia is commonly associated with acute and chronic inflammation, but the mechanisms of their interaction are not clear. We investigated whether microRNA 122 (MIR122), which is generated in the liver and is secreted into the blood, is involved in the development of anemia associated with inflammation. METHODS: We characterized the primary transcript of the human liver-specific MIR122 using northern blot, quantitative real-time PCR, and 3' and 5' RACE analyses. We studied regulation of MIR122 in human hepatocellular carcinoma (HCC) cell lines (Huh7 and HepG2) as well as in C57BL/6 and mice with disruption of the tumor necrosis factor gene (Tnf). Liver tissues were collected and analyzed by bioluminescence imaging or immunofluorescence. Inflammation in mice was induced by lipopolysaccharide (LPS) or by cerulein injections. Mice were given 4 successive injections of LPS, leading to inflammation-induced anemia. Steatohepatitis was induced with a choline-deficient high-fat diet. Hemolytic anemia was stimulated by phenylhydrazine injection. MIR122 was inhibited in mice by tail-vein injection of antogomiR-122 (an oligonucleotide antagonist of MIR122). MicroRNA and mRNA levels were determined by quantitative real time PCR. RESULTS: The primary transcript of MIR122 spanned 5 kb, comprising 3 exons; the third encodes MIR122. Within the MIR122 promoter region we identified a nuclear factor-κB (NF-κB) binding site and demonstrated that RELA, as well as activators of NF-κB (TNF and LPS), increased promoter activity of MIR122. Administration of LPS to mice induced secretion of MIR122 into blood, which required TNF. Secreted MIR122 reached the kidney and reduced expression of erythropoietin (Epo), which we identified as a MIR122 target gene. Injection of mice with antagomiR-122 increased blood levels of EPO, reticulocytes, and hemoglobin. We found an inverse relationship between blood levels of MIR122 and EPO in mice with acute pancreatitis or steatohepatitis, and also in patients with acute inflammation. CONCLUSION: In mice, we found that LPS-induced inflammation increases blood levels of MIR122, which reduces expression of Epo in the kidney; this is a mechanism of inflammation-induced anemia. Strategies to block MIR122 in patients with inflammation could reduce the development or progression of anemia. AU - Rivkin, M.* AU - Simerzin, A.* AU - Zorde-Khvalevsky, E.* AU - Chai, C.* AU - Yuval, J.B.* AU - Rosenberg, N.* AU - Harari-Steinfeld, R.* AU - Schneider, R.* AU - Amir, G.* AU - Condiotti, R.* AU - Heikenwälder, M. AU - Weber, A.* AU - Schramm, C.* AU - Wege, H.* AU - Kluwe, J.* AU - Galun, E.* AU - Giladi, H.* C1 - 49210 C2 - 40690 CY - Philadelphia SP - 999-1010.e3 TI - Inflammation-induced expression and secretion of microRNA 122 leads to reduced blood levels of kidney-derived erythropoietin and anemia. JO - Gastroenterology VL - 151 IS - 5 PB - W B Saunders Co-elsevier Inc PY - 2016 SN - 0016-5085 ER - TY - JOUR AB - BACKGROUND & AIMS: Cancer therapies are being developed based on our ability to direct T cells against tumor antigens. Glypican 3 (GPC3) is expressed by 75% of all hepatocellular carcinomas (HCC) but not in healthy liver tissue or other organs. We aimed to generate T cells with GPC3-specific receptors that recognize HCC and used them to eliminate GPC3-expressing xenograft tumors grown from human HCC cells in mice. METHODS: We used mass spectrometry and to obtain a comprehensive peptidome from GPC3-expressing hepatoma cells after immune-affinity purification of HLA-A2, and used bioinformatics to identify immunodominant peptides. To circumvent GPC3-tolerance resulting from fetal expression, dendritic cells from HLA-A2 negative donors were co-transfected with GPC3 and HLA-A2 RNA to stimulate and expand antigen-specific T cells. RESULTS: Peptide GPC3367 was identified as a predominant peptide on HLA-A2. We used A2-GPC3367 multimers to detect, select for, and clone GPC3-specific T cells. These clones bound the A2-GPC3367 multimer and secreted interferon-γ when cultured with GPC3367, but not with control peptide loaded cells. By genomic sequencing of these T-cell clones, we identified a gene encoding a dominant T-cell receptor. The gene was cloned, the sequence was codon optimized and expressed from a retroviral vector. Primary CD8(+) T cells that expressed the transgenic T-cell receptor specifically bound GPC3367 on HLA-A2. These T cells killed GPC3-expressing hepatoma cells in culture and slowed growth of HCC xenograft tumors in mice. CONCLUSION: We identified a GPC3367-specific T-cell receptor. Expression of this receptor by T cells allows them to recognize and kill GPC3-positive hepatoma cells. This finding could be used to advance development of adoptive T-cell therapy for HCC. AU - Dargel, C. AU - Bassani-Sternberg, M.* AU - Hasreiter, J. AU - Zani, F. AU - Bockmann, J.H. AU - Thiele, F. AU - Bohne, F. AU - Wisskirchen, K. AU - Wilde, S. AU - Sprinzl, M.F.* AU - Schendel, D.J. AU - Krackhardt, A.M. AU - Uckert, W.* AU - Wohlleber, D.* AU - Schiemann, M. AU - Stemmer, K. AU - Heikenwälder, M. AU - Busch, D.H. AU - Richter, G.* AU - Mann, M.* AU - Protzer, U. C1 - 45128 C2 - 37247 SP - 1042-1052 TI - T cells engineered to express a T-cell receptor specific for glypican-3 to recognize and kill hepatoma cells in vitro and in mice. JO - Gastroenterology VL - 149 IS - 4 PY - 2015 SN - 0016-5085 ER - TY - JOUR AB - BACKGROUND & AIMS: Little is known about the mechanisms of the progressive tissue destruction, inflammation, and fibrosis that occur during development of chronic pancreatitis. Autophagy is involved in multiple degenerative and inflammatory diseases, including pancreatitis, and requires the protein autophagy related 5 (ATG5). We created mice with defects in autophagy to determine its role in pancreatitis. METHODS: We created mice with pancreas-specific disruption of Atg5 (Ptf1aCre(ex1);Atg5(F/F) mice), and compared them to control mice. Pancreata were collected and histology, immunohistochemistry, transcriptome, and metabolome analyses were performed. ATG5-deficient mice were placed on diets containing 25% palm oil and compared to those on a standard diet. Another set of mice received the antioxidant N-acetylcysteine. Pancreatic tissues were collected from 8 patients with chronic pancreatitis (CP) and compared to pancreata from ATG5-deficient mice. RESULTS: Mice with pancreas-specific disruption of Atg5 developed atrophic CP, independent of β-cell function; a greater proportion of male mice developed CP than females. Pancreata from ATG5-deficient mice had signs of inflammation, necrosis, acinar-to-ductal metaplasia, and acinar-cell hypertrophy; this led to tissue atrophy and degeneration. Based on transcriptome and metabolome analyses, ATG5-deficient mice produced higher levels of reactive oxygen species than control mice, and had insufficient activation of glutamate-dependent metabolism. Pancreata from these mice had reduced autophagy, increased levels of p62, and increases in endoplasmic reticulum stress and mitochondrial damage, compared with tissues from control mice; p62 signaling to Nqo1 and p53 was also activated. Dietary antioxidants, especially in combination with palm oil-derived fatty acids, blocked progression to CP and pancreatic acinar atrophy. Tissues from patients with CP had many histologic similarities to those from ATG5-deficient mice. CONCLUSIONS: Mice with pancreas-specific disruption of Atg5 develop a form of CP similar to that of humans. CP development appears to involve defects in autophagy, glutamate-dependent metabolism, and increased production of reactive oxygen species. These mice might be used to identify therapeutic targets for CP. AU - Diakopoulos, K.N.* AU - Lesina, M.* AU - Wörmann, S.* AU - Song, L.* AU - Aichler, M. AU - Schild, L.* AU - Artati, A. AU - Römisch-Margl, W. AU - Wartmann, T.* AU - Fischer, R.* AU - Kabiri, Y. AU - Zischka, H. AU - Halangk, W.* AU - Demir, I.E.* AU - Pilsak, C.* AU - Walch, A.K. AU - Mantzoros, C.S.* AU - Steiner, J.M.* AU - Erkan, M.* AU - Schmid, R.M.* AU - Witt, H.* AU - Adamski, J. AU - Algül, H.* C1 - 42987 C2 - 35916 CY - Philadelphia SP - 626-638 TI - Impaired autophagy induces chronic atrophic pancreatitis in mice via sex- and nutrition-dependent processes. JO - Gastroenterology VL - 148 IS - 3 PB - W B Saunders Co-elsevier Inc PY - 2015 SN - 0016-5085 ER - TY - JOUR AB - BACKGROUND & AIMS: Viral clearance involves immune cell cytolysis of infected cells. However, studies of hepatitis B virus (HBV) infection in chimpanzees have indicated that cytokines released by T cells can also promote viral clearance via non-cytolytic processes. We investigated the non-cytolytic mechanisms by which T cells eliminate HBV from infected hepatocytes. METHODS: We performed cytokine ELISA of serum samples from patients with acute and chronic hepatitis B. Liver biopsies were analyzed by in situ hybridization. HepG2-H1.3 cells, HBV-infected HepaRG cells, and primary human hepatocytes were incubated with interferon-γ (IFNγ) or tumor necrosis factor-α (TNFα), or co-cultured with T cells. We measured markers of HBV replication, including the covalently closed circular DNA (cccDNA). RESULT: Levels of IFNγ and TNFα were increased in serum samples from patients with acute vs chronic hepatitis B and controls. In human hepatocytes with stably replicating HBV, as well as in HBV-infected primary human hepatocytes or HepaRG cells, IFNγ and TNFα each induced deamination of cccDNA and interfered with its stability; their effects were additive. HBV-specific T cells, through secretion of IFNγ and TNFα, inhibited HBV replication and reduced cccDNA in infected cells without the direct contact required for cytolysis. Blocking IFNγ and TNFα after T-cell stimulation prevented the loss of cccDNA. Deprivation of cccDNA required activation of nuclear APOBEC3 deaminases by the cytokines. In liver biopsies from patients with acute hepatitis B, but not chronic hepatitis B or controls, hepatocytes expressed APOBEC3A and APOBEC3B. CONCLUSION: IFNγ and TNFα, produced by T cells, reduce levels of HBV cccDNA in hepatocytes by inducing deamination and subsequent cccDNA decay. AU - Xia, Y. AU - Stadler, D. AU - Lucifora, J. AU - Reisinger, F. AU - Webb, D.* AU - Hösel, M.* AU - Michler, T. AU - Wisskirchen, K. AU - Cheng, X. AU - Zhang, K. AU - Chou, W.M. AU - Wettengel, J.M. AU - Malo, A.* AU - Bohne, F. AU - Hoffmann, D. AU - Eyer, F.* AU - Thimme, R.* AU - Falk, C.S.* AU - Thasler, W.E.* AU - Heikenwälder, M. AU - Protzer, U. C1 - 46945 C2 - 39075 SP - 194-205 TI - Interferon-γ and tumor necrosis factor-α produced by T cells reduce the HBV persistence form, cccDNA, without cytolysis. JO - Gastroenterology VL - 150 IS - 1 PY - 2015 SN - 0016-5085 ER - TY - JOUR AB - BACKGROUND & AIMS: Very early onset inflammatory bowel diseases (VEOIBD), including infant disorders, are a diverse group of diseases found in children less than 6 years of age. They have been associated with several gene variants. We aimed to identify genes that cause VEOIBD. METHODS: We performed whole-exome sequencing of DNA from 1 infants with severe enterocolitis and her parents. Candidate gene mutations were validated in 40 pediatric patients and functional studies were carried out using intestinal samples and human intestinal cell lines. RESULTS: We identified compound heterozygote mutations in the tetratricopeptide repeat domain 7 (TTC7A) gene in an infant from non-consanguineous parents with severe exfoliative apoptotic enterocolitis; we also detected the mutations in 2 unrelated families, each with 2 affected siblings. TTC7A interacts with EFR3 homolog B (EFR3B) to regulate phosphatidylinositol 4-kinase (PI4KA) at the plasma membrane. Functional studies demonstrated that TTC7A is expressed in human enterocytes. The mutations we identified in TTC7A result in either mislocalization or reduced expression of TTC7A. PI4KA was found to co-immunoprecipitate with TTC7A; the identified TTC7A mutations reduced this binding. Knockdown of TTC7A in human intestinal-like cell lines reduced their adhesion, increased apoptosis, and decreased production of phosphatidylinositol 4-phosphate. CONCLUSION: In a genetic analysis, we identified loss of function mutations in TTC7A in 5 infants with VEOIBD. Functional studies demonstrated that the mutations cause defects in enterocytes and T cells that lead to severe apoptotic enterocolitis. Defects in the PI4KA-TTC7A-EFR3B pathway are involved in the pathogenesis of VEOIBD. AU - Avitzur, Y.* AU - Guo, C.* AU - Mastropaolo, L.A.* AU - Bahrami, E.* AU - Chen, H.* AU - Zhao, Z.* AU - Elkadri, A.* AU - Dhillon, S.* AU - Murchie, R.* AU - Fattouh, R.* AU - Huynh, H.* AU - Walker, J.L.* AU - Wales, P.W.* AU - Cutz, E.* AU - Kakuta, Y.* AU - Dudley, J.* AU - Kammermeier, J.* AU - Powrie, F.* AU - Shah, N.* AU - Walz, C.* AU - Nathrath, M. AU - Kotlarz, D.* AU - Puchaka, J.* AU - Krieger, J.* AU - Racek, T.* AU - Kirchner, T.* AU - Walters, T.D.* AU - Brumell, J.H.* AU - Griffiths, A.M.* AU - Rezaei, N.* AU - Rashtian, P.* AU - Najafi, M.* AU - Monajemzadeh, M.* AU - Pelsue, S.* AU - McGovern, D.P.* AU - Uhlig, H.H.* AU - Schadt, E.* AU - Klein, C.* AU - Snapper, S.B.* AU - Muise, A.M.* C1 - 29289 C2 - 33637 CY - Philadelphia SP - 1028-1039 TI - Mutations in tetratricopeptide repeat domain 7A result in a severe form of very early onset inflammatory bowel disease. JO - Gastroenterology VL - 146 IS - 4 PB - W B Saunders Co-elsevier Inc PY - 2014 SN - 0016-5085 ER - TY - JOUR AB - Reductions in levels of the hunger-stimulating hormone ghrelin have been proposed to mediate part of the effects of vertical sleeve gastrectomy (VSG) and Roux-en-Y gastric bypass surgeries for obesity. We studied circulating levels of acyl and desacyl ghrelin in rats after these surgeries. We found that blood levels of ghrelin were reduced after VSG, but not after Roux-en-Y gastric bypass, based on enzyme-linked immunosorbent assay and mass-spectrometry analyses. We compared the effects of VSG in ghrelin-deficient mice and wild-type mice on food intake, body weight, dietary fat preference, and glucose tolerance. We found that VSG produced comparable outcomes in each strain. Reduced ghrelin signaling therefore does not appear to be required for these effects of VSG. AU - Chambers, A.P.* AU - Kirchner, H.* AU - Wilson-Perez, H.E.* AU - Willency, J.A.* AU - Hale, J.E.* AU - Gaylinn, B.D. AU - Thorner, M.O. AU - Pfluger, P.T. AU - Gutierrez, J.A.* AU - Tschöp, M.H. AU - Sandoval, D.A.* AU - Seeley, R.J.* C1 - 11123 C2 - 30508 SP - 50-52 TI - The effects of vertical sleeve gastrectomy in rodents are ghrelin independent. JO - Gastroenterology VL - 144 IS - 1 PB - Elsevier PY - 2013 SN - 0016-5085 ER - TY - JOUR AB - BACKGROUND & AIMS: Genome-wide association studies (GWAS) have identified 140 Crohn's disease (CD) susceptibility loci. For most loci, the variants that cause disease are not known and the genes affected by these variants have not been identified. We aimed to identify variants that cause CD through detailed sequencing, genetic association, expression, and functional studies. METHODS: We sequenced whole exomes of 42 unrelated subjects with CD and 5 healthy subjects (controls) and then filtered single nucleotide variants by incorporating association results from meta-analyses of CD GWAS and in silico mutation effect prediction algorithms. We then genotyped 9348 subjects with CD, 2868 subjects with ulcerative colitis, and 14,567 control subjects and associated variants analyzed in functional studies using materials from subjects and controls and in vitro model systems. RESULTS: We identified rare missense mutations in PR domain-containing 1 (PRDM1) and associated these with CD. These mutations increased proliferation of T cells and secretion of cytokines on activation and increased expression of the adhesion molecule L-selectin. A common CD risk allele, identified in GWAS, correlated with reduced expression of PRDM1 in ileal biopsy specimens and peripheral blood mononuclear cells (combined P = 1.6 × 10(-8)). We identified an association between CD and a common missense variant, Val248Ala, in nuclear domain 10 protein 52 (NDP52) (P = 4.83 × 10(-9)). We found that this variant impairs the regulatory functions of NDP52 to inhibit nuclear factor κB activation of genes that regulate inflammation and affect the stability of proteins in Toll-like receptor pathways. CONCLUSIONS: We have extended the results of GWAS and provide evidence that variants in PRDM1 and NDP52 determine susceptibility to CD. PRDM1 maps adjacent to a CD interval identified in GWAS and encodes a transcription factor expressed by T and B cells. NDP52 is an adaptor protein that functions in selective autophagy of intracellular bacteria and signaling molecules, supporting the role of autophagy in the pathogenesis of CD. AU - Ellinghaus, D.* AU - Zhang, H.* AU - Zeissig, S.* AU - Lipinski, S.* AU - Till, A.* AU - Jiang, T.* AU - Stade, B.* AU - Bromberg, Y.* AU - Ellinghaus, E.* AU - Keller, A.* AU - Rivas, M.A.* AU - Skieceviciene, J.* AU - Doncheva, N.T.* AU - Liu, X.* AU - Liu, Q.* AU - Jiang, F.* AU - Forster, M.* AU - Mayr, G.* AU - Albrecht, M.* AU - Häsler, R.* AU - Boehm, B.O.* AU - Goodall, J.* AU - Berzuini, C.R.* AU - Lee, J.* AU - Andersen, V.* AU - Vogel, U.* AU - Kupcinskas, L.* AU - Kayser, M.* AU - Krawczak, M.* AU - Nikolaus, S.* AU - Weersma, R.K.* AU - Ponsioen, C.Y.* AU - Sans, M.* AU - Wijmenga, C.* AU - Strachan, D.P.* AU - McArdle, W.L.* AU - Vermeire, S.* AU - Rutgeerts, P.* AU - Sanderson, J.D.* AU - Mathew, C.G.* AU - Vatn, M.H.* AU - Wang, J.* AU - Nöthen, M.M.* AU - Duerr, R.H.* AU - Büning, C.* AU - Brand, S.* AU - Glas, J.* AU - Winkelmann, J. AU - Illig, T. AU - Latiano, A.* AU - Annese, V.* AU - Halfvarson, J.* AU - D'Amato, M.* AU - Daly, M.J.* AU - Nothnagel, M.* AU - Karlsen, T.H.* AU - Subramani, S.* AU - Rosenstiel, P.* AU - Schreiber, S.* AU - Parkes, M.* AU - Franke, A.* C1 - 26316 C2 - 32170 SP - 339-347 TI - Association between variants of PRDM1 and NDP52 and Crohn's disease, based on exome sequencing and functional studies. JO - Gastroenterology VL - 145 IS - 2 PB - WB Saunders PY - 2013 SN - 0016-5085 ER - TY - JOUR AB - BACKGROUND & AIMS: Antiviral agents suppress hepatitis B virus (HBV) replication but do not clear the infection. A strong effector T-cell response is required to eradicate HBV, but this does not occur in patients with chronic infection. T cells might be directed toward virus-infected cells by expressing HBV-specific receptors, and thereby clear HBV and help to prevent development of liver cancer. In mice, we studied whether redirected T cells can engraft following adoptive transfer, without prior T-cell depletion, and whether the large amounts of circulating viral antigens inactivate the transferred T cells or lead to uncontrolled, immune-mediated damage. METHODS: CD8(+) T cells were isolated from mice and stimulated using an optimized protocol. Chimeric antigen receptors (CARs) that bind HBV envelope proteins (S-CAR) and activate T cells were expressed on the surface of cells using retroviral vectors. S-CAR-expressing CD8(+) T cells, which carried the marker CD45.1, were injected into CD45.2(+) HBV transgenic mice. We compared these mice with mice that received CD8(+) T cells induced by vaccination, cells that express a CAR without a proper signaling domain, or cells that express a CAR that does not bind HBV proteins (controls). RESULTS: CD8(+) T cells that expressed HBV-specific CARs recognized different HBV subtypes and were able to engraft and expand in immune-competent HBV transgenic mice. Following adoptive transfer, the S-CAR-expressing T cells localized to and functioned in the liver; they rapidly and efficiently controlled HBV replication, compared with controls, causing only transient liver damage. The large amount of circulating viral antigen did not impair or over-activate the S-CAR grafted T cells. CONCLUSION: T cells with a CAR specific for HBV envelop proteins localize to the livers of mice to reduce HBV replication, causing only transient liver damage. This immune-cell therapy might be developed for patients with chronic hepatitis B, regardless of their HLA-type. AU - Krebs, K.M. AU - Böttinger, N. AU - Huang, L.R.* AU - Chmielewski, M.* AU - Arzberger, S. AU - Gasteiger, G. AU - Jager, C. AU - Schmitt, E.* AU - Bohne, F. AU - Aichler, M. AU - Uckert, W.* AU - Abken, H.* AU - Heikenwälder, M. AU - Knolle, P.* AU - Protzer, U. C1 - 24348 C2 - 31514 SP - 456-465 TI - T cells expressing a chimeric antigen receptor that binds hepatitis B virus envelope proteins control virus replication in mice. JO - Gastroenterology VL - 145 IS - 2 PB - Saunders-Elsevier PY - 2013 SN - 0016-5085 ER - TY - JOUR AU - Tjalma, J.J.* AU - Garcia-Allende, P.* AU - van Scheltinga, A.T.* AU - Glatz, J.* AU - Hartmans, E.* AU - van Oosten, M.* AU - Koornstra, J.J.* AU - Kleibeuker, J.H.* AU - van Dam, G.M.* AU - Ntziachristos, V. AU - Nagengast, W.B.* C1 - 28019 C2 - 32905 SP - S177 TI - Towards instant detection of (pre)malignant lesions by ultrasensitive near-infrared fluorescence endoscopy, using molecular targeted fluorescent antibodies. JO - Gastroenterology VL - 144 PB - WB Saunders - Elsevier PY - 2013 SN - 0016-5085 ER - TY - JOUR AU - Balluff, B. AU - Rauser, S. AU - Ebert, M.P.* AU - Siveke, J.T.* AU - Höfler, H. AU - Walch, A.K. C1 - 8345 C2 - 30066 SP - 544-549 TI - Direct molecular tissue analysis by MALDI imaging mass spectrometry in the field of gastrointestinal disease. JO - Gastroenterology VL - 143 IS - 3 PB - Elsevier PY - 2012 SN - 0016-5085 ER - TY - JOUR AB - Studies of mechanisms responsible for the persistence of hepatitis B virus (HBV) infection have been hindered by a lack of appropriate animal models. HBV genomes can be delivered to livers of mice using hydrodynamic injection or high doses of an adenoviral vector; these lead to clearance of HBV. We found that infection of immunocompetent mice with low doses of an adenoviral vector resulted in persistent HBV infection; the mice neither underwent seroconversion to production of antibodies against HBV nor developed a strong HBV-specific effector T-cell response. As in patients with chronic HBV infection, DNA vaccination failed to generate T cells that cleared infection. This model of persistent HBV infection could be used to study the pathogenesis of chronic HBV infection and develop new therapeutic strategies. AU - Huang, L.R.* AU - Gabel, Y.A.* AU - Graf, S. AU - Arzberger, S. AU - Kurts, C.* AU - Heikenwälder, M. AU - Knolle, P.A.* AU - Protzer, U. C1 - 7574 C2 - 29866 SP - 1447-1450 TI - Transfer of HBV genomes using low doses of adenovirus vectors leads to persistent infection in immune competent mice. JO - Gastroenterology VL - 142 IS - 7 PB - HighWire Press PY - 2012 SN - 0016-5085 ER - TY - JOUR AB - BACKGROUND & AIMS: Little is known about the pathogenic mechanisms of autoimmune pancreatitis (AIP), an increasingly recognized, immune-mediated form of chronic pancreatitis. Current treatment options are limited and disease relapse is frequent. We investigated factors that contribute to the development of AIP and new therapeutic strategies. METHODS: We used quantitative polymerase chain reaction, immunohistochemical, and enzyme-linked immunosorbent analyses to measure the expression of cytokines and chemokines in tissue and serum samples from patients with and without AIP. We created a mouse model of human AIP by overexpressing lymphotoxin (LT)α and β specifically in acinar cells (Ela1-LTab mice). RESULTS: Messenger RNA levels of LTα and β were increased in pancreatic tissues from patients with AIP, compared with controls, and expression of chemokines (CXCL13, CCL19, CCL21, CCL1, and B-cell-activating factor) was increased in pancreatic and serum samples from patients. Up-regulation of these factors was not affected by corticosteroid treatment. Acinar-specific overexpression of LTαβ (Ela1-LTαβ) in mice led to an autoimmune disorder with various features of AIP. Chronic inflammation developed only in the pancreas but was sufficient to cause systemic autoimmunity. Acinar-specific overexpression of LTαβ did not cause autoimmunity in mice without lymphocytes (Ela1-LTab/Rag1(-/-)); moreover, lack of proinflammatory monocytes (Ela1-LTab/Ccr2(-/-)) failed to prevent AIP but prevented early pancreatic tissue damage. Administration of corticosteroids reduced pancreatitis but did not affect production of autoantibodies, such as antipancreatic secretory trypsin inhibitor in Ela1-LTab mice. In contrast, inhibition of LTβR signaling reduced chemokine expression, renal immune-complex deposition, and features of AIP in Ela1-LTab mice. CONCLUSIONS: Overexpression of LTαβ specifically in acinar cells of mice causes features of AIP. Reagents that neutralize LTβR ligands might be used to treat patients with AIP. AU - Seleznik, G.M.* AU - Reding, T.* AU - Romrig, F.* AU - Saito, Y.* AU - Mildner, A.* AU - Segerer, S.* AU - Sun, L.K.* AU - Regenass, S.* AU - Lech, M.* AU - Anders, H.J.* AU - McHugh, D.* AU - Kumagi, T.* AU - Hiasa, Y.* AU - Lackner, C.* AU - Haybaeck, J.* AU - Angst, E.* AU - Perren, A.* AU - Balmer, M.L.* AU - Slack, E.* AU - Macpherson, A.* AU - Manz, M.G.* AU - Weber, A.* AU - Browning, J.L.* AU - Arkan, M.C.* AU - Rülicke, T.* AU - Aguzzi, A.* AU - Prinz, M.* AU - Graf, R.* AU - Heikenwälder, M. C1 - 11198 C2 - 30547 SP - 1361-1374 TI - Lymphotoxin β receptor signaling promotes development of autoimmune pancreatitis. JO - Gastroenterology VL - 143 IS - 5 PB - Elsevier PY - 2012 SN - 0016-5085 ER - TY - JOUR AB - no Abstract AU - Bézière, N. AU - Ntziachristos, V. C1 - 6028 C2 - 29216 SP - 1979-1985 TI - Optoacoustic imaging: An emerging modality for the gastrointestinal tract. JO - Gastroenterology VL - 141 IS - 6 PB - AGA Institute PY - 2011 SN - 0016-5085 ER - TY - JOUR AB - Approved therapies for chronic hepatitis B include systemic administration of interferon (IFN)-alfa and inhibitors of hepatitis B virus (HBV) reverse-transcription. Systemic application of IFN-alfa is limited by side effects. Reverse-transcriptase inhibitors effectively control HBV replication, but rarely eliminate the virus and can select drug-resistant variants. We aimed to develop an alternative therapeutic approach that combines gene silencing with induction of IFN in the liver. METHODS:To stimulate an immune response while inhibiting HBV activity, we designed 3 small interfering (si)RNAs that target highly conserved sequences and multiple HBV transcripts of all genotypes. A 5'-triphosphate (3p) was added to the siRNAs, turning them into a ligand for the cytosolic helicase retinoic acid-inducible protein I, which becomes activated and induces expression of type-I IFNs. Antiviral activity was investigated in cell lines that replicate HBV, in HBV-infected primary human hepatocytes, and in HBV transgenic mice. RESULTS: 3p-double-stranded RNA (3p-RNA) activated retinoic acid-inducible protein I, induced a strong type I IFN response (expression of IFN-β) in liver cells and showed transient but strong antiviral activity. Bifunctional, HBV-specific, 3p-siRNAs controlled replication of HBV more efficiently and for longer periods of time than 3p-RNAs without silencing capacity or siRNAs that targeted identical sequences but did not contain 3p. CONCLUSIONS: HBV-specific 3p-siRNAs are bifunctional antiviral molecules that induce production of type I IFNs in the liver and target HBV RNAs to inhibit viral replication. AU - Ebert, G. AU - Poeck, H.* AU - Lucifora, J. AU - Baschuk, N.* AU - Esser, K. AU - Esposito, I. AU - Hartmann, G.* AU - Protzer, U. C1 - 6505 C2 - 28822 SP - 696-706 TI - 5' triphosphorylated small interfering RNAs control replication of hepatitis B virus and induce an interferon response in human liver cells and mice. JO - Gastroenterology VL - 141 IS - 2 PB - AGA Institute PY - 2011 SN - 0016-5085 ER - TY - JOUR AB - BACKGROUND & AIMS: An association between gastric cancer and the rs2294008 (C>T) polymorphism in the prostate stem cell antigen (PSCA) gene has been reported for several Asian populations. We set out to determine whether such an association exists in white individuals. METHODS: We genotyped 166 relatives of gastric cancer patients, including 43 Helicobacter pylori-infected subjects with hypochlorhydria and gastric atrophy, 65 infected subjects without these abnormalities, 58 H pylori-negative relatives, and 100 population controls. Additionally, a population-based study of chronic atrophic gastritis provided 533 cases and 1054 controls. We then genotyped 2 population-based, case-control studies of upper gastrointestinal cancer: the first included 312 gastric cancer cases and 383 controls; the second included 309 gastric cancer cases, 159 esophageal cancer cases, and 211 controls. Odds ratios were computed from logistic models and adjusted for confounding variables. RESULTS: Carriage of the risk allele (T) of rs2294008 in PSCA was associated with chronic atrophic gastritis (adjusted odds ratio [OR], 1.5; 95% confidence interval [CI]: 1.1-1.9) and noncardia gastric cancer (OR, 1.9; 95% CI: 1.3-2.8). The association was strongest for the diffuse histologic type (OR, 3.2; 95% CI: 1.2-10.7). An inverse association was observed between carriage of the risk allele and gastric cardia cancer (OR, 0.5; 95% CI: 0.3-0.9), esophageal adenocarcinoma (OR, 0.5; 95% CI: 0.3-0.9), and esophageal squamous cell carcinoma (OR, 0.4; 95% CI: 0.2-0.9) CONCLUSIONS: The rs2294008 polymorphism in PSCA increases the risk of noncardia gastric cancer and its precursors in white individuals but protects against proximal cancers. AU - Lochhead, P.* AU - Frank, B.* AU - Hold, G.L.* AU - Rabkin, C.S.* AU - Ng, M.T.H.* AU - Vaughan, T.L.* AU - Risch, H.A.* AU - Gammon, M.D.* AU - Lissowska, J.* AU - Weck, M.N.* AU - Raum, E.* AU - Müller, H.* AU - Illig, T. AU - Klopp, N. AU - Dawson, A.* AU - McColl, K.E.* AU - Brenner, H.* AU - Chow, W.H.* AU - El-Omar, E.M.* C1 - 5470 C2 - 28348 SP - 435-441 TI - Genetic variation in the prostate stem cell antigen gene and upper gastrointestinal cancer in white individuals. JO - Gastroenterology VL - 140 IS - 2 PB - Elsevier PY - 2011 SN - 0016-5085 ER - TY - JOUR AB - BACKGROUND & AIMS: The composition of the gastrointestinal microbiota is thought to have an important role in the etiology of inflammatory bowel diseases (IBDs) such as Crohn's disease (CD) and ulcerative colitis (UC). Interindividual variation and an inability to detect less abundant bacteria have made it difficult to correlate specific bacteria with disease. METHODS: We used 454 pyrotag sequencing to determine the compositions of microbial communities in feces samples collected from a cohort of 40 twin pairs who were concordant or discordant for CD or UC, and in mucosal samples from a subset of the cohort. The cohort primarily comprised patients who were in remission, but also some with active disease. RESULTS: The profiles of the microbial community differed with disease phenotypes; relative amounts of bacterial populations correlated with IBD phenotypes. The microbial compositions of individuals with CD differed from those of healthy individuals, but were similar between healthy individuals and individuals with UC. Profiles from individuals with CD that predominantly involved the ileum differed from those with CD that predominantly involved the colon; several bacterial populations increased or decreased with disease type. Changes specific to patients with ileal CD included the disappearance of core bacteria, such as Faecalibacterium and Roseburia, and increased amounts of Enterobacteriaceae and Ruminococcus gnavus. CONCLUSIONS: Bacterial populations differ in abundance among individuals with different phenotypes of CD. Specific species of bacteria are associated with ileal CD; further studies should investigate their role in pathogenesis. AU - Willing, B.P.* AU - Dicksved, J.* AU - Halfvarson, J.* AU - Andersson, A.F.* AU - Lucio, M. AU - Zheng, Z.* AU - Järnerot, G.* AU - Tysk, C.* AU - Jansson, J.K.* AU - Engstrand, L.* C1 - 5130 C2 - 27834 SP - 1844-1854 TI - A pyrosequencing study in twins shows that gastrointestinal microbial profiles vary with inflammatory bowel disease phenotypes. JO - Gastroenterology VL - 139 IS - 6 PB - Elsevier Inc. PY - 2010 SN - 0016-5085 ER - TY - JOUR AB - BACKGROUND & AIMS: Pancreatic cancers contain exclusively tumorigenic cancer stem cells (CSCs), which are highly resistant to chemotherapy, resulting in a relative increase in CSC numbers during gemcitabine treatment. Signaling through sonic hedgehog and mammalian target of rapamycin (mTOR), respectively, may be essential for CSC self-renewal and could represent putative targets for novel treatment modalities. METHODS: We used in vitro and in vivo models of pancreatic cancer to examine the effects of sonic hedgehog inhibition (cyclopamine/CUR199691) and mTOR blockade (rapamycin) on the tumorigenic CSC population. RESULTS: Surprisingly, neither cyclopamine nor rapamycin alone or as supplements to chemotherapy were capable of effectivety diminishing the CSC pool. Only the combined inhibition of both pathways together with chemotherapy reduced the number of CSCs to virtually undetectable levels in vitro and in vivo. Most importantly, in vivo administration of this triple combination in mice with established patient-derived pancreatic tumors was reasonably tolerated and translated into significantly prolonged long-term survival. CONCLUSIONS: The combined blockade of sonic hedgehog and mTOR signaling together with standard chemotherapy is capable of eliminating pancreatic CSCs. Further preclinical investigation of this promising approach may lead to the development of a novel therapeutic strategy to improve the devastating prognosis of patients with pancreatic cancer. AU - Mueller, M.T.* AU - Hermann, P.C.* AU - Witthauer, J.* AU - Rubio-Viqueira, B.* AU - Leicht, S.F.* AU - Huber, S.* AU - Ellwart, J.W. AU - Mustafa, M.* AU - Bartenstein, P.* AU - D'Haese, J.G.* AU - Schoenberg, M.H.* AU - Berger, F.* AU - Jauch, K.W.* AU - Hidalgo, M.* AU - Heeschen, C.* C1 - 1113 C2 - 26689 CY - Philadelphia SP - 1102-1113 TI - Combined targeted treatment to eliminate tumorigenic cancer stem cells in human pancreatic cancer. JO - Gastroenterology VL - 137 IS - 3 PB - W B Saunders Co-Elsevier Inc PY - 2009 SN - 0016-5085 ER - TY - JOUR AB - The final goal in hepatitis B therapy is eradication of the hepatitis B virus (HBV) replication template, the so-called covalently closed circular DNA (cccDNA). Current antiviral treatment of chronic hepatitis B depends on interferon alpha or nucleoside analogues inhibiting the viral reverse transcriptase. Despite treatment, cccDNA mostly persists in the host cell nucleus, continues to produce hepatitis B surface antigen (HBsAg), and causes relapsing disease. We therefore aimed at eliminating persistently infected hepatocytes carrying HBV cccDNA by redirecting cytolytic T cells toward HBsAg-producing cells. METHODS: We designed chimeric T-cell receptors directed against HBV surface proteins present on HBV-infected cells and used them to graft primary human T cells with antibody-like specificity. The receptors were composed of a single chain antibody fragment directed against HBV S or L protein fused to intracellular signalling domains of CD3xi and the costimulatory CD28 molecule. RESULTS: Our results show that these chimeric receptors, when retrovirally delivered and expressed on the cell surface, enable primary human T cells to recognize HBsAg-positive hepatocytes, release interferon gamma and interleukin 2, and, most importantly, lyse HBV replicating cells. When coincubated with HBV-infected primary human hepatocytes, these engineered, antigen-specific T cells selectively eliminated HBV-infected and thus cccDNA-positive target cells. CONCLUSIONS: Elimination of HBV cccDNA-positive hepatocytes following antiviral therapy is a major therapeutic goal in chronic hepatitis B, and adoptive transfer of grafted T cells provides a promising novel therapeutic approach. However, T-cell therapy may also cause liver damage and therefore needs further preclinical evaluation. AU - Bohne, F.* AU - Chmielewski, M.* AU - Ebert, G.* AU - Wiegmann, K.* AU - Kürschner, T.* AU - Schulze, A.* AU - Urban, S.* AU - Krönke, M.* AU - Abken, H.* AU - Protzer, U. C1 - 2278 C2 - 25462 SP - 239-247 TI - T cells redirected against hepatitis B virus surface proteins eliminate infected hepatocytes. JO - Gastroenterology VL - 134 IS - 1 PB - Elsevier PY - 2008 SN - 0016-5085 ER - TY - JOUR AB - Electronic (video) endoscopes are a significant new development in gastroenterology, offering the potential of enhanced teaching and permanent storage of pictorial data. The primary concern of gastroenterologists is the resolution and color performance of these instruments, as these parameters have important bearings on the ability to discern pathological changes in mucosa. We sought to determine the resolution and color capabilities of electronic colonscopes and compare them with a conventional fiber colonoscope. Resolution was determined using a standard test chart at various distances and the number of picture elements (a measure of resolution) was calculated. The mean number of picture elements was Fujinon (219), Fiber (172), Pentax (169), Toshiba (142), Olympus (140), and Welch Allyn (133). In close focus examination (target distances <1 cm), the Fujinon and Toshiba endoscopes had significantly higher resolution than the other instruments. Color was measured quantitatively using a standard color chart and a color analyzer. Color polygons were plotted for each endoscope on a reference chromaticity diagram. All systems had an acceptable overall performance but color was undersaturated with some systems. The optical performance of electronic endoscopes has improved considerably since the inception of electronic endoscopy. AU - Knyrim, K. AU - Seidlitz, H.K. AU - Vakil, N.B. AU - Hagenmüller, F. AU - Claßen, M. C1 - 42098 C2 - 0 SP - 776-782 TI - Optical performance of electronic imaging systems for the colon. JO - Gastroenterology VL - 96 IS - 3 PY - 1989 SN - 0016-5085 ER -