Effects of Helicobacter PYLORI Treatment on Incidence of Gastric Cancer in Older Individuals Gastroenterology (IF 18.392) Pub Date : 2018-03-14 Wai K. Leung, Irene OL. Wong, Ka Shing Cheung, Kar Fu Yeung, Esther W. Chan, Angel YS. Wong, Lijia Chen, Ian CK. Wong, David Y. Graham
Background & AimsAlthough eradication of Helicobacter pylori infection reduces the risk of gastric cancer, few data are available on its effects in older subjects. We compared the age-specific risk of gastric cancer in a large cohort of subjects who received H pylori eradication therapy vs a matched general population.MethodsWe searched the Hospital Authority database of Hong Kong to identify individuals with H pylori infection who had received a course of clarithromycin-containing eradication therapy from January 2003 through December 2012. We compared the gastric cancer incidence in this cohort with the expected incidence for the local general population by retrieving the gastric cancer incidence of the age- and sex-matched population from 2003 through 2014 (the latest available year) from the Hong Kong Cancer Registry. The primary outcome was the incidence of gastric cancer development in the cohort treated for H pylori infection vs the expected number of gastric cancer cases in the general population. Analyses were conducted by a priori age groups of less than 40 years, 40–59 years, and 60 years or older.ResultsAmong 73,237 subjects infected with H pylori who received eradication therapy, 200 (0.27%) developed gastric cancer during a median follow-up time of 7.6 years. Compared with the matched general population, the gastric cancer risk was significantly lower in subjects 60 years or older who had received H pylori treatment (standardized incidence ratio [SIR], 0.82; 95% CI, 0.69–0.97; P=.02) but not in younger groups. When data were stratified based on time from H pylori treatment (less than 5 years, 5–9 years, and 10 or more years), the risk of gastric cancer was significantly lower than the general population 10 or more years after eradication in the group 40–59 years old (SI, 0.32; 95% CI 0.08–0.88; P=.04) and the group 60 years or older (SIR, 0.42; 95% CI 0.42–0.84; p = 0.02) than the other age groups.ConclusionsIn an analysis of data from a public hospital database on Hong Kong, we associated treatment of H pylori infection with a lower risk of gastric cancer, particularly in older subjects, 10 or more years after treatment.
Non-lytic Lymphocytes Engineered to Express Virus-specific T-cell Receptors Limit HBV Infection by Activating APOBEC3 Gastroenterology (IF 18.392) Pub Date : 2018-03-14 Sarene Koh, Janine Kah, Christine Y.L. Tham, Ninghan Yang, Erica Ceccarello, Adeline Chia, Margaret Chen, Atefeh Khakpoor, Andrea Pavesi, Anthony T. Tan, Maura Dandri, Antonio Bertoletti
Background & AimsStrategies to develop virus-specific T cells against hepatic viral infections have been hindered by safety concerns. We engineered non-lytic human T cells to suppress replication of hepatitis B virus (HBV) and hepatitis C virus (HCV) without overt hepatotoxicity, and investigated their antiviral activity.MethodsWe electroporated resting T cells or T cells activated by anti-CD3 with mRNAs encoding HBV or HCV-specific T-cell receptors (TCRs) to create 2 populations of TCR-reprogrammed T cells. We tested their ability to suppress HBV or HCV replication without lysis in 2-dimensional and 3-dimensional cultures of HepG2.2.15 cells and HBV-infected HepG2-hNTCP cells. We also injected TCR-reprogrammed resting and activated T cells into HBV-infected uPA/SCID/IL2γ mice with humanized livers and measured levels of intrahepatic and serological viral parameters and serum alanine aminotransferase (ALT). Livers were collected for analysis of gene expression patterns to determine effects of the TCR-reprogrammed T cells.ResultsTCR-reprogrammed resting T cells produced comparable levels of interferon gamma (IFNG) but lower levels of perforin and granzyme than activated T cells and did not lyse HCV- or HBV-infected hepatoma cells. While T-cell secretion of IFNG was required to inhibit HCV replication, the HBV-specific TCR-reprogrammed resting T cells reduced HBV replication also through intracellular activation of apolipoprotein B mRNA editing enzyme, catalytic polypeptide 3 (APOBEC3). The mechanism of APOBEC3 intracellular activation involved temporal expression of lymphotoxin-β-receptor ligands on resting T cells after TCR-mediated antigen recognition and activation of lymphotoxin-β-receptor in infected cells.ConclusionsWe developed TCR-reprogrammed non-lytic T cells capable to activate APOBEC3 in hepatoma cells and in HBV-infected human hepatocytes in mice, limiting viral infection. These cells with limited hepatotoxicity might be developed for treatment of chronic HBV infection.
Patterns and trends of liver cancer incidence rates in Eastern and Southeastern Asian Countries (1983–2007) and predictions to 2030 Gastroenterology (IF 18.392) Pub Date : 2018-03-14 Jie Wu, Shigui Yang, Kaijin Xu, Cheng Ding, Yuqing Zhou, Xiaofang Fu, Yiping Li, Min Deng, Chencheng Wang, Xiaoxiao Liu, Lanjuan Li
Background & AimsWe examined temporal trends in liver cancer incidence rates overall and by histological type from the years 1983 through 2007. We predict trends in liver cancer incidence rates through 2030 for selected Eastern and Southeastern Asian countries.MethodsData on yearly liver cancer incident cases by age group and sex were drawn from 6 major selected Eastern and Southeastern Asian countries or regions with cancer registries available in the CI5plus database, including China, Japan, Hong Kong Special Administrative Region (SAR), the Philippines, Singapore, and Thailand. We also analyzed data for the United States and Australia for comparative purposes. Age-standardized incidence rates were calculated and plotted from 1983 through 2007. Numbers of new cases and incidence rates were predicted through 2030 by fitting and extrapolating age–period–cohort models.ResultsThe incidence rates of liver cancer have been decreasing, and decreases will continue in all selected Eastern and Southeastern Asian countries, except for Thailand, whose liver cancer incidence rate will increase due to the increasing incidence rate of intrahepatic cholangiocarcinomas. Even though the incidence rates of liver cancer are predicted to decrease in most Eastern and Southeastern Asian countries, the burden, in terms of new cases, will continue to increase because of population growth and aging.ConclusionsBased on an analysis of data from cancer registries from Asian countries, incidence rates of liver cancer are expected to decrease through 2030 in most Eastern and Southeastern Asian countries. However, in Thailand, the incidence rate of intrahepatic cholangiocarcinomas is predicted to increase, so health education programs are necessary.
Lamins and Lamin-associated Proteins in Gastrointestinal Health and Disease Gastroenterology (IF 18.392) Pub Date : 2018-03-13 Graham F. Brady, Raymond Kwan, Juliana Bragazzi Cunha, Jared S. Elenbaas, M. Bishr Omary
The nuclear lamina is a multi-protein lattice composed of A- and B-type lamins and their associated proteins. This protein lattice associates with heterochromatin and integral inner nuclear membrane proteins, providing a link between the genome, nucleoskeleton, and cytoskeleton. In the 1990s, mutations in EMD and LMNA were linked to Emery-Dreifuss muscular dystrophy. Since then, the number of diseases attributed to nuclear lamina defects, including laminopathies and other disorders, has increased to include more than 20 distinct genetic syndromes. Studies of patients and mouse genetic models have indicated the important roles for lamins and their associated proteins in the function of gastrointestinal organs including liver and pancreas. We review the interactions and functions of the lamina in relation to the nuclear envelope and genome, the ways in which its dysfunction is thought to contribute to human disease, and possible avenues for targeted therapies.
Dysregulated Response of Follicular Helper T cells to Hepatitis B Surface Antigen Promotes HBV Persistence in Mice and Associates With Outcomes of Patients Gastroenterology (IF 18.392) Pub Date : 2018-03-12 Xiaowen Wang, Qingyang Dong, Qian Li, Yuanyuan Li, Dianyuan Zhao, Jinjie Sun, Junliang Fu, Fanping Meng, Hu Lin, Junjie Luan, Biao Liu, Min Wang, Fusheng Wang, Fuchu He, Li Tang
Background & AimsProduction of neutralizing antibodies against hepatitis B surface antigen (HBsAg) is dysregulated in patients with persistent hepatitis V virus (HBV) infection. We investigated mechanisms by which this immune response to the virus is disrupted and whether it can be restored to promote clearance of HBV.MethodsImmune-competent C57BL/6N and C57BL/6J, as well as mice deficient in follicular helper T cells (Tfh cell-deficient), B cells, or Foxp3+ T-regulatory cells (Treg cell-deficient), were given hydrodynamic injections of pAAV/HBV1.2 plasmids. Some mice were given injections of sorted Tfh cells, pan-B cells, Treg cells, or a blocking antibody against CTLA4. Production of antibodies against HBsAg and clearance of HBV were assessed by flow cytometry, ELISA, PCR and immunohistochemical analyses. We obtained blood samples from patients with HBV infection and isolated Treg cells. We measured the ability of Treg cells to suppress production of interleukin 21 (IL21) in CD4+ T cells.ResultsImmune-competent C57BL/6N and C57BL/6J mice transfected with the plasmid encoding HBV had features of viral clearance and viral persistence observed in humans. A Tfh-cell response to HBsAg was required for clearance of HBV and was suppressed by Treg cells in mice with persistent HBV infection. Depletion of Treg cells or inhibition of Treg-cell function (with blocking antibody against CTLA4) restored the Tfh-cell response against HBsAg and clearance of HBV in mice. Impaired Tfh cell response to HBsAg was observed in blood from patients with chronic HBV infection, responsiveness was restored by depletion of Treg cells or blocking antibody against CTLA4.ConclusionsIn studies of HBV-infected mice and blood from patients with chronic HBV infection, we found a Tfh-cell response to HBsAg of to be required for HBV clearance, and that this response was blocked by Treg cells. Inhibiting Treg cell activity using neutralizing antibody against CTLA4 restored the ability of Tfh cells to clear HBV infection; this approach might be developed for treatment of patients with chronic HBV infection.
Secretion of Hepatitis C Virus Replication Intermediates Reduces Activation of Toll Like Receptor 3 in Hepatocytes Gastroenterology (IF 18.392) Pub Date : 2018-03-11 Oliver Grünvogel, Ombretta Colasanti, Ji-Young Lee, Volker Klöss, Sandrine Belouzard, Anna Reustle, Katharina Esser-Nobis, Jasper Hesebeck-Brinckmann, Pascal Mutz, Katrin Hoffmann, Arianeb Mehrabi, Ronald Koschny, Florian W.R. Vondran, Daniel Gotthardt, Paul Schnitzler, Christoph Neumann-Haefelin, Robert Thimme, Marco Binder, Ralf Bartenschlager, Jean Dubuisson, Alexander H. Dalpke, Volker Lohmann
Background & aims Hepatitis C virus (HCV) infections most often result in chronic outcomes, although the virus constantly produces replication intermediates, in particular double-stranded RNA (dsRNA), representing potent inducers of innate immunity. We aimed to characterize the fate of HCV dsRNA in hepatocyte cultures to identify mechanisms contributing to viral persistence in presence of an active innate immune response. Methods We analyzed hepatocyte-based culture models for HCV for induction of innate immunity, secretion of virus positive- or negative-strand RNA, and viral replication using different quantification methods and microscopy techniques. Expression of pattern recognition receptors was reconstituted in hepatoma cells by lentiviral transduction. Results HCV-infected cells secrete substantial amounts of virus positive- and negative-strand RNAs in extracellular vesicles (EVs), toward the apical and basolateral domain of hepatocytes. Secretion of negative-strand RNA was independent from virus production, and viral RNA secreted in EVs contained higher relative amounts of negative strands, indicating that mostly virus dsRNA is released. A substantial part of viral replication complexes and dsRNA was found in the endosomal compartment and multi-vesicular bodies, indicating that secretion of HCV replication intermediates is mediated by the exosomal pathway. Block of vesicle release in HCV-positive cells increased intracellular dsRNA levels and increased activation of toll like receptor 3 (TLR3), inhibiting HCV replication. Conclusions Using hepatocyte-based culture models for HCV, we found a portion of HCV dsRNA intermediates to be released from infected cells in EVs, which reduces activation of TLR3. This represents a novel mechanism how HCV evades host immune responses, potentially contributing to viral persistence.
Baseline Factors Associated with Improvements in Decompensated Cirrhosis After Direct-acting Antiviral Therapy for HCV Infection Gastroenterology (IF 18.392) Pub Date : 2018-03-11 O. El-Sherif, Z.G. Jiang, E.B. Tapper, K.C. Huang, A. Zhong, A. Osinusi, M. Charlton, M. Manns, N.H. Afdhal, K. Mukamal, J. McHutchison, D.M. Brainard, N. Terrault, M.P. Curry
Background & Aims Treatment with direct-acting antiviral (DAA) agents can reduce model for end-stage liver disease and Child-Pugh-Turcotte (CPT) scores in patients with decompensated cirrhosis caused by hepatitis C virus (HCV). However, many of these patients still die or require liver transplantation. We collected data on baseline features of patients and aimed to develop a scoring system to predict response to DAA therapy. Methods We performed a retrospective analysis of data from 4 trials of the effects of sofosbuvir-based therapy in patients with HCV-associated decompensated cirrhosis (502 of CPT class B and 120 of CPT class C). In these trials, patients were given 12 or 24 weeks treatment with ledipasvir, sofosbuvir, and ribavirin or velpatasvir, sofosbuvir, and/or ribavirin, or 48 weeks treatment with sofosbuvir and ribavirin. We collected demographic, clinical, treatment response, and laboratory data from patients and tested their associations with patient outcomes at 36 weeks. The primary outcome was factors associated with reduction of CPT score to class A. Results The presence of ascites or encephalopathy, serum level of albumin below 3.5g/dL or alanine aminotransferase (ALT) below 60 U/L, and body mass index (BMI) above 25 kg/m2 associated with an increased risk of not achieving a reduction in CPT to class A, independent of sustained viral response to therapy. Serum level of albumin below 2.8g/dL and abnormal level of bilirubin associated with an increased risk of liver transplantation or death. We developed a scoring system based on 5 baseline factors (BMI, encephalopathy, ascites, and serum levels of ALT and albumin), which we called the BE3A; scores associated significantly with patient outcomes. For patients with scores of 4–5, the hazard ratio for reduction of CPT score to class A was 52.3 (95% CI, 15.2–179.7). Conclusions We identified 5 baseline factors (BMI, encephalopathy, ascites, and serum levels of ALT and albumin) associated with a reduction of CPT score to class A in patients with HCV-associated decompensated cirrhosis receiving DAA therapy. We developed a predictive score using these factors, called the BE3A score, that can be used as a shared decision making tool, quantifying the potential benefits of DAA therapy for patients with decompensated cirrhosis.
Analysis of Genes Associated with Monogenic Primary Immunodeficiency Identifies Rare Variants in XIAP in Patients With Crohn’s disease Gastroenterology (IF 18.392) Pub Date : 2018-03-06 Leila Amininejad, Benoit Charloteaux, Emilie Theatre, Claire Liefferinckx, Julia Dmitrieva, Pierre Hayard, Vincianne Muls, Jean-Marc Maisin, Michael Schapira, Jean-Michel Ghislain, Pierre Closset, Mehdi Talib, Marc Abramowicz, Yukihide Momozawa, Valerie Deffontaine, François Crins, Myriam Mni, Latifa Karim, Nadine Cambisano, Sandra Ornemese, Alessandro Zucchi, Charlotte Minsart, Jacques Deviere, Jean-Pierre Hugot, Martine De vos, Edouard Louis, Severine Vermeire, Andre Van Gossum, Wouter Coppieters, Jean-Claude Twizere, Michel Georges, Denis Franchimont
Background & AimsA few rare monogenic primary immunodeficiencies (PID) are characterized by chronic intestinal inflammation that resembles Crohn’s disease (CD). We investigated whether 23 genes associated with 10 of these monogenic disorders contain common, low-frequency or rare variants that increase risk for CD.MethodsCommon and low frequency variants in 1 Mb loci centered on the candidate genes were analyzed using meta-data corresponding to genotypes of approximately 17,000 patients with CD or without CD (controls) in Europe. The contribution of rare variants was assessed by high-throughput sequencing of 4750 individuals, including 660 early-onset and/or familial cases among the 2390 patients with CD. Variants were expressed from vectors in SW480 or HeLa cells and functions of their products were analyzed in immunofluorescence, luciferase, immunoprecipitation, and immunoblot assays.ResultsWe reproduced the association of the IL10 locus with CD (P=.007), although none of the significantly associated variants modified the coding sequence of IL10. We found XIAP to be significantly enriched for rare coding mutations in patients with CD vs controls (P=.02). We identified 4 previously unreported missense variants associated with CD. Variants in XIAP cause PID X-linked lymphoproliferative disease type 2 (XLP2), yet none of the carriers of these variants had all the clinical features of XLP2. Identified XIAP variants S123N, R233Q and P257A were associated with an impaired activation of NOD2 signaling following MDP stimulation.ConclusionsIn a systematic analysis of variants in 23 PID-associated genes, we confirmed the association of variants in XIAP with CD. Further screens for CD-associated variants and analyses of their functions could increase our understanding of the relationship between PID-associated genes and CD pathogenesis.
Change in Populations of Macrophages Promotes Development of Delayed Gastric Emptying in Mice Gastroenterology (IF 18.392) Pub Date : 2018-03-06 Gianluca Cipriani, Simon J. Gibbons, Katie E. Miller, Daniel S. Yang, Matthew L. Terhaar, Seth T. Eisenman, Tamas Ördög, David R. Linden, Gabriela B. Gajdos, Joseph H. Szurszewski, Gianrico Farrugia
Background & AimsMuscularis propria macrophages lie close to cells that regulate gastrointestinal motor function, including interstitial cells of Cajal (ICC) and myenteric neurons. In animal models of diabetic gastroparesis, development of delayed gastric emptying has been associated with loss of macrophages that express cytoprotective markers and reduced networks of ICC. Mice with long-term diabetes and normal gastric emptying have macrophages that express anti-inflammatory markers and have normal gastric ICC. Mice homozygous for the osteopetrosis spontaneous mutation in the colony stimulating factor 1 gene (Csf1op/op) do not have macrophages; when they are given streptozotocin to induce diabetes, they do not develop delayed gastric emptying. We investigated whether population of the gastric muscularis propria of diabetic Csf1op/op mice with macrophages is necessary to change gastric emptying, ICC, and myenteric neurons and investigated the macrophage-derived factors that determine whether diabetic mice do or do not develop delayed gastric emptying.MethodsWild-type and Csf1op/op mice were given streptozotocin to induce diabetes. Some Csf1op/op mice were given daily intraperitoneal injections of CSF1 for 7 weeks; gastric tissues were collected and cellular distributions were analyzed by immunohistochemistry. CD45+, CD11b+, F4/80+ macrophages were dissociated from gastric muscularis propria, isolated by flow cytometry and analyzed by quantitative real-time PCR. Cultured gastric muscularis propria from Csf1op/op mice was exposed to medium that was conditioned by culture with bone marrow derived macrophages from wild type mice.ResultsGastric muscularis propria from Csf1op/op mice given CSF1 contained macrophages; 11/15 diabetic mice given CSF1 developed delayed gastric emptying and had damaged ICC. In non-diabetic Csf1op/op mice, administration of CSF1 reduced numbers of gastric myenteric neurons but did not affect the proportion of nitrergic neurons or ICC. In diabetic Csf1op/op mice given CSF1 that developed delayed gastric emptying the proportion of nitrergic neurons was the same as in non-diabetic wild type controls. Medium conditioned by macrophages previously exposed to oxidative injury caused damage to ICC in cultured gastric muscularis propria from Csf1op/op mice; neutralizing antibodies against IL6R or TNF prevented this damage to ICC. CD45+, CD11b+, F4/80+ macrophages isolated from diabetic wild type mice with delayed gastric emptying expressed higher levels of mRNAs encoding inflammatory markers (IL6 and inducible nitric oxide synthase) and lower levels of mRNAs encoding markers of anti-inflammatory cells (heme oxygenase 1, arginase 1, and FIZZ1) than macrophages isolated from diabetic mice with normal gastric emptying.ConclusionsIn studies of Csf1op/op and wild-type mice with diabetes, we found delayed gastric emptying to be associated with increased production of inflammatory factors, and reduced production of anti-inflammatory factors, by macrophages, leading to loss of ICC.
Neuronal Transforming Growth Factor beta Signaling via SMAD3 Contributes to Pain in Animal Models of Chronic Pancreatitis Gastroenterology (IF 18.392) Pub Date : 2018-03-02 Liansheng Liu, Yaohui Zhu, Michaël Noë, Qian Li, Pankaj Jay Pasricha
Background & Aims Chronic pancreatitis (CP) is characterized by pancreatic inflammation and fibrosis, associated with increased pancreatic expression of transforming growth factor beta (TGFB). It is not clear how these might contribute to pain. We investigated whether TGFB signaling via SMAD induces sensitization of pancreatic sensory neurons to increase nociception. Methods CP was induced in Sprague-Dawley rats by infusion of trinitrobenzene sulfonic acid; some rats were given intrathecal infusions of TGFB1. CP was induced in control mice by administration of cerulein; we also studied β1glo/Ptf1acre-ER mice, which upon induction overexpress TGFB1 in pancreatic acinar cells, and TGFBr1f/f-CGRPcreER mice, which have inducible disruption of TGFBr1 in calcitonin gene-related peptide- positive neurons. Dominant negative forms of human TGFBR2 and SMAD3 were overexpressed from viral vectors in rat pancreas. Some rats were given the SMAD3 inhibitors SIS3 or halofuginone. After induction of CP, mice were analyzed in for pain in behavior tests or electrophysiologic studies of sensory neurons. Pancreatic nociceptor excitability was examined by patch clamp techniques and nociception was measured by Von Frey Filament tests for referred somatic hyperalgesia and behavioral responses to pancreatic electrical stimulation. Pancreata were collected from mice and rats and analyzed histologically and by ELISA and immunohistochemistry. Results Over-expression of TGFB in pancreatic acinar cells of mice and infusion of TGFB1 into rats resulted in sensory neuron hyperexcitability, SMAD3 activation, and increased nociception. This was accompanied by a reduction in the transient A-type current in pancreas-specific sensory neurons in rats, a characteristic of nociceptive sensitization in animal models of CP. Conversely, pancreata from TGFBr1f/f-CGRPcreER mice, rats with pancreatic expression of dominant-negative forms of human TGFBR2 or SMAD3, and rats given small molecule inhibitors of SMAD3 had attenuated neuronal sensitization and pain behavior following induction of CP. In contrast to findings from peripheral administration of TGFB1, intrathecal infusion of TGFB1 reduced hyperalgesia in rats with CP. Conclusions In pancreata of mice and rats, TGFB promotes peripheral nociceptive sensitization via a direct effect on primary sensory neurons mediated by intra-neuronal SMAD3. This is distinct from the central nervous system, where TGFB reduces nociception. These results provide an explanation for the link between fibrosis and pain and pain in patients with CP. This signaling pathway might be targeted therapeutically to reduce pain in patients with CP.
Loss of Chromatin Remodeling Proteins and/or CDKN2A Associates With Metastasis of Pancreatic Neuroendocrine Tumors and Reduced Patient Survival Times Gastroenterology (IF 18.392) Pub Date : 2018-03-02 Somak Roy, William A. LaFramboise, Ta-Chiang Liu, Dengfeng Cao, Alyssa Luvison, Caitlyn Miller, Maureen A. Lyons, Roderick J. O’Sullivan, Amer H. Zureikat, Melissa E. Hogg, Allan Tsung, Kenneth K. Lee, Nathan Bahary, Randall E. Brand, Jennifer S. Chennat, Kenneth E. Fasanella, Kevin McGrath, Marina N. Nikiforova, Georgios I. Papachristou, Adam Slivka, Herbert J. Zeh, Aatur D. Singhi
Despite prognostic grading and staging systems, it is a challenge to predict outcomes for patients with pancreatic neuroendocrine tumors (PanNETs). Sequencing studies of PanNETs have identified alterations in death domain-associated protein (DAXX) and ATRX chromatin remodeler (ATRX). In tumors, mutations in DAXX or ATRX and corresponding loss of protein expression correlate with shorter times of disease-free survival and disease-specific survival of patients. However, DAXX or ATRX proteins were lost in only 50% of distant metastases analyzed. We performed whole-exome sequencing analyses of 20 distant metastases from 20 patients with a single non-syndrome, non-functional PanNET. We found distant metastases contained alterations in MEN1 (n=8), ATRX (n=5), DAXX (n=5), TSC2 (n=3), and DEPDC5 (n=3). We found copy number loss of CDKN2A in 15 metastases (75%) and alterations in genes that regulate chromatin remodeling including SETD2 (n=4), ARID1A (n=2), CHD8 (n=2), and DNMT1 (n=2). In a separate analysis of 347 primary PanNETs, we found loss or deletion of DAXX and ATRX, disruption of SETD2 function (based on loss of H3K36me3), loss of ARID1A expression or deletions in CDKN2A in 81% of primary PanNETs with distant metastases. Among patients with loss or deletion of at least 1 of these proteins or genes, 39% survived disease free for 5 years and 44% had disease-specific survival times of 10 years. Among patients without any of these alterations, 98% survived disease free for 5 years and 95% had disease-specific survival times of 10 years. Therefore, primary PanNETs with loss of DAXX, ATRX, H3K36me3, ARID1A, and/or CDKN2A associate with shorter survival times of patients. Our findings indicate that alterations in chromatin remodeling genes and CDKN2A contribute to metastasis of PanNETs.
Dietary Patterns After the Weaning and Lactation Period Associate with Celiac Disease Autoimmunity in Children Gastroenterology (IF 18.392) Pub Date : 2018-03-02 Monica Barroso, Sytske A. Beth, Trudy Voortman, Vincent W.V. Jaddoe, Menno C. van Zelm, Henriette A. Moll, Jessica C. Kiefte-de Jong
Background & Aims There have been many studies of associations between infant feeding practices and development of celiac disease during childhood, but few studies have focused on overall diets of young children following the weaning period. We aimed to examine the association between common dietary patterns in infants and the occurrence of celiac disease autoimmunity during childhood. Methods We performed a prospective analysis of data from the Generation R Study that comprised 1997 children born from April 2002 through January 2006 in Rotterdam, the Netherlands. Food consumption around 1 year of age was assessed with a validated food-frequency questionnaire. Dietary data were examined using a priori (based on existing guidelines) and a posteriori (Principal Component Analysis and Reduced Rank Regression) dietary pattern analyses. Five dietary patterns were compared. Celiac disease autoimmunity, determined based on serum concentration of transglutaminase-2 autoantibody (TG2A) below or above 7 U/mL, was evaluated at 6 years. Associations between dietary pattern adherence scores and celiac disease autoimmunity were examined using multivariable logistic regression models. Results Higher adherence to the a posteriori-derived prudent dietary pattern (high intake of vegetables, vegetable oils, pasta, and grains and low consumption of refined cereals and sweet beverages) at 1 year was significantly associated with lower odds of celiac disease autoimmunity at 6 years (odds ratio, 0.67; 95% CI, 0.54–0.84). No significant associations were found for other 4 dietary patterns. Conclusions In a prospective study of dietary patterns of young children in the Netherlands, we associated a diet with high consumption of vegetables and grains, and low consumption of refined cereals and sweet beverages, with lower odds of celiac disease autoimmunity. Early-life dietary patterns might therefore be involved in the development of celiac disease during childhood.
Methotrexate Reduces DNA Integrity in Sperm From Men with Inflammatory Bowel Disease Gastroenterology (IF 18.392) Pub Date : 2018-03-02 Dana Ley, Jeffrey Jones, John Parrish, Sana Salih, Freddy Caldera, Edna Tirado, Benjamin Leader, Sumona Saha
There are few data on the effects of methotrexate on reproductive capacity in men with inflammatory bowel diseases (IBD). We performed a case–control study to determine the effects of methotrexate on sperm quality and genetic integrity. We compared sperm samples from 7 men with IBD who had been exposed to methotrexate for at least 3 months with sperm samples collected from 1912 age-matched men at fertility centers (controls), where sperm parameters would be expected to be worse than those of the general population. Sperm were evaluated by basic semen analysis and advanced sperm integrity testing. In samples from men with IBD, all basic semen analysis parameters were within normal limits. However, these samples had reduced sperm integrity, based on significant increases in levels of DNA fragmentation and damage from oxidative stress compared with controls. Our findings indicate that methotrexate can reduce DNA integrity in sperm and cause damage via oxidative stress.
Endoscopic Full Thickness Resection Gastroenterology (IF 18.392) Pub Date : 2018-03-02 Elizabeth Rajan, Louis M. Wong Kee Song
Recent advances in minimally invasive endoscopic approaches have pushed the boundaries of well-established resection techniques for therapeutic and diagnostic applications. Endoscopic full thickness resection techniques are a key development in the management of challenging epithelial and subepithelial lesions that are not amenable to conventional endoscopic resection methods and previously required a surgical approach. Endoscopic full thickness biopsy represents a paradigm shift in tissue acquisition and will enhance our understanding of the pathophysiology, and guide therapy of gastrointestinal neuromuscular diseases, as well as other inflammatory and neoplastic conditions. This review highlights current tools and techniques available for endoscopic full thickness resection and biopsy, as well as outcomes from such interventions.
“Endoscopic submucosal dissection: a critical appraisal of its role in Western endoscopy practice” Gastroenterology (IF 18.392) Pub Date : 2018-03-02 M.J. Bourke, H. Neuhaus, J.J. Bergman
Endoscopic submucosal dissection (ESD) was developed in Japan, early in this century, to provide a minimally invasive yet curative treatment for the large numbers of patients with early gastric cancer identified by the national screening program. Previously the majority of these patients were treated surgically at substantial cost with the significant risk of short and long-term morbidity. Enbloc excision of these early cancers, most with a limited risk of nodal metastasis, allowed complete staging of the tumor, stratification of the subsequent therapeutic approach and potential cure. This transformative innovation changed the nature of endoscopic treatment for superficial mucosal neoplasia and ultimately, for the first time, allowed endoscopists to assert that the early cancer had been definitively cured. Subsequently western endoscopists have increasingly embraced the therapeutic possibilities offered by ESD, but with some justifiable scientific caution. Herein we provide an evidence based critical appraisal of the role of ESD in advanced endoscopic tissue resection.
Novel developments in endoscopic mucosal imaging Gastroenterology (IF 18.392) Pub Date : 2018-02-17 F. van der Sommen, W.L. Curvers, W.B. Nagengast
Endoscopic techniques such as High-definition and optical-chromoendoscopy have had enormous impact on endoscopy practice. Since these techniques allow assessment of most subtle morphological mucosal abnormalities, further improvements in endoscopic practice lay in increasing the detection efficacy of endoscopists. Several new developments could assist in this. First, web based training tools could improve the skills of the endoscopist for enhancing the detection and classification of lesions. Secondly, incorporation of computer aided detection will be the next step to raise endoscopic quality of the captured data. These systems will aid the endoscopist in interpreting the increasing amount of visual information in endoscopic images providing real-time objective second reading. In addition, developments in the field of molecular imaging open opportunities to add functional imaging data, visualizing biological parameters, of the gastrointestinal tract to white-light morphology imaging. For the successful implementation of abovementioned techniques, a true multi-disciplinary approach is of vital importance.
Determining Risk of Colorectal Cancer and Starting Age of Screening Based on Lifestyle, Environmental, and Genetic Factors Gastroenterology (IF 18.392) Pub Date : 2018-02-17 Jihyoun Jeon, Mengmeng Du, Robert E. Schoen, Michael Hoffmeister, Polly A. Newcomb, Sonja I. Berndt, Bette Caan, Peter T. Campbell, Andrew T. Chan, Jenny Chang-Claude, Graham G. Giles, Jian Gong, Tabitha A. Harrison, Jeroen R. Huyghe, Eric J. Jacobs, Li Li, Yi Lin, Loïc Le Marchand, John D. Potter, Flora Qu, Stephanie A. Bien, Niha Zubair, Robert J. Macinnis, Daniel D. Buchanan, John L. Hopper, Yin Cao, Reiko Nishihara, Gad Rennert, Martha L. Slattery, Duncan C. Thomas, Michael O. Woods, Ross L. Prentice, Stephen B. Gruber, Yingye Zheng, Hermann Brenner, Richard B. Hayes, Emily White, Ulrike Peters, Li Hsu
Background & Aims Guidelines for initiating colorectal cancer (CRC) screening are based on family history but do not consider lifestyle, environmental, or genetic risk factors. We developed models to determine risk of CRC, based on lifestyle and environmental factors and genetic variants, and to identify an optimal age to begin screening. Methods We collected data from 9748 CRC cases and 10,590 controls in the Genetics and Epidemiology of Colorectal Cancer Consortium and the Colorectal Transdisciplinary study, from 1992 through 2005. Half of the participants were used to develop the risk determination model and the other half were used to evaluate the discriminatory accuracy (validation set). Models of CRC risk were created based on family history, 19 lifestyle and environmental factors (E-score), and 63 CRC-associated single-nucleotide polymorphisms identified in genome-wide association studies (G-score). We evaluated the discriminatory accuracy of the models by calculating area under the receiver operating characteristic curve (AUC) values, adjusting for study, age, and endoscopy findings for the validation set. We used the models to project the 10-year absolute risk of CRC for a given risk profile and recommend ages to begin screening, in comparison to CRC risk for an average individual at 50 years of age, using external population incidence rates for non-Hispanic whites from the Surveillance, Epidemiology, and End Results Program registry. Results In our models, E-score and G-score each determined risk of CRC with greater accuracy than family history. A model that combined both scores and family history estimated CRC risk with an AUC value of 0.63 (95% CI, 0.62–0.64) for men and 0.62 (95% CI, 0.61–0.63) for women; AUC values based on only family history ranged from 0.53 to 0.54 and those based only E-score or G-score ranged from 0.59 to 0.60. Although screening is recommended to begin at age 50 years for individuals with no family history of CRC, starting ages calculated based on combined E-score and G-score differed by 12 years for men and 14 for women, for individuals with the highest vs the lowest 10% of risk. Conclusions We used data from 2 large international consortia to develop CRC risk calculation models that included genetic and environmental factors along with family history. These determine risk of CRC and starting ages for screening with greater accuracy than the family history only model, which is based on the current screening guideline. These scoring systems might serve as a first step toward developing individualized CRC prevention strategies.
EUS-guided Transluminal Interventions Gastroenterology (IF 18.392) Pub Date : 2018-02-16 Uzma D. Siddiqui, Michael J. Levy
The role of endoscopic ultrasound (EUS) has transitioned from a diagnostic to a therapeutic one over the past 40 years. With the advent of curvilinear array echoendoscopes in the 1990’s with an accessory channel, multiple tools and devices have been developed and utilized for a variety of transluminal interventions. EUS provides a viable option and is becoming the procedure of choice for many interventions including bile and pancreatic duct drainage, guiding angiotherapy, pancreatic fluid collection management, gallbladder drainage, and creating a gastrojejunostomy. Although reports demonstrate the technical success of these interventions, there is tremendous study heterogeneity and a relative lack of controlled randomized trials, which may limit our understanding of their role and utility. Furthermore, adverse events are relatively common and occasionally severe. Despite the limitations, available data strongly indicate the efficacy of EUS interventions when performed by well-trained endosonographers in carefully selected patients and managed in a multi-disciplinary setting.
The Effect of Endoscopic Surveillance in Patients with Barrett's Esophagus: A Systematic Review and Meta-analysis Gastroenterology (IF 18.392) Pub Date : 2018-02-16 Don Chamil Codipilly, Apoorva Krishna Chandar, Siddharth Singh, Sachin Wani, Nicholas J. Shaheen, John M. Inadomi, Amitabh Chak, Prasad G. Iyer
Background & Aims Guidelines recommend endoscopic surveillance of patients with Barrett’s esophagus (BE) to identify those with dysplasia (a precursor of carcinoma) or early-stage esophageal adenocarcinoma (EAC), who can be treated endoscopically. However, it is unclear whether surveillance increases survival times of patients with BE. We performed a systematic review and meta-analysis to qualitatively and quantitatively examine evidence for the association of endoscopic surveillance in patients with BE with survival and other outcomes. Methods We searched publication databases for studies reporting the effects of endoscopic surveillance on mortality and other EAC-related outcomes. We reviewed randomized controlled trials, case–control studies, studies comparing patients with BE who received regular surveillance to those who did not receive regular surveillance, as well as studies comparing outcomes of patients with surveillance-detected EAC vs. symptom-detected EACs. We performed a meta-analysis of surveillance studies to generate summary estimates using a random effects model. The primary aim was to examine the association of BE surveillance on EAC-related mortality. Secondary aims were to examine the association of BE surveillance with all-cause mortality and EAC stage at time of diagnosis. Results A single case-control study did not reveal any association between surveillance and EAC-related mortality. A meta-analysis of 4 cohort studies found lower EAC-related and all-cause mortality associated with regular surveillance (relative risk, 0.60; 95% CI, 0.50–0.71 and hazard ratio, 0.75; 95% CI, 0.59–0.94). Meta-analysis of 12 cohort studies demonstrated lower EAC-related and all-cause mortality among patients with surveillance-detected EAC vs. symptom-detected EAC (relative risk, 0.73; 95% CI, 0.57–0.94 and hazard ratio, 0.59; 95% CI, 0.45–0.76). Lead- and length-time bias adjustment substantially attenuated/eliminated the observed benefits. Surveillance was associated with detection of EAC at earlier stages. A randomized trial is underway to evaluate the effects of endoscopic surveillance on mortality in patients with BE. Conclusions In a systematic review and meta-analysis of the effects of surveillance in patients with BE, surveillance as currently performed was associated with detection of earlier stage EAC and may provide a small survival benefit. However, the effects of confounding biases on these estimates are not fully defined, and may completely or partially explain the observed differences between surveyed and unsurveyed patients.
Endoscopic management of pancreatobiliary neoplasms Gastroenterology (IF 18.392) Pub Date : 2018-02-16 Andrew Y. Wang, Patrick S. Yachimski
Endoscopic ultrasound (EUS) and endoscopic retrograde cholangiopancreatography (ERCP) are the mainstays of interventional endoscopic practice. EUS occupies a central role in the diagnosis of pancreatobiliary neoplasms and offers a platform for a wide range of direct tumor therapies. Initial steps have demonstrated the feasibility of such applications in animal models and pilot studies. Larger clinical trials and incorporation of EUS-based therapies into cooperative cancer studies might demonstrate an impact in the clinical prognosis of patients with pancreatic cancer. ERCP plays an important role in elucidating indeterminate biliary strictures and in treating patients with malignant biliary obstruction who are symptomatic or have borderline resectable or unresectable disease. ERCP-directed ablative therapies enable neoadjuvant and palliative intervention in patients with malignant biliary obstruction, in particular perihilar cholangiocarcinoma. Additional comparative, multicenter studies are needed to better understand the safety and efficacy of endobiliary brachytherapy, photodynamic therapy, and radiofrequency ablation in patients with pancreatobiliary malignancies.
Controversies in Endoscopic Eradication Therapy for Barrett’s Esophagus Gastroenterology (IF 18.392) Pub Date : 2018-02-16 Srinadh Komanduri, V. Raman Muthusamy, Sachin Wani
Advances in Endoscopic Eradication Therapy for Barrett’s Esophagus associated neoplasia have resulted in a significant paradigm shift in diagnosis and management of this complex disease. A robust body of literature critically evaluating outcomes of resection and ablative strategies has allowed gastroenterologists to make quality, evidence based decisions for their patients. Despite this progress, there are still many unanswered questions and challenges that remain. Ultimately, identification of a cost-effective screening modality, biomarkers for risk stratification, and strides to eliminate post surveillance endoscopy after EET are essential to reach our long term goal for eradication of esophageal adenocarcinoma.
HCV Genotype 6a Escape from and Resistance to Velpatasvir, Pibrentasvir, and Sofosbuvir in Robust Infectious Cell Culture Models Gastroenterology (IF 18.392) Pub Date : 2018-02-15 Long V. Pham, Santseharay Ramirez, Judith M. Gottwein, Ulrik Fahnøe, Yi-Ping Li, Jannie Pedersen, Jens Bukh
Background & Aims Chronic liver diseases caused by hepatitis C virus (HCV) genotype 6 are prevalent in Asia, and millions of people require treatment with direct-acting antiviral regimens, such as NS5A inhibitor velpatasvir combined with the NS5B polymerase inhibitor sofosbuvir. We developed infectious cell culture models of HCV genotype 6a infection to study the effects of these inhibitors and the development of resistance. Methods The consensus sequences of prototype strains HK2 (MG717925) and HK6a (MG717928), originating from serum of patients with chronic HCV infection, were determined by Sanger sequencing of genomes amplified by reverse transcription-PCR. In vitro non-infectious full-length clones of these 6a strains were subsequently adapted in Huh7.5 cells, primarily by using substitutions identified in JFH1-based core-NS5A and core-NS5B genotype 6a recombinants. We studied the efficacy of NS5A- and NS5B- inhibitors in concentration–response assays. We examined the effects of long-term culture of Huh7.5 cells incubated with velpatasvir and sofosbuvir singly or combined following infection with passaged full-length HK2 or HK6a recombinant viruses. Resistance-associated substitutions (RAS) were identified by Sanger and next-generation sequencing, and their effects on viral fitness and in drug susceptibility were determined in reverse-genetic experiments. Results Adapted full-length HCV genotype 6a recombinants HK2cc and HK6acc had fast propagation kinetics and high infectivity titers. Compared to a HCV genotype 1a recombinant, HCV genotype 6a recombinants of strains HK2 and HK6a were equally sensitive to daclatasvir, elbasvir, velpatasvir, pibrentasvir, and sofosbuvir, but less sensitive to ledipasvir, ombitasvir, and dasabuvir. Long-term exposure of HCV genotype 6a-infected Huh7.5 cells with a combination of velpatasvir and sofosbuvir resulted in clearance of the virus, but the virus escaped the effects of single inhibitors via emergence of the RAS L31V in NS5A (conferring resistance to velpatasvir) and S282T in NS5B (conferring resistance to sofosbuvir). Engineered recombinant genotype 6a viruses with single RAS mediated resistance to velpatasvir or sofosbuvir. HCV genotype 6a viruses with RAS NS5A-L31V or NS5B-S282T was however able to propagate and escape in Huh7.5 cells exposed to the combination of velpatasvir and sofosbuvir. Further, HCV genotype 6a with NS5A-L31V was able to propagate and escape in the presence of pibrentasvir with emergence of NS5A-L28S, conferring a high level of resistance to this inhibitor. Conclusions Strains of HCV genotype 6a isolated from patients can be adapted to propagate in cultured cells, permitting studies of the complete life cycle for this important genotype. The combination of velpatasvir and sofosbuvir is required to block propagation of original HCV genotype 6a, which quickly becomes resistant to single inhibitors via the rapid emergence and persistence of RAS. These features of HCV genotype 6a could compromise treatment.
Endoscopic Management of Transmural Defects (Leaks/Perforations/Fistulae) Gastroenterology (IF 18.392) Pub Date : 2018-02-15 Willem A. Bemelman, Todd H. Baron
Transmural defects of the gastrointestinal tract can be classified into three distinct entities that include leaks, perforations, and fistulae. Each arise from different mechanisms and are managed accordingly. Leaks occur most often following surgery, while perforations occur most often following flexible endoscopic maneuvers. Fistula arise from a variety of mechanisms, such as an evolution from surgical leaks as well as from specific disease states. Endoscopic management plays a vital role in the treatment of transmural defects as long as the region of interest can be accessed with the appropriate endoscopic accessories. Endoscopic approaches can be broadly classified into those that provide closure and those that provide diversion of luminal contents. With advances in technology, a myriad of devices and accessories are available that allow a tailored approach. Endoscopic approaches to leaks, perforations, and fistulae are discussed in this review.
Clinical and Genomic Correlates of Neutrophil Reactive Oxygen Species Production in Pediatric Patients with Crohn’s Disease Gastroenterology (IF 18.392) Pub Date : 2018-02-15 Lee A. Denson, Ingrid Jurickova, Rebekah Karns, Kelly A. Shaw, David J. Cutler, David Okou, Anne Dodd, Kathryn Quinn, Kajari Mondal, Bruce J. Aronow, Yael Haberman, Aaron Linn, Adam Price, Ramona Bezold, Kathleen Lake, Kimberly Jackson, Thomas D. Walters, Anne Griffiths, Robert N. Baldassano, Joshua D. Noe, Jeffrey S. Hyams, Wallace V. Crandall, Barbara S. Kirschner, Melvin B. Heyman, Scott Snapper, Stephen L. Guthery, Marla C. Dubinsky, Neal S. Leleiko, Anthony R. Otley, Ramnik J. Xavier, Christine Stevens, Mark J. Daly, Michael E. Zwick, Subra Kugathasan
Background & Aims Individuals with monogenic disorders of phagocyte function develop chronic colitis that resembles Crohn’s Disease (CD). We tested for associations between mutations in genes encoding NADPH oxidases, neutrophil function, and phenotypes of CD in pediatric patients. Methods We performed whole-exome sequence analysis to identify mutations in genes encoding NADPH oxidases (such as CYBA, CYBB, NCF1, NCF2, NCF4, RAC1, and RAC2) using DNA from 543 pediatric patients with inflammatory bowel diseases. Blood samples were collected from an additional 129 pediatric patients with CD and 26 children without IBD (controls); we performed assays for neutrophil activation, reactive oxygen species (ROS) production, and bacteria uptake and killing. Whole-exome sequence analysis was performed using DNA from 46 of the children with CD to examine associations with NADPH gene mutations; RNA sequence analyses were performed using blood cells from 46 children with CD to test for variations in neutrophil gene expression associated with ROS production. Results We identified 26 missense mutations in CYBA, CYBB, NCF1, NCF2, and NCF4. Patients with CD who carried mutations in these genes were 3-fold more likely to have perianal disease (P=.0008) and stricturing complications (P=.002) than children with CD without these mutations. Among patients with CD with none of these mutations, 9% had undergone abdominal surgery; among patients with mutations in these NADPH oxidase genes, 31% had undergone abdominal surgery (P=.0004). A higher proportion of neutrophils from children with CD had low ROS production (47%) than from controls (15%) among the 129 patients tested for ROS (P=.002). Minor alleles of the NADPH genes were detected in 7% of children with CD whose neutrophils produced normal levels of ROS vs 38% of children whose neutrophils produced low levels of ROS (P=.009). Neutrophils that produced low levels of ROS had specific alterations in genes that regulate glucose metabolism and anti-microbial responses. Conclusions We identified missense mutations in genes that encode NADPH oxidases in children with CD; these associated with a more aggressive disease course and reduced ROS production by neutrophils from the patients.
Advances and challenges in endoscopic training Gastroenterology (IF 18.392) Pub Date : 2018-02-15 Kevin A. Waschke, Walter Coyle
One of the challenges of the current era is ensuring that endoscopic training is accomplished effectively in the face of multiple competing demands. As healthcare delivery evolves, with rising patient complexity and increasing productivity requirements, there is mounting pressure on the time available for training in the clinical setting. The practice of endoscopy itself continues to expand to include increasingly complex procedures (e.g. therapeutic endoscopic ultrasound (EUS), endoscopic submucosal dissection (ESD), peroral endoscopic myotomy (POEM), etc.), all of which require dedicated endoscopy training. The rapid pace of progress in the field of endoscopy means that the demand for endoscopy training is not limited to the formal period of training but instead spans the spectrum to include physicians already in practice. In the light of recent advances in our understanding of endoscopy training, the following review will serve to highlight the current state of affairs with respect to endoscopic training and how we can consider approaching these challenges.
Endoscopic Myotomy for Foregut Motility Disorders Gastroenterology (IF 18.392) Pub Date : 2018-02-15 Mouen A. Khashab, Petros C. Benias, Lee L. Swanstrom
Peroral endoscopic myotomy (POEM) is an advanced endoscopic procedure classically performed for the treatment of achalasia. The procedure is based on principles of submucosal endoscopy and is comprised of a mucosal incision, submucosal tunneling, myotomy and mucosal closure. Multiple published studies that collectively include more than 6000 patients reported clinical success in more than 80-90% of patients. Recent literature also suggested durability of response over a medium-term follow-up. POEM is associated with a low rate of adverse events when performed by experienced operators. Gastroesophageal reflux is not infrequent after POEM but does not seem significantly different from reflux which occurs after Heller myotomy. POEM also seems to be effective in the treatment of spastic esophageal disorders (e.g. Jackhammer and diffuse esophageal spasm). Lastly, the role of gastric POEM (G-POEM) in the treatment of gastroparesis has been investigated in recent studies with promising results.
Increased Expression of the Long Non-coding RNA LINC01503, Regulated by TP63, in Squamous Cell Carcinoma and Effects on Oncogenic Activities of Cancer Cell Lines Gastroenterology (IF 18.392) Pub Date : 2018-02-15 Jian-Jun Xie, Yan-Yi Jiang, Yuan Jiang, Chun-Quan Li, Lim-Mei Chee, Omer An, Anand Mayakonda, Ling-Wen Ding, Lin Long, Chun Sun, Le-Hang Lin, Li Chen, Jian-Yi Wu, Zhi-Yong Wu, Qi Cao, Wang-Kai Fang, Wei Yang, Stephen J. Meltzer, Henry Yang, Melissa Fullwood, Li-Yan Xu, En-Min Li, De-Chen Lin, H. Phillip Koeffler
Background & Aims Long non-coding RNAs (lncRNAs) are expressed in tissue-specific pattern, but it is not clear how these are regulated. We aimed to identify squamous cell carcinoma (SCC)-specific lncRNAs and investigate mechanisms that control their expression and function. Methods We studied expression patterns and functions of 4 SCC-specific lncRNAs. We obtained 113 esophageal SCC (ESCC) and matched non-tumor esophageal tissues from a hospital in Shantou City, China, and performed quantitative reverse transcription PCR assays to measure expression levels of LINC01503. We collected clinical data from patients and compared expression levels with survival times. LINC01503 was knocked down using small interfering RNAs and oligonucleotides in TE7, TE5 and KYSE510 cell lines and overexpressed in KYSE30 cells. Cells were analyzed by chromatin immunoprecipitation sequencing, luciferase reporter assays, colony formation, migration and invasion, and mass spectrometry analyses. Cells were injected into nude mice and growth of xenograft tumors was measured. LINC01503 interaction with proteins was studied using fluorescence in situ hybridization, RNA pulldown, and RNA immunoprecipitation analyses. Results We identified a lncRNA, LINC01503, which is regulated by a super-enhancer and is expressed at significantly higher levels in esophageal and head and neck SCCs than in non-tumor tissues. High levels in SCCs correlated with shorter survival times of patients. The transcription factor TP63 bound to the super-enhancer at the LINC01503 locus and activated its transcription. Expression of LINC01503 in ESCC cell lines increased their proliferation, colony formation, migration and invasion. Knockdown of LINC01503 in SCC cells reduced their proliferation, colony formation, migration and invasion, and growth of xenograft tumors in nude mice. Expression of LINC01503 in ESCC cell lines reduced ERK2 dephosphorylation by DUSP6, leading to activation of ERK signaling via MAPK. LINC01503 disrupted the interaction between EBP1 and the p85 subunit of PI3K, increasing AKT signaling. Conclusions We identified a lncRNA, LINC01503, that is increased in SCC cells compared with non-tumor cells. Increased expression of LINC01503 promotes ESCC cell proliferation, migration, invasion, and growth of xenograft tumors. It might be developed as a biomarker of aggressive SCCs in patients.
Alterations in Intestinal Microbiota Lead to Production of Interleukin 17 by Intrahepatic γδ T-cell Receptor-positive Cells and Pathogenesis of Cholestatic Liver Disease Gastroenterology (IF 18.392) Pub Date : 2018-02-15 Dana Tedesco, Manoj Thapa, Chui-Yoke Chin, Yong Ge, Minghao Gong, Jing Li, Sanjeev Gumber, Patrick Speck, Elizabeth J. Elrod, Eileen M. Burd, William H. Kitchens, Joseph F. Magliocca, Andrew B. Adams, David S. Weiss, Mansour Mohamadzadeh, Arash Grakoui
Background & Aims Variants at the ATP binding cassette subfamily B member 4 gene (ABCB4 or MDR2) locus, which encodes a biliary transport protein, are associated with a spectrum of cholestatic liver diseases. Exacerbation of liver disease has been linked to increased hepatic levels of interleukin 17 (IL17), yet the mechanisms of this increase are not understood. We studied mice with disruption of Mdr2 to determine how defects in liver and alteration in the microbiota contribute to production of IL17 by intrahepatic γδ T cells. Methods We performed studies with Mdr2-/- and littermate FVB/NJ (control) mice. IL17 was measured in serum samples by an ELISA. Mice were injected with neutralizing antibodies against the γδ T-cell receptor (TCR; anti-γδ TCR) or mouse IL17A (anti-IL17A). Livers were collected and bacteria were identified in homogenates by culture procedures; TCRγδ+ cells were isolated by flow cytometry. Fecal samples were collected from mice and analyzed by 16s ribosomal DNA sequencing. Cells were stimulated with antibodies or bacteria, and cytokine production was measured. We obtained tissues from 10 patients undergoing liver transplantation for primary sclerosing cholangitis or chronic hepatitis C virus (HCV) infection. Tissues were analyzed for cytokine production by γδ TCR+ cells. Results Mdr2-/- mice had collagen deposition around hepatic bile ducts and periportal–bridging fibrosis with influx of inflammatory cells and increased serum levels of IL17 compared with control mice. Administration of anti-IL17A reduced hepatic fibrosis. Livers from Mdr2–/– mice had increased numbers of IL17A+ γδTCR+ cells—particularly of IL17A+ Vγ6Jγ1 γδ TCR+ cells. Fecal samples from Mdr2-/- mice were enriched in Lactobacillus, and liver tissues were enriched in Lactobacillus gasseri compared with control mice. Mdr2-/- mice also had increased intestinal permeability. The γδ TCR+ cells isolated from Mdr2-/- livers produced IL17 in response to heat-killed L gasseri. Intraperitoneal injection of control mice with L gasseri led to increased serum levels of IL17 and liver infiltration by inflammatory cells; injection of these mice with anti-γδ TCR reduced serum level of IL17. Intravenous injections of Mdr2-/- mice with anti-γδ TCR reduced fibrosis; liver levels of IL17, and inflammatory cells; and serum levels of IL17. γδTCR+ cells isolated from livers of patients with primary sclerosing cholangitis, but not HCV infection, produced IL17. Conclusions In Mdr2-/- mice, we found development of liver fibrosis and inflammation to require hepatic activation of γδ TCR+ cells and production of IL17 production, mediated by exposure to L gasseri. This pathway appears to contribute to development of cholestatic liver disease in patients.
Less is more: A minimalist approach to endoscopy Gastroenterology (IF 18.392) Pub Date : 2018-02-15 Nicholas J. Shaheen, M. Brian Fennerty, Jacques J. Bergman
A substantial literature documents inappropriate usage of gastrointestinal endoscopy in a variety of clinical settings. Overusage of endoscopy appears to be common, and 30% or more of procedures performed in some clinical settings have questionable indications. The potential reasons for overuse of endoscopy are multiple, and include cancer phobia, fear of medical malpractice litigation, profit motive, the investigation of “incidentalomas” found on other imaging, and under-appreciation of the delayed harms of endoscopy, among other reasons. Clinical guidelines, which should limit overuse of endoscopy, may instead serve to promote it, if authors opt to be “conservative,” recommending endoscopy in situations of unclear utility. Several strategies may decrease overuse of endoscopy, including careful attention to risk stratification when choosing patients to screen, adherence to guidelines for surveillance intervals for colonoscopy, the use of quality indicators to identify outliers in endoscopy utilization, and education on appropriate indications and the risks of overuse at the medical student, residency and fellowship levels.
P300 Acetyltransferase Mediates Stiffness-Induced Activation of Hepatic Stellate Cells Into Tumor-promoting Myofibroblasts Gastroenterology (IF 18.392) Pub Date : 2018-02-15 Changwei Dou, Zhikui Liu, Kangsheng Tu, Hongbin Zhang, Chen Chen, Usman Yaqoob, Yuanguo Wang, Jialing Wen, Jan van Deursen, Delphine Sicard, Daniel Tschumperlin, Hongzhi Zou, Wei-Chien Huang, Raul Urrutia, Vijay H. Shah, Ningling Kang
Background & Aims Hepatic stellate cells (HSCs) contribute to desmoplasia and stiffness of liver metastases by differentiating into matrix-producing myofibroblasts. We investigated whether stiffness due to the presence of tumors increases activation of HSCs into myofibroblasts and their tumor-promoting effects, as well as the role of E1A binding protein p300 (EP300 or p300), a histone acetyltransferase that regulates transcription, in these processes. Methods HSCs were isolated from liver tissues of patients, mice in which the p300 gene was flanked by 2 loxP sites (p300F/F mice), and p300+/+ mice (controls). The HSCs were placed on polyacrylamide gels with precisely defined stiffness, and their activation (differentiation into myofibroblasts) was assessed by immunofluorescence and immunoblot analyses for alpha-smooth muscle actin. In HSCs from mice, the p300 gene was disrupted by cre recombinase. In human HSCs, levels of p300 were knocked down with small hairpin RNAs or a mutant form of p300 that is not phosphorylated by AKT (p300S1834A) was overexpressed. Human HSCs were also cultured with inhibitors of p300 (C646), PI3K signaling to AKT (LY294002), or RHOA (C3 transferase) and effects on stiffness-induced activation were measured. RNA sequencing and chromatin immunoprecipitation quantitative PCR were used to identified HSC genes that changed expression levels in response to stiffness. We measured effects of HSC-conditioned media on proliferation of HT29 colon cancer cells and growth of tumors following subcutaneous injection of these cells into mice. MC38 colon cancer cells were injected into portal veins of p300F/Fcre and control mice and liver metastases were measured. p300F/Fcre and control mice were given intraperitoneal injections of CCl4 to induce liver fibrosis. Liver tissues were collected and analyzed by immunofluorescence, immunoblot, and histology. Result Substrate stiffness was sufficient to activate HSCs, leading to nuclear accumulation of p300. Disrupting p300 level or activity blocked stiffness-induced activation of HSC. In HSCs, substrate stiffness activated AKT signaling via RHOA to induce phosphorylation of p300 at serine 1834; this caused p300 to translocate to the nucleus, where it upregulated transcription of genes that increase activation of HSCs and metastasis, including CXCL12. MC38 cells, injected into portal veins, formed fewer metastases in livers of p300F/Fcre mice than control mice. Expression of p300 was increased in livers of mice following injection of CCl4; HSC activation and collagen deposition were reduced in livers of p300F/Fcre mice compared with control mice. Conclusions In studies of mice, we found liver stiffness to activate HSC differentiation into myofibroblasts, which required nuclear accumulation of p300. P300 increases HSC expression of genes that promote metastasis.
Advances in CRC prevention: screening and surveillance Gastroenterology (IF 18.392) Pub Date : 2018-02-15 Evelien Dekker, Douglas K. Rex
Colorectal cancer (CRC) is amongst the most commonly diagnosed cancers and causes of death from cancer across the world. CRC can, however, be detected in asymptomatic patients at a curable stage, and several studies have shown lower mortality among patients who undergo screening compared to those who do not. Using colonoscopy in CRC screening also results in the detection of precancerous polyps that can be directly removed during the procedure, thereby reducing the incidence of cancer. In the past decade, convincing evidence has appeared that the effectiveness of colonoscopy as CRC prevention tool is associated with the quality of the procedure. This review aims to provide an up-to-date overview of recent efforts to improve colonoscopy effectiveness of by enhancing detection and improving the completeness and safety of resection of colorectal lesions.
Functional Bowel Disorders Gastroenterology’s 75th anniversary Gastroenterology (IF 18.392) Pub Date : 2018-02-15 John W. Wiley, Lin Chang
Articles appearing in Gastroenterology have played an integral role in the evolution of our understanding of Functional Bowel Disorders (FBD), including Irritable Bowel Syndrome (IBS), beginning with the prescient contributions of Almy and Tulin in 1947 and 1949 that highlighted the role of stress to enhance perception of abdominal pain and promote colon contractions. Subsequent publications have codified diagnostic criteria and stratified subpopulations of FBD (Manning and ROME I-IV), which resulted in improved symptom-based therapeutic interventions. Advances in our understanding of the pathophysiology of FBD, particularly IBS, published in Gastroenterology has led to our current appreciation that FBD represent dysfunction in the bidirectional brain-gut axis, intestinal barrier dysfunction and interactions with the microbiota and dietary factors. Team science and the application of next-generation -omics methods are leading the way to improved diagnostic criteria and targeted therapeutic interventions. As the field evolves, publications appearing in Gastroenterology will continue to be at the forefront of these advances.
Acute Pancreatitis and Pancreatic Cancer Risk: A Nationwide Matched-cohort Study in Denmark Gastroenterology (IF 18.392) Pub Date : 2018-02-09 Jakob Kirkegård, Deirdre Cronin-Fenton, Uffe Heide-Jørgensen, Frank Viborg Mortensen
Background & Aims Acute pancreatitis may be a risk factor for pancreatic cancer. However, findings from studies on this association are conflicting. We investigated the association between acute pancreatitis and increased risk of pancreatic cancer. Methods We conducted a nationwide, population-based, matched cohort study of all patients admitted to a hospital in Denmark with a diagnosis of acute pancreatitis from January 1, 1980 through October 31, 2012. As many as 5 individuals from the general population without acute pancreatitis were matched for age and sex to each patient with acute pancreatitis. Pancreatic cancer risk was expressed as hazard ratios (HRs) with 95% CIs, calculated using the Cox proportional hazards model. Cox models were stratified by age, sex, and year of pancreatitis diagnosis and adjusted for alcohol- and smoking-related conditions, and Charlson Comorbidity Index score. Results We included 41,669 patients diagnosed with incident acute pancreatitis and 208,340 comparison individuals. Patients with acute pancreatitis had an increased risk of pancreatic cancer compared with the age- and sex-matched general population throughout the follow-up period. The risk decreased over time but remained high after more than 5 years of follow up (adjusted HR, 2.02; 95% CI, 1.57–2.61). Two- and 5-year absolute risks of pancreatic cancer among patients with acute pancreatitis were 0.68% (95% CI, 0.61%–0.77%) and 0.85% (95% CI, 0.76%–0.94), respectively. Conclusions In a nationwide, population-based, matched cohort study, we observed an association between diagnosis of acute pancreatitis and long-term risk of pancreatic cancer.
Biotechnology Challenges to In Vitro Maturation of Hepatic Stem Cells Gastroenterology (IF 18.392) Pub Date : 2018-02-08 Chen Chen, Alejandro Soto-Gutierrez, Pedro M. Baptista, Bart Spee
The incidence of liver disease is increasing globally. The only curative therapy for severe end-stage liver disease, liver transplantation, is limited by the shortage of organ donors. In vitro models of liver physiology have been developed and new technologies and approaches are rapidly progressing. Stem cells might be used as a source of liver tissue for development of models, therapies, and tissue engineering applications. However, we have been unable to generate and maintain stable and mature adult liver cells ex vivo. We review factors that promote hepatocyte differentiation and maturation, including growth factors, transcription factors, microRNAs, small molecules, and the microenvironment. We discuss how the hepatic circulation, microbiome, and nutrition affect liver function, and the criteria for considering cells derived from stem cells to be fully mature hepatocytes. We explain the challenges to cell transplantation and consider future technologies for use in hepatic stem cell maturation, including 3-dimensional biofabrication and genome modification.
Surveillance Imaging and Alpha Fetoprotein for Early Detection of Hepatocellular Carcinoma in Patients With Cirrhosis: A Meta-analysis Gastroenterology (IF 18.392) Pub Date : 2018-02-06 Kristina Tzartzeva, Joseph Obi, Nicole E. Rich, Neehar D. Parikh, Jorge A. Marrero, Adam Yopp, Akbar Waljee, Amit G. Singal
Background & Aims Society guidelines differ in their recommendations for surveillance to detect early-stage hepatocellular carcinoma (HCC) in patients with cirrhosis. We compared the performance of surveillance imaging, with or without alpha fetoprotein (AFP), for early detection of HCC in patients with cirrhosis Methods Two reviewers searched MEDLINE and SCOPUS from January 1990 through August 2016 to identify published sensitivity and specificity of surveillance strategies for overall and early detection of HCC. Pooled estimates were calculated and compared using the DerSimonian and Laird method for a random effects model. The study was conducted in accordance with Preferred Reporting Items for Systematic Review and Meta-analysis guidelines. Results Thirty-two studies (comprising 13367 patients) characterized sensitivity of imaging with or without AFP measurement for detection of HCC in patients with cirrhosis. Ultrasound detected any stage HCC with 84% sensitivity (95% CI, 76%–92%), but early-stage HCC with only 47% sensitivity (95% CI, 33%–61%). In studies comparing ultrasound with vs without AFP measurement, ultrasound detected any stage HCC with a lower level of sensitivity than ultrasound plus AFP measurement (relative risk [RR], 0.88; 95% CI, 0.83–0.93) and early-stage HCC with a lower level of sensitivity than ultrasound plus AFP measurement (RR, 0.81; 95% CI, 0.71–0.93). However, ultrasound alone detected HCC with a higher level of specificity than ultrasound plus AFP measurement (RR, 1.08; 95% CI, 1.05–1.09). Ultrasound with vs without AFP detected early-stage HCC with 63% sensitivity (95% CI, 48%–75%) and 45% sensitivity (95% CI, 30%–62%), respectively (P=.002). Only 4 studies evaluated computed tomography or magnetic resonance image-based surveillance, which detected HCC with 84% sensitivity (95% CI, 70%–92%). Conclusions In a meta-analysis of publications, we found ultrasound alone to detect early-stage HCC with a low level of sensitivity in patients with cirrhosis. Addition of AFP to ultrasound analysis significantly increases the sensitivity of HCC detection in clinical practice.
Effects of Training and Feedback on Accuracy of Predicting Rectosigmoid Neoplastic Lesions and Selection of Surveillance Intervals by Endoscopists Performing Optical Diagnosis of Diminutive Polyps Gastroenterology (IF 18.392) Pub Date : 2018-02-06 Jasper L.A. Vleugels, Marcel G.W. Dijkgraaf, Yark Hazewinkel, Linda K. Wanders, Paul Fockens, Evelien Dekker, M.C.J.M. Becx, A.M. van Berkel, W. Bruins Slot, M. Cazemier, A.C.T.M. Depla, J.M.J. Geesing, T.A. Grool, G.M.P. Houben, J.M. Jansen, K. Kessels, M.E. van Leerdam, N. van Lelyveld, R.C. Mallant – Hent, W.A. Marsman, E. Schnekenburger, E.J. van Soest, B.W. van der Spek, P.C.F. Stokkers, J. Tenthof van Noorden, S.A.C. van Tuyl, K.M.A.J. Tytgat, M.T. Uiterwaal, R.C. Verdonk, M.A.M.T. Verhagen, M. van der Vlugt, A. Voorburg, C.A. Wientjes
Background & Aims Real-time differentiation of diminutive polyps (1–5 mm) during endoscopy could replace histopathology analysis. According to guidelines, implementation of optical diagnosis into routine practice would require it to identify rectosigmoid neoplastic lesions with a negative predictive value (NPV) of over 90%, using histologic findings as a reference, and agreement with histology-based surveillance intervals for more than 90% of cases. Methods We performed a prospective study with 39 endoscopists accredited to perform colonoscopies on participants with positive results from fecal immunochemical tests in the Bowel Cancer Screening Program at 13 centers in the Netherlands. Endoscopists were trained in optical diagnosis using a validated module (WASP). After meeting predefined performance thresholds in the training program, the endoscopists started a 1-year program (continuation phase) in which they performed narrow-band imaging analyses during colonoscopies of participants in the screening program and predicted histological findings with confidence levels. The endoscopists were randomly assigned to groups that received feedback or no feedback on the accuracy of their predictions. Primary outcome measures were endoscopists’ abilities to identify rectosigmoid neoplastic lesions (using histology as a reference) with NPVs of 90% or more, and selecting surveillance intervals that agreed with those determined by histology for at least 90% of cases. Results Of 39 endoscopists initially trained, 27 (69%) completed the training program. During the continuation phase, these 27 endoscopists performed 3144 colonoscopies in which 4504 diminutive polyps were removed. The endoscopists identified neoplastic lesions with a pooled NPV of 90.8% (95% CI, 88.6–92.6); their proposed surveillance intervals agreed with those determined by histologic analysis for 95.4% of cases (95% CI, 94.0–96.6). Findings did not differ between the group that did versus did not receive feedback. Sixteen endoscopists (59%) identified rectosigmoid neoplastic lesions with NPVs greater than 90% and selected surveillance intervals in agreement with those determined from histology for more than 90% of patients. Conclusions In a prospective study following a validated training module, we found that a selected group of endoscopists identified rectosigmoid neoplastic lesions with pooled NPVs greater than 90% and accurately selected surveillance intervals for over 90% of patients over the course of 1 year. Providing regular interim feedback on the accuracy of neoplastic lesion prediction and surveillance interval selection did not lead to differences in those endpoints. Monitoring is suggested as individual performance varied. ClinicalTrials.gov no: NCT02516748.
STING Signaling Promotes Inflammation in Experimental Acute Pancreatitis Gastroenterology (IF 18.392) Pub Date : 2018-02-06 Qinglan Zhao, Yi Wei, Stephen J. Pandol, Lingyin Li, Aida Habtezion
Background & Aims Acute pancreatitis (AP) is characterized by severe inflammation and acinar cell death. Transmembrane protein 173 (TMEM173 or STING) is a pattern recognition receptor on immune cells that recognizes cytosolic nucleic acids and transmits signals that activate production of interferons and the innate immune response. We investigated whether leukocyte STING signaling mediates inflammation mice with AP. Methods We induced AP in C57BL/6J mice (control) and C57BL/6J-Tmem173gt/J mice (STING-knockout mice) by injection of cerulein or placement on choline-deficient DL-ethionine supplemented diet. In some mice, STING signaling was induced by administration of a pharmacologic agonist. AP was also induced in C57BL/6J mice with bone marrow transplants from control or STING-knockout mice and in mice with disruption of the cyclic GMP-AMP synthase (Cgas) gene. Pancreata were collected, analyzed by histology, and acini were isolated and analyzed by flow cytometry, quantitative PCR, immunoblots, and ELISAs. Bone-marrow-derived macrophages were collected from mice and tested for their ability to detect DNA from dying acinar cells in the presence and absence of deoxyribonuclease (DNaseI). Results STING signaling was activated in pancreata from mice with AP but not mice without AP. STING-knockout mice developed less-severe AP (less edema, inflammation, and markers of pancreatic injury) than control mice, whereas mice given a STING agonist developed more severe AP than controls. In immune cells collected from pancreata, STING was expressed predominantly in macrophages. Levels of CGAS were increased in mice with vs without AP, and CGAS-knockout mice had decreased edema, inflammation, and other markers of pancreatic injury upon induction of AP than control mice. Wildtype mice given bone marrow transplants from STING-knockout mice had less pancreatic injury and lower serum levels of lipase and pancreatic trypsin activity following induction of AP than mice given wildtype bone marrow. DNA from dying acinar cells activated STING signaling in macrophages, which was inhibited by addition of DNaseI. Conclusions In mice with AP, STING senses acinar cell death (by detecting DNA from dying acinar cells) and activates a signaling pathway that promotes inflammation. Macrophages express STING and activate pancreatic inflammation in AP.
Prevalence of and Factors Associated with Fecal Incontinence—Results From a Population-based Survey Gastroenterology (IF 18.392) Pub Date : 2018-02-03 Stacy B. Menees, Christopher V. Almario, Brennan M.R. Spiegel, William D. Chey
Background & Aims Fecal incontinence (FI) is characterized by uncontrolled passage of solid or liquid stool. We aimed to determine the prevalence and severity of FI in a large sample of United States residents. Methods We recruited a representative sample of patients in October 2015 to complete the National Gastrointestinal (GI) Survey; a mobile app called MyGiHealth was used to systematically collect data on GI symptoms. FI was defined as accidental leakage of solid or liquid stool. Severity of FI was determined by responses to the National Institutes of Health FI PROMIS questionnaire. Multivariable regression models were used to identify factors associated with FI prevalence and severity. Results Among 71,812 individuals who completed the National GI Survey, 14.4% reported FI in the past; of these, 33.3% had FI within the last 7 days. Older age, male sex, and Hispanic ethnicity increased the likelihood of having FI within the past week. Individuals with Crohn’s disease, ulcerative colitis, celiac disease, irritable bowel syndrome, or diabetes were more likely; to report FI. Non-Hispanic blacks and Hispanics and individuals with Crohn’s disease, celiac; disease, diabetes, HIV/AIDS, or chronic idiopathic constipation had more severe symptoms of FI than individuals without these features. Conclusions In a large population-based survey, 1 in 7 people reported previous FI. FI is age-related and more prevalent among individuals with inflammatory bowel disease, celiac disease, irritable bowel syndrome, or diabetes than people without these disorders. Proactive screening for FI among these groups is warranted.
Pharmacokinetics and Exposure Response Relationships of Ustekinumab in Patients with Crohn's Disease Gastroenterology (IF 18.392) Pub Date : 2018-02-01 Omoniyi J. Adedokun, Zhenhua Xu, Christopher Gasink, Douglas Jacobstein, Philippe Szapary, Jewel Johanns, Long-Long Gao, Hugh M. Davis, Steve Hanauer, Brian G. Feagan, Subrata Ghosh, William J. Sandborn
Background and Aims Ustekinumab is a monoclonal antibody that binds with high affinity to the p40 subunit of human interleukin 12 (IL12 and IL23) that has been approved for treatment of patients with moderate to severe Crohn’s disease (CD). However, there are few data on its pharmacokinetic properties or the relationship between drug exposure levels and patient response. We collected data from 2 phase 3 induction studies and 1 maintenance study to determine ustekinumab’s pharmacokinetic features, relationship between exposure and response, and optimal serum concentrations for efficacy. Methods We collected data on serum concentrations of ustekinumab and efficacy from induction studies of patients with moderate to severe CD given ustekinumab for 8 weeks following a single intravenous dose (either 130 mg or approximately 6 mg/kg). We collected the same data from a maintenance study of patients with a response to ustekinumab in the induction study who then received subcutaneous injections (90 mg) every 8 or 12 weeks for 44 weeks. At week 44 of the maintenance study (52 weeks after treatment began) patients were evaluated for the primary endpoint of clinical remission (defined as a CD activity index score below 150 points), endoscopic markers of efficacy, and serum level of C-reactive protein. Ustekinumab concentration data were categorized into quartiles and relationships between exposure and response were assessed. Optimal concentration cut-off values were evaluated using receiver operating characteristic curve analysis. Results Serum concentrations of ustekinumab over time were proportional to dose and did not differ significantly between the induction studies. In the maintenance study, ustekinumab concentration reached the steady state by the second maintenance dose; the median trough concentration was approximately 3-fold higher in patients given ustekinumab at 8-week intervals compared with 12-week intervals. Ustekinumab serum concentrations associated with rates of clinical remission and endoscopic efficacy endpoints, correlated inversely with level of C-reactive protein, and did not associate with use of immunomodulators. Trough concentrations of ustekinumab of 0.8 (or even up to 1.4 μg/mL) or greater were associated with maintenance of clinical remission in a higher proportion of patients than patients with lower trough concentrations. Conclusions In an analysis of data from phase 3 studies of patients with moderate to severe CD, we found serum concentrations of ustekinumab to be proportional to dose and associate with treatment efficacy. Concentrations of ustekinumab did not seem to be affected by co-treatment with immunomodulators. Clinicaltrials.gov no: NCT01369329 (UNITI 1), NCT01369342 (UNITI 2), and NCT01369355 (IM UNITI)
Microbiota-derived Metabolic Factors Reduce Campylobacteriosis in Mice Gastroenterology (IF 18.392) Pub Date : 2018-02-01 Xiaolun Sun, Kathryn Winglee, Raad Z. Gharaibeh, Josee Gauthier, Zhen He, Prabhanshu Tripathi, Dorina Avram, Steven Bruner, Anthony Fodor, Christian Jobin
Background & Aims Campylobacter jejuni, a prevalent food-borne bacterial pathogen, exploits the host innate response to induce colitis. Little is known about the roles of microbiota in C jejuni-induced intestinal inflammation. We investigated interactions between microbiota and intestinal cells during C jejuni infection of mice. Methods Germ-free C57BL/6 Il10–/– mice were colonized with conventional microbiota and infected with a single dose of C jejuni (109 colony-forming units/mouse), via gavage. Conventional microbiota was cultured under aerobic, microaerobic, or anaerobic conditions and orally transplanted into GF Il10–/– mice. Colon tissues were collected from mice and analyzed by histology, real-time PCR, and immunoblotting. Fecal microbiota and bile acids were analyzed with 16S sequencing and high-performance liquid chromatography with mass spectrometry, respectively. Results Introduction of conventional microbiota reduced C jejuni-induced colitis in previously germ-free Il10–/– mice, independent of fecal load of C jejuni, accompanied by reduced activation of mTOR. Microbiota transplantation and 16S rDNA sequencing experiments showed that Clostridium XI, Bifidobacterium, and Lactobacillus were enriched in fecal samples from mice colonized with microbiota cultured in anaerobic conditions (which reduce colitis) compared to mice fed microbiota cultured under aerobic conditions (susceptible to colitis). Oral administration to mice of microbiota-derived secondary bile acid sodium deoxycholate, but not ursodeoxycholic acid or lithocholic acid, reduced C jejuni-induced colitis. Depletion of secondary bile acid-producing bacteria with antibiotics that kill anaerobic bacteria (clindamycin) promoted C jejuni-induced colitis in SPF Il10–/– mice compared to the non-specific antibiotic nalidixic acid; colitis induction by antibiotics was associated with reduced level of luminal deoxycholate. Conclusions We identified a mechanism by which the microbiota controls susceptibility to C jejuni infection in mice, via bacteria-derived secondary bile acids.
HIF1-alpha Regulates Acinar Cell Function and Response to Injury in Mouse Pancreas Gastroenterology (IF 18.392) Pub Date : 2018-02-01 Min-Jung Park, Sapna Iyer, Xiang Xue, Juliana Bragazzi Cunha, Shufang Gu, David Moons, Steven W. Pipe, John A. Williams, Diane M. Simeone, Yatrik M. Shah, M. Bishr Omary
We investigated whether intra-pancreatic coagulation, with deposition of the fibrinogen-γ dimer (Fib-γD) and hypoxia, affect the severity of acute pancreatitis (AP) in mice. Pancreata of mice with AP induced by administration of cerulein or by L-arginine, or from patients with AP, had increased deposition of Fib-γD compared to control pancreata. Heparin administration protected mice from cerulein-induced AP and prevented Fib-γD formation. Cerulein administration resulted in activation and stabilization of HIF1-alpha in pancreata of ODD-luc HIF1-alpha reporter mice. Cerulein also led to induction of genes regulated by HIF1-alpha, including VEGFA and ERO1A, before evidence of Fib-γD deposition or histologic features of AP. Expression of tissue factor, which is regulated by VEGF, also increased following cerulein administration. Mice with acinar cell-specific disruption of Hif1a (Hif1aAc–/–) developed spontaneous endoplasmic reticulum stress and less severe AP, but did not accumulate Fib-γD following administration of cerulein. Feeding mice increased pancreatic expression of HIF1-alpha, indicating a physiologic role in the exocrine pancreas. Therefore, HIF1-alpha has bifunctional roles, in exocrine pancreas homeostasis and progression of AP that is promoted by intra-pancreatic coagulation.
Consensus Recommendations for Evaluation, Interpretation, and Utilization of Computed Tomography and Magnetic Resonance Enterography in Patients With Small Bowel Crohn’s Disease Gastroenterology (IF 18.392) Pub Date : 2018-01-10
Computed tomography and magnetic resonance enterography have become routine small bowel imaging tests to evaluate patients with established or suspected Crohn’s disease, but the interpretation and use of these imaging modalities can vary widely. A shared understanding of imaging findings, nomenclature, and utilization will improve the utility of these imaging techniques to guide treatment options, as well as assess for treatment response and complications. Representatives from the Society of Abdominal Radiology Crohn’s Disease-Focused Panel, the Society of Pediatric Radiology, the American Gastroenterological Association, and other experts, systematically evaluated evidence for imaging findings associated with small bowel Crohn’s disease enteric inflammation and established recommendations for the evaluation, interpretation, and use of computed tomography and magnetic resonance enterography in small bowel Crohn’s disease. This work makes recommendations for imaging findings that indicate small bowel Crohn’s disease, how inflammatory small bowel Crohn’s disease and its complications should be described, elucidates potential extra-enteric findings that may be seen at imaging, and recommends that cross-sectional enterography should be performed at diagnosis of Crohn’s disease and considered for small bowel Crohn’s disease monitoring paradigms. A useful morphologic construct describing how imaging findings evolve with disease progression and response is described, and standard impressions for radiologic reports that convey meaningful information to gastroenterologists and surgeons are presented.
Glycome Patterns of Perfusate in Livers Before Transplantation Associate With Primary Non-function Gastroenterology (IF 18.392) Pub Date : 2018-01-06 Xavier Verhelst, Anja Geerts, Ina Jochmans, Dieter Vanderschaeghe, Agnes Paradissis, Aude Vanlander, Frederik Berrevoet, Géraldine Dahlqvuist, Frederik Nevens, Jacques Pirenne, Xavier Rogiers, Nico Callewaert, Roberto I Troisi, Hans Van Vlierberghe
Background & Aims Primary non-function (PNF) is a rare complication after liver transplantation that requires urgent re-transplantation. PNF is associated with livers from extended-criteria donors. Clinical and biochemical factors have not been identified that reliably associate with graft function after liver transplantation. Serum patterns of N-glycans associate with changes in the liver. We analyzed perfusate from grafted liver to identify protein glycosylation patterns associated with PNF. Methods We performed a prospective study of consecutive patients who underwent liver transplantation (66 patients, from 1 center, in the derivation set and 56 patients, from 2 centers, in the validation set) in Belgium, from October 1, 2011 through July 31, 2013. All donor grafts were transported using cold static storage, and perfusate samples were collected from the livers by flushing of hepatic veins before transplantation. Protein-linked N-glycans were isolated from perfusate samples and analyzed with a multi-capillary electrophoresis-based ABI3130 sequencer. We compared glycan patterns between patients with vs without PNF of transplanted livers. PNF was defined as the need for urgent re-transplantation when a graft had no evidence of function, after exclusion of other causes such as hepatic artery thrombosis or acute cellular rejection. Results The relative abundance of a single glycan, agalacto core-alpha-1,6-fucosylated biantennary glycan (NGA2F) was significantly increased in perfusate of livers given to 4 patients who developed PNF after liver transplantation compared to livers given to patients who did not develop PNF. Level of NGA2F could identified patients with PNF with 100% accuracy. This glycomarker was the only factor associated with PNF in multivariate analysis in the derivation and the validation sets (P<.0001). Conclusions In an analysis of patients who underwent liver transplantation, we associated graft perfusate level of the glycosylated protein NGA2F with development of PNF with 100% accuracy, and validated this finding in a separate cohort of patients. This biomarker might be used to assess grafts before transplantation—especially when high-risk organs are under consideration.
Glycosylation of Immunoglobulin G Associates With Clinical Features of Inflammatory Bowel Diseases Gastroenterology (IF 18.392) Pub Date : 2018-01-06 Mirna Šimurina, Noortje de Haan, Frano Vučković, Nicholas A. Kennedy, Jerko Štambuk, David Falck, Irena Trbojević-Akmačić, Florent Clerc, Genadij Razdorov, Anna Khon, Anna Latiano, Renata D'Incà, Silvio Danese, Stephan Targan, Carol Landers, Marla Dubinsky, Dermot P.B. McGovern, Vito Annese, Manfred Wuhrer, Gordan Lauc
Background and aims Causes of inflammatory bowel diseases are not well understood and the most prominent forms Crohn’s disease (CD) and ulcerative colitis (UC) are sometimes hard to distinguish. Glycosylation of immunoglobulin G (IgG) has been associated with CD and UC. IgG Fc-glycosylation affects IgG effector functions. We evaluated changes in IgG Fc-glycosylation associated with UC and CD, as well as with disease characteristics in different patient groups. Methods We analyzed 3441 plasma samples, obtained from 2 independent cohorts of patients with CD (874 patients in Italy and 391 in the United States [US]) or UC (1056 in Italy and 253 in the US and healthy individuals (controls) (427 in Italy and 440 in the US). IgG Fc-glycosylation (tryptic glycopeptides) was analyzed by liquid chromatography coupled to mass spectrometry. We analyzed associations between disease status (UC vs controls, CD vs controls, and UC vs CD) and glycopeptide traits, and associations between clinical characteristics and glycopeptide traits, using a logistic regression model with age and sex included as covariates. Results Patients with CD or UC had lower levels of IgG galactosylation than controls. For example, the odds ratio (OR) for IgG1 galactosylation in patients with CD was 0.59 (95% CI, 0.51–0.69) and for patients with UC was 0.81 (95% CI, 0.71–0.92). Fucosylation of IgG was increased in patients with CD vs controls (for IgG1: OR, 1.27; 95% CI, 1.12–1.44) but decreased in patients with UC vs controls (for IgG23: OR, 0.72; 95% CI, 0.63–0.82). Decreased galactosylation associated with more severe CD or UC, including the need for surgery in patients with UC vs controls (for IgG1: OR, 0.69; 95% CI, 0.54–0.89) and in patients with CD vs controls (for IgG23: OR, 0.78; 95% CI, 0.66–0.91). Conclusion In a retrospective analysis of plasma samples from patients with CD or UC, we associated levels of IgG Fc-glycosylation with disease (compared to controls) and its clinical features. These findings could increase our understanding of mechanisms of CD and UC pathogenesis and be used to develop diagnostics or guide treatment.
Increasing Rates of Surgery for Patients With Non-Malignant Colorectal Polyps in the United States Gastroenterology (IF 18.392) Pub Date : 2018-01-06 Anne F. Peery, Katherine S. Cools, Paula D. Strassle, Sarah K. McGill, Seth D. Crockett, Aubrey Barker, Mark Koruda, Ian S. Grimm
Background & Aims Despite the availability of endoscopic therapy, many patients in the United States undergo surgical resection for non-malignant colorectal polyps. We aimed to quantify and examine trends in the use of surgery for non-malignant colorectal polyps in a nationally representative sample. Methods We analyzed data from the Healthcare Cost and Utilization Project National Inpatient Sample for the years 2000 through 2014. We included all adult patients who underwent elective colectomy or proctectomy and had a diagnosis of either non-malignant colorectal polyp or colorectal cancer. We compared trends in surgery for non-malignant colorectal polyps with surgery for colorectal cancer and calculated age, sex, race, region, and teaching status/bed-size specific incidence rates of surgery for non-malignant colorectal polyps. Results From 2000 through 2014, there were 1,230,458 surgeries for non-malignant colorectal polyps and colorectal cancer in the United States. Among those surgeries, 25% were performed for non-malignant colorectal polyps. The incidence of surgery for non-malignant colorectal polyps has increased significantly, from 5.9 in 2000 to 9.4 in 2014 per 100,000 adults (incidence rate difference, 3.56; 95% CI 3.40–3.72), while the incidence of surgery for colorectal cancer has significantly decreased, from 31.5 to 24.7 surgeries per 100,000 adults (incidence rate difference, –6.80; 95% CI, –7.11 to –6.49). The incidence of surgery for non-malignant colorectal polyps has been increasing among individuals 20–79, in men and women and including all races and ethnicities. Conclusions In an analysis of a large, nationally representative sample, we found that surgery for non-malignant colorectal polyps is common and has significantly increased over the last 14 years.
Accuracy of the Enhanced Liver Fibrosis Test vs Fibrotest, Elastography and Indirect Markers in Detection of Advanced Fibrosis in Patients with Alcoholic Liver Disease Gastroenterology (IF 18.392) Pub Date : 2018-01-06 Maja Thiele, Bjørn Stæhr Madsen, Janne Fuglsang Hansen, Sönke Detlefsen, Steen Antonsen, Aleksander Krag
Background & Aims Alcohol is the leading cause of cirrhosis and liver-related mortality, but we lack serum markers to detect compensated disease. We compared the accuracy of the Enhanced Liver Fibrosis test (ELF), the FibroTest, liver stiffness measurements (made by transient elastography and 2-dimensional shear-wave elastography), and 6 indirect marker tests in detection of advanced liver fibrosis (Kleiner stage ≥F3). Methods We performed a prospective study of 10 liver fibrosis markers (patented and not), all performed on the same day. Patients were recruited from primary centers (municipal alcohol rehabilitation, n=128; 6% with advanced fibrosis) and secondary healthcare centers (hospital outpatient clinics, n=161; 36% with advanced fibrosis) in the Region of Southern Denmark from 2013 through 2016. Biopsy-verified fibrosis stage was used as the reference standard. The primary aim was to validate ELF in detection of advanced fibrosis in patients with alcoholic liver disease recruited from primary and secondary healthcare centers, using the literature-based cut-off value of 10.5. Secondary aims were to assess the diagnostic accuracy of ELF for significant fibrosis and cirrhosis and to determine whether combinations of fibrosis markers increase diagnostic yield. Results The ELF identified patients with advanced liver fibrosis with an area under the receiver operating characteristic curve (AUROC) of 0.92 (95% CI, 0.89–0.96); findings did not differ significantly between patients from primary vs secondary care (P=.917). ELF more accurately identified patients with advanced liver fibrosis than indirect marker tests, but ELF and FibroTest had comparable diagnostic accuracies (AUROC of FibroTest, 0.90) (P=.209 for comparison with ELF). Results from the ELF and FibroTest did not differ significantly from those of liver stiffness measurement in intention-to-diagnose analyses (AUROC for transient elastography, 0.90), but did differ in the per-protocol analysis (AUROC for transient elastography, 0.97) (P=.521 and .004 for comparison with ELF). Adding a serum marker to transient elastography analysis did not increase accuracy. For patients in primary care, ELF values below 10.5 and FibroTest values below 0.58 had negative predictive values for advanced liver fibrosis of 98% and 94%, respectively. Conclusion In a prospective, direct comparison of tests, ELF and FibroTest identified advanced liver fibrosis in alcoholic patients from primary and secondary care with high diagnostic accuracy (AUROC values of 0.90 or higher using biopsy as reference). Advanced fibrosis can be ruled out in primary healthcare patients based on an ELF value below 10.5 or a FibroTest value below 0.58.
Hedgehog–YAP Signaling Pathway Regulates Glutaminolysis to Control Hepatic Stellate Cell Activation Gastroenterology (IF 18.392) Pub Date : 2018-01-03 Kuo Du, Jeongeun Hyun, Richard T. Premont, Steve S. Choi, Gregory A. Michelotti, Marzena Swiderska-Syn, George D. Dalton, Eric Thelen, Bahar Salimian Rizi, Youngmi Jung, Anna Mae Diehl
Background & Aims Cirrhosis results from accumulation of myofibroblasts derived from quiescent hepatic stellate cells (Q-HSCs); it regresses when myofibroblastic HSCs are depleted. Hedgehog signaling promotes transdifferentiation of HSCs by activating Yes-associated protein 1 (YAP1 or YAP) and inducing aerobic glycolysis. However, increased aerobic glycolysis alone cannot meet the high metabolic demands of myofibroblastic HSCs. Determining the metabolic processes of these cells could lead to strategies to prevent progressive liver fibrosis, so we investigated whether glutaminolysis (conversion of glutamine to alpha-ketoglutarate) sustains energy metabolism and permits anabolism when Q-HSCs become myofibroblastic, and whether this is controlled by hedgehog signaling to YAP. Methods Primary HSCs were isolated from C57BL/6 or Smoflox/flox mice; we also performed studies with rat and human myofibroblastic HSCs. We measured changes of glutaminolytic genes during culture-induced primary HSC transdifferentiation. Glutaminolysis was disrupted in cells by glutamine deprivation or pathway inhibitors (BPTES, CB-839, EGCG and AOA), and effects on mitochondrial respiration, cell growth and migration, and fibrogenesis were measured. Hedgehog signaling to YAP was disrupted in cells by adenovirus expression of Cre-recombinase or by small hairpin RNA knockdown of YAP. Hedgehog and YAP activity were inhibited by incubation cells with cyclopamine or verteporfin, and effects on glutaminolysis were measured. Acute and chronic liver fibrosis were induced in mice by intraperitoneal injection of CCl4 or methionine choline-deficient diet. Some mice were then given injections of BPTES to inhibit glutaminolysis, and myofibroblast accumulation was measured. We also performed mRNA and immunohistochemical analyses of percutaneous liver biopsies from healthy human and 4 patients with no fibrosis, 6 patients with mild fibrosis, and 3 patients with severe fibrosis. Results Expression of genes that regulate glutaminolysis increased during transdifferentiation of primary Q-HSCs into myofibroblastic HSCs, and inhibition of glutaminolysis disrupted transdifferentitation. Blocking glutaminolysis in myofibroblastic HSCs suppressed mitochondrial respiration, cell growth and migration, and fibrogenesis; replenishing glutaminolysis metabolites to these cells restored these activities. Knockout of the hedgehog signaling intermediate SMO or knockdown of YAP inhibited expression of glutaminase, the rate limiting enzyme in glutaminolysis. Hedgehog and YAP inhibitors blocked glutaminolysis and suppressed myofibroblastic activities in HSCs. In livers of patients and of mice with acute or chronic fibrosis, glutaminolysis was induced in myofibroblastic HSCs. In mice with liver fibrosis, inhibition of glutaminase blocked accumulation of myofibroblasts and fibrosis progression. Conclusions Glutaminolysis controls accumulation of myofibroblast HSCs in mice and might be a therapeutic target for cirrhosis.
Effects of Weight Loss Medications on Cardiometabolic Risk Profiles: A Systematic Review and Network Meta-Analysis Gastroenterology (IF 18.392) Pub Date : 2018-01-03 Rohan Khera, Ambarish Pandey, Apoorva K. Chandar, Mohammad H. Murad, Larry J. Prokop, Ian J. Neeland, Jarett Berry, Michael Camilleri, Siddharth Singh
Background & Aims We performed a systematic review and network meta-analysis to evaluate the overall and comparative effects of weight-loss medications, approved by the Food and Drug Administration (FDA) for long-term use, on cardiometabolic risk profiles of obese adults. Methods We performed a systematic literature review through February 28, 2017 to identify randomized clinical trials of the effects of FDA-approved weight loss medications (orlistat, lorcaserin, naltrexone-bupropion, phentermine-topiramate, and liraglutide), administered to obese adults for 1 year or more, compared with placebo or another active agent. Outcomes of interest included changes in blood glucose (fasting blood glucose [FBG] and hemoglobin A1c [A1c]), cholesterol profile (low-density lipoprotein and high-density lipoproteins [HDL]), blood pressure (systolic/diastolic) and waist circumference (WC). We performed pair-wise and network meta-analyses with outcomes reported as weighted and standardized mean differences. Quality of evidence was rated using GRADE. Results In a meta-analysis of 28 randomized controlled trials (29018 participants; median body mass index, 36.1kg/m2), we associated weight-loss medications with a modest decrease in FBG (weighted mean difference, 4.0 mg/dL; 95% CI, –4.4 to –3.6) and WC (weighted mean difference, reduction of 3.3 cm; 95% CI, –3.5 to –3.1), without clinically meaningful changes in systolic/diastolic blood pressure or cholesterol profile vs placebo (standardized mean difference below 0.2); effects varied among drugs. Phentermine-topiramate use was associated with a substantial decrease in WC and a modest decrease in FBG, A1c, and blood pressure and had minimal effect on cholesterol. Liraglutide use was associated with a substantial decrease in FBG, A1c, and WC and a minimal effect on blood pressure and cholesterol. Naltrexone-bupropion use was associated with moderate increase in HDL cholesterol but had a minimal effect on FBG and WC. Orlistat use was associated with a decrease in low-density lipoprotein and HDL-cholesterol. No drug improved all cardiometabolic risk factors. Conclusions In a systematic review and network meta-analysis, we found FDA-approved weight loss medications to have only modest positive effects on cardiometabolic risk profile. Further research is needed to evaluate the long-term cardio-metabolic benefits of these medications. PROSPERO: CRD42016039486
Efficacy of Laparoscopic Nissen Fundoplication vs Transoral Incisionless Fundoplication or Proton Pump Inhibitors in Patients With Gastroesophageal Reflux Disease: a Systematic Review and Network Meta-analysis Gastroenterology (IF 18.392) Pub Date : 2018-01-03 Joel Richter, Ambuj Kumar, Seth Lipka, Branko Miladinovic, Vic Velanovich
Background & Aims The effects of transoral incisionless fundoplication (TIF) and laparoscopic Nissen fundoplication (LNF) have been compared with those of proton pump inhibitors (PPIs) or a sham procedure in patients with gastroesophageal reflux disease (GERD), but there has been no direct comparison of TIF vs LNF. We performed a systematic review and network meta-analysis of randomized controlled trials to compare the relative efficacies of TIF vs LNF in patients with GERD. Methods We searched publication databases and conference abstracts through May 10, 2017 for randomized controlled trials that compared the efficacy of TIF or LNF with that of a sham procedure or PPIs in patients with GERD. We performed a network meta-analysis using Bayesian methods under random-effects multiple treatment comparisons. We assessed ranking probability by surface under the cumulative ranking curve. Results Our search identified 7 trials comprising 1128 patients. Surface under the cumulative ranking curve ranking indicated TIF had highest probability of increasing patients’ health-related quality of life (HRQOL; 0.96), followed by LNF (0.66), a sham procedure (0.35), and PPIs (0.042). LNF had the highest probability of increasing percent time at pH<4 (0.99), followed by PPIs (0.64), TIF (0.32), and the sham procedure (0.05). LNF also had the highest probability of increasing LES pressure (0.78), followed by TIF (0.72) and PPIs (0.01). Patients who underwent the sham procedure had the had the highest probability for persistent esophagitis (0.74), followed by those receiving TIF (0.69), LNF (0.38), and PPIs (0.19). Meta-regression showed a shorter follow-up time as a significant confounder for the outcome of HRQOL in studies of TIF. Conclusions In a systematic review and network meta-analysis of trials of patients with GERD, we found LNF to have the greatest ability to improve physiologic parameters of GERD, including increased LES pressure and percent time a pH<4. Although TIF produced the largest increase in HRQOL, this could be due to the shorter follow-up time of patients treated with TIF vs LNF or PPIs. TIF is a minimally invasive endoscopic procedure, yet based on evaluation of benefits vs risks, we do not recommend it as a long-term alternative to PPI or LNF treatment of GERD.
Cystic Fibrosis Colorectal Cancer Screening Consensus Recommendations Gastroenterology (IF 18.392) Pub Date : 2017-12-29
Background & Aims Improved therapy has substantially increased survival of persons with cystic fibrosis (CF). But the risk of colorectal cancer (CRC) in adults with CF is 5−10 times greater compared to the general population, and 25−30 times greater in CF patients after an organ transplantation. To address this risk, the CF Foundation convened a multi-stakeholder task force to develop CRC screening recommendations. Methods The 18-member task force consisted of experts including pulmonologists, gastroenterologists, a social worker, nurse coordinator, surgeon, epidemiologist, statistician, CF adult, and a parent. The committee comprised 3 workgroups: Cancer Risk, Transplant, and Procedure and Preparation. A guidelines specialist at the CF Foundation conducted an evidence synthesis February−March 2016 based on PubMed literature searches. Task force members conducted additional independent searches. A total of 1159 articles were retrieved. After initial screening, the committee read 198 articles in full and analyzed 123 articles to develop recommendation statements. An independent decision analysis evaluating the benefits of screening relative to harms and resources required was conducted by the Department of Public Health at Erasmus Medical Center, Netherlands using the Microsimulation Screening Analysis model from the Cancer Innervation and Surveillance Modeling Network. The task force included recommendation statements in the final guideline only if they reached an 80% acceptance threshold. Results The task force makes 10 CRC screening recommendations that emphasize shared, individualized decision-making and familiarity with CF-specific gastrointestinal challenges. We recommend colonoscopy as the preferred screening method, initiation of screening at age 40 years, 5-year re-screening and 3-year surveillance intervals (unless shorter interval is indicated by individual findings), and a CF-specific intensive bowel preparation. Organ transplant recipients with CF should initiate CRC screening at age 30 years within 2 years of the transplantation because of the additional risk for colon cancer associated with immunosuppression. Conclusions These recommendations aim to help CF adults, families, primary care physicians, gastroenterologists, and CF and transplantation centers address the issue of CRC screening. They differ from guidelines developed for the general population with respect to the recommended age of screening initiation, screening method, preparation, and the interval for repeat screening and surveillance.
Functional Bowel Disorders: A Roadmap to Guide the Next Generation of Research Gastroenterology (IF 18.392) Pub Date : 2017-12-27 Lin Chang, Carlo Di Lorenzo, Gianrico Farrugia, Frank A. Hamilton, Gary M. Mawe, Pankaj J. Pasricha, John W. Wiley
In June 2016, the National Institutes of Health hosted a workshop on functional bowel disorders (FBDs), particularly irritable bowel syndrome, with the objective of elucidating gaps in current knowledge and recommending strategies to address these gaps. The workshop aimed to provide a roadmap to help strategically guide research efforts during the next decade. Attendees were a diverse group of internationally recognized leaders in basic and clinical FBD research. This document summarizes the results of their deliberations, including the following general conclusions and recommendations. First, the high prevalence, economic burden, and impact on quality of life associated with FBDs necessitate an urgent need for improved understanding of FBDs. Second, preclinical discoveries are at a point that they can be realistically translated into novel diagnostic tests and treatments. Third, FBDs are broadly accepted as bidirectional disorders of the brain−gut axis, differentially affecting individuals throughout life. Research must integrate each component of the brain−gut axis and the influence of biological sex, early-life stressors, and genetic and epigenetic factors in individual patients. Fourth, research priorities to improve diagnostic and management paradigms include enhancement of the provider−patient relationship, longitudinal studies to identify risk and protective factors of FBDs, identification of biomarkers and endophenotypes in symptom severity and treatment response, and incorporation of emerging “-omics” discoveries. These paradigms can be applied by well-trained clinicians who are familiar with multimodal treatments. Fifth, essential components of a successful program will include the generation of a large, validated, broadly accessible database that is rigorously phenotyped; a parallel, linkable biorepository; dedicated resources to support peer-reviewed, hypothesis-driven research; access to dedicated bioinformatics expertise; and oversight by funding agencies to review priorities, progress, and potential synergies with relevant stakeholders.
Cost-Effectiveness of Screening Individuals With Cystic Fibrosis for Colorectal Cancer Gastroenterology (IF 18.392) Pub Date : 2017-12-27 Andrea Gini, Ann G. Zauber, Dayna R. Cenin, Amir-Houshang Omidvari, Sarah E. Hempstead, Aliza K. Fink, Albert B. Lowenfels, Iris Lansdorp-Vogelaar
Background & Aims Individuals with cystic fibrosis are at increased risk of colorectal cancer (CRC) compared to the general population, and risk is higher among those who received an organ transplant. We performed a cost-effectiveness analysis to determine optimal CRC screening strategies for patients with cystic fibrosis. Methods We adjusted the existing Microsimulation Screening Analysis-Colon microsimulation model to reflect increased CRC risk and lower life expectancy in patients with cystic fibrosis. Modeling was performed separately for individuals who never received an organ transplant and patients who had received an organ transplant. We modeled 76 colonoscopy screening strategies that varied the age range and screening interval. The optimal screening strategy was determined based on a willingness to pay threshold of $100,000 per life-year gained. Sensitivity and supplementary analyses were performed, including fecal immunochemical test (FIT) as an alternative test, earlier ages of transplantation, and increased rates of colonoscopy complications, to assess whether optimal screening strategies would change. Results Colonoscopy every 5 years, starting at age 40 years, was the optimal colonoscopy strategy for patients with cystic fibrosis who never received an organ transplant; this strategy prevented 79% of deaths from CRC. Among patients with cystic fibrosis who had received an organ transplant, optimal colonoscopy screening should start at an age of 30 or 35 years, depending on the patient’s age at time of transplantation. Annual FIT screening was predicted to be cost-effective for patients with cystic fibrosis. However, the level of accuracy of the FIT in population is not clear. Conclusions Using a Microsimulation Screening Analysis-Colon microsimulation model, we found screening of patients with cystic fibrosis for CRC to be cost-effective. Due to the higher risk in these patients for CRC, screening should start at an earlier age with a shorter screening interval. The findings of this study (especially those on FIT screening) may be limited by restricted evidence available for patients with cystic fibrosis.
Reshaping the Tumor Stroma for Treatment of Pancreatic Cancer Gastroenterology (IF 18.392) Pub Date : 2017-12-26 Claire Vennin, Kendelle J. Murphy, Jennifer P. Morton, Thomas R. Cox, Marina Pajic, Paul Timpson
Pancreatic cancer is accompanied with a fibrotic reaction that alters interactions between tumor cells and the stroma to promote tumor progression. Consequently, strategies to target the tumor stroma might be used to treat patients with pancreatic cancer. We review recently developed approaches for re-shaping the pancreatic tumor stroma and discuss how these might improve patient outcomes. We also describe relationships between the pancreatic tumor extracellular matrix, the vasculature, the immune system, and metabolism and discuss the implications for the development of stromal compartment-specific therapies.
Pan-Genotype Hepatitis E Virus Replication in Stem Cell-derived Hepatocellular Systems Gastroenterology (IF 18.392) Pub Date : 2017-12-24 Xianfang Wu, Viet Loan Dao Thi, Peng Liu, Constantin N. Takacs, Kuanhui Xiang, Linda Andrus, Jérôme Gouttenoire, Darius Moradpour, Charles M. Rice
Background & Aims The 4 genotypes of hepatitis E virus (HEV) that infect humans (genotypes 1–4) vary in geographical distribution, transmission, and pathogenesis. Little is known about the properties of HEV or its hosts that contribute to these variations. Primary isolates grow poorly in cell culture; most studies have relied on variants adapted to cancer cell lines, which likely alter virus biology. We investigated the infection and replication of primary isolates of HEV in hepatocyte-like cells (HLCs) derived from human embryonic and induced pluripotent stem cells. Methods Using a cell culture-adapted genotype 3 strain and primary isolates of genotypes 1–4, we compared viral replication kinetics, sensitivity to drugs, and ability of HEV to activate the innate immune response. We studied HLCs using quantitative reverse-transcriptase PCR and immunofluorescence assay and ELISAs. We used an embryonic stem cell line that can be induced to express the CRISPR-Cas9 machinery to disrupt the peptidylprolyl isomerase A gene (PPIA), encoding cyclophilin A (CYPA)—a protein reported to inhibit replication of cell culture-adapted HEV. We further modified this line to rescue expression of CYPA before terminal differentiation to HLCs and performed HEV infection studies. Results HLCs were permissive for infection by non-adapted, primary isolates of HEV genotypes 1–4. HEV infection of HLCs induced a replication-dependent type III interferon response. Replication of primary HEV isolates, unlike the cell culture-adapted strain, was not affected by disruption of PPIA or exposure to the CYPA inhibitor cyclosporine A. Conclusions Cell culture adaptations alter the replicative capacities of HEV. HLCs offer an improved, physiologically relevant, and genetically tractable system for studying the replication of primary HEV isolates. HLCs could provide a model to aid development of HEV drugs and a system to guide personalized regimens, especially for patients with chronic hepatitis E who have developed resistance to ribavirin.
Nuclear Receptor Subfamily 1 Group D Member 1 Regulates Circadian Activity Of NLRP3 Inflammasome to Reduce the Severity of Fulminant Hepatitis in Mice Gastroenterology (IF 18.392) Pub Date : 2017-12-24 B. Pourcet, M. Zecchin, L. Ferri, J. Beauchamp, S. Sitaula, C. Billon, S. Delhaye, J. Vanhoutte, A. Mayeuf-Louchart, Q. Thorel, J. Haas, J. Eeckhoute, D. Dombrowicz, C. Duhem, A. Boulinguiez, S. Lancel, Y. Sebti, T. Burris, B. Staels, H. Duez
Background & Aims The innate immune system responds not only to bacterial signals, but also to non-infectious danger-associated molecular patterns that activate the NLRP3 inflammasome complex after tissue injury. Immune functions vary over the course of the day, but it is not clear whether these changes affect the activity of the NLRP3 inflammasome. We investigated whether the core clock component nuclear receptor subfamily 1 group D member 1 (NR1D1, also called Rev-erbα) regulates expression, activity of the NLRP3 inflammasome, and its signaling pathway. Methods We collected naïve peritoneal macrophages and plasma, at multiple times of day, from Nr1d1–/– mice and their Nr1d1+/+ littermates (controls) and analyzed expression of NLR family pyrin domain containing 3 (NLRP3), interleukin 1 beta (IL1B, in plasma), and IL18 (in plasma). We also collected bone marrow-derived primary macrophages from these mice. Levels of NR1D1 were knocked down with small hairpin RNAs in human primary macrophages. Bone marrow-derived primary macrophages from mice and human primary macrophages were incubated with lipopolysaccharide (LPS) to induce expression of NLRP3, IL1B and IL18; cells were incubated with LPS and ATP to activate the NLRP3 complex. We analyzed caspase 1 activity and cytokine secretion. NR1D1 was activated in primary mouse and human macrophages by incubation with SR9009; some of the cells were also incubated with an NLRP3 inhibitor or inhibitors of caspase 1. Nr1d1–/– mice and control mice were given intraperitoneal injections of LPS to induce peritoneal inflammation; plasma samples were isolated and levels of cytokines were measured. Nr1d1–/– mice, control mice, and control mice given injections of SR9009 were given LPS and galactosamine to induce fulminant hepatitis and MCC950 to specifically inhibit NLRP3; plasma was collected to measure cytokines and a marker of liver failure (ALAT); liver tissues were collected and analyzed by quantitative PCR, immunohistochemistry, and flow cytometry. Results In peritoneal macrophages, expression of NLRP3 and activation of its complex varied with time of day (circadian rhythm)—this regulation required NR1D1. Primary macrophages from Nr1d1–/– mice and human macrophages with knockdown of NR1D1 had altered expression patterns of NLRP3, compared to macrophages that expressed NR1D1, and altered patterns of IL1B and 1L18 production. Mice with disruption of Nr1d1 developed more-severe acute peritoneal inflammation and fulminant hepatitis than control mice. Incubation of macrophage with the NR1D1 activator SR9009 reduced expression of NLRP3 and secretion of cytokines. Mice given SR9009 developed less-severe liver failure and had longer survival times than mice given saline (control). Conclusions In studies of Nr1d1–/– mice and human macrophages with pharmacologic activation of NR1D1, we found NR1D1 to regulate the timing of NLRP3 expression and production of inflammatory cytokines by macrophages. Activation of NR1D1 reduced the severity of peritoneal inflammation and fulminant hepatitis in mice.
Neuromodulators for Functional GI Disorders (Disorders of Gut-Brain Interaction): A Rome Foundation Working Team Report Gastroenterology (IF 18.392) Pub Date : 2017-12-22 Douglas A. Drossman, Jan Tack, Alexander C. Ford, Eva Szigethy, Hans Törnblom, Lukas Van Oudenhove
Background and Aims Central neuromodulators (antidepressants, antipsychotics and other CNS targeted medications) are increasingly used for treatment of functional GI disorders (FGIDs), now recognized as disorders of gut-brain interaction. However, the available evidence and guidance for the use of central neuromodulators in these conditions is scanty and incomplete. In this Rome Foundation Working Team report, a multidisciplinary team summarized available research evidence and clinical experience to provide guidance and treatment recommendations. Methods The working team summarized the literature on the pharmacology of central neuromodulators and their effects on gastrointestinal sensorimotor function and conducted an evidence based review on their use for treating FGID syndromes. Because of paucity of data in FGIDs, we included data for non-GI painful disorders and specific symptoms of pain, nausea and vomiting. This information was combined into a final document comprising a synthesis of available evidence and recommendations for clinical use guided by the research and clinical experience of the experts on the committee. Results The evidence based review on neuromodulators in FGID, restricted by the limited available controlled trials, was integrated with open label studies and case series along with the experience of experts to create recommendations using a consensus (Delphi) approach. Due to the diversity of conditions and complexity of treatment options, specific recommendations were generated for different functional GI disorders. However, some general recommendations include: 1) Low to modest doses of tricyclic antidepressants provide the most convincing evidence of benefit for treating chronic gastrointestinal pain and painful FGIDs and serotonin noradrenergic reuptake inhibitors can also be recommended though further studies are needed; 2) Augmentation, i.e., adding a second treatment (adding quetiapine, aripiprazole, buspirone α2δ [Delta] ligand agents) is recommended when a single medication is unsuccessful or produces side effects in higher dosages; 3) Treatment should be continued for 6-12 months to potentially prevent relapse, and 4) Implementation of successful treatment requires effective communication skills to improve patient acceptance, adherence and to optimize the patient provider relationship. Conclusion Based on systematic and selectively focused review and the consensus of a multidisciplinary panel, we have provided summary information and guidelines for the use of central neuromodulators in the treatment of chronic GI symptoms and FGIDs. Further studies are needed to confirm and refine these recommendations.
Efficacy of NS5A Inhibitors Against Hepatitis C Virus Genotypes 1–7 and Escape Variants Gastroenterology (IF 18.392) Pub Date : 2017-12-22 Judith M. Gottwein, Long V. Pham, Lotte S. Mikkelsen, Lubna Ghanem, Santseharay Ramirez, Troels K.H. Scheel, Thomas H.R. Carlsen, Jens Bukh
Background & Aims Inhibitors of the hepatitis C virus (HCV) NS5A protein are a key component of effective treatment regimens, but the genetic heterogeneity of HCV has limited the efficacy of these agents and mutations lead to resistance. We directly compared the efficacy of all clinically relevant NS5A inhibitors against HCV genotype 1–7 prototype isolates and resistant escape variants, and investigated the effects of pre-existing resistance-associated substitutions (RAS) on HCV escape from treatment. Methods We measured the efficacy of different concentrations of daclatasvir, ledipasvir, ombitasvir, elbasvir, ruzasvir, velpatasvir, and pibrentasvir in cultured cells infected with HCV recombinants expressing genotype 1–7 NS5A proteins with or without RAS. We engineered HCV variants that included RAS identified in escape experiments, using recombinants with or without T/Y93H and daclatasvir, or that contained RAS previously reported from patients. Results NS5A inhibitors had varying levels of efficacy against original and resistant viruses. Only velpatasvir and pibrentasvir had uniform high activity against all HCV genotypes tested. RAS hotspots in NS5A were found at amino acids 28, 30, 31, and 93. Engineered escape variants had high levels of fitness. Pibrentasvir had the highest level of efficacy against variants; viruses with RAS at amino acids 28, 30, or 31 had no apparent resistance to pibrentasvir, and HCV with RAS at amino acid 93 had a low level of resistance to this drug. However, specific combinations of RAS and deletion of amino acid 32 led to significant resistance to pibrentasvir. For the remaining NS5A inhibitors tested, RAS at amino acids 28 and 93 led to high levels of resistance. Among these inhibitors, velpatasvir was more effective against variants with RAS at amino acid 30 and some variants with RAS at amino acid 31 than the other agents. Variants with the pre-existing RAS T/Y93H acquired additional NS5A changes during escape experiments, resulting in HCV variants with specific combinations of RAS, showing high fitness and high resistance. Conclusions We performed a comprehensive comparison of the efficacy of the 7 clinically relevant inhibitors of HCV NS5A and identified variants associated with resistance to each agent. These findings could improve treatment of patients with HCV infection.
Villin-1 and Gelsolin Regulate Changes in Actin Dynamics That Affect Cell Survival Signaling Pathways and Intestinal Inflammation Gastroenterology (IF 18.392) Pub Date : 2017-12-21 Swati Roy, Amin Esmaeilniakooshkghazi, Srinivas Patnaik, Yaohong Wang, Sudeep P. George, Afzal Ahrorov, Jason K. Hou, Allan J. Herron, Hiromi Sesaki, Seema Khurana
Background & Aims Cell stress signaling pathways result in phosphorylation of the eukaryotic translation initiation factor 2 subunit alpha (EIF2S1 or EIF2A), which affects regulation of protein translation. Translation reprogramming mitigates stress by activating pathways that result in autophagy and cell death, to eliminate damaged cells. Actin is modified during stress and EIF2A is dephosphorylated to restore homeostasis. It is not clear how actin affects EIF2A signaling. We studied the actin-binding proteins villin 1 (VIL1) and gelsolin (GSN) in intestinal epithelial cells (IECs) to determine whether they respond to cell stress response and affect signaling pathways. Methods We performed studies with mice with disruptions in Vil1 and Gsn (double-knockout mice). Wild-type mice either were or were not (controls) exposed to cell stressors such as tumor necrosis factor and adherent-invasive Escherichia coli. Distal ileum tissues were collected from mice; IECs and enteroids were cultured and analyzed by histology, immunoblots, phalloidin staining, immunohistochemistry, electron microscopy, and flow cytometry. HT-29 cells were incubated with cell stressors such as dithiothreitol, interferon, and adherent-invasive E coli or control agents; cells were analyzed by immunoblots and quantitative PCR. Full-length and mutant EIF2A were expressed from a lentiviral vector. The mouse immunity related GTPase (IRGM1) was overexpressed in embryonic fibroblasts from dynamin1 like (DNML1) protein-knockout mice or their wild-type littermates. IRGM1 was overexpressed in embryonic fibroblasts from receptor interacting serine/threonine kinase 1-knockout mice or their wild-type littermates. Human IRGM was overexpressed in human epithelial cell lines incubated with the DNML1-specific inhibitor Mdivi-1. Mitochondria were analyzed by semi-quantitative confocal imaging. We performed immunohistochemical analyses of distal ileum tissues from 6–8 patients with Crohn’s disease (CD) and 6–8 individuals without CD (controls). Results In IECs exposed to cell stressors, EIF2A signaling reduced expression of VIL1 and GSN. However, VIL1 and GSN were required for dephosphorylation of EIF2A and recovery from cell stress. In mouse and human IECs, prolonged, unresolved stress was accompanied by continued downregulation of VIL1 and GSN, resulting in constitutive phosphorylation of EIF2A and overexpression of IRGM1 (or IRGM), which regulates autophagy. Overexpression of IRGM1 (or IRGM) induced cell death by necroptosis, accompanied by release of damage associated molecular patterns (DAMPs). In double-knockout mice, constitutive phosphorylation of EIF2A and over-expression of IRGM1 resulted in spontaneous ileitis that resembled human CD in symptoms and histology. Distal ileum tissues from patients with CD had lower levels of VIL1 and GSN, increased phosphorylation of EIF2A, increased levels of IRGM and necroptosis, and increased release of nuclear DAMPs compared to controls. Conclusions In studies of intestinal epithelial tissues from patients with CD and embryonic fibroblasts from mice, along with enteroids and human IEC lines, we found that induction of cell stress alters the cytoskeleton in IECs, via changes in the actin-binding proteins VIL1 and GSN. Acute changes in actin dynamics increase IEC survival, whereas long-term changes in actin dynamics lead to IEC death and intestinal inflammation. IRGM regulates necroptosis and release of DAMPs to induce gastrointestinal inflammation, linking IRGM activity with CD.
Galectin-3 Mediates Tumor Cell–Stroma Interactions by Activating Pancreatic Stellate Cells to Produce Cytokines via Integrin Signaling Gastroenterology (IF 18.392) Pub Date : 2017-12-21 Wei Zhao, Jaffer A. Ajani, Guha Sushovan, N. Ochi, Rosa Hwang, Margarete Hafley, Randy L. Johnson, Robert S. Bresalier, Craig D. Logsdon, Zhiqian Zhang, Shumei Song
Background & Aims Pancreatic ductal adenocarcinoma (PDAC) is characterized by activated pancreatic stellate cells (PSCs), abundance of extracellular matrix (ECM), and production of cytokines and chemokines. Galectin 3 (GAL3), a β galactoside-specific lectin, contributes to PDAC development but its effects on the stroma and cytokine production are unclear. Methods The effect of recombinant human GAL3 (rGAL3) on activation of PSC, production of cytokines, and ECM proteins were determined by proliferation, invasion, cytokine array, and quantitative PCR. We assessed co-cultures of PDAC cells with GAL3 genetic alterations with PSCs. Production of interleukin 8 (IL8) and activities of nuclear factor (NF)-κB were determined by ELISA and luciferase reporter analyses. We studied the effects of inhibitors of NF-κB and integrin linked kinase (ILK) on pathways activated by rGAL3. Results In analyses of the GEO database and our dataset, we observed higher levels of GAL3, IL8, and other cytokines in PDAC than in non-tumor tissues. Production of IL8, GMCSF, CXCL1, and CCL2 increased in PSCs exposed to rGAL3 compared to control. Culture of PSCs with PDAC cells that express different levels of GAL3 resulted in proliferation and invasion of PSCs that increased with level of GAL3. GAL3 stimulated transcription of IL8 through integrin subunit beta 1 (ITGB1) on PSCs, which activates NF-κB through ILK. Inhibitors of ILK or NF-κB or a neutralizing antibody against ITGB1 blocked transcription and production of IL8 from PSCs induced by rGAL3. The GAL3 inhibitor significantly reduced growth and metastases of orthotopic tumors that formed from PDAC and PSC cells co-implanted in mice. Conclusion GAL3 activates PSC cells to produce inflammatory cytokines via ITGB1signaling to ILK and activation of NF-κB. Inhibition of this pathway reduced growth and metastases of pancreatic orthotopic tumors in mice.
Knockdown of Anillin Actin Binding Protein Blocks Cytokinesis in Hepatocytes and Reduces Liver Tumor Development in Mice without Affecting Regeneration Gastroenterology (IF 18.392) Pub Date : 2017-12-21 Shuyuan Zhang, Liem H. Nguyen, Kejin Zhou, Ho-Chou Tu, Alfica Sehgal, Ibrahim Nassour, Lin Li, Purva Gopal, Joshua Goodman, Amit G. Singal, Adam Yopp, Yu Zhang, Daniel J. Siegwart, Hao Zhu
Background & Aims Cytokinesis can fail during normal post-natal liver development, leading to polyploid hepatocytes. We investigated whether inhibiting cytokinesis in the liver; slows tumor growth without compromising the health of normal hepatocytes. We inhibited cytokinesis in cancer cells by knocking down anillin actin binding protein (ANLN), a cytoskeletal scaffolding protein that regulates cytokinesis and might promote tumorigenesis, in mice with liver disease. Methods We analyzed clinical and gene expression data from The Cancer Genome Atlas, Oncomine, PrognoScan, and a hepatocellular carcinoma (HCC) tissue microarray. We; knocked down ANLN with small interfering RNAs (siRNAs) in H2.35 liver cells and performed; image analyses of cells undergoing cytokinesis. siRNAs were delivered to LAP-MYC mice, which develop hepatoblastoma, using lipid nanoparticles. H2.35 cells with knockdown of ANLN or control cells were injected into FRG mice, which develop chronic liver damage, and tumor growth was monitored. We also developed mice with inducible expression of; transgenes encoding small hairpin RNAs (shRNAs) against Anln mRNA and studied liver; tumorigenesis following administration of diethylnitrosamine and carbon tetrachloride. siRNAs against Anln mRNA were conjugated to N-acetylgalactosamine to reduce toxicity and; increase hepatocyte tropism; their effects were studied in mouse models of liver cancer; and regeneration. Results Levels of ANLN mRNA were increased in human HCC tissues compared to non-tumor liver tissues. siRNA knockdown of ANLN blocked cytokinesis in H2.35 liver cells. Administration of siRNA against ANLN increased survival times of LAP-MYC mice, compared to mice given a control siRNA. H2.35 liver cells with shRNA knockdown of ANLN formed tumors more slowly in FRG mice than control H2.35 cells. Mice with inducible expression of shRNAs against Anln mRNA developed fewer liver tumors after administration of diethylnitrosamine and carbon tetrachloride than control mice. Knockdown of ANLN did not affect liver regeneration after acute and chronic liver injuries. Conclusions Knockdown of ANLN in liver cells blocks cytokinesis and inhibits development of liver tumors in mice. Agents that inhibit ANLN in the liver might be effective for prevention; or treatment of HCC.
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