Texas Medical Center Digestive Diseases Center

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Texas Medical Center Digestive Diseases Center

Transcript Of Texas Medical Center Digestive Diseases Center

Texas Medical Center Digestive Diseases Center
4th Annual “Frontiers in Digestive Diseases”: Stem Cells in GI Health & Disease
Saturday, January 12, 2013 The Marriot Medical Center Hotel
Houston, Texas Table of Contents................................................................................................................................2 Agenda................................................................................................................................................ 3 List of Abstracts .............................................................................................................................4 - 5 Abstracts ......................................................................................................................................6 - 36 List of Participants......................................................................................................................37 - 40 Acknowledgements ........................................................................................................................... 41

On the cover: 1 2



(1) P-10. Kahalil Ettayebi. Villin expression in (A) induced human intestinal organoids vs. (B) human jejunal intestinal enteroids. Detection of villin expression (green) on brush border membrane of apical absorptive cells; E-cadherin (red); Nuclear staining (blue).
(2) P-29. Xuemei Shi. GLP-2 augments cell proliferation in the mini-gut after 5-fluorouracil treatment. Organoids (cultured from the mouse jejunal crypts) were treated with glucagon-like peptide 2 (GLP-2) and then 5-fluorouracil. Cell proliferation was estimated by BrdU incorporation (in green). The nucleus was counterstained with TOPRO-3 (in blue).
(3) P-30. Zhongcheng Shi. Colocalization of SOX9 and IGFBP-4 in the intestinal epithelium is visualized by immunofluorescent staining. This supports our in vitro data that igfbp4 is directly regulated by SOX9 (and that SOX9 suppresses cellular proliferation through IGFBP-4).
(4) P-34. Petri Urvil. Bipartite network showing metabolite cluster associations in patients with Clostridium difficile infection. The red metabolite cluster is associated with patients susceptible to disease recurrence and represent potential diagnostic biomarkers.

Texas Medical Center Digestive Diseases Center
4th Annual “Frontiers in Digestive Diseases”: Stem Cells in GI Health & Disease
A G E N D A Saturday, January 12, 2013 The Marriot Medical Center Hotel
Houston, Texas


Coffee and Continental Breakfast

Session I. Theme: 8:15- 8:45am Digestive 8:45 – 9:25am
9:25 – 10:05am
10:05 – 10:35am

GI Stem Cells
Welcome. Moderator: Mary K. Estes, PhD, Director, Texas Medical Center Diseases Center; Professor, Baylor College of Medicine (BCM).
"Intestinal stem cells during regeneration and homeostasis" Calvin Kuo, MD, PhD, Associate Professor, Medicine, Stanford University School of Medicine
“Human intestinal organoids from pluripotent stem cells” Noah Shroyer, PhD, Assistant Professor, Pediatrics, Cincinnati Children’s Hospital Medical Center.
“Function & mechanism of LGR4/5 in GI stem cells and carcinogenesis” Qingyun (Jim) Liu, PhD, Professor, Texas Therapeutics Institute, UTHealth

10:35 – 11:00am Morning Coffee Break

Session II. Theme:

Liver Stem Cells & Therapeutics Moderator: Tor Savidge, PhD, Associate Professor, Pathology, BCM

11:00 – 11:40am

“Developmental morphogens, progenitors and adult liver repair” Anna Mae Diehl, MD, Professor, Medicine; Chief, Division of Gastroenterology, Duke University Medical School

11:40 – 12:10pm

“Exploring human cell therapy for metabolic liver disease” Karl-Dimiter Bissig, MD, PhD, Assistant Professor, Center for Cell and Gene Therapy, Stem Cells and Regenerative Medicine, BCM

12:10 – 1:45pm Lunch & Poster Viewing/Judging

1:45 – 2:15pm 2:15pm

"Transplantation of human stem cells for translational ID studies" Gustavo Valbuena, MD, PhD, Assistant Professor, Pathology, University of Texas Medical Branch, Galveston TX.
Discussion and Closing Remarks


Texas Medical Center Digestive Diseases Center
4th Annual “Frontiers in Digestive Diseases”: Stem Cells in GI Health & Disease


Page Name


6 Yanna Cao, MD

Human mesenchymal stem cells attenuate

Assistant Professor

pancreatic acinar injury in acute pancreatitis in mice

Surgery, UTHealth

7 Chi-I Chiang, M.S.

Identification of target genes in colorectal cancer

Research Assistant

cells directly regulated by the transcription factor



8 Sue Crawford

Autophagy hijacked through viroporin-activated

Research Associate

CaMKK-β signaling is required for rotavirus

Molecular Virology & Microbiology, BCM


9 Moreshwar Desai MD

Bile acids induce myocardial dysfunction: candidate

Assistant Professor, Pediatrics-Critical Care

mechanism for cirrhotic cardiomyopathy


10 Khalil Ettayebi

Human jejunal enteroid cultures as a functional

Sr Staff Scientist, Molecular Virology & Microbiology, model of human small intestine to study infection


with human enteric microbes

11 Peera Hemarajata

Putative histidine decarboxylase regulator (PhdR)

Research Assistant (GS)

modulates histamine production and

Pathology, BCM

immunomodulation by Lactobacillus reuteri

12 Anne Hutson, PhD

Increased miR-143 and decreased miR-31

Assistant Professor

expression in Apc(min/+) mouse adenomas and


human FAP polyps


13 Joseph M. Hyser, PhD

Multiple calcium entry pathways are activated by the

Assistant Professor

rotavirus viroporin NSP4

Molecular Virology and Microbiology, BCM

14 Li Jiao, MD, PhD

Instrumental variable analysis on soluble receptor

Assistant Professor

for advanced glycation end-products and risk of

Medicine - GI and Hepatology, BCM

pancreatic cancer - a pilot study

15 Claudia Kettlun-Leyton, PhD

Autologous cell therapy for tyrosinemia type 1:

Postdoctoral Associate

exploring human cell therapy in a mouse

Center for Gene Therapy, BCM

16 Daniel J. Kota, PhD

Investigating the immune modulatory properties of

Postdoctoral Fellow

human Mesenchymal Stem/Stromal cells

Pediatric Surgery, UTHealth

17 Daniel H. Leung, MD

High prevalence of HBV non-immunity in vaccinated

Assistant Professor

pediatric liver transplant recipients

Pediatrics - GI, Hepatology, and Nutrition


18 Yuying Liu, PhD

Lactobacillus reuteri DSM 17938 inversely changes

Assistant Professor

the effector and regulatory T cell population in a

Pediatrics - Gastroenterology

mouse model of necrotizing enterocolitis


19 Janielle P Maynard

Purinergic signaling in hepatocellular carcinoma

Research Assistant, Translational Biology and

Molecular Medicine BCM

20 Amber Miller

Lymphotoxin signaling is important for rotavirus

Research Assistant (GS)

specific IgA induction

Molecular Virology and Microbiology, BCM


Texas Medical Center Digestive Diseases Center
4th Annual “Frontiers in Digestive Diseases”: Stem Cells in GI Health & Disease

21 Dorottya Nagy-Szakal, MD Postdoctoral Associate Pediatric – Gastroenterology, BCM
22 Numan Oezguen, PhD Instructor, Pathology & Immunology BCM
23 Luis Ojeda, PhD Postdoc Fellow Center for Cell and Gene Therapy, BCM
24 Narayan Sastri Palla, MS Postdoctoral Associate Molecular Virology and Microbiology, BCM
25 Shujuan Pan, PhD Assistant Professor Pathology, BCM
26 Karina Pokusaeva, PhD Postdoctoral Associate Pathology and Immunology, BCM
27 Muralidhar Premkumar, MBBS Assistant Professor Neonatology, BCM
28 Sasirekha Ramani, PhD Postdoctoral Associate Molecular Virology and Microbiology, BCM
29 Xuemei Shi,MD Research Associate Pediatrics-Nutrition, BCM
30 Zhongcheng Shi, MD Postdoctoral Associate Immunology and Pediatrics, BCM
31 Shumei Song, MD, PhD Assistant Professor Gastrointestinal Medical Oncology MDACC
32 Jennifer K. Spinler, PhD Instructor, Microbial Genetics & Genomics Texas Children's Microbiome Center, BCM
33 Bryan Tackett Graduate Student Texas Children's Liver Center, BCM
34 Petri Urvil Laboratory Director Pathology, BCM
35 Donna L. White, PhD, MPH Assistant Professor Medicine - GI and Hepatology, BCM
36 Lisa D. White, PhD Associate Professor Mol. & Human Gen
Denotes past Pilot/Feasibility awardee
Denotes 2012 Pilot/Feasibility awardee

DNA methylation and microbiome separation of ulcerative colitis in treatment naïve children
S-nitrosothiols: novel regulators of gut-microbial crosstalk
Generation of human iPSC by RNA reprogramming and novel 3D culture and differentiation of iPS cells into hepatocyte like cells
Biochemical and biophysical evidence of calcium binding to coiled coil domain of rotavirus NSP4
Roles of ERManI in liver disease associated with alpha1-antitrypsin deficiency
Production of gamma-aminobutyric acid by intestinal microbe bifidobacterium dentium and other commensal bacteria Loss of enterocyte derived argininosuccinate lyase results in increased incidence of necrotizing enterocolitis The VP8* of neonatal rotavirus strain G10P[11] binds to type II precursor glycans
GLP-2 receptor is required for the growth of intestinal crypts ex vivo
SOX9 directly activates IGFBP-4 and suppresses proliferation in Apcmin/+ adenomas and colorectal cancer cells
Loss of TGF- adaptor 2SP activates notch
signaling and SOX9 in esophageal adenocarcinoma
Pangenomic analysis of Lactobacillus reuteri highlights the evolution of a human-specific ecotype
P2Y2 purinergic receptor signaling plays a key role in diet-Induced obesity in mice
A multiomics approach to understanding the gutmicrobial ecology and mechanisms of recurrent Clostridium difficile infection Comparative evaluation of bioimpedance and anthropometric measurements as predictors of HCV-related fibrosis and inflammation risk Whole Transcriptome RNA Sequencing: From Library Preparation to Sequencing


Texas Medical Center Digestive Diseases Center
4th Annual “Frontiers in Digestive Diseases”: Stem Cells in GI Health & Disease
Human Mesenchymal Stem Cells Attenuate Pancreatic Acinar Injury in Acute Pancreatitis in Mice
Michael Gonzales1, Xuxia Gao1, Yanna Cao1,2, Hasen Xue1, Judith F. Aronson3, Shibani Pati1, Tien C. Ko1,2
1Department of Surgery, The University of Texas Health Science Center at Houston, Texas; 2Department of Surgery and 3Department of Pathology, The University of Texas Medical Branch at
Galveston, Texas
Introduction. Human mesenchymal stem cells (hMSCs) can modulate the host immune response against various diseases in animal models and in human with improved survival rate and organ function. Recently, the study on the rat acute pancreatitis models has shown a beneficial effect towards controlling pancreas organ damage. However, the underlying mechanisms of hMSCs’ effects are unclear. Therefore, the objective of this study is to use a mouse acute pancreatitis (AP) model to evaluate hMSCs’ effects for further investigation of the underlying mechanisms.
Experimental Design and Methods. In the mouse AP model, C57BL/6 mice (male, 8 wks old) were divided into 3 groups: control (PBS only, n=2), AP (cerulein and PBS, n=6), AP+hMSCs (cerulein and hMSCs, n=6). Cerulein was given via 12 hourly intraperitoneal injections (50 µg/kg), while the same amount and frequency of PBS was injected for control. The hMSC (1 million/0.1 ml, obtained from Lonza Inc.) or PBS was injected via tail vein at 24 and 48 hours following the first cerulein injection. The mice were euthanized 4 days after the first cerulein injections. The pancreas was harvested for H&E staining and histological assessment on edema, necrosis, inflammation, acinar injury (including distribution of lesions, glandular atrophy, pseudotubular complexes) and fibrosis.
Results. In respective to the groups of control, AP, AP+hMSCs, there are no significant differences in edema (1.25, 1.7, 2.3, p=0.14), necrosis (0, 0.3, 0.7, p=0.6), inflammation (0.25, 1.0, 0.9, p=0.06), and fibrosis scores (0, 1.7, 1.0, p=0.12). However, there are significant differences in combined acinar injury scores (0, 7.3, 3.2, p<0.001) including distribution of lesions (0, 3.2, 1.5, p<0.001), glandular atrophy (0, 2.0, 1.0, p=0.002), and pseudotubular complexes (0, 2.2, 0.7, p=0.001).
Conclusions. hMSCs treatment after AP induction showed no effect on edema, necrosis, inflammation, and fibrosis, but significantly reduced acinar injury shown by the decreased acinar injury scores. Our data demonstrate that hMSC treatment attenuates AP injury to pancreatic acini in a mouse AP model. The protective effects of hMSCs in AP may function through a reduced acinar injury or an accelerated acinar regeneration after AP induction. Future experimentation is required for the underlying mechanisms of hMSCs’ effects, which may ultimately lead to the development of therapeutic strategies using hMSCs in AP and in other inflammatory diseases.

Texas Medical Center Digestive Diseases Center
4th Annual “Frontiers in Digestive Diseases”: Stem Cells in GI Health & Disease
Identification of target genes in colorectal cancer cells directly regulated by the transcription factor SOX9
Chi-I Chiang, Zhongcheng Shi, Toni-Ann Mistretta, Angela Major, and Yuko Mori-Akiyama Department of Pathology and Immunology, Baylor College of Medicine and Texas Children’s Hospital
BACKGROUND & AIMS: The transcription factor SOX9 is expressed in diverse cancers including most human primary colorectal cancer (CRC). The role of SOX9 in cancers has been a focus of many recent studies and there have been reports of both oncogenic and suppressive roles of SOX9 in various cancers, including breast, prostate, ovarian cancers, and melanomas. In CRC, some reports suggested an anti-oncogenic role for SOX9, while others demonstrated an oncogenic role. A recent genome-scale analysis of human CRC identified SOX9 as one of the frequently mutated genes and the mutations of SOX9 were either frame shift or nonsense mutations. Nevertheless, role of SOX9 in CRC has not been well studied and to date only a few direct targets of SOX9 in intestinal epithelial cells have been identified. In this study, we identified target genes of SOX9 and in vivo regulations of these genes by SOX9 were confirmed using two mouse models. METHODS: Mice that ectopically express SOX9 throughout in the intestinal epithelium were used for identifying RNA profile. Chromatin immunoprecipitation (ChIP)-based assays that involve genome-wide mapping validated the direct binding of SOX9 on the potential target genes. The regulation of these genes by SOX9 was further confirmed in adenoma cells of Sox9-deficient Apcmin mice. RESULTS: Combined analysis of RNA profiling by microarray using SOX9-overexpressing mouse intestine and genome-wide ChIP sequencing using human colorectal cancer cells revealed direct target genes of SOX9 that includeed anti-oncogenic as well as oncogenic genes. Our data suggested that SOX9 may potentially support cancer stem cells, however, SOX9 may suppress proliferation and progression.

Texas Medical Center Digestive Diseases Center
4th Annual “Frontiers in Digestive Diseases”: Stem Cells in GI Health & Disease
Autophagy hijacked through viroporin-activated CaMKK-β signaling is required for rotavirus replication
Sue E. Crawford, Joseph M. Hyser, Budi Utama, and Mary K. Estes Baylor College of Medicine
Autophagy is a cellular degradation process involving an intracellular membrane trafficking pathway that recycles cellular components or eliminates intracellular microbes in lysosomes. Many pathogens subvert autophagy to enhance their replication but the mechanisms these pathogens use to initiate the autophagy process have not been elucidated. This study identifies rotavirus (RV) as the first pathogen that uses its viroporin, NSP4, which releases calcium from the endoplasmic reticulum (ER) into the cytoplasm to activate a CaMKK-β and AMPK-dependent signaling pathway to initiate the autophagy process. RV hijacks this membrane trafficking pathway to transport viral proteins from the ER to sites of viral replication to produce infectious virus. This process requires PI3-kinase activity, autophagy-initiation proteins Atg3 and Atg5, and is abrogated by chelating cytoplasmic calcium or inhibiting CaMKK-β. These studies identify a novel mechanism of virusmediated calcium-activated signaling that initiates autophagy and hijacks this membrane trafficking pathway to transport viral proteins to sites of viral assembly.

Texas Medical Center Digestive Diseases Center
4th Annual “Frontiers in Digestive Diseases”: Stem Cells in GI Health & Disease

Bile acids induce myocardial dysfunction: candidate mechanism for cirrhotic cardiomyopathy

*Moreshwar Desai1, *Sayeepriyadarshini Anakk2, Zeena Eblimit1, Renan Orellana1, Hernan Vasquez3, Henry Cheng-Ju Wu3, Heinrich Taegtmeyer3, Daniel Penny4, Saul Karpen5, David D. Moore2

1Section of Pediatric Critical Care and Liver Center, 2Dept. Molecular and Cellular Biology, 4Dept. of Pediatric Cardiology, Baylor College of Medicine; 3 Dept. of Cardiology UTHSC; 5 Dept of Pediatric
Gastroenterology, Emory, Atlanta, GA. [*equal contribution].

Introduction: Cardiac dysfunction in cirrhosis - manifesting itself as cirrhotic cardiomyopathy, is a life threatening complication of end-stage cirrhotic liver diseases with a prevalence approaching 50% of cirrhotic adults and children. Hearts of cirrhotic patients show electrocardiographic abnormalities and systolic dysfunction under stress. The pathophysiology is poorly understood, impairing prevention and treatment. Bile acid (BA) excess (cholanemia) is a pathognomonic feature of cirrhosis. It drives pathological processes in several extra-hepatic organs, but the cardiac effects are unknown. Using an established mouse model of cholanemia, FXR-/-/SHP-/- (DKO) along with ex vivo BA perfusion of the whole heart, we tested the hypothesis that cholanemia impairs myocardial contractility, catecholamine response, metabolism and alters cell signaling pathways. Methods: 12 week old male DKO mice (n=6) and their age matched littermate (WT) controls underwent echocardiography (ECHO), electrocardiography (ECG), treadmill stress tests and acute catecholamine (isoprenaline) challenge with stress ECHO. Hearts were analyzed for protein expression of various signaling pathways. To evaluate the direct effects of BA excess on contractility and metabolism, whole hearts (n=5/grp) were challenged with Taurocholic acid (TCA) [500μmol for 20 minutes] or vehicle ex vivo using Langendorff system. Results: DKO mice showed evidence of liver injury, cholestasis and cholanemia (Anakk et al . J Clin Invest. 2011 Jan;121(1):86-95). DKO mice showed significant bradycardia and prolonged PR interval on ECG. ECHO revealed a 30% decrease in cardiac output [CO] compared to WT controls (Table). On challenge with isoprenaline (200mcg/kg i.p), heart rate, ejection fraction and CO was significantly attenuated in the DKO mice and the time taken for peak response was twice that of WT controls. On the treadmill, DKO mice showed exercise intolerance as evidenced by lesser distance covered and earlier fatigue (Table). DKO hearts showed a robust 3 fold increase in AKT phosphorylation and a modest 1.5 fold increase in JNK phosphorylation. At the RNA level, there was a 50% upregulation of GLUT-1 and 75% reduction of PDK4, suggesting increased glucose uptake and oxidation, and a 50% downregulation of UCP3, m-CPT1, mCPT-2 and h-FABP suggesting decreased fatty acid oxidation. A 50% decrease in contractility with concomitant decrease in glucose and fatty acid oxidation was demonstrated in TCA perfused hearts ex vivo. Conclusions: Cholanemia induces contractile dysfunction, catecholamine resistance, metabolic derangement and induction of stress mediated signaling in the heart. Further studies into myocardial remodeling and consequences of “pathologic” bile acid-myocardial interaction as chief driver for cirrhotic cardiomyopathy are warranted. [TCH startup funds (MD), CPRIT RP120138 (DDM)]

Heart Rate (bpm) PR interval (m-sec) Cardiac Output (ml/min) Peak Response (min) Distance on treadmill (m) Time to exhaustion (min)

WT (n=6) [Mean±SD]
750±25 22±2 24±3.5 20±10 425±50 18.5±2

DKO (n=6) [Mean±SD]
700±50 28±3 17±3.2 40±12 300±25 15±1.5

P value (t-test)
0.04 0.002 0.03 0.007 0.007 0.04


Texas Medical Center Digestive Diseases Center
4th Annual “Frontiers in Digestive Diseases”: Stem Cells in GI Health & Disease
Human Jejunal Enteroid Cultures as a Functional Model of Human Small Intestine to Study Infection with Human Enteric Microbes
Khalil Ettayebi1, Xi-Lei Zeng1, Sue E. Crawford1, Joseph M. Hyser1, James Broughman1, Antone R. Opekum2,3, David Y. Graham3, Vadim Sherman4, and Mary K. Estes1 (1) Department of Molecular Virology, Baylor College of Medicine (2) Department of Pediatrics, Baylor College of Medicine (3) Department of Medicine, Baylor College of Medicine (4) Bariatric surgery at Methodist Hospital
One rate-limiting step in translational research is the absence of reliable pre-clinical models that adequately reflect relevant human physiology and disease pathology. The goal of the current study was to establish human enteroid cultures and validate them as functional pre-clinical models of clinically important diarrheal models. Methods: We successfully cultured human enteroids from jejunal tissues obtained from patients undergoing bariatric surgery. Jejunal human intestinal enteroids (jHIEs) have been established from 11 different individuals, maintained in culture for 5 months, and can be frozen and recultured. The enteroids contain multiple cell types, including enterocytes, goblet cells, enteroendocrine cells, and Paneth cells. The epithelial cells, containing a number of ion channels, are organized around luminal cavities located within the core of the jHIEs. We tested the jHIEs as a new model for the cultivation and study of the gastrointestinal virus, rotavirus. Proof-of-principle analyses showed that jHIEs support replication of three rotaviruses, a lab strain RRV and two clinical isolates RV-35 and RV-08-42, based on detection of the nonstructural viral protein NSP4 by immunofluorescence, increased levels of viral RNA by quantitative reverse transcription-PCR (qRT-PCR), and production of infectious progeny virus. This work demonstrates that jHIEs offer a promising new model to study rotaviruses and they may be useful to study a variety of gastrointestinal-microbe interactions.
This study was supported by NIH grants (P30DK056338; PO1 AI057788, RO1 AI080656 and U18 NS080763).