scholarly journals HDAC6/HNF4α loop mediated by miR-1 promotes bile acids-induced gastric intestinal metaplasia

2020 ◽  
Vol 24 (1) ◽  
pp. 103-116
Author(s):  
Na Wang ◽  
Min Chen ◽  
Zhen Ni ◽  
Ting Li ◽  
Jiaoxia Zeng ◽  
...  
Keyword(s):  
Mouse Model ◽  
Intestinal Metaplasia ◽  
Bile Acids ◽  
Transgenic Mouse ◽  
Cell Model ◽  
Critical Role ◽  
Precancerous Lesion ◽  
Transgenic Mouse Model ◽  
Luciferase Reporter ◽  
Gastric Intestinal Metaplasia

Abstract Background Gastric intestinal metaplasia (IM) is considered a precancerous lesion, and bile acids (BA) play a critical role in the induction of IM. Ectopic expression of HNF4α was observed in a BA-induced IM cell model. However, the mechanisms underlying the upregulation of the protein in IM cells remains to be elucidated. Methods The effects of HNF4α on gastric mucosal cells in vivo were identified by a transgenic mouse model and RNA-seq was used to screen downstream targets of deoxycholic acid (DCA). The expression of pivotal molecules and miR-1 was detected by immunohistochemistry and in situ hybridization in normal, gastritis and IM tissue slides or microarrays. The transcriptional regulation of HDAC6 was investigated by chromatin immunoprecipitation (ChIP) and luciferase reporter assays. Results The transgenic mouse model validated that HNF4α stimulated the HDAC6 expression and mucin secretion in gastric mucosa. Increased HDAC6 and HNF4α expression was also detected in the gastric IM cell model and patient specimens. HNF4α could bind to and activate HDAC6 promoter. In turn, HDAC6 enhanced the HNF4α protein level in GES-1 cells. Furthermore, miR-1 suppressed the expression of downstream intestinal markers by targeting HDAC6 and HNF4α. Conclusions Our findings show that the HDAC6/HNF4α loop regulated by miR-1 plays a critical role in gastric IM. Blocking the activation of this loop could be a potential approach to preventing BA-induced gastric IM or even gastric cancer (GC).

scholarly journals HDAC6/FOXP3/HNF4α Axis Promotes Bile Acids Induced Gastric Intestinal Metaplasia

2021 ◽  
Author(s):  
Na Wang ◽  
Siran Wu ◽  
Luyao Zhang ◽  
Min Chen ◽  
Jiaoxia Zeng ◽  
...  
Keyword(s):  
Transgenic Mice ◽  
Intestinal Metaplasia ◽  
Bile Acids ◽  
Epigenetic Modification ◽  
Bile Reflux ◽  
Precancerous Lesion ◽  
Ectopic Expression ◽  
Histone Deacetylation ◽  
Luciferase Reporter ◽  
Luciferase Reporter Gene

Abstract Background: Gastric intestinal metaplasia (IM) is an important precancerous lesion. Our previous study has shown that ectopic expression of HDAC6 promotes the activation of intestinal markers in bile acids (BA) induced gastric IM cells; however, the mechanism underlying how HDAC6-mediated epigenetic modifications regulate intestinal markers is not clear.Methods: RNA-sequencing (RNA-seq) was used to detect the molecular changes in GES-1 cells after HDAC6 overexpression. The potential binding sites of FOXP3 with the promoter region of HNF4α were verified by ChIP and luciferase reporter gene assays. The ChIP assay was also used to detect the histone deacetylation. The levels of mucin in gastric or intestinal mucosa were detected by AB-PAS staining. Transgenic mice were used to explore the pro-metaplastic function of DCA and HNF4α in vivo.Results: Deoxycholic acid (DCA) upregulated HDAC6 in gastric cells, which further inhibited the transcription of FOXP3. Then, FOXP3 transcriptionally inhibited HNF4α, which further inhibits the expression of downstream intestinal markers. These molecules have been shown to be clinically relevant, as FOXP3 levels were negatively correlated with HDAC6 and HNF4α in IM tissues. Transgenic mice experiments confirmed that HNF4α overexpression combined with DCA induced gastric mucosa to secrete intestinal mucus and caused an abnormal mucosal structure. Conclusions: Our findings suggest that HDAC6 reduces FOXP3 through epigenetic modification, thus forming HDAC6/FOXP3/HNF4α axis to promote gastric IM. Inhibition of HDAC6 may be a potential approach to prevent gastric IM in patients with bile reflux.

scholarly journals Specific overexpression of IL-7 in the intestinal mucosa: the role in intestinal intraepithelial lymphocyte development

2008 ◽  
Vol 294 (6) ◽  
pp. G1421-G1430 ◽  
Author(s):  
Hua Yang ◽  
Blair Madison ◽  
Deborah L. Gumucio ◽  
Daniel H. Teitelbaum
Keyword(s):  
T Cell ◽  
Mouse Model ◽  
Transgenic Mice ◽  
Transgenic Mouse ◽  
Critical Role ◽  
Transgenic Mouse Model ◽  
Intraepithelial Lymphocytes ◽  
Specific Expression ◽  
And Function

IL-7 plays a crucial role in controlling T cell development and homeostasis. Since IL-7 may be derived from extraintestinal sources, and exogenous IL-7 broadly affects lymphoid populations, the actions of epithelial cell (EC)-derived IL-7 are not fully understood. The effect of intestinal specific expression of IL-7 on intestinal mucosal lymphocytes was investigated by using an IL-7 transgenic mouse model. We generated an intestinal EC-specific overexpressing IL-7 transgenic mouse model (IL-7vill) and compared their phenotype and function to wild-type C57BL/6J mice. EC-derived IL-7 overexpression was found to be exclusively in the small and large intestine. Numbers and subtypes of mucosal lymphocytes, including intraepithelial lymphocytes (IEL) and lamina propria lymphocytes (LPL), significantly changed in IL-7vill mice. From a functional standpoint, IEL proliferation also significantly increased in IL-7vill mice. IEL cytokine expression significantly changed in both T cell receptor (TCR)-αβ+ and TCR-γδ+ IEL subpopulations, including a significant increase in IFN-γ and TNF-α as well as an increase in keratinocyte growth factor expression. EC expression of CD103 (integrin αEβ7), the ligand of E-cadherin, markedly upregulated and may account for a mechanism of the massive expansion of IEL in transgenic mice. Systemic lymphoid populations did not change in transgenic mice. IL-7 overexpression by intestinal EC significantly affected IEL phenotype and function. These results offer insight into the role of IL-7 in IEL development and suggest a critical role of EC-derived expression of IL-7 in the phenotype and function of IEL.

scholarly journals The CRE luc Bioluminescence Transgenic Mouse Model for Detecting Ligand Activation of GPCRs

2013 ◽  
Vol 19 (2) ◽  
pp. 232-241 ◽  
Author(s):  
Holly Dressler ◽  
Kyriakos Economides ◽  
Sarah Favara ◽  
Nancy N. Wu ◽  
Zhen Pang ◽  
...  
Keyword(s):  
Mouse Model ◽  
Transgenic Mouse ◽  
Expression Profiles ◽  
Tissue Expression ◽  
Transgenic Mouse Model ◽  
Luciferase Reporter ◽  
Primary Cells ◽  
Compound Libraries ◽  
Cellular Environment

Numerous assays have been developed to investigate the interactions between G-protein–coupled receptors (GPCRs) and their ligands since GPCRs are key therapeutic targets. Reporter-based assays using the cAMP response element (CRE) coupled with bioluminescence from a luciferase reporter have been used extensively in vitro with high-throughput screens (HTS) of large chemical compound libraries. We have generated a transgenic mouse model (CRE luc) with a luciferase reporter under the control of a synthetic promoter that contains several CREs, which supports real-time bioimaging of GPCR ligand activity in whole animals, tissues, or primary cells. In the CRE luc model, GPCR signaling through the cAMP pathway can be detected from the target GPCR that is in a native cellular environment with a full complement of associated receptors and membrane constituents. Multiple independent lines have been produced by random integration of the transgene, resulting in tissue expression profiles covering the major organs. The goal of the CRE luc model is to accelerate the transition from HTS to profiling of GPCR small-molecule leads in preclinical animal disease models, as well as define the mechanism of action of GPCR drugs in three experimental formats: primary cells, tissue homogenates, and whole animals.

scholarly journals HDAC6/FOXP3/HNF4α axis promotes bile acids induced gastric intestinal metaplasia

2021 ◽  
Author(s):  
Na Wang ◽  
Siran Wu ◽  
Luyao Zhang ◽  
Min Chen ◽  
Jiaoxia Zeng ◽  
...  
Keyword(s):  
Transgenic Mice ◽  
Intestinal Metaplasia ◽  
Bile Acids ◽  
Epigenetic Modification ◽  
Bile Reflux ◽  
Precancerous Lesion ◽  
Ectopic Expression ◽  
Histone Deacetylation ◽  
Luciferase Reporter ◽  
Luciferase Reporter Gene

Abstract Background: Gastric intestinal metaplasia (IM) is an important precancerous lesion. Our previous study has shown that ectopic expression of HDAC6 promotes the activation of intestinal markers in bile acids (BA) induced gastric IM cells; however, the mechanism underlying how HDAC6-mediated epigenetic modifications regulate intestinal markers is not clear.Methods: RNA-sequencing (RNA-seq) was used to detect the molecular changes in GES-1 cells after HDAC6 overexpression. The potential binding sites of FOXP3 with the promoter region of HNF4α were verified by ChIP and luciferase reporter gene assays. The ChIP assay was also used to detect the histone deacetylation. The levels of mucin in gastric or intestinal mucosa were detected by AB-PAS staining. Transgenic mice were used to explore the pro-metaplastic function of DCA and HNF4α in vivo.Results: Deoxycholic acid (DCA) upregulated HDAC6 in gastric cells, which further inhibited the transcription of FOXP3. Then, FOXP3 transcriptionally inhibited HNF4α, which further inhibits the expression of downstream intestinal markers. These molecules have been shown to be clinically relevant, as FOXP3 levels were negatively correlated with HDAC6 and HNF4α in IM tissues. Transgenic mice experiments confirmed that HNF4α overexpression combined with DCA induced gastric mucosa to secrete intestinal mucus and caused an abnormal mucosal structure. Conclusions: Our findings suggest that HDAC6 reduces FOXP3 through epigenetic modification, thus forming HDAC6/FOXP3/HNF4α axis to promote gastric IM. Inhibition of HDAC6 may be a potential approach to prevent gastric IM in patients with bile reflux.

scholarly journals In Vivo Optical Imaging of Myelination Events in a Myelin Basic Protein Promoter-Driven Luciferase Transgenic Mouse Model

ASN NEURO ◽  
2018 ◽  
Vol 10 ◽  
pp. 175909141877732 ◽  
Author(s):  
James Cao ◽  
Yanping Hu ◽  
Mohammed Salman Shazeeb ◽  
Carlos E. Pedraza ◽  
Nilesh Pande ◽  
...  
Keyword(s):  
Mouse Model ◽  
Myelin Basic Protein ◽  
Real Time ◽  
Transgenic Mouse ◽  
Basic Protein ◽  
Neuronal Cell ◽  
Transgenic Mouse Model ◽  
Luciferase Reporter ◽  
Transcriptional Level

The compact myelin sheath is important for axonal function, and its loss can lead to neuronal cell death and irreversible functional deficits. Myelin is vulnerable to a variety of metabolic, toxic, and autoimmune insults. In diseases like multiple sclerosis, there is currently no therapy to stop myelin loss, underscoring the need for neuroprotective and remyelinating therapies. Noninvasive, robust techniques are also needed to confirm the effect of such therapies in animal models. This article describes the generation, characterization, and potential uses for a myelin basic protein-luciferase (MBP-luci) transgenic mouse model, in which the firefly luciferase reporter gene is selectively controlled by the MBP promoter. In vivo bioluminescence imaging can be used to visualize and quantify demyelination and remyelination at the transcriptional level, noninvasively, and in real time. Transgenic mice were assessed in the cuprizone-induced model of demyelination, and luciferase activity highly correlated with demyelination and remyelination events as confirmed by both magnetic resonance imaging and postmortem histological analysis. Furthermore, MBP-luci mice demonstrated enhanced luciferase signal and remyelination in the cuprizone model after treatment with a peroxisome proliferator activated receptor-delta selective agonist and quetiapine. Imaging sensitivity was further enhanced by using CycLuc 1, a luciferase substrate, which has greater blood–brain barrier penetration. We demonstrated the utility of MBP-luci model in tracking myelin changes in real time and supporting target and therapeutic validation efforts.

A TRANSGENIC MOUSE MODEL WITH A LUCIFERASE REPORTER FOR STUDYING IN VIVO TRANSCRIPTIONAL REGULATION OF THE HUMAN CYP3A4 GENE

2003 ◽  
Vol 31 (8) ◽  
pp. 1054-1064 ◽  
Author(s):  
Weisheng Zhang ◽  
Anthony F. Purchio ◽  
Kevin Chen ◽  
Jianming Wu ◽  
Li Lu ◽  
...  
Keyword(s):  
Transcriptional Regulation ◽  
Mouse Model ◽  
Transgenic Mouse ◽  
Transgenic Mouse Model ◽  
Luciferase Reporter ◽  
Cyp3a4 Gene
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