THOC5, a member of the mRNA export complex, contributes to processing of a subset of wingless/integrated (Wnt) target mRNAs and integrity of the gut epithelial barrier
Abstract:BackgroundTHO (Suppressors of the transcriptional defects of hpr1 delta by overexpression) complex 5 (THOC5), an mRNA export protein, is involved in the expression of only 1% of all genes. Using an interferon inducible knockout mouse system, we have previously shown that THOC5 is an essential element in the maintenance of hematopoietic stem cells and cytokine-mediated hematopoiesis in adult mice. Here we interrogate THOC5 function in cell differentiation beyond the hematopoietic system and study pathological c… Show more
“…This upregulation was less evident after the treatment with antibiotic and may thus be a particular effect of the yeast. Conversely, a cluster of upregulated gene sets related to antioxidant activity matched with the antibiotic treatment, including particularly IGF-1, a key gene capable of reducing the induction of cell apoptosis while maintaining cell growth under oxidative stress (Baregamian et al, 2006), and THOC5 which reduces bacteria translocation and is a major regulator of homeostatic self-renewal within the intestinal crypt (Saran et al, 2013). Moreover, the CM diet was able to reduce the ETEC effect, increasing the expression of PPAR-γ, a gene widely recognized for its anti-inflammatory effect (Ortuño Sahagún et al, 2012) and confirming in vitro data (Zanello et al, 2011).…”
The ability of live yeasts to modulate pig intestinal cell signals in response to infection with Escherichia coli F4ac (ETEC) has not been studied in-depth. The aim of this trial was to evaluate the effect of Saccharomyces cerevisiae CNCM I-4407 (Sc), supplied at different times, on the transcriptome profile of the jejunal mucosa of pigs 24 h after infection with ETEC. In total, 20 piglets selected to be ETEC-susceptible were weaned at 24 days of age (day 0) and allotted by litter to one of following groups: control (CO), CO + colistin (AB), CO + 5 × 10 10 colony-forming unit (CFU) Sc/kg feed, from day 0 (PR) and CO + 5 × 10 10 CFU Sc/kg feed from day 7 (CM). On day 7, the pigs were orally challenged with ETEC and were slaughtered 24 h later after blood sampling for haptoglobin (Hp) and C-reactive protein (CRP) determination. The jejunal mucosa was sampled (1) for morphometry; (2) for quantification of proliferation, apoptosis and zonula occludens (ZO-1); (3) to carry out the microarray analysis. A functional analysis was carried out using Gene Set Enrichment Analysis. The normalized enrichment score (NES) was calculated for each gene set, and statistical significance was defined when the False Discovery Rate % was <25 and P-values of NES were <0.05. The blood concentration of CRP and Hp, and the score for ZO-1 integrity on the jejunal villi did not differ between groups. The intestinal crypts were deeper in the AB (P = 0.05) and the yeast groups (P < 0.05) than in the CO group. Antibiotic treatment increased the number of mitotic cells in intestinal villi as compared with the control group (P < 0.05). The PR group tended to increase the mitotic cells in villi and crypts and tended to reduce the cells in apoptosis as compared with the CM group. The transcriptome profiles of the AB and PR groups were similar. In both groups, the gene sets involved in mitosis and in mitochondria development ranked the highest, whereas in the CO group, the gene sets related to cell junction and anion channels were affected. In the CM group, the gene sets linked to the metabolic process, and transcription ranked the highest; a gene set linked with a negative effect on growth was also affected. In conclusion, the constant supplementation in the feed with the strain of yeast tested was effective in counteracting the detrimental effect of ETEC infection in susceptible pigs limits the early activation of the gene sets related to the impairment of the jejunal mucosa.
“…This upregulation was less evident after the treatment with antibiotic and may thus be a particular effect of the yeast. Conversely, a cluster of upregulated gene sets related to antioxidant activity matched with the antibiotic treatment, including particularly IGF-1, a key gene capable of reducing the induction of cell apoptosis while maintaining cell growth under oxidative stress (Baregamian et al, 2006), and THOC5 which reduces bacteria translocation and is a major regulator of homeostatic self-renewal within the intestinal crypt (Saran et al, 2013). Moreover, the CM diet was able to reduce the ETEC effect, increasing the expression of PPAR-γ, a gene widely recognized for its anti-inflammatory effect (Ortuño Sahagún et al, 2012) and confirming in vitro data (Zanello et al, 2011).…”
The ability of live yeasts to modulate pig intestinal cell signals in response to infection with Escherichia coli F4ac (ETEC) has not been studied in-depth. The aim of this trial was to evaluate the effect of Saccharomyces cerevisiae CNCM I-4407 (Sc), supplied at different times, on the transcriptome profile of the jejunal mucosa of pigs 24 h after infection with ETEC. In total, 20 piglets selected to be ETEC-susceptible were weaned at 24 days of age (day 0) and allotted by litter to one of following groups: control (CO), CO + colistin (AB), CO + 5 × 10 10 colony-forming unit (CFU) Sc/kg feed, from day 0 (PR) and CO + 5 × 10 10 CFU Sc/kg feed from day 7 (CM). On day 7, the pigs were orally challenged with ETEC and were slaughtered 24 h later after blood sampling for haptoglobin (Hp) and C-reactive protein (CRP) determination. The jejunal mucosa was sampled (1) for morphometry; (2) for quantification of proliferation, apoptosis and zonula occludens (ZO-1); (3) to carry out the microarray analysis. A functional analysis was carried out using Gene Set Enrichment Analysis. The normalized enrichment score (NES) was calculated for each gene set, and statistical significance was defined when the False Discovery Rate % was <25 and P-values of NES were <0.05. The blood concentration of CRP and Hp, and the score for ZO-1 integrity on the jejunal villi did not differ between groups. The intestinal crypts were deeper in the AB (P = 0.05) and the yeast groups (P < 0.05) than in the CO group. Antibiotic treatment increased the number of mitotic cells in intestinal villi as compared with the control group (P < 0.05). The PR group tended to increase the mitotic cells in villi and crypts and tended to reduce the cells in apoptosis as compared with the CM group. The transcriptome profiles of the AB and PR groups were similar. In both groups, the gene sets involved in mitosis and in mitochondria development ranked the highest, whereas in the CO group, the gene sets related to cell junction and anion channels were affected. In the CM group, the gene sets linked to the metabolic process, and transcription ranked the highest; a gene set linked with a negative effect on growth was also affected. In conclusion, the constant supplementation in the feed with the strain of yeast tested was effective in counteracting the detrimental effect of ETEC infection in susceptible pigs limits the early activation of the gene sets related to the impairment of the jejunal mucosa.
“…1. (A) Paraffin sections of liver slices from before (0 min) and after (15,60, and 120 min) injury were prepared and stained with pErk1/2 specific antibody. Bars represent 50 μm.…”
Section: Stimulation Of Primary Hepatocytes With the Conditioned Medimentioning
confidence: 99%
“…Immunohistochemical studies were performed as detailed previously [15]. Anti phosphoErk1/2 rabbit antibody and mouse monoclonal antibody against proliferating-cell-nuclear antigen (PCNA) were from Cell Signaling Technology.…”
“…We then quantified cancer cell adhesion to PCLS.Figure 5
Tumor cell adhesion assay using PCLS derived from genetically manipulated THOC5 KO mice. (A) : Rosa26-CreER T2 :THOC5 (flox/flox) mice were injected with Tamoxifen as described previously [31]. Protein was extracted from organs on 0, 4 and 5 days after the first injection, and subjected to THOC5 and actin specific immunoblot.…”
Section: Resultsmentioning
confidence: 99%
“…(A) : Rosa26-CreER T2 :THOC5 (flox/flox) mice were injected with Tamoxifen as described previously [31]. Protein was extracted from organs on 0, 4 and 5 days after the first injection, and subjected to THOC5 and actin specific immunoblot.…”
BackgroundOne of the most insidious characteristics of cancer is its spread to and ability to compromise distant organs via the complex process of metastasis. Communication between cancer cells and organ-resident cells via cytokines/chemokines and direct cell-cell contacts are key steps for survival, proliferation and invasion of metastasized cancer cells in organs. Precision-cut liver slices (PCLS) are considered to closely reflect the in vivo situation and are potentially useful for studying the interaction of cancer cells with liver-resident cells as well as being a potentially useful tool for screening anti-cancer reagents. Application of the PCLS technique in the field of cancer research however, has not yet been well developed.ResultsWe established the mouse PCLS system using perfluorodecalin (PFD) as an artificial oxygen carrier. Using this system we show that the adherence of green fluorescent protein (GFP) labeled MDA-MB-231 (highly invasive) cells to liver tissue in the PCLS was 5-fold greater than that of SK-BR-3 (less invasive) cells. In addition, we generated PCLS from THOC5, a member of transcription/export complex (TREX), knockout (KO) mice. The PCLS still expressed Gapdh or Albumin mRNAs at normal levels, while several chemokine/growth factor or metalloprotease genes, such as Cxcl12, Pdgfa, Tgfb, Wnt11, and Mmp1a genes were downregulated more than 2-fold. Interestingly, adhesion of cancer cells to THOC5 KO liver slices was far less (greater than 80% reduction) than to wild-type liver slices.ConclusionMouse PCLS cultures in the presence of PFD may serve as a useful tool for screening local adherence and invasiveness of individual cancer cells, since single cells can be observed. This method may also prove useful for identification of genes in liver-resident cells that support cancer invasion by using PCLS from transgenic liver.
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