Limits...
Gut microbiome derived metabolites modulate intestinal epithelial cell damage and mitigate Graft-versus-Host Disease

View Article: PubMed Central - PubMed

ABSTRACT

The impact of alterations in intestinal microbiota on microbial metabolites and on disease processes, such as graft-versus-host disease (GVHD), is not known. Here we performed unbiased analysis to identify novel alterations in gastrointestinal microbiota-derived short chain fatty acids (SCFA) after allogeneic bone marrow transplant (allo-BMT). Alterations in the amounts of only one SCFA, butyrate, were observed only within the intestinal tissue. The reduced butyrate in CD326+ intestinal epithelial cells (IECs) after allo-BMT resulted in decreased histone acetylation, which was restored upon local administration of exogenous butyrate. Butyrate restoration improved IEC junctional integrity, decreased apoptosis, and mitigated GVHD. Furthermore, alteration of the indigenous microbiota with 17 rationally selected strains of high butyrate producing Clostridia also decreased GVHD. These data demonstrate a heretofore unrecognized role of microbial metabolites and suggest that local and specific alteration of microbial metabolites has direct salutary effects on GVHD target tissues and can mitigate its severity.

No MeSH data available.


Related in: MedlinePlus

Intragastric butyrate gavage enhances its uptake and histone acetylation in IECsImmunoblot of CD326+ IECs from syngeneic (BALB/c → BALB/c) or allogeneic (C57BL/6J → BALB/c) BMT recipients that received vehicle or butyrate (10mg/kg). (a) Acetylated histone-H4 21 days post-BMT with densitometric analysis of three experiments combined, shown below blots; representative immunoblots shown. (b) 13C label incorporation of butyrate in luminal contents and intestinal tissue of the large intestine in mice gavaged with 13C-Butyrate or non-labeled 12C-Butyrate, n=6 mice per group. (c) 13C label incorporation in tricarboxylic acid metabolite pools of the large intestine of mice fed a bolus (2 g/kg) of 13C-Butyrate or 12C-Butyrate, n=6 mice per group. ND = not detected. (d) Levels of SLC5A8 in IECs of syngeneic and allogeneic BMT recipients 21 days following transplant, from a. (e) ChIP of butyrate treated IECs (CD326+) and binding of acetylated histone H4 in the promoter region of Slc5a8. (f) Weight loss on day 21 with (g) GVHD clinical score, (h) survival, and (i) intestinal histopathology 21 days post-BMT of recipients treated with intragastric vehicle or butyrate; syngeneic n=6, n=12 mice per allogeneic group. (j) Transmission electron microscopy (TEM) of intestines, isolated from BMT recipients with or without intragastric gavage of butyrate for duration of experiment; all samples were isolated 7 days following BMT and stained with ruthenium red (0.1%); arrows indicate cell-cell interface. (k) Level of FITC-dextran translocation across the GI-barrier into blood serum in butyrate treated allogeneic BMT recipients, compared to vehicle control 21 days post BMT. *P < .05; **P < .01; ***P < .0001 of ANOVA a - d, i, k; students t-test e; Mantel-Cox log-rank test g, h. Bars and error bars represent the means and standard errors of the mean, respectively.
© Copyright Policy
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4836986&req=5

Figure 3: Intragastric butyrate gavage enhances its uptake and histone acetylation in IECsImmunoblot of CD326+ IECs from syngeneic (BALB/c → BALB/c) or allogeneic (C57BL/6J → BALB/c) BMT recipients that received vehicle or butyrate (10mg/kg). (a) Acetylated histone-H4 21 days post-BMT with densitometric analysis of three experiments combined, shown below blots; representative immunoblots shown. (b) 13C label incorporation of butyrate in luminal contents and intestinal tissue of the large intestine in mice gavaged with 13C-Butyrate or non-labeled 12C-Butyrate, n=6 mice per group. (c) 13C label incorporation in tricarboxylic acid metabolite pools of the large intestine of mice fed a bolus (2 g/kg) of 13C-Butyrate or 12C-Butyrate, n=6 mice per group. ND = not detected. (d) Levels of SLC5A8 in IECs of syngeneic and allogeneic BMT recipients 21 days following transplant, from a. (e) ChIP of butyrate treated IECs (CD326+) and binding of acetylated histone H4 in the promoter region of Slc5a8. (f) Weight loss on day 21 with (g) GVHD clinical score, (h) survival, and (i) intestinal histopathology 21 days post-BMT of recipients treated with intragastric vehicle or butyrate; syngeneic n=6, n=12 mice per allogeneic group. (j) Transmission electron microscopy (TEM) of intestines, isolated from BMT recipients with or without intragastric gavage of butyrate for duration of experiment; all samples were isolated 7 days following BMT and stained with ruthenium red (0.1%); arrows indicate cell-cell interface. (k) Level of FITC-dextran translocation across the GI-barrier into blood serum in butyrate treated allogeneic BMT recipients, compared to vehicle control 21 days post BMT. *P < .05; **P < .01; ***P < .0001 of ANOVA a - d, i, k; students t-test e; Mantel-Cox log-rank test g, h. Bars and error bars represent the means and standard errors of the mean, respectively.

Mentions: We next determined if the reduced amount of butyrate in IECs could be restored in vivo and further, whether this would have a functional impact on histone acetylation. In addition to utilizing transporters, butyrate can also diffuse across the mucosal barrier into IECs when present in high concentrations10,22. Therefore, we hypothesized that administration of high amounts of butyrate locally would restore histone acetylation of IECs, in vivo. To test this, C57BL/6J cells were transferred to Balb/c mice and administered vehicle or butyrate via daily intragastric gavage. Daily butyrate administration for 21 days significantly restored acetylation of histone H4 compared with untreated allo-BMT recipients (Fig. 3a).


Gut microbiome derived metabolites modulate intestinal epithelial cell damage and mitigate Graft-versus-Host Disease
Intragastric butyrate gavage enhances its uptake and histone acetylation in IECsImmunoblot of CD326+ IECs from syngeneic (BALB/c → BALB/c) or allogeneic (C57BL/6J → BALB/c) BMT recipients that received vehicle or butyrate (10mg/kg). (a) Acetylated histone-H4 21 days post-BMT with densitometric analysis of three experiments combined, shown below blots; representative immunoblots shown. (b) 13C label incorporation of butyrate in luminal contents and intestinal tissue of the large intestine in mice gavaged with 13C-Butyrate or non-labeled 12C-Butyrate, n=6 mice per group. (c) 13C label incorporation in tricarboxylic acid metabolite pools of the large intestine of mice fed a bolus (2 g/kg) of 13C-Butyrate or 12C-Butyrate, n=6 mice per group. ND = not detected. (d) Levels of SLC5A8 in IECs of syngeneic and allogeneic BMT recipients 21 days following transplant, from a. (e) ChIP of butyrate treated IECs (CD326+) and binding of acetylated histone H4 in the promoter region of Slc5a8. (f) Weight loss on day 21 with (g) GVHD clinical score, (h) survival, and (i) intestinal histopathology 21 days post-BMT of recipients treated with intragastric vehicle or butyrate; syngeneic n=6, n=12 mice per allogeneic group. (j) Transmission electron microscopy (TEM) of intestines, isolated from BMT recipients with or without intragastric gavage of butyrate for duration of experiment; all samples were isolated 7 days following BMT and stained with ruthenium red (0.1%); arrows indicate cell-cell interface. (k) Level of FITC-dextran translocation across the GI-barrier into blood serum in butyrate treated allogeneic BMT recipients, compared to vehicle control 21 days post BMT. *P < .05; **P < .01; ***P < .0001 of ANOVA a - d, i, k; students t-test e; Mantel-Cox log-rank test g, h. Bars and error bars represent the means and standard errors of the mean, respectively.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4836986&req=5

Figure 3: Intragastric butyrate gavage enhances its uptake and histone acetylation in IECsImmunoblot of CD326+ IECs from syngeneic (BALB/c → BALB/c) or allogeneic (C57BL/6J → BALB/c) BMT recipients that received vehicle or butyrate (10mg/kg). (a) Acetylated histone-H4 21 days post-BMT with densitometric analysis of three experiments combined, shown below blots; representative immunoblots shown. (b) 13C label incorporation of butyrate in luminal contents and intestinal tissue of the large intestine in mice gavaged with 13C-Butyrate or non-labeled 12C-Butyrate, n=6 mice per group. (c) 13C label incorporation in tricarboxylic acid metabolite pools of the large intestine of mice fed a bolus (2 g/kg) of 13C-Butyrate or 12C-Butyrate, n=6 mice per group. ND = not detected. (d) Levels of SLC5A8 in IECs of syngeneic and allogeneic BMT recipients 21 days following transplant, from a. (e) ChIP of butyrate treated IECs (CD326+) and binding of acetylated histone H4 in the promoter region of Slc5a8. (f) Weight loss on day 21 with (g) GVHD clinical score, (h) survival, and (i) intestinal histopathology 21 days post-BMT of recipients treated with intragastric vehicle or butyrate; syngeneic n=6, n=12 mice per allogeneic group. (j) Transmission electron microscopy (TEM) of intestines, isolated from BMT recipients with or without intragastric gavage of butyrate for duration of experiment; all samples were isolated 7 days following BMT and stained with ruthenium red (0.1%); arrows indicate cell-cell interface. (k) Level of FITC-dextran translocation across the GI-barrier into blood serum in butyrate treated allogeneic BMT recipients, compared to vehicle control 21 days post BMT. *P < .05; **P < .01; ***P < .0001 of ANOVA a - d, i, k; students t-test e; Mantel-Cox log-rank test g, h. Bars and error bars represent the means and standard errors of the mean, respectively.
Mentions: We next determined if the reduced amount of butyrate in IECs could be restored in vivo and further, whether this would have a functional impact on histone acetylation. In addition to utilizing transporters, butyrate can also diffuse across the mucosal barrier into IECs when present in high concentrations10,22. Therefore, we hypothesized that administration of high amounts of butyrate locally would restore histone acetylation of IECs, in vivo. To test this, C57BL/6J cells were transferred to Balb/c mice and administered vehicle or butyrate via daily intragastric gavage. Daily butyrate administration for 21 days significantly restored acetylation of histone H4 compared with untreated allo-BMT recipients (Fig. 3a).

View Article: PubMed Central - PubMed

ABSTRACT

The impact of alterations in intestinal microbiota on microbial metabolites and on disease processes, such as graft-versus-host disease (GVHD), is not known. Here we performed unbiased analysis to identify novel alterations in gastrointestinal microbiota-derived short chain fatty acids (SCFA) after allogeneic bone marrow transplant (allo-BMT). Alterations in the amounts of only one SCFA, butyrate, were observed only within the intestinal tissue. The reduced butyrate in CD326+ intestinal epithelial cells (IECs) after allo-BMT resulted in decreased histone acetylation, which was restored upon local administration of exogenous butyrate. Butyrate restoration improved IEC junctional integrity, decreased apoptosis, and mitigated GVHD. Furthermore, alteration of the indigenous microbiota with 17 rationally selected strains of high butyrate producing Clostridia also decreased GVHD. These data demonstrate a heretofore unrecognized role of microbial metabolites and suggest that local and specific alteration of microbial metabolites has direct salutary effects on GVHD target tissues and can mitigate its severity.

No MeSH data available.


Related in: MedlinePlus