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Hyaluronan Synthase 3 Null Mice Exhibit Decreased Intestinal Inflammation and Tissue Damage in the DSS-Induced Colitis Model.

Kessler SP, Obery DR, de la Motte C - Int J Cell Biol (2015)

Bottom Line: Hyaluronan (HA) overproduction is a hallmark of multiple inflammatory diseases, including inflammatory bowel disease (IBD).Therefore, we hypothesized that, during a pathologic challenge, HA promotes inflammation.Our data suggest, HAS3 plays a crucial role in driving gut inflammation.

View Article: PubMed Central - PubMed

Affiliation: Department of Pathobiology, Lerner Research Institute Cleveland Clinic, 9500 Euclid Avenue, Cleveland, OH 44195, USA.

ABSTRACT
Hyaluronan (HA) overproduction is a hallmark of multiple inflammatory diseases, including inflammatory bowel disease (IBD). Hyaluronan can act as a leukocyte recruitment molecule and in the most common mouse model of intestinal inflammation, the chemically induced dextran sodium sulfate (DSS) experimental colitis model, we previously determined that changes in colon distribution of HA occur before inflammation. Therefore, we hypothesized that, during a pathologic challenge, HA promotes inflammation. In this study, we tested the progression of inflammation in mice for the hyaluronan synthase genes (HAS1, HAS3, or both HAS1 and HAS3) in the DSS-colitis model. Our data demonstrate that both the HAS1/HAS3 double and the HAS3 mice are protected from colitis, compared to wild-type and HAS1 mice, as determined by measurement of weight loss, disease activity, serum IL-6 levels, histologic scoring, and immunohistochemistry. Most notable is the dramatic increase in submucosal microvasculature, hyaluronan deposition, and leukocyte infiltration in the inflamed colon tissue of wild-type and HAS1 mice. Our data suggest, HAS3 plays a crucial role in driving gut inflammation. Developing a temporary targeted therapeutic intervention of HAS3 expression or function in the microcirculation may emerge as a desirable strategy toward tempering colitis in patients undergoing flares of IBD.

No MeSH data available.


Related in: MedlinePlus

Immunohistochemistry of DSS-treated mice. (a) Adjacent rectal sections to the H&E stained tissues presented in Figure 6 were stained for hyaluronan (green) with the biotinylated hyaluronan binding protein (HABP) and Alexa Fluor streptavidin-488. Nuclei were stained with DAPI (blue). Images are 10x magnification captured with a Leica digital fluorescent microscope and ImageProPlus software. (b) Semiquantitative densitometric analysis of HA staining in longitudinal folds in the images presented in panel (a). The untreated wild-type rectal section illustrates the regions measured (yellow). Lamina propria HA staining (LP) is the region above the muscularis mucosae while submucosa HA staining (SM) is the area below the muscularis mucosae and above the muscularis externa.
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fig7: Immunohistochemistry of DSS-treated mice. (a) Adjacent rectal sections to the H&E stained tissues presented in Figure 6 were stained for hyaluronan (green) with the biotinylated hyaluronan binding protein (HABP) and Alexa Fluor streptavidin-488. Nuclei were stained with DAPI (blue). Images are 10x magnification captured with a Leica digital fluorescent microscope and ImageProPlus software. (b) Semiquantitative densitometric analysis of HA staining in longitudinal folds in the images presented in panel (a). The untreated wild-type rectal section illustrates the regions measured (yellow). Lamina propria HA staining (LP) is the region above the muscularis mucosae while submucosa HA staining (SM) is the area below the muscularis mucosae and above the muscularis externa.

Mentions: The profound changes that occur in the mouse distal colon over the 10 days of DSS-induced colitis can be seen in Figure 6. Distal colon (rectal) sections were selected for specific analysis since this model is bacterially driven and the rectal area has the highest microbe burden. Once the epithelial layers have been effaced and crypt damage ensues due to continuous chemical exposure, direct microbe interaction with host immune cells occurs. Similar pathological changes are routinely observed in human IBD patient intestinal tissue after repeated flares of inflammation. While submucosal swelling and epithelial damage commence at day 3 in wild-type and the HAS1 groups, this effect is delayed or dramatically reduced in both HAS1/3 and HAS3 mouse groups even at 10 days of treatment (Figure 6). In addition, leukocyte infiltration, muscularis mucosae hyperplasia, and increased angiogenesis are all evident in day 7 and day 10 in the wild-type and HAS1 groups but not in HAS1/3 or HAS3 mice. To determine the extent of submucosal expansion in the longitudinal rectal folds in each tissue over the course of the experimental colitis in Figure 6(a), we measured the area of swelling using imaging software (Figure 6(b)). The representative submucosal region measured is indicated in the untreated wild-type mouse. Both the H&E images and the expansion measurements indicate the dramatic changes in the submucosal region of wild-type and HAS1 mice, especially at day 7 and day 10, compared to the HAS3 and HAS1/3 mice. Immunostaining of these adjacent matched rectal sections for hyaluronan deposition and clearance over the 10-day time course also reveals distinctly different HA staining patterns in the mice lacking the intact HAS3 allele. Figure 7 demonstrates that in all groups there is increased deposition of hyaluronan at day 3 in the submucosa and mucosal/lamina propria regions of the colon. By day 5, in the wild-type and HAS1 groups, submucosal swelling is very pronounced, whereas this swelling did not occur in animals with HAS3 deletion. At days 7 and 10, the mice that possess HAS3 (the wild-type and HAS1 groups) exhibit a very noticeable persistence and increase in hyaluronan deposition in the damaged mucosa as well as around the highly vascularized areas in the submucosa. Semiquantitative densitometric analysis of the HA staining in the longitudinal folds of the lamina propria and submucosal regions was performed to track the changes in hyaluronan in both compartments in each genotype over the course of the colitis (Figure 7(b)). The regions measured for each subcompartment of the rectal fold are illustrated as yellow masked areas on the image from the untreated wild-type mouse. In unchallenged animals, homeostatic levels of HA in the lamina propria extracellular matrix of distal colon tissue are typically decreased with deletion of enzymes HAS1 (~50%) and especially HAS3 (~75%) compared to wild type. In the submucosa, overall lower levels of HA are produced per area, but the relationship of decreased levels of HA with deletion of HAS1 and HAS3 is also observed. Unexpectedly, HA levels in the HAS3 tissue are lower than in the HAS1/HAS3 double deletion intestinal tissue.


Hyaluronan Synthase 3 Null Mice Exhibit Decreased Intestinal Inflammation and Tissue Damage in the DSS-Induced Colitis Model.

Kessler SP, Obery DR, de la Motte C - Int J Cell Biol (2015)

Immunohistochemistry of DSS-treated mice. (a) Adjacent rectal sections to the H&E stained tissues presented in Figure 6 were stained for hyaluronan (green) with the biotinylated hyaluronan binding protein (HABP) and Alexa Fluor streptavidin-488. Nuclei were stained with DAPI (blue). Images are 10x magnification captured with a Leica digital fluorescent microscope and ImageProPlus software. (b) Semiquantitative densitometric analysis of HA staining in longitudinal folds in the images presented in panel (a). The untreated wild-type rectal section illustrates the regions measured (yellow). Lamina propria HA staining (LP) is the region above the muscularis mucosae while submucosa HA staining (SM) is the area below the muscularis mucosae and above the muscularis externa.
© Copyright Policy - open-access
Related In: Results  -  Collection

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fig7: Immunohistochemistry of DSS-treated mice. (a) Adjacent rectal sections to the H&E stained tissues presented in Figure 6 were stained for hyaluronan (green) with the biotinylated hyaluronan binding protein (HABP) and Alexa Fluor streptavidin-488. Nuclei were stained with DAPI (blue). Images are 10x magnification captured with a Leica digital fluorescent microscope and ImageProPlus software. (b) Semiquantitative densitometric analysis of HA staining in longitudinal folds in the images presented in panel (a). The untreated wild-type rectal section illustrates the regions measured (yellow). Lamina propria HA staining (LP) is the region above the muscularis mucosae while submucosa HA staining (SM) is the area below the muscularis mucosae and above the muscularis externa.
Mentions: The profound changes that occur in the mouse distal colon over the 10 days of DSS-induced colitis can be seen in Figure 6. Distal colon (rectal) sections were selected for specific analysis since this model is bacterially driven and the rectal area has the highest microbe burden. Once the epithelial layers have been effaced and crypt damage ensues due to continuous chemical exposure, direct microbe interaction with host immune cells occurs. Similar pathological changes are routinely observed in human IBD patient intestinal tissue after repeated flares of inflammation. While submucosal swelling and epithelial damage commence at day 3 in wild-type and the HAS1 groups, this effect is delayed or dramatically reduced in both HAS1/3 and HAS3 mouse groups even at 10 days of treatment (Figure 6). In addition, leukocyte infiltration, muscularis mucosae hyperplasia, and increased angiogenesis are all evident in day 7 and day 10 in the wild-type and HAS1 groups but not in HAS1/3 or HAS3 mice. To determine the extent of submucosal expansion in the longitudinal rectal folds in each tissue over the course of the experimental colitis in Figure 6(a), we measured the area of swelling using imaging software (Figure 6(b)). The representative submucosal region measured is indicated in the untreated wild-type mouse. Both the H&E images and the expansion measurements indicate the dramatic changes in the submucosal region of wild-type and HAS1 mice, especially at day 7 and day 10, compared to the HAS3 and HAS1/3 mice. Immunostaining of these adjacent matched rectal sections for hyaluronan deposition and clearance over the 10-day time course also reveals distinctly different HA staining patterns in the mice lacking the intact HAS3 allele. Figure 7 demonstrates that in all groups there is increased deposition of hyaluronan at day 3 in the submucosa and mucosal/lamina propria regions of the colon. By day 5, in the wild-type and HAS1 groups, submucosal swelling is very pronounced, whereas this swelling did not occur in animals with HAS3 deletion. At days 7 and 10, the mice that possess HAS3 (the wild-type and HAS1 groups) exhibit a very noticeable persistence and increase in hyaluronan deposition in the damaged mucosa as well as around the highly vascularized areas in the submucosa. Semiquantitative densitometric analysis of the HA staining in the longitudinal folds of the lamina propria and submucosal regions was performed to track the changes in hyaluronan in both compartments in each genotype over the course of the colitis (Figure 7(b)). The regions measured for each subcompartment of the rectal fold are illustrated as yellow masked areas on the image from the untreated wild-type mouse. In unchallenged animals, homeostatic levels of HA in the lamina propria extracellular matrix of distal colon tissue are typically decreased with deletion of enzymes HAS1 (~50%) and especially HAS3 (~75%) compared to wild type. In the submucosa, overall lower levels of HA are produced per area, but the relationship of decreased levels of HA with deletion of HAS1 and HAS3 is also observed. Unexpectedly, HA levels in the HAS3 tissue are lower than in the HAS1/HAS3 double deletion intestinal tissue.

Bottom Line: Hyaluronan (HA) overproduction is a hallmark of multiple inflammatory diseases, including inflammatory bowel disease (IBD).Therefore, we hypothesized that, during a pathologic challenge, HA promotes inflammation.Our data suggest, HAS3 plays a crucial role in driving gut inflammation.

View Article: PubMed Central - PubMed

Affiliation: Department of Pathobiology, Lerner Research Institute Cleveland Clinic, 9500 Euclid Avenue, Cleveland, OH 44195, USA.

ABSTRACT
Hyaluronan (HA) overproduction is a hallmark of multiple inflammatory diseases, including inflammatory bowel disease (IBD). Hyaluronan can act as a leukocyte recruitment molecule and in the most common mouse model of intestinal inflammation, the chemically induced dextran sodium sulfate (DSS) experimental colitis model, we previously determined that changes in colon distribution of HA occur before inflammation. Therefore, we hypothesized that, during a pathologic challenge, HA promotes inflammation. In this study, we tested the progression of inflammation in mice for the hyaluronan synthase genes (HAS1, HAS3, or both HAS1 and HAS3) in the DSS-colitis model. Our data demonstrate that both the HAS1/HAS3 double and the HAS3 mice are protected from colitis, compared to wild-type and HAS1 mice, as determined by measurement of weight loss, disease activity, serum IL-6 levels, histologic scoring, and immunohistochemistry. Most notable is the dramatic increase in submucosal microvasculature, hyaluronan deposition, and leukocyte infiltration in the inflamed colon tissue of wild-type and HAS1 mice. Our data suggest, HAS3 plays a crucial role in driving gut inflammation. Developing a temporary targeted therapeutic intervention of HAS3 expression or function in the microcirculation may emerge as a desirable strategy toward tempering colitis in patients undergoing flares of IBD.

No MeSH data available.


Related in: MedlinePlus