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A p38 MAPK-mediated alteration of COX-2/PGE2 regulates immunomodulatory properties in human mesenchymal stem cell aging.

Yu KR, Lee JY, Kim HS, Hong IS, Choi SW, Seo Y, Kang I, Kim JJ, Lee BC, Lee S, Kurtz A, Seo KW, Kang KS - PLoS ONE (2014)

Bottom Line: In this study, we evaluated the effect of replicative senescence on the immunomodulatory ability of hMSCs in vitro and in vivo.Among the anti-inflammatory cytokines, the production of prostaglandin E2 (PGE2) and the expression of its primary enzyme, cyclooxygenase-2 (COX-2), were profoundly increased by pre-stimulation with interferon gamma (IFN-γ) and tumor necrosis factor alpha (TNF-α), and this response was significantly decreased with consecutive passages.In conclusion, our data indicate that the immunomodulatory ability of hMSCs gradually declines with consecutive passages via a p38-mediated alteration of COX-2 and PGE2 levels.

View Article: PubMed Central - PubMed

Affiliation: Adult Stem Cell Research Center, College of Veterinary Medicine, Seoul National University, Seoul, South Korea; Research Institute for Veterinary Medicine, College of Veterinary Medicine, Seoul National University, Seoul, Korea.

ABSTRACT
Because human mesenchymal stem cells (hMSC) have profound immunomodulatory effects, many attempts have been made to use hMSCs in preclinical and clinical trials. For hMSCs to be used in therapy, a large population of hMSCs must be generated by in vitro expansion. However, the immunomodulatory changes following the in vitro expansion of hMSCs have not been elucidated. In this study, we evaluated the effect of replicative senescence on the immunomodulatory ability of hMSCs in vitro and in vivo. Late-passage hMSCs showed impaired suppressive effect on mitogen-induced mononuclear cell proliferation. Strikingly, late-passage hMSCs had a significantly compromised protective effect against mouse experimental colitis, which was confirmed by gross and histologic examination. Among the anti-inflammatory cytokines, the production of prostaglandin E2 (PGE2) and the expression of its primary enzyme, cyclooxygenase-2 (COX-2), were profoundly increased by pre-stimulation with interferon gamma (IFN-γ) and tumor necrosis factor alpha (TNF-α), and this response was significantly decreased with consecutive passages. We demonstrated that the impaired phosphorylation activity of p38 MAP kinase (p38 MAPK) in late-passage hMSCs led to a compromised immunomodulatory ability through the regulation of COX-2. In conclusion, our data indicate that the immunomodulatory ability of hMSCs gradually declines with consecutive passages via a p38-mediated alteration of COX-2 and PGE2 levels.

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Administration of late-passage hMSCs reduces the protective effects against DSS-induced colitis in mice.(A–F) 3% DSS water was administered to mice for seven days to induce colitis. Early- or late-passage hMSCs were injected intraperitoneally one day after the administration of DSS. The percentage of body weight loss (A), the Mantel Cox analysis of survival rate (B) and the disease activity index for colitis severity (C) were monitored as clinical progression. Mice were sacrificed ten days after the induction of colitis with DSS, and the length (D), weight-to-length ratios of the colons (E) and colon sections were stained with H&E and histopathologic evaluation was investigated by determining lymphocyte infiltration and intestinal damage (F, G). (*P<0.05, **P<0.01).
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pone-0102426-g003: Administration of late-passage hMSCs reduces the protective effects against DSS-induced colitis in mice.(A–F) 3% DSS water was administered to mice for seven days to induce colitis. Early- or late-passage hMSCs were injected intraperitoneally one day after the administration of DSS. The percentage of body weight loss (A), the Mantel Cox analysis of survival rate (B) and the disease activity index for colitis severity (C) were monitored as clinical progression. Mice were sacrificed ten days after the induction of colitis with DSS, and the length (D), weight-to-length ratios of the colons (E) and colon sections were stained with H&E and histopathologic evaluation was investigated by determining lymphocyte infiltration and intestinal damage (F, G). (*P<0.05, **P<0.01).

Mentions: To support the observation that the decrease in the hMSC immunosuppressive properties correlates with replicative senescence in vitro, we subsequently investigated the potential therapeutic efficacy of early- and late-hMSCs in the experimental model of acute colitis that was induced by oral dextran sulfate sodium (DSS) administration. It has been reported that DSS causes intestinal inflammation due to the exposure of the submucosa to exterior antigens (intestinal bacteria and food), leading to the recruitment or activation of inflammatory cells associated with innate immunity [23]. Mice were randomly assigned with similar body weight to investigate the therapeutic effect of early- and late-passage hMSC in intestinal inflammation. Oral administration of a 3% DSS solution induced acute colitis, characterized by clinical symptoms (diarrhea and bloody stool) with sustained weight loss resulting in 50% mortality (9/18). Intraperitoneal injection of early-passage hMSCs reduced the loss of body-weight and decreased the mortality of mice compared to PBS- or late-passage hMSC injections (Figure 3A and B). Strikingly, the transplantation of early-passage hMSCs rescued 100% (n = 15) of the mice from colitis-induced lethality (Figure 3B). Treatment of late-passage hMSCs, however, did not exert these beneficial effects. On day 7, the disease activity index was significantly decreased by treatment with early-passage MSCs. In contrast, the administration of late-passage hMSCs did not show beneficial effects on the disease activity index (Figure 3C). The disease activity index was measured according to standards for the quantification of symptoms of patients with Crohn's disease [24]. On day 10, the mice were sacrificed, and the entire colon from the caecum to the anus was acquired. The length, mass weight and histopathology of the colon were investigated to determine the inflammation status. The colon length decreased in mice treated with PBS or late-passage hMSCs. However, the colon length was restored by an injection of early-passage hMSCs (Figure 3D). The increased mass-to-length ratio revealed that both hyperemia and inflammation were exhibited in the colons of colitis-induced mice. Early-passage hMSCs reduced the mass-to-length ratio significantly compared to late-passage hMSCs (Figure 3E). Colon samples were processed and stained with H&E for histopathological evaluation. Histologic examination showed the destruction of the entire epithelium and submucosal edema and the infiltration of inflammatory cells into the lamina propria and submucosa in the colon of DSS-treated mice (Figure 3F). Importantly, the administration of early-passage hMSCs greatly reduced histologic damage and the histologic score in the colon. In contrast, late-passage hMSCs did not prevent histologic damage or decrease the histologic score (Figure 3F and G). These in vivo data suggest that replicative senescence significantly impairs the ability of hMSCs to deactivate the colonic inflammatory response.


A p38 MAPK-mediated alteration of COX-2/PGE2 regulates immunomodulatory properties in human mesenchymal stem cell aging.

Yu KR, Lee JY, Kim HS, Hong IS, Choi SW, Seo Y, Kang I, Kim JJ, Lee BC, Lee S, Kurtz A, Seo KW, Kang KS - PLoS ONE (2014)

Administration of late-passage hMSCs reduces the protective effects against DSS-induced colitis in mice.(A–F) 3% DSS water was administered to mice for seven days to induce colitis. Early- or late-passage hMSCs were injected intraperitoneally one day after the administration of DSS. The percentage of body weight loss (A), the Mantel Cox analysis of survival rate (B) and the disease activity index for colitis severity (C) were monitored as clinical progression. Mice were sacrificed ten days after the induction of colitis with DSS, and the length (D), weight-to-length ratios of the colons (E) and colon sections were stained with H&E and histopathologic evaluation was investigated by determining lymphocyte infiltration and intestinal damage (F, G). (*P<0.05, **P<0.01).
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4121064&req=5

pone-0102426-g003: Administration of late-passage hMSCs reduces the protective effects against DSS-induced colitis in mice.(A–F) 3% DSS water was administered to mice for seven days to induce colitis. Early- or late-passage hMSCs were injected intraperitoneally one day after the administration of DSS. The percentage of body weight loss (A), the Mantel Cox analysis of survival rate (B) and the disease activity index for colitis severity (C) were monitored as clinical progression. Mice were sacrificed ten days after the induction of colitis with DSS, and the length (D), weight-to-length ratios of the colons (E) and colon sections were stained with H&E and histopathologic evaluation was investigated by determining lymphocyte infiltration and intestinal damage (F, G). (*P<0.05, **P<0.01).
Mentions: To support the observation that the decrease in the hMSC immunosuppressive properties correlates with replicative senescence in vitro, we subsequently investigated the potential therapeutic efficacy of early- and late-hMSCs in the experimental model of acute colitis that was induced by oral dextran sulfate sodium (DSS) administration. It has been reported that DSS causes intestinal inflammation due to the exposure of the submucosa to exterior antigens (intestinal bacteria and food), leading to the recruitment or activation of inflammatory cells associated with innate immunity [23]. Mice were randomly assigned with similar body weight to investigate the therapeutic effect of early- and late-passage hMSC in intestinal inflammation. Oral administration of a 3% DSS solution induced acute colitis, characterized by clinical symptoms (diarrhea and bloody stool) with sustained weight loss resulting in 50% mortality (9/18). Intraperitoneal injection of early-passage hMSCs reduced the loss of body-weight and decreased the mortality of mice compared to PBS- or late-passage hMSC injections (Figure 3A and B). Strikingly, the transplantation of early-passage hMSCs rescued 100% (n = 15) of the mice from colitis-induced lethality (Figure 3B). Treatment of late-passage hMSCs, however, did not exert these beneficial effects. On day 7, the disease activity index was significantly decreased by treatment with early-passage MSCs. In contrast, the administration of late-passage hMSCs did not show beneficial effects on the disease activity index (Figure 3C). The disease activity index was measured according to standards for the quantification of symptoms of patients with Crohn's disease [24]. On day 10, the mice were sacrificed, and the entire colon from the caecum to the anus was acquired. The length, mass weight and histopathology of the colon were investigated to determine the inflammation status. The colon length decreased in mice treated with PBS or late-passage hMSCs. However, the colon length was restored by an injection of early-passage hMSCs (Figure 3D). The increased mass-to-length ratio revealed that both hyperemia and inflammation were exhibited in the colons of colitis-induced mice. Early-passage hMSCs reduced the mass-to-length ratio significantly compared to late-passage hMSCs (Figure 3E). Colon samples were processed and stained with H&E for histopathological evaluation. Histologic examination showed the destruction of the entire epithelium and submucosal edema and the infiltration of inflammatory cells into the lamina propria and submucosa in the colon of DSS-treated mice (Figure 3F). Importantly, the administration of early-passage hMSCs greatly reduced histologic damage and the histologic score in the colon. In contrast, late-passage hMSCs did not prevent histologic damage or decrease the histologic score (Figure 3F and G). These in vivo data suggest that replicative senescence significantly impairs the ability of hMSCs to deactivate the colonic inflammatory response.

Bottom Line: In this study, we evaluated the effect of replicative senescence on the immunomodulatory ability of hMSCs in vitro and in vivo.Among the anti-inflammatory cytokines, the production of prostaglandin E2 (PGE2) and the expression of its primary enzyme, cyclooxygenase-2 (COX-2), were profoundly increased by pre-stimulation with interferon gamma (IFN-γ) and tumor necrosis factor alpha (TNF-α), and this response was significantly decreased with consecutive passages.In conclusion, our data indicate that the immunomodulatory ability of hMSCs gradually declines with consecutive passages via a p38-mediated alteration of COX-2 and PGE2 levels.

View Article: PubMed Central - PubMed

Affiliation: Adult Stem Cell Research Center, College of Veterinary Medicine, Seoul National University, Seoul, South Korea; Research Institute for Veterinary Medicine, College of Veterinary Medicine, Seoul National University, Seoul, Korea.

ABSTRACT
Because human mesenchymal stem cells (hMSC) have profound immunomodulatory effects, many attempts have been made to use hMSCs in preclinical and clinical trials. For hMSCs to be used in therapy, a large population of hMSCs must be generated by in vitro expansion. However, the immunomodulatory changes following the in vitro expansion of hMSCs have not been elucidated. In this study, we evaluated the effect of replicative senescence on the immunomodulatory ability of hMSCs in vitro and in vivo. Late-passage hMSCs showed impaired suppressive effect on mitogen-induced mononuclear cell proliferation. Strikingly, late-passage hMSCs had a significantly compromised protective effect against mouse experimental colitis, which was confirmed by gross and histologic examination. Among the anti-inflammatory cytokines, the production of prostaglandin E2 (PGE2) and the expression of its primary enzyme, cyclooxygenase-2 (COX-2), were profoundly increased by pre-stimulation with interferon gamma (IFN-γ) and tumor necrosis factor alpha (TNF-α), and this response was significantly decreased with consecutive passages. We demonstrated that the impaired phosphorylation activity of p38 MAP kinase (p38 MAPK) in late-passage hMSCs led to a compromised immunomodulatory ability through the regulation of COX-2. In conclusion, our data indicate that the immunomodulatory ability of hMSCs gradually declines with consecutive passages via a p38-mediated alteration of COX-2 and PGE2 levels.

Show MeSH
Related in: MedlinePlus