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Exosomal miR-223 Contributes to Mesenchymal Stem Cell-Elicited Cardioprotection in Polymicrobial Sepsis.

Wang X, Gu H, Qin D, Yang L, Huang W, Essandoh K, Wang Y, Caldwell CC, Peng T, Zingarelli B, Fan GC - Sci Rep (2015)

Bottom Line: However, WT-MSCs were able to provide protection which was associated with exosome release.Conversely, WT-MSC-derived-exosomes displayed protective effects.By contrast, WT-exosomes encased higher levels of miR-223, which could be delivered to cardiomyocytes, resulting in down-regulation of Sema3A and Stat3.

View Article: PubMed Central - PubMed

Affiliation: Department of Pharmacology and Cell Biophysics, University of Cincinnati College of Medicine, Cincinnati, OH, USA.

ABSTRACT
Mesenchymal stem cells (MSCs) have been shown to elicit cardio-protective effects in sepsis. However, the underlying mechanism remains obscure. While recent studies have indicated that miR-223 is highly enriched in MSC-derived exosomes, whether exosomal miR-223 contributes to MSC-mediated cardio-protection in sepsis is unknown. In this study, loss-of-function approach was utilized, and sepsis was induced by cecal ligation and puncture (CLP). We observed that injection of miR-223-KO MSCs at 1 h post-CLP did not confer protection against CLP-triggered cardiac dysfunction, apoptosis and inflammatory response. However, WT-MSCs were able to provide protection which was associated with exosome release. Next, treatment of CLP mice with exosomes released from miR-223-KO MSCs significantly exaggerated sepsis-induced injury. Conversely, WT-MSC-derived-exosomes displayed protective effects. Mechanistically, we identified that miR-223-KO exosomes contained higher levels of Sema3A and Stat3, two known targets of miR-223 (5p &3p), than WT-exosomes. Accordingly, these exosomal proteins were transferred to cardiomyocytes, leading to increased inflammation and cell death. By contrast, WT-exosomes encased higher levels of miR-223, which could be delivered to cardiomyocytes, resulting in down-regulation of Sema3A and Stat3. These data for the first time indicate that exosomal miR-223 plays an essential role for MSC-induced cardio-protection in sepsis.

No MeSH data available.


Related in: MedlinePlus

The effects of WT-exosomes and miR-223-KO exosomes on CLP-induced inflammatory response, cardiac dysfunction and animal mortality.(A–C) CLP-mice treated with WT-exosomes (n = 11) showed lower levels of serum TNF-α (A), IL-1β (B), and IL-6 (C), whereas CLP-mice injected with KO-exosomes (n = 11) exhibited higher levels of circulating TNF-α (A), IL-1β (B), and IL-6 (C), compared with those treated with incomplete DMEM medium (n = 10) (^p < 0.05 vs. shams; *p < 0.05 vs. CLP + medium; #p < 0.05 vs. CLP + medium). (D) Results of echocardiography measurement showed that values of the left ventricular ejection fraction (EF%, E) and the fractional shortening (FS%, F) were significantly decreased in CLP mice injected with incomplete DMEM medium (n = 10), compared with shams (n = 8). Remarkably, the reduction of EF% and FS% was attenuated in WT-exosome-treated CLP mice (n = 11); whereas it was aggravated in CLP mice administrated with miR-223-KO exosomes (n = 11) (^p < 0.05 vs. shams; *p < 0.05 vs. CLP + medium; #p < 0.05 vs. CLP + medium). (G) The survival of CLP-mice was significantly improved by WT-exosome treatment, whereas it was worse by miR-223-KO exosome injection (n = 8, *p < 0.05 vs. CLP + medium).
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f6: The effects of WT-exosomes and miR-223-KO exosomes on CLP-induced inflammatory response, cardiac dysfunction and animal mortality.(A–C) CLP-mice treated with WT-exosomes (n = 11) showed lower levels of serum TNF-α (A), IL-1β (B), and IL-6 (C), whereas CLP-mice injected with KO-exosomes (n = 11) exhibited higher levels of circulating TNF-α (A), IL-1β (B), and IL-6 (C), compared with those treated with incomplete DMEM medium (n = 10) (^p < 0.05 vs. shams; *p < 0.05 vs. CLP + medium; #p < 0.05 vs. CLP + medium). (D) Results of echocardiography measurement showed that values of the left ventricular ejection fraction (EF%, E) and the fractional shortening (FS%, F) were significantly decreased in CLP mice injected with incomplete DMEM medium (n = 10), compared with shams (n = 8). Remarkably, the reduction of EF% and FS% was attenuated in WT-exosome-treated CLP mice (n = 11); whereas it was aggravated in CLP mice administrated with miR-223-KO exosomes (n = 11) (^p < 0.05 vs. shams; *p < 0.05 vs. CLP + medium; #p < 0.05 vs. CLP + medium). (G) The survival of CLP-mice was significantly improved by WT-exosome treatment, whereas it was worse by miR-223-KO exosome injection (n = 8, *p < 0.05 vs. CLP + medium).

Mentions: Next, we asked whether MSC-derived exosomes had immune-suppressive effects, similar to MSCs in vivo, and if so, whether miR-223 is required for such effects. To address these issues, we infused WT-exosomes, KO-exosomes, or incomplete culture medium into mice via the tail vein at 1 h post-CLP surgery. 12 h later, sera were collected from those mice for cytokine assays. As shown in Fig. 6A–C, the serum levels of TNF-α, IL-1β, and IL-6 were greatly reduced in WT-exosome-treated CLP-mice, whereas the serum levels of these pro-inflammatory factors were significantly increased in KO-exosome-injected septic mice, compared with medium-treated control samples (n = 8–11). These data suggest that: 1) MSC-derived exosomes can be used in place of intact stem cells to inhibit sepsis-induced inflammatory response; and 2) MSC-exosome-induced anti-inflammatory effects are dependent on miR-223, at least in part.


Exosomal miR-223 Contributes to Mesenchymal Stem Cell-Elicited Cardioprotection in Polymicrobial Sepsis.

Wang X, Gu H, Qin D, Yang L, Huang W, Essandoh K, Wang Y, Caldwell CC, Peng T, Zingarelli B, Fan GC - Sci Rep (2015)

The effects of WT-exosomes and miR-223-KO exosomes on CLP-induced inflammatory response, cardiac dysfunction and animal mortality.(A–C) CLP-mice treated with WT-exosomes (n = 11) showed lower levels of serum TNF-α (A), IL-1β (B), and IL-6 (C), whereas CLP-mice injected with KO-exosomes (n = 11) exhibited higher levels of circulating TNF-α (A), IL-1β (B), and IL-6 (C), compared with those treated with incomplete DMEM medium (n = 10) (^p < 0.05 vs. shams; *p < 0.05 vs. CLP + medium; #p < 0.05 vs. CLP + medium). (D) Results of echocardiography measurement showed that values of the left ventricular ejection fraction (EF%, E) and the fractional shortening (FS%, F) were significantly decreased in CLP mice injected with incomplete DMEM medium (n = 10), compared with shams (n = 8). Remarkably, the reduction of EF% and FS% was attenuated in WT-exosome-treated CLP mice (n = 11); whereas it was aggravated in CLP mice administrated with miR-223-KO exosomes (n = 11) (^p < 0.05 vs. shams; *p < 0.05 vs. CLP + medium; #p < 0.05 vs. CLP + medium). (G) The survival of CLP-mice was significantly improved by WT-exosome treatment, whereas it was worse by miR-223-KO exosome injection (n = 8, *p < 0.05 vs. CLP + medium).
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Related In: Results  -  Collection

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f6: The effects of WT-exosomes and miR-223-KO exosomes on CLP-induced inflammatory response, cardiac dysfunction and animal mortality.(A–C) CLP-mice treated with WT-exosomes (n = 11) showed lower levels of serum TNF-α (A), IL-1β (B), and IL-6 (C), whereas CLP-mice injected with KO-exosomes (n = 11) exhibited higher levels of circulating TNF-α (A), IL-1β (B), and IL-6 (C), compared with those treated with incomplete DMEM medium (n = 10) (^p < 0.05 vs. shams; *p < 0.05 vs. CLP + medium; #p < 0.05 vs. CLP + medium). (D) Results of echocardiography measurement showed that values of the left ventricular ejection fraction (EF%, E) and the fractional shortening (FS%, F) were significantly decreased in CLP mice injected with incomplete DMEM medium (n = 10), compared with shams (n = 8). Remarkably, the reduction of EF% and FS% was attenuated in WT-exosome-treated CLP mice (n = 11); whereas it was aggravated in CLP mice administrated with miR-223-KO exosomes (n = 11) (^p < 0.05 vs. shams; *p < 0.05 vs. CLP + medium; #p < 0.05 vs. CLP + medium). (G) The survival of CLP-mice was significantly improved by WT-exosome treatment, whereas it was worse by miR-223-KO exosome injection (n = 8, *p < 0.05 vs. CLP + medium).
Mentions: Next, we asked whether MSC-derived exosomes had immune-suppressive effects, similar to MSCs in vivo, and if so, whether miR-223 is required for such effects. To address these issues, we infused WT-exosomes, KO-exosomes, or incomplete culture medium into mice via the tail vein at 1 h post-CLP surgery. 12 h later, sera were collected from those mice for cytokine assays. As shown in Fig. 6A–C, the serum levels of TNF-α, IL-1β, and IL-6 were greatly reduced in WT-exosome-treated CLP-mice, whereas the serum levels of these pro-inflammatory factors were significantly increased in KO-exosome-injected septic mice, compared with medium-treated control samples (n = 8–11). These data suggest that: 1) MSC-derived exosomes can be used in place of intact stem cells to inhibit sepsis-induced inflammatory response; and 2) MSC-exosome-induced anti-inflammatory effects are dependent on miR-223, at least in part.

Bottom Line: However, WT-MSCs were able to provide protection which was associated with exosome release.Conversely, WT-MSC-derived-exosomes displayed protective effects.By contrast, WT-exosomes encased higher levels of miR-223, which could be delivered to cardiomyocytes, resulting in down-regulation of Sema3A and Stat3.

View Article: PubMed Central - PubMed

Affiliation: Department of Pharmacology and Cell Biophysics, University of Cincinnati College of Medicine, Cincinnati, OH, USA.

ABSTRACT
Mesenchymal stem cells (MSCs) have been shown to elicit cardio-protective effects in sepsis. However, the underlying mechanism remains obscure. While recent studies have indicated that miR-223 is highly enriched in MSC-derived exosomes, whether exosomal miR-223 contributes to MSC-mediated cardio-protection in sepsis is unknown. In this study, loss-of-function approach was utilized, and sepsis was induced by cecal ligation and puncture (CLP). We observed that injection of miR-223-KO MSCs at 1 h post-CLP did not confer protection against CLP-triggered cardiac dysfunction, apoptosis and inflammatory response. However, WT-MSCs were able to provide protection which was associated with exosome release. Next, treatment of CLP mice with exosomes released from miR-223-KO MSCs significantly exaggerated sepsis-induced injury. Conversely, WT-MSC-derived-exosomes displayed protective effects. Mechanistically, we identified that miR-223-KO exosomes contained higher levels of Sema3A and Stat3, two known targets of miR-223 (5p &3p), than WT-exosomes. Accordingly, these exosomal proteins were transferred to cardiomyocytes, leading to increased inflammation and cell death. By contrast, WT-exosomes encased higher levels of miR-223, which could be delivered to cardiomyocytes, resulting in down-regulation of Sema3A and Stat3. These data for the first time indicate that exosomal miR-223 plays an essential role for MSC-induced cardio-protection in sepsis.

No MeSH data available.


Related in: MedlinePlus