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Prostacyclin-producing human mesenchymal cells target H19 lncRNA to augment endogenous progenitor function in hindlimb ischaemia.

Deng Y, Yang Z, Terry T, Pan S, Woodside DG, Wang J, Ruan K, Willerson JT, Dixon RA, Liu Q - Nat Commun (2016)

Bottom Line: Here we develop an innovative strategy to enhance the paracrine effects of hMSCs.Transplanted PGI2-hMSCs do not incorporate long term into host tissue, but rather they mediate host regeneration and muscle mass gain in a paracrine manner.Mechanistically, this involves long noncoding RNA H19 in promoting PGI2-hMSC-associated survival and proliferation of host progenitor cells under hypoxic conditions.

View Article: PubMed Central - PubMed

Affiliation: Wafic Said Molecular Cardiology Research Laboratory, Texas Heart Institute, P.O. Box 20345, MC 2-255, Houston, Texas 77225-0345, USA.

ABSTRACT
Promoting the paracrine effects of human mesenchymal stem cell (hMSC) therapy may contribute to improvements in patient outcomes. Here we develop an innovative strategy to enhance the paracrine effects of hMSCs. In a mouse hindlimb ischaemia model, we examine the effects of hMSCs in which a novel triple-catalytic enzyme is introduced to stably produce prostacyclin (PGI2-hMSCs). We show that PGI2-hMSCs facilitate perfusion recovery and enhance running capability as compared with control hMSCs or iloprost (a stable PGI2 analogue). Transplanted PGI2-hMSCs do not incorporate long term into host tissue, but rather they mediate host regeneration and muscle mass gain in a paracrine manner. Mechanistically, this involves long noncoding RNA H19 in promoting PGI2-hMSC-associated survival and proliferation of host progenitor cells under hypoxic conditions. Together, our data reveal the novel ability of PGI2-hMSCs to stimulate host regenerative processes and improve physical function by regulating long noncoding RNA in resident progenitor cells.

No MeSH data available.


Related in: MedlinePlus

Significantly higher numbers of endogenous proliferating Ki67+Sca-1+ cells were located adjacent to the PGI2-hMSC site 3 days after cell injections.(a) Representative confocal images illustrate the distribution of endogenous Ki67+Sca-1+ (yellow arrows) and Ki67+Sca-1−cells (purple arrows) in ischaemic gastrocnemius muscle areas surrounding injected hMSCs. (b) Quantitative analysis indicated a significantly higher percentage of Ki67+Sca-1+cells surrounding PGI2-hMSCs injection sites as compared with 3.1-MSCs+ILO or to 3.1-hMSCs sites (**P<0.01; one-way ANOVA with Newman–Keuls post hoc test). (c) Significantly higher percentage of Ki67+Sca-1−cells surrounded 3.1-MSCs+ILO injection sites than 3.1-hMSC injection sites (*P<0.05; one-way ANOVA with Newman–Keuls post hoc test). No significant differences were seen between 3.1-MSCs+ILO and PGI2-hMSC treatment or between PGI2-hMSC and 3.1-hMSC treatment. Data are shown as mean±s.e.m. A total of 30 high-power fields (HPFs) from three mice (male and female) per treatment group (10 HPFs per mouse, see details in Method) was used. Mice were sex matched among the three treatment groups. Scale bar, 25 μm.
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f5: Significantly higher numbers of endogenous proliferating Ki67+Sca-1+ cells were located adjacent to the PGI2-hMSC site 3 days after cell injections.(a) Representative confocal images illustrate the distribution of endogenous Ki67+Sca-1+ (yellow arrows) and Ki67+Sca-1−cells (purple arrows) in ischaemic gastrocnemius muscle areas surrounding injected hMSCs. (b) Quantitative analysis indicated a significantly higher percentage of Ki67+Sca-1+cells surrounding PGI2-hMSCs injection sites as compared with 3.1-MSCs+ILO or to 3.1-hMSCs sites (**P<0.01; one-way ANOVA with Newman–Keuls post hoc test). (c) Significantly higher percentage of Ki67+Sca-1−cells surrounded 3.1-MSCs+ILO injection sites than 3.1-hMSC injection sites (*P<0.05; one-way ANOVA with Newman–Keuls post hoc test). No significant differences were seen between 3.1-MSCs+ILO and PGI2-hMSC treatment or between PGI2-hMSC and 3.1-hMSC treatment. Data are shown as mean±s.e.m. A total of 30 high-power fields (HPFs) from three mice (male and female) per treatment group (10 HPFs per mouse, see details in Method) was used. Mice were sex matched among the three treatment groups. Scale bar, 25 μm.

Mentions: To determine whether PGI2-hMSCs induced an endogenous cellular response, we examined their effects in situ during hindlimb ischaemia. Immunofluorescence staining of gastrocnemius muscle obtained 3 days after cell delivery showed the presence of cells positive for the proliferation-associated nuclear protein Ki67. Ki67+ cells tended to localize in areas adjacent to the location of PGI2-hMSCs (Fig. 4) and were rarely observed in regions further away from PGI2-hMSCs (>250 μm distance). Notably, most Ki67+ cells expressed stem cell antigen-1 (Sca-1; Fig. 5a), a common marker on a variety of stem/progenitor cells, including haematopoietic cells, mesenchymal stem cells, endothelial progenitor cells (EPCs) and a muscle side population of stem cells1011121314. In addition, we found a similar anatomical distribution of endogenous Ki67+ cells in 3.1-hMSCs+ILO- and 3.1-hMSC-treated mice. When we quantified cell numbers in areas surrounding injected hMSCs (125 × 125 μm2), the accumulation of Ki67+Sca-1+cells was about twofold higher in PGI2-hMSC-treated mice than in mice treated with 3.1-hMSCs+ILO or 3.1-hMSCs (P<0.01; one-way ANOVA; Fig. 5b). Because Sca-1− cell populations contain satellite cells that contribute to muscle regeneration15, we quantified Ki67+Sca-1−cells in the same samples. In contrast to our results with Ki67+Sca-1+ cells, we found significantly higher numbers of Ki67+Sca-1−cells at 3.1-hMSCs+ILO injection sites than at 3.1-hMSC sites (P<0.05; one-way ANOVA; Fig. 5c). Thus, injections of PGI2-hMSCs and 3.1-hMSCs+ILO helped to create a focal area to foster proliferation of endogenous progenitor cells that led to muscle mass gain.


Prostacyclin-producing human mesenchymal cells target H19 lncRNA to augment endogenous progenitor function in hindlimb ischaemia.

Deng Y, Yang Z, Terry T, Pan S, Woodside DG, Wang J, Ruan K, Willerson JT, Dixon RA, Liu Q - Nat Commun (2016)

Significantly higher numbers of endogenous proliferating Ki67+Sca-1+ cells were located adjacent to the PGI2-hMSC site 3 days after cell injections.(a) Representative confocal images illustrate the distribution of endogenous Ki67+Sca-1+ (yellow arrows) and Ki67+Sca-1−cells (purple arrows) in ischaemic gastrocnemius muscle areas surrounding injected hMSCs. (b) Quantitative analysis indicated a significantly higher percentage of Ki67+Sca-1+cells surrounding PGI2-hMSCs injection sites as compared with 3.1-MSCs+ILO or to 3.1-hMSCs sites (**P<0.01; one-way ANOVA with Newman–Keuls post hoc test). (c) Significantly higher percentage of Ki67+Sca-1−cells surrounded 3.1-MSCs+ILO injection sites than 3.1-hMSC injection sites (*P<0.05; one-way ANOVA with Newman–Keuls post hoc test). No significant differences were seen between 3.1-MSCs+ILO and PGI2-hMSC treatment or between PGI2-hMSC and 3.1-hMSC treatment. Data are shown as mean±s.e.m. A total of 30 high-power fields (HPFs) from three mice (male and female) per treatment group (10 HPFs per mouse, see details in Method) was used. Mice were sex matched among the three treatment groups. Scale bar, 25 μm.
© Copyright Policy - open-access
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4835554&req=5

f5: Significantly higher numbers of endogenous proliferating Ki67+Sca-1+ cells were located adjacent to the PGI2-hMSC site 3 days after cell injections.(a) Representative confocal images illustrate the distribution of endogenous Ki67+Sca-1+ (yellow arrows) and Ki67+Sca-1−cells (purple arrows) in ischaemic gastrocnemius muscle areas surrounding injected hMSCs. (b) Quantitative analysis indicated a significantly higher percentage of Ki67+Sca-1+cells surrounding PGI2-hMSCs injection sites as compared with 3.1-MSCs+ILO or to 3.1-hMSCs sites (**P<0.01; one-way ANOVA with Newman–Keuls post hoc test). (c) Significantly higher percentage of Ki67+Sca-1−cells surrounded 3.1-MSCs+ILO injection sites than 3.1-hMSC injection sites (*P<0.05; one-way ANOVA with Newman–Keuls post hoc test). No significant differences were seen between 3.1-MSCs+ILO and PGI2-hMSC treatment or between PGI2-hMSC and 3.1-hMSC treatment. Data are shown as mean±s.e.m. A total of 30 high-power fields (HPFs) from three mice (male and female) per treatment group (10 HPFs per mouse, see details in Method) was used. Mice were sex matched among the three treatment groups. Scale bar, 25 μm.
Mentions: To determine whether PGI2-hMSCs induced an endogenous cellular response, we examined their effects in situ during hindlimb ischaemia. Immunofluorescence staining of gastrocnemius muscle obtained 3 days after cell delivery showed the presence of cells positive for the proliferation-associated nuclear protein Ki67. Ki67+ cells tended to localize in areas adjacent to the location of PGI2-hMSCs (Fig. 4) and were rarely observed in regions further away from PGI2-hMSCs (>250 μm distance). Notably, most Ki67+ cells expressed stem cell antigen-1 (Sca-1; Fig. 5a), a common marker on a variety of stem/progenitor cells, including haematopoietic cells, mesenchymal stem cells, endothelial progenitor cells (EPCs) and a muscle side population of stem cells1011121314. In addition, we found a similar anatomical distribution of endogenous Ki67+ cells in 3.1-hMSCs+ILO- and 3.1-hMSC-treated mice. When we quantified cell numbers in areas surrounding injected hMSCs (125 × 125 μm2), the accumulation of Ki67+Sca-1+cells was about twofold higher in PGI2-hMSC-treated mice than in mice treated with 3.1-hMSCs+ILO or 3.1-hMSCs (P<0.01; one-way ANOVA; Fig. 5b). Because Sca-1− cell populations contain satellite cells that contribute to muscle regeneration15, we quantified Ki67+Sca-1−cells in the same samples. In contrast to our results with Ki67+Sca-1+ cells, we found significantly higher numbers of Ki67+Sca-1−cells at 3.1-hMSCs+ILO injection sites than at 3.1-hMSC sites (P<0.05; one-way ANOVA; Fig. 5c). Thus, injections of PGI2-hMSCs and 3.1-hMSCs+ILO helped to create a focal area to foster proliferation of endogenous progenitor cells that led to muscle mass gain.

Bottom Line: Here we develop an innovative strategy to enhance the paracrine effects of hMSCs.Transplanted PGI2-hMSCs do not incorporate long term into host tissue, but rather they mediate host regeneration and muscle mass gain in a paracrine manner.Mechanistically, this involves long noncoding RNA H19 in promoting PGI2-hMSC-associated survival and proliferation of host progenitor cells under hypoxic conditions.

View Article: PubMed Central - PubMed

Affiliation: Wafic Said Molecular Cardiology Research Laboratory, Texas Heart Institute, P.O. Box 20345, MC 2-255, Houston, Texas 77225-0345, USA.

ABSTRACT
Promoting the paracrine effects of human mesenchymal stem cell (hMSC) therapy may contribute to improvements in patient outcomes. Here we develop an innovative strategy to enhance the paracrine effects of hMSCs. In a mouse hindlimb ischaemia model, we examine the effects of hMSCs in which a novel triple-catalytic enzyme is introduced to stably produce prostacyclin (PGI2-hMSCs). We show that PGI2-hMSCs facilitate perfusion recovery and enhance running capability as compared with control hMSCs or iloprost (a stable PGI2 analogue). Transplanted PGI2-hMSCs do not incorporate long term into host tissue, but rather they mediate host regeneration and muscle mass gain in a paracrine manner. Mechanistically, this involves long noncoding RNA H19 in promoting PGI2-hMSC-associated survival and proliferation of host progenitor cells under hypoxic conditions. Together, our data reveal the novel ability of PGI2-hMSCs to stimulate host regenerative processes and improve physical function by regulating long noncoding RNA in resident progenitor cells.

No MeSH data available.


Related in: MedlinePlus