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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

PGI2-hMSCs showed superior acute retention in ischaemic hindlimbs.(a) Representative in vivo bioluminescent images of NOD–SCID mice at indicated intervals after injection of vehicle (PBS), 3.1-hMSCs, PGI2-hMSCs or 3.1-hMSCs+ILO into the gastrocnemius muscle of the ischaemic hindlimbs. The minimal noninvasive visualized value was 1 × 106 photons s−1 cm−2 sr−1. (b) Quantification of maximal bioluminescence in cell-treated ischaemic hindlimbs at the indicated time. *P<0.05, PGI2-hMSCs versus 3.1-hMSCs or versus 3.1-hMSCs+ILO by a one-way ANOVA with Newman–Keuls post hoc test. Data are shown as mean±s.e.m. N=5 mice (male and female) per treatment group. All treatment groups were sex matched. (c–e) hMSCs were not detected in the heart, spleen, lung, kidney or liver at 14 days after cell delivery.
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f2: PGI2-hMSCs showed superior acute retention in ischaemic hindlimbs.(a) Representative in vivo bioluminescent images of NOD–SCID mice at indicated intervals after injection of vehicle (PBS), 3.1-hMSCs, PGI2-hMSCs or 3.1-hMSCs+ILO into the gastrocnemius muscle of the ischaemic hindlimbs. The minimal noninvasive visualized value was 1 × 106 photons s−1 cm−2 sr−1. (b) Quantification of maximal bioluminescence in cell-treated ischaemic hindlimbs at the indicated time. *P<0.05, PGI2-hMSCs versus 3.1-hMSCs or versus 3.1-hMSCs+ILO by a one-way ANOVA with Newman–Keuls post hoc test. Data are shown as mean±s.e.m. N=5 mice (male and female) per treatment group. All treatment groups were sex matched. (c–e) hMSCs were not detected in the heart, spleen, lung, kidney or liver at 14 days after cell delivery.

Mentions: The benefit of PGI2-hMSC therapy could stem from their long-term superior engraftment in ischaemic hindlimbs when compared with 3.1-hMSCs. To track the fate of exogenously delivered cells longitudinally, we transduced hMSCs with a lentiviral construct that contains the triple-fusion reporter genes of human herpes simplex virus type-1-thymidine kinase, mCherry fluorophore and firefly luciferase (Supplementary Fig. 2). Luciferase catalyses light-emitting photochemical reactions of luciferin in live cells, which enables whole-body imaging to track the distribution and engraftment of transplanted cells. We performed a lentiviral infection of hMSCs 3 days before producing PGI2-hMSCs and 3.1-hMSCs, in parallel, to ensure that the lentiviral transduction efficiency was the same in both cell types. In vitro luciferase assays with lentivirus-infected hMSCs indicated that bioluminescent intensity readings correlated positively with cell numbers (Supplementary Fig. 2). Immediately after injecting luciferin into the mice, we recorded sequential readings until the maximal signal intensity was reached. During the measuring time after cell administration, whole-body images showed that bioluminescence was seen only in ischaemic hindlimbs. No signals above background level were detected in other organs/tissues or in vehicle-treated mice (Fig. 2a). At day 1 after cell injection, we found a higher maximal bioluminescent intensity in the ischaemic hindlimbs of PGI2-hMSC-treated mice compared with those that received 3.1-hMSCs or 3.1-hMSCs+ILO (5.14±2.16 × 107 versus 1.75±0.29 × 107 or 1.63±0.50 × 107 photons s−1 cm−2 sr−1, respectively; Fig. 2b). The bioluminescent signal in the PGI2-hMSC-treated group peaked at day 3 (12.20±3.05 × 107 photons s−1 cm−2 sr−1) and was significantly higher than that seen in mice treated with 3.1-hMSCs or 3.1-hMSCs+ILO (2.49±0.58 × 107 and 2.40±1.10 × 107 photons s−1 cm−2 sr−1, respectively; P<0.05; one-way ANOVA). The signal from the PGI2-hMSC-treated group began to decrease by day 5 (5.74±2.82 × 107 versus 1.08±0.11 × 107 or versus 0.80±0.24 × 107 photons s−1 cm−2 sr−1 in PGI2-hMSC, 3.1-hMSC and 3.1-hMSCs+ILO-treated mice, respectively). At day 14, we detected a large reduction in the signal to close to background levels (1.00 × 105 photons s−1 cm−2 sr−1) in the readings in each group. This exponential decline of bioluminescence indicated the sharp loss of hMSCs at 14 days post injection.


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)

PGI2-hMSCs showed superior acute retention in ischaemic hindlimbs.(a) Representative in vivo bioluminescent images of NOD–SCID mice at indicated intervals after injection of vehicle (PBS), 3.1-hMSCs, PGI2-hMSCs or 3.1-hMSCs+ILO into the gastrocnemius muscle of the ischaemic hindlimbs. The minimal noninvasive visualized value was 1 × 106 photons s−1 cm−2 sr−1. (b) Quantification of maximal bioluminescence in cell-treated ischaemic hindlimbs at the indicated time. *P<0.05, PGI2-hMSCs versus 3.1-hMSCs or versus 3.1-hMSCs+ILO by a one-way ANOVA with Newman–Keuls post hoc test. Data are shown as mean±s.e.m. N=5 mice (male and female) per treatment group. All treatment groups were sex matched. (c–e) hMSCs were not detected in the heart, spleen, lung, kidney or liver at 14 days after cell delivery.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

f2: PGI2-hMSCs showed superior acute retention in ischaemic hindlimbs.(a) Representative in vivo bioluminescent images of NOD–SCID mice at indicated intervals after injection of vehicle (PBS), 3.1-hMSCs, PGI2-hMSCs or 3.1-hMSCs+ILO into the gastrocnemius muscle of the ischaemic hindlimbs. The minimal noninvasive visualized value was 1 × 106 photons s−1 cm−2 sr−1. (b) Quantification of maximal bioluminescence in cell-treated ischaemic hindlimbs at the indicated time. *P<0.05, PGI2-hMSCs versus 3.1-hMSCs or versus 3.1-hMSCs+ILO by a one-way ANOVA with Newman–Keuls post hoc test. Data are shown as mean±s.e.m. N=5 mice (male and female) per treatment group. All treatment groups were sex matched. (c–e) hMSCs were not detected in the heart, spleen, lung, kidney or liver at 14 days after cell delivery.
Mentions: The benefit of PGI2-hMSC therapy could stem from their long-term superior engraftment in ischaemic hindlimbs when compared with 3.1-hMSCs. To track the fate of exogenously delivered cells longitudinally, we transduced hMSCs with a lentiviral construct that contains the triple-fusion reporter genes of human herpes simplex virus type-1-thymidine kinase, mCherry fluorophore and firefly luciferase (Supplementary Fig. 2). Luciferase catalyses light-emitting photochemical reactions of luciferin in live cells, which enables whole-body imaging to track the distribution and engraftment of transplanted cells. We performed a lentiviral infection of hMSCs 3 days before producing PGI2-hMSCs and 3.1-hMSCs, in parallel, to ensure that the lentiviral transduction efficiency was the same in both cell types. In vitro luciferase assays with lentivirus-infected hMSCs indicated that bioluminescent intensity readings correlated positively with cell numbers (Supplementary Fig. 2). Immediately after injecting luciferin into the mice, we recorded sequential readings until the maximal signal intensity was reached. During the measuring time after cell administration, whole-body images showed that bioluminescence was seen only in ischaemic hindlimbs. No signals above background level were detected in other organs/tissues or in vehicle-treated mice (Fig. 2a). At day 1 after cell injection, we found a higher maximal bioluminescent intensity in the ischaemic hindlimbs of PGI2-hMSC-treated mice compared with those that received 3.1-hMSCs or 3.1-hMSCs+ILO (5.14±2.16 × 107 versus 1.75±0.29 × 107 or 1.63±0.50 × 107 photons s−1 cm−2 sr−1, respectively; Fig. 2b). The bioluminescent signal in the PGI2-hMSC-treated group peaked at day 3 (12.20±3.05 × 107 photons s−1 cm−2 sr−1) and was significantly higher than that seen in mice treated with 3.1-hMSCs or 3.1-hMSCs+ILO (2.49±0.58 × 107 and 2.40±1.10 × 107 photons s−1 cm−2 sr−1, respectively; P<0.05; one-way ANOVA). The signal from the PGI2-hMSC-treated group began to decrease by day 5 (5.74±2.82 × 107 versus 1.08±0.11 × 107 or versus 0.80±0.24 × 107 photons s−1 cm−2 sr−1 in PGI2-hMSC, 3.1-hMSC and 3.1-hMSCs+ILO-treated mice, respectively). At day 14, we detected a large reduction in the signal to close to background levels (1.00 × 105 photons s−1 cm−2 sr−1) in the readings in each group. This exponential decline of bioluminescence indicated the sharp loss of hMSCs at 14 days post injection.

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