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The Role of Genetically Modified Mesenchymal Stem Cells in Urinary Bladder Regeneration.

Snow-Lisy DC, Diaz EC, Bury MI, Fuller NJ, Hannick JH, Ahmad N, Sharma AK - PLoS ONE (2015)

Bottom Line: Muscle content decreased to ~25% at 10 weeks in Cyr61KD groups.Wnt5aOX resulted in high numbers of vessels and muscle content (~206 vessels/mm2 and ~51%, respectively) at 4 weeks.Over-expressing cell constructs resulted in peripheral nerve regeneration while Cyr61KD animals were devoid of peripheral nerve regeneration at 4 weeks.

View Article: PubMed Central - PubMed

Affiliation: Ann & Robert H. Lurie Children's Hospital of Chicago, Division of Pediatric Urology, Chicago, IL, United States of America.

ABSTRACT
Recent studies have demonstrated that mesenchymal stem cells (MSCs) combined with CD34+ hematopoietic/stem progenitor cells (HSPCs) can function as surrogate urinary bladder cells to synergistically promote multi-faceted bladder tissue regeneration. However, the molecular pathways governing these events are unknown. The pleiotropic effects of Wnt5a and Cyr61 are known to affect aspects of hematopoiesis, angiogenesis, and muscle and nerve regeneration. Within this study, the effects of Cyr61 and Wnt5a on bladder tissue regeneration were evaluated by grafting scaffolds containing modified human bone marrow derived MSCs. These cell lines were engineered to independently over-express Wnt5a or Cyr61, or to exhibit reduced expression of Cyr61 within the context of a nude rat bladder augmentation model. At 4 weeks post-surgery, data demonstrated increased vessel number (~250 vs ~109 vessels/mm2) and bladder smooth muscle content (~42% vs ~36%) in Cyr61OX (over-expressing) vs Cyr61KD (knock-down) groups. Muscle content decreased to ~25% at 10 weeks in Cyr61KD groups. Wnt5aOX resulted in high numbers of vessels and muscle content (~206 vessels/mm2 and ~51%, respectively) at 4 weeks. Over-expressing cell constructs resulted in peripheral nerve regeneration while Cyr61KD animals were devoid of peripheral nerve regeneration at 4 weeks. At 10 weeks post-grafting, peripheral nerve regeneration was at a minimal level for both Cyr61OX and Wnt5aOX cell lines. Blood vessel and bladder functionality were evident at both time-points in all animals. Results from this study indicate that MSC-based Cyr61OX and Wnt5aOX cell lines play pivotal roles with regards to increasing the levels of functional vasculature, influencing muscle regeneration, and the regeneration of peripheral nerves in a model of bladder augmentation. Wnt5aOX constructs closely approximated the outcomes previously observed with the co-transplantation of MSCs with CD34+ HSPCs and may be specifically targeted as an alternate means to achieve functional bladder regeneration.

No MeSH data available.


Related in: MedlinePlus

Effects of Cyr61 and Wnt5a on regenerating vasculature and musculature.(A) Cyr61OX and Wnt5aOX showed early and persistent increased vasculature, comparably greater than unmanipulated MSCs, with numbers of vessels/mm2 similar to MSC/CD34+ HSPC grafts at 4 weeks. (§ Unseeded, MSC and MSC/CD34+ HSPC data, shown as solid and dotted lines, previously reported [7]). At both 4 and 10 weeks, Cyr61KD demonstrated significantly decreased vasculature as compared to Cyr61OX. Data shown as means ± SEM; ***P<0.001 for Cyr61KD vs Cyr61OX. (B) Wnt5aOX demonstrated mean muscle content most comparable to MSC/CD34+ HSPC grafts [percent red (muscle): (red (muscle) pixels / red (muscle) + blue (collagen) pixels)]. (§ Unseeded, MSC and MSC/CD34+ HSPC data, shown as solid and dotted lines, previously reported [7]). Cyr61KD demonstrated a significant decrease in muscle percentage from 4 to 10 weeks (p<0.05), with levels significantly lower than Cyr61OX at both time points. Data shown as means ± SEM; *P<0.05 and **P<0.01 for Cyr61KD vs Cyr61OX. (C) Sample photomicrographs of Masson’s trichrome staining demonstrate differences in muscle/collagen content and level of vasculature (marked with black arrows) between Cyr61KD and the Cyr61OX and Wnt5aOX groups. Density score was greatest for Wnt5aOX at 4 weeks. Scale bar, 50 μm. (Unseeded, MSC and MSC/CD34+ HSPC images shown in S1 Fig).
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pone.0138643.g002: Effects of Cyr61 and Wnt5a on regenerating vasculature and musculature.(A) Cyr61OX and Wnt5aOX showed early and persistent increased vasculature, comparably greater than unmanipulated MSCs, with numbers of vessels/mm2 similar to MSC/CD34+ HSPC grafts at 4 weeks. (§ Unseeded, MSC and MSC/CD34+ HSPC data, shown as solid and dotted lines, previously reported [7]). At both 4 and 10 weeks, Cyr61KD demonstrated significantly decreased vasculature as compared to Cyr61OX. Data shown as means ± SEM; ***P<0.001 for Cyr61KD vs Cyr61OX. (B) Wnt5aOX demonstrated mean muscle content most comparable to MSC/CD34+ HSPC grafts [percent red (muscle): (red (muscle) pixels / red (muscle) + blue (collagen) pixels)]. (§ Unseeded, MSC and MSC/CD34+ HSPC data, shown as solid and dotted lines, previously reported [7]). Cyr61KD demonstrated a significant decrease in muscle percentage from 4 to 10 weeks (p<0.05), with levels significantly lower than Cyr61OX at both time points. Data shown as means ± SEM; *P<0.05 and **P<0.01 for Cyr61KD vs Cyr61OX. (C) Sample photomicrographs of Masson’s trichrome staining demonstrate differences in muscle/collagen content and level of vasculature (marked with black arrows) between Cyr61KD and the Cyr61OX and Wnt5aOX groups. Density score was greatest for Wnt5aOX at 4 weeks. Scale bar, 50 μm. (Unseeded, MSC and MSC/CD34+ HSPC images shown in S1 Fig).

Mentions: Both increasing and decreasing Cyr61 expression resulted in greater levels of graft vasculature than previously observed with unmanipulated MSCs [7] in which increasing Cyr61 expression produced a much stronger effect. At 4 weeks, Cyr61KD vessel number and percent vasculature were 1.3x and 1.5x higher than MSC (108.8±5.5 vs 83.4±15.8 vessels/mm2 and 2.25±0.27% vs 1.46±0.16%) while Cyr61OX vessel number and percent vasculature were 3x and 4x higher than MSC (249.9±22.3 vs 83.4±15.8 vessels/mm2 and 5.78±0.29% vs 1.46±0.16%) (Fig 2A). For unmanipulated MSCs and Cyr61KD, there was no observed effect of graft duration (4 vs 10 weeks) on the level of vascularization. At 10 weeks, the number of vessels/mm2 remained stable for Cyr61OX but percentage vasculature increased (8.49±0.62% vs 5.78±0.24%, p<0.05), reflecting a shift towards larger vessels. Cyr61OX vessel number and percent vasculature were significantly greater than Cyr61KD at both time-points (p<0.001; p<0.001).


The Role of Genetically Modified Mesenchymal Stem Cells in Urinary Bladder Regeneration.

Snow-Lisy DC, Diaz EC, Bury MI, Fuller NJ, Hannick JH, Ahmad N, Sharma AK - PLoS ONE (2015)

Effects of Cyr61 and Wnt5a on regenerating vasculature and musculature.(A) Cyr61OX and Wnt5aOX showed early and persistent increased vasculature, comparably greater than unmanipulated MSCs, with numbers of vessels/mm2 similar to MSC/CD34+ HSPC grafts at 4 weeks. (§ Unseeded, MSC and MSC/CD34+ HSPC data, shown as solid and dotted lines, previously reported [7]). At both 4 and 10 weeks, Cyr61KD demonstrated significantly decreased vasculature as compared to Cyr61OX. Data shown as means ± SEM; ***P<0.001 for Cyr61KD vs Cyr61OX. (B) Wnt5aOX demonstrated mean muscle content most comparable to MSC/CD34+ HSPC grafts [percent red (muscle): (red (muscle) pixels / red (muscle) + blue (collagen) pixels)]. (§ Unseeded, MSC and MSC/CD34+ HSPC data, shown as solid and dotted lines, previously reported [7]). Cyr61KD demonstrated a significant decrease in muscle percentage from 4 to 10 weeks (p<0.05), with levels significantly lower than Cyr61OX at both time points. Data shown as means ± SEM; *P<0.05 and **P<0.01 for Cyr61KD vs Cyr61OX. (C) Sample photomicrographs of Masson’s trichrome staining demonstrate differences in muscle/collagen content and level of vasculature (marked with black arrows) between Cyr61KD and the Cyr61OX and Wnt5aOX groups. Density score was greatest for Wnt5aOX at 4 weeks. Scale bar, 50 μm. (Unseeded, MSC and MSC/CD34+ HSPC images shown in S1 Fig).
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pone.0138643.g002: Effects of Cyr61 and Wnt5a on regenerating vasculature and musculature.(A) Cyr61OX and Wnt5aOX showed early and persistent increased vasculature, comparably greater than unmanipulated MSCs, with numbers of vessels/mm2 similar to MSC/CD34+ HSPC grafts at 4 weeks. (§ Unseeded, MSC and MSC/CD34+ HSPC data, shown as solid and dotted lines, previously reported [7]). At both 4 and 10 weeks, Cyr61KD demonstrated significantly decreased vasculature as compared to Cyr61OX. Data shown as means ± SEM; ***P<0.001 for Cyr61KD vs Cyr61OX. (B) Wnt5aOX demonstrated mean muscle content most comparable to MSC/CD34+ HSPC grafts [percent red (muscle): (red (muscle) pixels / red (muscle) + blue (collagen) pixels)]. (§ Unseeded, MSC and MSC/CD34+ HSPC data, shown as solid and dotted lines, previously reported [7]). Cyr61KD demonstrated a significant decrease in muscle percentage from 4 to 10 weeks (p<0.05), with levels significantly lower than Cyr61OX at both time points. Data shown as means ± SEM; *P<0.05 and **P<0.01 for Cyr61KD vs Cyr61OX. (C) Sample photomicrographs of Masson’s trichrome staining demonstrate differences in muscle/collagen content and level of vasculature (marked with black arrows) between Cyr61KD and the Cyr61OX and Wnt5aOX groups. Density score was greatest for Wnt5aOX at 4 weeks. Scale bar, 50 μm. (Unseeded, MSC and MSC/CD34+ HSPC images shown in S1 Fig).
Mentions: Both increasing and decreasing Cyr61 expression resulted in greater levels of graft vasculature than previously observed with unmanipulated MSCs [7] in which increasing Cyr61 expression produced a much stronger effect. At 4 weeks, Cyr61KD vessel number and percent vasculature were 1.3x and 1.5x higher than MSC (108.8±5.5 vs 83.4±15.8 vessels/mm2 and 2.25±0.27% vs 1.46±0.16%) while Cyr61OX vessel number and percent vasculature were 3x and 4x higher than MSC (249.9±22.3 vs 83.4±15.8 vessels/mm2 and 5.78±0.29% vs 1.46±0.16%) (Fig 2A). For unmanipulated MSCs and Cyr61KD, there was no observed effect of graft duration (4 vs 10 weeks) on the level of vascularization. At 10 weeks, the number of vessels/mm2 remained stable for Cyr61OX but percentage vasculature increased (8.49±0.62% vs 5.78±0.24%, p<0.05), reflecting a shift towards larger vessels. Cyr61OX vessel number and percent vasculature were significantly greater than Cyr61KD at both time-points (p<0.001; p<0.001).

Bottom Line: Muscle content decreased to ~25% at 10 weeks in Cyr61KD groups.Wnt5aOX resulted in high numbers of vessels and muscle content (~206 vessels/mm2 and ~51%, respectively) at 4 weeks.Over-expressing cell constructs resulted in peripheral nerve regeneration while Cyr61KD animals were devoid of peripheral nerve regeneration at 4 weeks.

View Article: PubMed Central - PubMed

Affiliation: Ann & Robert H. Lurie Children's Hospital of Chicago, Division of Pediatric Urology, Chicago, IL, United States of America.

ABSTRACT
Recent studies have demonstrated that mesenchymal stem cells (MSCs) combined with CD34+ hematopoietic/stem progenitor cells (HSPCs) can function as surrogate urinary bladder cells to synergistically promote multi-faceted bladder tissue regeneration. However, the molecular pathways governing these events are unknown. The pleiotropic effects of Wnt5a and Cyr61 are known to affect aspects of hematopoiesis, angiogenesis, and muscle and nerve regeneration. Within this study, the effects of Cyr61 and Wnt5a on bladder tissue regeneration were evaluated by grafting scaffolds containing modified human bone marrow derived MSCs. These cell lines were engineered to independently over-express Wnt5a or Cyr61, or to exhibit reduced expression of Cyr61 within the context of a nude rat bladder augmentation model. At 4 weeks post-surgery, data demonstrated increased vessel number (~250 vs ~109 vessels/mm2) and bladder smooth muscle content (~42% vs ~36%) in Cyr61OX (over-expressing) vs Cyr61KD (knock-down) groups. Muscle content decreased to ~25% at 10 weeks in Cyr61KD groups. Wnt5aOX resulted in high numbers of vessels and muscle content (~206 vessels/mm2 and ~51%, respectively) at 4 weeks. Over-expressing cell constructs resulted in peripheral nerve regeneration while Cyr61KD animals were devoid of peripheral nerve regeneration at 4 weeks. At 10 weeks post-grafting, peripheral nerve regeneration was at a minimal level for both Cyr61OX and Wnt5aOX cell lines. Blood vessel and bladder functionality were evident at both time-points in all animals. Results from this study indicate that MSC-based Cyr61OX and Wnt5aOX cell lines play pivotal roles with regards to increasing the levels of functional vasculature, influencing muscle regeneration, and the regeneration of peripheral nerves in a model of bladder augmentation. Wnt5aOX constructs closely approximated the outcomes previously observed with the co-transplantation of MSCs with CD34+ HSPCs and may be specifically targeted as an alternate means to achieve functional bladder regeneration.

No MeSH data available.


Related in: MedlinePlus