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Defective Self-Renewal and Differentiation of GBA-Deficient Neural Stem Cells Can Be Restored By Macrophage Colony-Stimulating Factor.

Lee H, Bae JS, Jin HK - Mol. Cells (2015)

Bottom Line: We found that neural stem cells (NSCs) derived from a neuronopathic GD model exhibited decreased ability for self-renewal and neuronal differentiation.Enhanced proliferation and neuronal differentiation of GBA-deficient NSCs was associated with elevated release of macrophage colony-stimulating factor (M-CSF) from BM-MSCs.Our findings suggest that soluble M-CSF derived from BM-MSCs can modulate GBA-deficient NSCs, resulting in their improved proliferation and neuronal differentiation.

View Article: PubMed Central - PubMed

Affiliation: Stem Cell Neuroplasticity Research Group, Cell and Matrix Research Institute, College of Veterinary Medicine, Kyungpook National University, Daegu 702-701, Korea.

ABSTRACT
Gaucher disease (GD) is an autosomal recessive lysosomal storage disorder caused by mutations in the glucocerebrosidase gene (GBA), which encodes the lysosomal enzyme glucosylceramidase (GCase). Deficiency in GCase leads to characteristic visceral pathology and lethal neurological manifestations in some patients. Investigations into neurogenesis have suggested that neurodegenerative disorders, such as GD, could be overcome or at least ameliorated by the generation of new neurons. Bone marrow-derived mesenchymal stem cells (BM-MSCs) are potential candidates for use in the treatment of neurodegenerative disorders because of their ability to promote neurogenesis. Our objective was to examine the mechanism of neurogenesis by BM-MSCs in GD. We found that neural stem cells (NSCs) derived from a neuronopathic GD model exhibited decreased ability for self-renewal and neuronal differentiation. Co-culture of GBA-deficient NSCs with BM-MSCs resulted in an enhanced capacity for self-renewal, and an increased ability for differentiation into neurons or oligodendrocytes. Enhanced proliferation and neuronal differentiation of GBA-deficient NSCs was associated with elevated release of macrophage colony-stimulating factor (M-CSF) from BM-MSCs. Our findings suggest that soluble M-CSF derived from BM-MSCs can modulate GBA-deficient NSCs, resulting in their improved proliferation and neuronal differentiation.

No MeSH data available.


Related in: MedlinePlus

BM-MSCs increased neurotrophic factor signaling. (A) A cytokine antibody array was incubated with conditioned media recovered from three-dimensional cultures of BM-MSC-treated NSCs or untreated NSCs. Co-culture of Gba−/− NSCs with BM-MSCs resulted in the upregulation of M-CSF, IL-1ra, MCP-1, MMP-2, and MMP-3 (red squared spots). Densitometry analyses of cytokine/chemokine signals were normalized to positive and negative antibody array controls for Gba+/− NSCs. (B) Analysis using qPCR assays revealed greater transcriptional down-regulation of M-CSF in NSCs from Gba−/− mice compared with that in Gba+/− mice (n = 4 per group). *p < 0.05 compared with the Gba+/− controls. (C) ELISA analysis was used to assess M-CSF expression in NSCs. M-CSF expression was reduced to a greater extent in NSCs of Gba−/− mice compared with that in Gba+/− NSCs. Gba−/− NSCs co-cultured with BM-MSCs exhibited greater M-CSF expression levels compared with those in Gba−/− NSCs that were not co-cultured with BM-MSCs (n = 5 mice per group). All data are presented as the mean ± SEM. *p < 0.05.
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f4-molce-38-9-806: BM-MSCs increased neurotrophic factor signaling. (A) A cytokine antibody array was incubated with conditioned media recovered from three-dimensional cultures of BM-MSC-treated NSCs or untreated NSCs. Co-culture of Gba−/− NSCs with BM-MSCs resulted in the upregulation of M-CSF, IL-1ra, MCP-1, MMP-2, and MMP-3 (red squared spots). Densitometry analyses of cytokine/chemokine signals were normalized to positive and negative antibody array controls for Gba+/− NSCs. (B) Analysis using qPCR assays revealed greater transcriptional down-regulation of M-CSF in NSCs from Gba−/− mice compared with that in Gba+/− mice (n = 4 per group). *p < 0.05 compared with the Gba+/− controls. (C) ELISA analysis was used to assess M-CSF expression in NSCs. M-CSF expression was reduced to a greater extent in NSCs of Gba−/− mice compared with that in Gba+/− NSCs. Gba−/− NSCs co-cultured with BM-MSCs exhibited greater M-CSF expression levels compared with those in Gba−/− NSCs that were not co-cultured with BM-MSCs (n = 5 mice per group). All data are presented as the mean ± SEM. *p < 0.05.

Mentions: Our observations indicate that soluble bioactive factors secreted from BM-MSCs could promote the proliferation of Gba−/− NSCs. To identify these factors, we screened and compared the conditioned media of the various Gba+/− and Gba−/− NSCs cultures for 50 different secreted cytokines, using an antibody-based mouse cytokine array (Fig. 4A). Expression levels of M-CSF, IL-1ra, MCP-1, MMP-2 and MMP-3 were greater in the conditioned media of Gba−/− NSCs co-cultured with BM-MSCs than in the conditioned media of NSCs without co-culture (Fig. 4A). These results suggest the involvement of M-CSF, IL-1ra, MCP-1, MMP-2, and MMP-3 with respect to the self-renewal of Gba−/− NSCs co-cultured with BM-MSCs. Using qPCR assays, we examined mRNA expression levels of Mcsf, IL-1ra, MCP-1, Mmp2, and Mmp3 in Gba+/− and Gba−/− NSCs. Compared with controls, expression of Mcsf was significantly decreased in Gba−/− NSCs (Fig. 4B). Levels of IL-1ra, MCP-1, Mmp2, and Mmp3 were increased in Gba−/− NSCs compared with those in Gba+/− NSCs. We suspect that reduced expression of M-CSF might lead to the impaired self-renewal observed in Gba−/− NSCs. M-CSF derived from BM-MSCs is a potential candidate molecule for alleviating this defect. To confirm the secretion of this factor, we performed ELISAs and found that M-CSF levels were significantly elevated in the conditioned media of Gba−/− NSCs co-cultured with BM-MSCs (Fig. 4C). We also found that M-CSF levels were significantly decreased in Gba−/− NSCs compared with those in Gba+/− NSCs (Fig. 4C), neither of which were co-cultured with BM-MSCs.


Defective Self-Renewal and Differentiation of GBA-Deficient Neural Stem Cells Can Be Restored By Macrophage Colony-Stimulating Factor.

Lee H, Bae JS, Jin HK - Mol. Cells (2015)

BM-MSCs increased neurotrophic factor signaling. (A) A cytokine antibody array was incubated with conditioned media recovered from three-dimensional cultures of BM-MSC-treated NSCs or untreated NSCs. Co-culture of Gba−/− NSCs with BM-MSCs resulted in the upregulation of M-CSF, IL-1ra, MCP-1, MMP-2, and MMP-3 (red squared spots). Densitometry analyses of cytokine/chemokine signals were normalized to positive and negative antibody array controls for Gba+/− NSCs. (B) Analysis using qPCR assays revealed greater transcriptional down-regulation of M-CSF in NSCs from Gba−/− mice compared with that in Gba+/− mice (n = 4 per group). *p < 0.05 compared with the Gba+/− controls. (C) ELISA analysis was used to assess M-CSF expression in NSCs. M-CSF expression was reduced to a greater extent in NSCs of Gba−/− mice compared with that in Gba+/− NSCs. Gba−/− NSCs co-cultured with BM-MSCs exhibited greater M-CSF expression levels compared with those in Gba−/− NSCs that were not co-cultured with BM-MSCs (n = 5 mice per group). All data are presented as the mean ± SEM. *p < 0.05.
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Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4588724&req=5

f4-molce-38-9-806: BM-MSCs increased neurotrophic factor signaling. (A) A cytokine antibody array was incubated with conditioned media recovered from three-dimensional cultures of BM-MSC-treated NSCs or untreated NSCs. Co-culture of Gba−/− NSCs with BM-MSCs resulted in the upregulation of M-CSF, IL-1ra, MCP-1, MMP-2, and MMP-3 (red squared spots). Densitometry analyses of cytokine/chemokine signals were normalized to positive and negative antibody array controls for Gba+/− NSCs. (B) Analysis using qPCR assays revealed greater transcriptional down-regulation of M-CSF in NSCs from Gba−/− mice compared with that in Gba+/− mice (n = 4 per group). *p < 0.05 compared with the Gba+/− controls. (C) ELISA analysis was used to assess M-CSF expression in NSCs. M-CSF expression was reduced to a greater extent in NSCs of Gba−/− mice compared with that in Gba+/− NSCs. Gba−/− NSCs co-cultured with BM-MSCs exhibited greater M-CSF expression levels compared with those in Gba−/− NSCs that were not co-cultured with BM-MSCs (n = 5 mice per group). All data are presented as the mean ± SEM. *p < 0.05.
Mentions: Our observations indicate that soluble bioactive factors secreted from BM-MSCs could promote the proliferation of Gba−/− NSCs. To identify these factors, we screened and compared the conditioned media of the various Gba+/− and Gba−/− NSCs cultures for 50 different secreted cytokines, using an antibody-based mouse cytokine array (Fig. 4A). Expression levels of M-CSF, IL-1ra, MCP-1, MMP-2 and MMP-3 were greater in the conditioned media of Gba−/− NSCs co-cultured with BM-MSCs than in the conditioned media of NSCs without co-culture (Fig. 4A). These results suggest the involvement of M-CSF, IL-1ra, MCP-1, MMP-2, and MMP-3 with respect to the self-renewal of Gba−/− NSCs co-cultured with BM-MSCs. Using qPCR assays, we examined mRNA expression levels of Mcsf, IL-1ra, MCP-1, Mmp2, and Mmp3 in Gba+/− and Gba−/− NSCs. Compared with controls, expression of Mcsf was significantly decreased in Gba−/− NSCs (Fig. 4B). Levels of IL-1ra, MCP-1, Mmp2, and Mmp3 were increased in Gba−/− NSCs compared with those in Gba+/− NSCs. We suspect that reduced expression of M-CSF might lead to the impaired self-renewal observed in Gba−/− NSCs. M-CSF derived from BM-MSCs is a potential candidate molecule for alleviating this defect. To confirm the secretion of this factor, we performed ELISAs and found that M-CSF levels were significantly elevated in the conditioned media of Gba−/− NSCs co-cultured with BM-MSCs (Fig. 4C). We also found that M-CSF levels were significantly decreased in Gba−/− NSCs compared with those in Gba+/− NSCs (Fig. 4C), neither of which were co-cultured with BM-MSCs.

Bottom Line: We found that neural stem cells (NSCs) derived from a neuronopathic GD model exhibited decreased ability for self-renewal and neuronal differentiation.Enhanced proliferation and neuronal differentiation of GBA-deficient NSCs was associated with elevated release of macrophage colony-stimulating factor (M-CSF) from BM-MSCs.Our findings suggest that soluble M-CSF derived from BM-MSCs can modulate GBA-deficient NSCs, resulting in their improved proliferation and neuronal differentiation.

View Article: PubMed Central - PubMed

Affiliation: Stem Cell Neuroplasticity Research Group, Cell and Matrix Research Institute, College of Veterinary Medicine, Kyungpook National University, Daegu 702-701, Korea.

ABSTRACT
Gaucher disease (GD) is an autosomal recessive lysosomal storage disorder caused by mutations in the glucocerebrosidase gene (GBA), which encodes the lysosomal enzyme glucosylceramidase (GCase). Deficiency in GCase leads to characteristic visceral pathology and lethal neurological manifestations in some patients. Investigations into neurogenesis have suggested that neurodegenerative disorders, such as GD, could be overcome or at least ameliorated by the generation of new neurons. Bone marrow-derived mesenchymal stem cells (BM-MSCs) are potential candidates for use in the treatment of neurodegenerative disorders because of their ability to promote neurogenesis. Our objective was to examine the mechanism of neurogenesis by BM-MSCs in GD. We found that neural stem cells (NSCs) derived from a neuronopathic GD model exhibited decreased ability for self-renewal and neuronal differentiation. Co-culture of GBA-deficient NSCs with BM-MSCs resulted in an enhanced capacity for self-renewal, and an increased ability for differentiation into neurons or oligodendrocytes. Enhanced proliferation and neuronal differentiation of GBA-deficient NSCs was associated with elevated release of macrophage colony-stimulating factor (M-CSF) from BM-MSCs. Our findings suggest that soluble M-CSF derived from BM-MSCs can modulate GBA-deficient NSCs, resulting in their improved proliferation and neuronal differentiation.

No MeSH data available.


Related in: MedlinePlus