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Aberrant gene expression profiles, during in vitro osteoblast differentiation, of telomerase deficient mouse bone marrow stromal stem cells (mBMSCs).

Saeed H, Iqtedar M - J. Biomed. Sci. (2015)

Bottom Line: However, the effect of telomerase deficiency on differential regulation of osteoblast specific genes, based on functional gene grouping, during in vitro osteoblast differentiation has not been reported before.To examine these effects, Terc (-/-) BMSCs (bone marrow stromal stem cells) were employed which exhibited reduced proliferation during in vitro osteogenesis along with increased population doubling time and level compared to wild type (WT) BMSCs during the normal culture.More profound changes were observed in genes engaged in extracellular matrix production: Col1a1, Col1a2, Mmp10, Serpinh1 and Col4a1.

View Article: PubMed Central - PubMed

Affiliation: Endocrine Research Laboratory, KMEB, Department of Endocrinology and Metabolism, Odense University Hospital, Odense, Denmark. hamid.pharmacy@pu.edu.pk.

ABSTRACT

Background: Telomerase deficiency has been associated with inadequate differentiation of mesenchymal stem cells. However, the effect of telomerase deficiency on differential regulation of osteoblast specific genes, based on functional gene grouping, during in vitro osteoblast differentiation has not been reported before.

Results: To examine these effects, Terc (-/-) BMSCs (bone marrow stromal stem cells) were employed which exhibited reduced proliferation during in vitro osteogenesis along with increased population doubling time and level compared to wild type (WT) BMSCs during the normal culture. Osteogenic super array at day 10 of osteoblast differentiation revealed that telomerase deficiency strongly affected the osteoblast commitment by down-regulating Runx2, Twist and Vdr - known transcription regulators of osteogenesis. Similarly, in Terc (-/-) BMSCs a marked reduction in other genes engaged in various phases of osteoblast differentiation were observed, such as Fgfr2 involved in bone mineralization, Phex and Dmp1 engaged in ossification, and Col11a1 and Col2a1 involved in cartilage condensation. A similar trend was observed for genes involved in osteoblast proliferation (Tgfb1, Fgfr2 and Pdgfa) and bone mineral metabolism (Col1a1, Col2a1, Col1a2 and Col11a1). More profound changes were observed in genes engaged in extracellular matrix production: Col1a1, Col1a2, Mmp10, Serpinh1 and Col4a1.

Conclusion: Taken together, these data suggest that telomerase deficiency causes impairment of BMSCs differentiation into osteoblasts affecting commitment, proliferation, matrix mineralization and maturation. Thus, modulating telomerase in BMSCs with advanced aging could improve BMSCs responsiveness towards osteoblast differentiation signals, optimal for osteoblast commitment, proliferation and maturation processes.

No MeSH data available.


Related in: MedlinePlus

Differential expression of genetic molecules involved in bone mineral metabolism and cell growth during differentiation inTerc-/-BMSCs. A & B) Differential expression of genes involved in bone mineral metabolism affecting calcium ion binding and phosphate transport in Terc-/- BMSCs compared to WT controls. C-D) Differential transcriptional profiles of Terc-/- BMSCs during in vitro osteoblast differentiation. Several genes with known involvement in osteoblast cell cycle, proliferation and differentiation were up-regulated (red bars) and down-regulated (green bars) in Terc-/- BMSCs compared to control. Genes that were not amplified or not detectable in the PCR array were marked as ‘n.d’ (not detectable). Osteogenic super array data are represented as fold change relative to WT controls of three independent biological replicates pooled together.
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Fig3: Differential expression of genetic molecules involved in bone mineral metabolism and cell growth during differentiation inTerc-/-BMSCs. A & B) Differential expression of genes involved in bone mineral metabolism affecting calcium ion binding and phosphate transport in Terc-/- BMSCs compared to WT controls. C-D) Differential transcriptional profiles of Terc-/- BMSCs during in vitro osteoblast differentiation. Several genes with known involvement in osteoblast cell cycle, proliferation and differentiation were up-regulated (red bars) and down-regulated (green bars) in Terc-/- BMSCs compared to control. Genes that were not amplified or not detectable in the PCR array were marked as ‘n.d’ (not detectable). Osteogenic super array data are represented as fold change relative to WT controls of three independent biological replicates pooled together.

Mentions: To study further, the effects of telomerase deficiency during in vitro osteoblast differentiation we performed Mouse Osteogenesis RT2 Profiler™ PCR Array as described in material and methods. Data analysis revealed defective osteoblast differentiation with differential expression of several genes at various phases of osteoblast differentiation. Briefly, major changes were observed in genes controlling extracellular matrix production which accounted for 40% of the genes, followed by cell growth and differentiation (24%), bone mineralization (16%), skeletal development (14%) and transcription factors and regulators (6%) respectively (Figure 2A). Moreover, during PCR super array several of the genes were not amplified or detectable and were marked as ‘n.d’ (not detectable) in the figures (Figures 2, 3, 4 and 5). Moreover, results are arranged and described according to functional gene grouping specified by super array bioscience cooperation.Figure 2


Aberrant gene expression profiles, during in vitro osteoblast differentiation, of telomerase deficient mouse bone marrow stromal stem cells (mBMSCs).

Saeed H, Iqtedar M - J. Biomed. Sci. (2015)

Differential expression of genetic molecules involved in bone mineral metabolism and cell growth during differentiation inTerc-/-BMSCs. A & B) Differential expression of genes involved in bone mineral metabolism affecting calcium ion binding and phosphate transport in Terc-/- BMSCs compared to WT controls. C-D) Differential transcriptional profiles of Terc-/- BMSCs during in vitro osteoblast differentiation. Several genes with known involvement in osteoblast cell cycle, proliferation and differentiation were up-regulated (red bars) and down-regulated (green bars) in Terc-/- BMSCs compared to control. Genes that were not amplified or not detectable in the PCR array were marked as ‘n.d’ (not detectable). Osteogenic super array data are represented as fold change relative to WT controls of three independent biological replicates pooled together.
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4318164&req=5

Fig3: Differential expression of genetic molecules involved in bone mineral metabolism and cell growth during differentiation inTerc-/-BMSCs. A & B) Differential expression of genes involved in bone mineral metabolism affecting calcium ion binding and phosphate transport in Terc-/- BMSCs compared to WT controls. C-D) Differential transcriptional profiles of Terc-/- BMSCs during in vitro osteoblast differentiation. Several genes with known involvement in osteoblast cell cycle, proliferation and differentiation were up-regulated (red bars) and down-regulated (green bars) in Terc-/- BMSCs compared to control. Genes that were not amplified or not detectable in the PCR array were marked as ‘n.d’ (not detectable). Osteogenic super array data are represented as fold change relative to WT controls of three independent biological replicates pooled together.
Mentions: To study further, the effects of telomerase deficiency during in vitro osteoblast differentiation we performed Mouse Osteogenesis RT2 Profiler™ PCR Array as described in material and methods. Data analysis revealed defective osteoblast differentiation with differential expression of several genes at various phases of osteoblast differentiation. Briefly, major changes were observed in genes controlling extracellular matrix production which accounted for 40% of the genes, followed by cell growth and differentiation (24%), bone mineralization (16%), skeletal development (14%) and transcription factors and regulators (6%) respectively (Figure 2A). Moreover, during PCR super array several of the genes were not amplified or detectable and were marked as ‘n.d’ (not detectable) in the figures (Figures 2, 3, 4 and 5). Moreover, results are arranged and described according to functional gene grouping specified by super array bioscience cooperation.Figure 2

Bottom Line: However, the effect of telomerase deficiency on differential regulation of osteoblast specific genes, based on functional gene grouping, during in vitro osteoblast differentiation has not been reported before.To examine these effects, Terc (-/-) BMSCs (bone marrow stromal stem cells) were employed which exhibited reduced proliferation during in vitro osteogenesis along with increased population doubling time and level compared to wild type (WT) BMSCs during the normal culture.More profound changes were observed in genes engaged in extracellular matrix production: Col1a1, Col1a2, Mmp10, Serpinh1 and Col4a1.

View Article: PubMed Central - PubMed

Affiliation: Endocrine Research Laboratory, KMEB, Department of Endocrinology and Metabolism, Odense University Hospital, Odense, Denmark. hamid.pharmacy@pu.edu.pk.

ABSTRACT

Background: Telomerase deficiency has been associated with inadequate differentiation of mesenchymal stem cells. However, the effect of telomerase deficiency on differential regulation of osteoblast specific genes, based on functional gene grouping, during in vitro osteoblast differentiation has not been reported before.

Results: To examine these effects, Terc (-/-) BMSCs (bone marrow stromal stem cells) were employed which exhibited reduced proliferation during in vitro osteogenesis along with increased population doubling time and level compared to wild type (WT) BMSCs during the normal culture. Osteogenic super array at day 10 of osteoblast differentiation revealed that telomerase deficiency strongly affected the osteoblast commitment by down-regulating Runx2, Twist and Vdr - known transcription regulators of osteogenesis. Similarly, in Terc (-/-) BMSCs a marked reduction in other genes engaged in various phases of osteoblast differentiation were observed, such as Fgfr2 involved in bone mineralization, Phex and Dmp1 engaged in ossification, and Col11a1 and Col2a1 involved in cartilage condensation. A similar trend was observed for genes involved in osteoblast proliferation (Tgfb1, Fgfr2 and Pdgfa) and bone mineral metabolism (Col1a1, Col2a1, Col1a2 and Col11a1). More profound changes were observed in genes engaged in extracellular matrix production: Col1a1, Col1a2, Mmp10, Serpinh1 and Col4a1.

Conclusion: Taken together, these data suggest that telomerase deficiency causes impairment of BMSCs differentiation into osteoblasts affecting commitment, proliferation, matrix mineralization and maturation. Thus, modulating telomerase in BMSCs with advanced aging could improve BMSCs responsiveness towards osteoblast differentiation signals, optimal for osteoblast commitment, proliferation and maturation processes.

No MeSH data available.


Related in: MedlinePlus