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Huntington disease iPSCs show early molecular changes in intracellular signaling, the expression of oxidative stress proteins and the p53 pathway.

Szlachcic WJ, Switonski PM, Krzyzosiak WJ, Figlerowicz M, Figiel M - Dis Model Mech (2015)

Bottom Line: Several molecular and developmental effects of HD have been identified using neural stem cells (NSCs) and differentiated cells, such as neurons and astrocytes.Surprisingly, we found that a number of changes affecting cellular processes in HD were also present in undifferentiated pluripotent HD iPSCs, including the dysregulation of the MAPK and Wnt signaling pathways and the dysregulation of the expression of genes related to oxidative stress, such as Sod1.Interestingly, a common protein interactor of the huntingtin protein and the proteins in the above pathways is p53, and the expression of p53 was dysregulated in HD YAC128 iPSCs and human HD iPSCs.

View Article: PubMed Central - PubMed

Affiliation: Institute of Bioorganic Chemistry, Polish Academy of Sciences, Noskowskiego 12/14, Poznań 61-704, Poland.

No MeSH data available.


Related in: MedlinePlus

The level of β-catenin phosphorylation was increased in YAC128 iPSCs. (A) The levels of total β-catenin, p-β-catenin (S33/37) and GAPDH were investigated in clonal lines of HD YAC128 (n=5) and wild-type (WT) iPSCs (n=6) using western blotting. (B) Quantification of the level of p-β-catenin (S33/37) showed increased phosphorylation in the HD YAC128 iPSC lines without changes in (C) the level of total β-catenin protein. (D) qPCR analysis revealed a trend toward the increased expression of Gsk3β, but the extent of the increase was not significant. * indicates the YAC128/Oct-eGFP lines.
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DMM019406F4: The level of β-catenin phosphorylation was increased in YAC128 iPSCs. (A) The levels of total β-catenin, p-β-catenin (S33/37) and GAPDH were investigated in clonal lines of HD YAC128 (n=5) and wild-type (WT) iPSCs (n=6) using western blotting. (B) Quantification of the level of p-β-catenin (S33/37) showed increased phosphorylation in the HD YAC128 iPSC lines without changes in (C) the level of total β-catenin protein. (D) qPCR analysis revealed a trend toward the increased expression of Gsk3β, but the extent of the increase was not significant. * indicates the YAC128/Oct-eGFP lines.

Mentions: The Wnt pathway is another transduction cascade that has been implicated in the pathogenesis of HD (Godin et al., 2010); therefore, we questioned whether YAC128 iPSCs also showed dysregulation of the Wnt pathway. We monitored the phosphorylation of β-catenin at serines 33/37, 41/45 and 675 using phospho-specific antibodies. Western blotting analysis revealed an increase in the level of β-catenin phosphorylation at serines 33/37 in YAC128 iPSCs versus WT iPSCs (Fig. 4A,B), whereas the levels of serines 41/45 and 675 phosphorylation were similar in the YAC128 and WT iPSCs (supplementary material Fig. S5). However, there was no difference in levels of total β-catenin protein in these cells. Interestingly, a trend toward an increased expression of Gsk3β mRNA in the YAC128 iPSCs was observed (Fig. 4C); however, this change was not significant.Fig. 4.


Huntington disease iPSCs show early molecular changes in intracellular signaling, the expression of oxidative stress proteins and the p53 pathway.

Szlachcic WJ, Switonski PM, Krzyzosiak WJ, Figlerowicz M, Figiel M - Dis Model Mech (2015)

The level of β-catenin phosphorylation was increased in YAC128 iPSCs. (A) The levels of total β-catenin, p-β-catenin (S33/37) and GAPDH were investigated in clonal lines of HD YAC128 (n=5) and wild-type (WT) iPSCs (n=6) using western blotting. (B) Quantification of the level of p-β-catenin (S33/37) showed increased phosphorylation in the HD YAC128 iPSC lines without changes in (C) the level of total β-catenin protein. (D) qPCR analysis revealed a trend toward the increased expression of Gsk3β, but the extent of the increase was not significant. * indicates the YAC128/Oct-eGFP lines.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4582098&req=5

DMM019406F4: The level of β-catenin phosphorylation was increased in YAC128 iPSCs. (A) The levels of total β-catenin, p-β-catenin (S33/37) and GAPDH were investigated in clonal lines of HD YAC128 (n=5) and wild-type (WT) iPSCs (n=6) using western blotting. (B) Quantification of the level of p-β-catenin (S33/37) showed increased phosphorylation in the HD YAC128 iPSC lines without changes in (C) the level of total β-catenin protein. (D) qPCR analysis revealed a trend toward the increased expression of Gsk3β, but the extent of the increase was not significant. * indicates the YAC128/Oct-eGFP lines.
Mentions: The Wnt pathway is another transduction cascade that has been implicated in the pathogenesis of HD (Godin et al., 2010); therefore, we questioned whether YAC128 iPSCs also showed dysregulation of the Wnt pathway. We monitored the phosphorylation of β-catenin at serines 33/37, 41/45 and 675 using phospho-specific antibodies. Western blotting analysis revealed an increase in the level of β-catenin phosphorylation at serines 33/37 in YAC128 iPSCs versus WT iPSCs (Fig. 4A,B), whereas the levels of serines 41/45 and 675 phosphorylation were similar in the YAC128 and WT iPSCs (supplementary material Fig. S5). However, there was no difference in levels of total β-catenin protein in these cells. Interestingly, a trend toward an increased expression of Gsk3β mRNA in the YAC128 iPSCs was observed (Fig. 4C); however, this change was not significant.Fig. 4.

Bottom Line: Several molecular and developmental effects of HD have been identified using neural stem cells (NSCs) and differentiated cells, such as neurons and astrocytes.Surprisingly, we found that a number of changes affecting cellular processes in HD were also present in undifferentiated pluripotent HD iPSCs, including the dysregulation of the MAPK and Wnt signaling pathways and the dysregulation of the expression of genes related to oxidative stress, such as Sod1.Interestingly, a common protein interactor of the huntingtin protein and the proteins in the above pathways is p53, and the expression of p53 was dysregulated in HD YAC128 iPSCs and human HD iPSCs.

View Article: PubMed Central - PubMed

Affiliation: Institute of Bioorganic Chemistry, Polish Academy of Sciences, Noskowskiego 12/14, Poznań 61-704, Poland.

No MeSH data available.


Related in: MedlinePlus