Elevation of phosphate levels impairs skeletal myoblast differentiation.

Adalbert Raimann, Alexander Dangl, Alireza Javanmardi, Susanne Greber-Platzer, Monika Egerbacher, Peter Pietschmann, Gabriele Haeusler

Cell and tissue research 2020 Nov

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Hyperphosphatemic conditions such as chronic kidney disease are associated with severe muscle wasting and impaired life quality. While regeneration of muscle tissue is known to be reliant on recruitment of myogenic progenitor cells, the effects of elevated phosphate loads on this process have not been investigated in detail so far. This study aims to clarify the direct effects of hyperphosphatemic conditions on skeletal myoblast differentiation in a murine in vitro model. C2C12 murine muscle progenitor cells were supplemented with phosphate concentrations resembling moderate to severe hyperphosphatemia (1.4-2.9 mmol/l). Phosphate-induced effects were quantified by RT-PCR and immunoblotting. Immunohistochemistry was performed to count nuclear positive cells under treatment. Cell viability and metabolic activity were assessed by XTT and BrdU incorporation assays. Inorganic phosphate directly induced ERK-phosphorylation in pre-differentiated C2C12 myoblast cells. While phosphate concentrations resembling the upper normal range significantly reduced Myogenin expression (- 22.5%, p = 0.015), severe hyperphosphatemic conditions further impaired differentiation (Myogenin - 61.0%, p < 0.0001; MyoD - 51.0%; p < 0.0001). Analogue effects were found on the protein level (Myogenin - 42.0%, p = 0.004; MyoD - 25.7%, p = 0.002). ERK inhibition strongly attenuated phosphate-induced effects on Myogenin expression (p = 0.002). Metabolic activity was unaffected by the treatments. Our data point to a phosphate-induced inhibition of myoblast differentiation without effects on cell viability. Serum phosphate levels as low as the upper normal serum range significantly impaired marker gene expression in vitro. Investigation of cellular effects of hyperphosphatemia may help to better define serum cutoffs and modify existing treatment approaches of phosphate binders, especially in patients at risk of sarcopenia.

Citation

Adalbert Raimann, Alexander Dangl, Alireza Javanmardi, Susanne Greber-Platzer, Monika Egerbacher, Peter Pietschmann, Gabriele Haeusler. Elevation of phosphate levels impairs skeletal myoblast differentiation. Cell and tissue research. 2020 Nov;382(2):427-432