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Histone lysine crotonylation during acute kidney injury in mice

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ABSTRACT

Acute kidney injury (AKI) is a potentially lethal condition for which no therapy is available beyond replacement of renal function. Post-translational histone modifications modulate gene expression and kidney injury. Histone crotonylation is a recently described post-translational modification. We hypothesized that histone crotonylation might modulate kidney injury. Histone crotonylation was studied in cultured murine proximal tubular cells and in kidneys from mice with AKI induced by folic acid or cisplatin. Histone lysine crotonylation was observed in tubular cells from healthy murine and human kidney tissue. Kidney tissue histone crotonylation increased during AKI. This was reproduced by exposure to the protein TWEAK in cultured tubular cells. Specifically, ChIP-seq revealed enrichment of histone crotonylation at the genes encoding the mitochondrial biogenesis regulator PGC-1α and the sirtuin-3 decrotonylase in both TWEAK-stimulated tubular cells and in AKI kidney tissue. To assess the role of crotonylation in kidney injury, crotonate was used to increase histone crotonylation in cultured tubular cells or in the kidneys in vivo. Crotonate increased the expression of PGC-1α and sirtuin-3, and decreased CCL2 expression in cultured tubular cells and healthy kidneys. Systemic crotonate administration protected from experimental AKI, preventing the decrease in renal function and in kidney PGC-1α and sirtuin-3 levels as well as the increase in CCL2 expression. For the first time, we have identified factors such as cell stress and crotonate availability that increase histone crotonylation in vivo. Overall, increasing histone crotonylation might have a beneficial effect on AKI. This is the first observation of the in vivo potential of the therapeutic manipulation of histone crotonylation in a disease state.

No MeSH data available.


Crotonate increases histone crotonylation in mouse kidney. Mice were treated with crotonate at different doses and for different times. Histone crotonylation was measured by western blotting (anti-Kcr). (A) Dose-response curve at 48 h. Mice received vehicle, or 3, 6 or 12 mmol/kg body weight crotonate by intraperitoneal injection. Mean±s.e.m. of four mice per group. *P<0.05 vs control (non-parametric Mann–Whitney U-test). (B) Timecourse curve. Mice were killed 24, 48 or 72 h following intraperitoneal injection injection of 12 mmol/kg body weight crotonate or vehicle (control). Mean±s.e.m. of four mice per group. *P<0.05 vs control (non-parametric Mann–Whitney U-test).
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DMM024455F4: Crotonate increases histone crotonylation in mouse kidney. Mice were treated with crotonate at different doses and for different times. Histone crotonylation was measured by western blotting (anti-Kcr). (A) Dose-response curve at 48 h. Mice received vehicle, or 3, 6 or 12 mmol/kg body weight crotonate by intraperitoneal injection. Mean±s.e.m. of four mice per group. *P<0.05 vs control (non-parametric Mann–Whitney U-test). (B) Timecourse curve. Mice were killed 24, 48 or 72 h following intraperitoneal injection injection of 12 mmol/kg body weight crotonate or vehicle (control). Mean±s.e.m. of four mice per group. *P<0.05 vs control (non-parametric Mann–Whitney U-test).

Mentions: We explored whether crotonate modulates kidney histone crotonylation in vivo. Systemic administration of crotonate increased histone crotonylation in mouse kidney in a dose- and time-dependent manner (Fig. 4A,B). The 6 mmol/kg body weight crotonate dose did not significantly change kidney histone crotonylation (Fig. S6A) or PGC-1α mRNA expression (Fig. S6B) at 24 h. Thus, 12 mmol/kg body weight crotonate was used for further experiments and was found to increase whole kidney histone crotonylation (Fig. 4), PGC-1α mRNA levels (Fig. 5A) and PGC-1α protein (Fig. 5B), and to decrease kidney CCL2 mRNA levels (Fig. 5C). Thus, the potential nephroprotective actions of crotonate observed in cultured tubular cells (increased expression of the nephroprotective gene PGC-1α and decreased inflammatory gene expression) was reproduced in vivo.Fig. 4.


Histone lysine crotonylation during acute kidney injury in mice
Crotonate increases histone crotonylation in mouse kidney. Mice were treated with crotonate at different doses and for different times. Histone crotonylation was measured by western blotting (anti-Kcr). (A) Dose-response curve at 48 h. Mice received vehicle, or 3, 6 or 12 mmol/kg body weight crotonate by intraperitoneal injection. Mean±s.e.m. of four mice per group. *P<0.05 vs control (non-parametric Mann–Whitney U-test). (B) Timecourse curve. Mice were killed 24, 48 or 72 h following intraperitoneal injection injection of 12 mmol/kg body weight crotonate or vehicle (control). Mean±s.e.m. of four mice per group. *P<0.05 vs control (non-parametric Mann–Whitney U-test).
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Related In: Results  -  Collection

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DMM024455F4: Crotonate increases histone crotonylation in mouse kidney. Mice were treated with crotonate at different doses and for different times. Histone crotonylation was measured by western blotting (anti-Kcr). (A) Dose-response curve at 48 h. Mice received vehicle, or 3, 6 or 12 mmol/kg body weight crotonate by intraperitoneal injection. Mean±s.e.m. of four mice per group. *P<0.05 vs control (non-parametric Mann–Whitney U-test). (B) Timecourse curve. Mice were killed 24, 48 or 72 h following intraperitoneal injection injection of 12 mmol/kg body weight crotonate or vehicle (control). Mean±s.e.m. of four mice per group. *P<0.05 vs control (non-parametric Mann–Whitney U-test).
Mentions: We explored whether crotonate modulates kidney histone crotonylation in vivo. Systemic administration of crotonate increased histone crotonylation in mouse kidney in a dose- and time-dependent manner (Fig. 4A,B). The 6 mmol/kg body weight crotonate dose did not significantly change kidney histone crotonylation (Fig. S6A) or PGC-1α mRNA expression (Fig. S6B) at 24 h. Thus, 12 mmol/kg body weight crotonate was used for further experiments and was found to increase whole kidney histone crotonylation (Fig. 4), PGC-1α mRNA levels (Fig. 5A) and PGC-1α protein (Fig. 5B), and to decrease kidney CCL2 mRNA levels (Fig. 5C). Thus, the potential nephroprotective actions of crotonate observed in cultured tubular cells (increased expression of the nephroprotective gene PGC-1α and decreased inflammatory gene expression) was reproduced in vivo.Fig. 4.

View Article: PubMed Central - PubMed

ABSTRACT

Acute kidney injury (AKI) is a potentially lethal condition for which no therapy is available beyond replacement of renal function. Post-translational histone modifications modulate gene expression and kidney injury. Histone crotonylation is a recently described post-translational modification. We hypothesized that histone crotonylation might modulate kidney injury. Histone crotonylation was studied in cultured murine proximal tubular cells and in kidneys from mice with AKI induced by folic acid or cisplatin. Histone lysine crotonylation was observed in tubular cells from healthy murine and human kidney tissue. Kidney tissue histone crotonylation increased during AKI. This was reproduced by exposure to the protein TWEAK in cultured tubular cells. Specifically, ChIP-seq revealed enrichment of histone crotonylation at the genes encoding the mitochondrial biogenesis regulator PGC-1&alpha; and the sirtuin-3 decrotonylase in both TWEAK-stimulated tubular cells and in AKI kidney tissue. To assess the role of crotonylation in kidney injury, crotonate was used to increase histone crotonylation in cultured tubular cells or in the kidneys in vivo. Crotonate increased the expression of PGC-1&alpha; and sirtuin-3, and decreased CCL2 expression in cultured tubular cells and healthy kidneys. Systemic crotonate administration protected from experimental AKI, preventing the decrease in renal function and in kidney PGC-1&alpha; and sirtuin-3 levels as well as the increase in CCL2 expression. For the first time, we have identified factors such as cell stress and crotonate availability that increase histone crotonylation in vivo. Overall, increasing histone crotonylation might have a beneficial effect on AKI. This is the first observation of the in vivo potential of the therapeutic manipulation of histone crotonylation in a disease state.

No MeSH data available.