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Histone lysine crotonylation during acute kidney injury in mice

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-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.

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Crotonate prevents downregulation of kidney PGC-1α and SIRT3 as well as causing CCL2 upregulation in experimental AKI. AKI was induced by a folic acid overdose in mice pretreated with or without 12 mmol/kg body weight crotonate. All mice were killed at 72 h. (A) Crotonate prevented the increase in serum BUN and creatinine levels and in Kim-1 mRNA expression observed in AKI. Mean±s.e.m. of five mice per group. *P<0.05 vs vehicle control, #P<0.05 vs vehicle AKI (non-parametric Mann–Whitney U-test). (B-D) The kidney PGC-1α mRNA decrease (B), SIRT3 mRNA decrease (C), and CCL2 mRNA increase (D) were prevented by pretreatment with crotonate. Mean±s.e.m. of five animals per group. *P<0.05 vs vehicle control, #P<0.05 vs vehicle AKI (non-parametric Mann–Whitney U-test). (E,F) Kidney PGC-1α (E) and SIRT3 (F) protein levels decreased in AKI and this was prevented by pretreatment with crotonate. Western blot of whole kidney protein. Mean±s.e.m. of five animals per group. *P<0.05 vs vehicle control, #P<0.05 vs vehicle AKI (non-parametric Mann–Whitney U-test).
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DMM024455F6: Crotonate prevents downregulation of kidney PGC-1α and SIRT3 as well as causing CCL2 upregulation in experimental AKI. AKI was induced by a folic acid overdose in mice pretreated with or without 12 mmol/kg body weight crotonate. All mice were killed at 72 h. (A) Crotonate prevented the increase in serum BUN and creatinine levels and in Kim-1 mRNA expression observed in AKI. Mean±s.e.m. of five mice per group. *P<0.05 vs vehicle control, #P<0.05 vs vehicle AKI (non-parametric Mann–Whitney U-test). (B-D) The kidney PGC-1α mRNA decrease (B), SIRT3 mRNA decrease (C), and CCL2 mRNA increase (D) were prevented by pretreatment with crotonate. Mean±s.e.m. of five animals per group. *P<0.05 vs vehicle control, #P<0.05 vs vehicle AKI (non-parametric Mann–Whitney U-test). (E,F) Kidney PGC-1α (E) and SIRT3 (F) protein levels decreased in AKI and this was prevented by pretreatment with crotonate. Western blot of whole kidney protein. Mean±s.e.m. of five animals per group. *P<0.05 vs vehicle control, #P<0.05 vs vehicle AKI (non-parametric Mann–Whitney U-test).

Mentions: We next explored whether crotonate was nephroprotective in vivo. Mice were pretreated with 12 mmol/kg body weight crotonate, and 24 h later, AKI was induced by a folic acid overdose and mice were killed at 72 h, when renal failure peaks (Sanz et al., 2010b). First, we observed that crotonate resulted in lower serum levels of BUN and creatinine, markers of renal dysfunction severity, and in lower KIM-1 mRNA levels, a marker of kidney injury (Fig. 6A). PAS-stained kidney sections revealed a trend towards decreased tubular injury in crotonate-treated mice (Fig. S7). AKI was associated with increased CCL2 expression (Sanz et al., 2010b) and reduced whole kidney SIRT3 expression (Fig. S8) within the time points studied. In this line, systemic administration of crotonate prevented the decrease in kidney PGC-1α and SIRT3 levels in AKI (Fig. 6B,C,E,F) as well as the increase in CCL2 mRNA expression (Fig. 6D). This suggests a protective effect of crotonate, and thereby of histone crotonylation, against inflammation and mitochondrial stress during AKI.Fig. 6.


Histone lysine crotonylation during acute kidney injury in mice
Crotonate prevents downregulation of kidney PGC-1α and SIRT3 as well as causing CCL2 upregulation in experimental AKI. AKI was induced by a folic acid overdose in mice pretreated with or without 12 mmol/kg body weight crotonate. All mice were killed at 72 h. (A) Crotonate prevented the increase in serum BUN and creatinine levels and in Kim-1 mRNA expression observed in AKI. Mean±s.e.m. of five mice per group. *P<0.05 vs vehicle control, #P<0.05 vs vehicle AKI (non-parametric Mann–Whitney U-test). (B-D) The kidney PGC-1α mRNA decrease (B), SIRT3 mRNA decrease (C), and CCL2 mRNA increase (D) were prevented by pretreatment with crotonate. Mean±s.e.m. of five animals per group. *P<0.05 vs vehicle control, #P<0.05 vs vehicle AKI (non-parametric Mann–Whitney U-test). (E,F) Kidney PGC-1α (E) and SIRT3 (F) protein levels decreased in AKI and this was prevented by pretreatment with crotonate. Western blot of whole kidney protein. Mean±s.e.m. of five animals per group. *P<0.05 vs vehicle control, #P<0.05 vs vehicle AKI (non-parametric Mann–Whitney U-test).
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DMM024455F6: Crotonate prevents downregulation of kidney PGC-1α and SIRT3 as well as causing CCL2 upregulation in experimental AKI. AKI was induced by a folic acid overdose in mice pretreated with or without 12 mmol/kg body weight crotonate. All mice were killed at 72 h. (A) Crotonate prevented the increase in serum BUN and creatinine levels and in Kim-1 mRNA expression observed in AKI. Mean±s.e.m. of five mice per group. *P<0.05 vs vehicle control, #P<0.05 vs vehicle AKI (non-parametric Mann–Whitney U-test). (B-D) The kidney PGC-1α mRNA decrease (B), SIRT3 mRNA decrease (C), and CCL2 mRNA increase (D) were prevented by pretreatment with crotonate. Mean±s.e.m. of five animals per group. *P<0.05 vs vehicle control, #P<0.05 vs vehicle AKI (non-parametric Mann–Whitney U-test). (E,F) Kidney PGC-1α (E) and SIRT3 (F) protein levels decreased in AKI and this was prevented by pretreatment with crotonate. Western blot of whole kidney protein. Mean±s.e.m. of five animals per group. *P<0.05 vs vehicle control, #P<0.05 vs vehicle AKI (non-parametric Mann–Whitney U-test).
Mentions: We next explored whether crotonate was nephroprotective in vivo. Mice were pretreated with 12 mmol/kg body weight crotonate, and 24 h later, AKI was induced by a folic acid overdose and mice were killed at 72 h, when renal failure peaks (Sanz et al., 2010b). First, we observed that crotonate resulted in lower serum levels of BUN and creatinine, markers of renal dysfunction severity, and in lower KIM-1 mRNA levels, a marker of kidney injury (Fig. 6A). PAS-stained kidney sections revealed a trend towards decreased tubular injury in crotonate-treated mice (Fig. S7). AKI was associated with increased CCL2 expression (Sanz et al., 2010b) and reduced whole kidney SIRT3 expression (Fig. S8) within the time points studied. In this line, systemic administration of crotonate prevented the decrease in kidney PGC-1α and SIRT3 levels in AKI (Fig. 6B,C,E,F) as well as the increase in CCL2 mRNA expression (Fig. 6D). This suggests a protective effect of crotonate, and thereby of histone crotonylation, against inflammation and mitochondrial stress during AKI.Fig. 6.

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.


Related in: MedlinePlus