Histone acetyltransferase P300 deficiency promotes ferroptosis of vascular smooth muscle cells by activating the HIF-1α/HMOX1 axis

Abstract

Background E1A-associated 300-kDa protein (P300), an endogenous histone acetyltransferase, contributes to modifications of the chromatin landscape of genes involved in multiple cardiovascular diseases. Ferroptosis of vascular smooth muscle cells (VSMCs) is a novel pathological mechanism of aortic dissection. However, whether P300 regulates VSMC ferroptosis remains unknown. Methods Cystine deprivation (CD) and imidazole ketone erastin (IKE) were used to induce VSMC ferroptosis. Two different knockdown plasmids targeting P300 and A-485 (a specific inhibitor of P300) were used to investigate the function of P300 in the ferroptosis of human aortic smooth muscle cells (HASMCs). Cell counting kit-8, lactate dehydrogenase and flow cytometry with propidium iodide staining were performed to assess the cell viability and death under the treatment of CD and IKE. BODIPY-C11 assay, immunofluorescence staining of 4-hydroxynonenal and malondialdehyde assay were conducted to detect the level of lipid peroxidation. Furthermore, co-immunoprecipitation was utilized to explore the interaction between P300 and HIF-1α, HIF-1α and P53. Results Compared with normal control, the protein level of P300 was significantly decreased in HASMCs treated with CD and IKE, which was largely nullified by the ferroptosis inhibitor ferrostatin-1 but not by the autophagy inhibitor or apoptosis inhibitor. Knockdown of P300 by short-hairpin RNA or inhibition of P300 activity by A-485 promoted CD- and IKE-induced HASMC ferroptosis, as evidenced by a reduction in cell viability and aggravation of lipid peroxidation of HASMCs. Furthermore, we found that hypoxia-inducible factor-1α (HIF-1α)/heme oxygenase 1 (HMOX1) pathway was responsible for the impacts of P300 on ferroptosis of HASMCs. The results of co-immunoprecipitation demonstrated that P300 and P53 competitively bound HIF-1α to regulate the expression of HMOX1. Under normal conditions, P300 interacted with HIF-1α to inhibit HMOX1 expression, while reduced expression of P300 induced by ferroptosis inducers would favor HIF-1α binding to P53 to trigger HMOX1 overexpression. Furthermore, the aggravated effects of P300 knockdown on HASMC ferroptosis were largely nullified by HIF-1α knockdown or the HIF-1α inhibitor BAY87-2243. Conclusion Thus, our results revealed that P300 deficiency or inactivation facilitated CD- and IKE-induced VSMC ferroptosis by activating the HIF-1α/HMOX1 axis, which may contribute to the development of diseases related to VSMC ferroptosis.

Oxidative stress drives vascular smooth muscle cell damage in acute Stanford type A aortic dissection through HIF-1α/HO-1 mediated ferroptosis

“…In vitro experiments verified that classical ferroptosis inhibitor liproxstatin-1 could suppress the expression of HO-1 in H 2 O 2 -induced ferroptosis model. Our findings were in line with previous in vitro experiments, which demonstrated that HO-1 was overexpressed in both cystine deprivation and imidazole ketone erastin induced VSMC ferroptosis models [ 45 ].…”

“…Over the decades, HIF-1α has been proved to be induced under hypoxia, oxidative stress and other transient stimulus [ 53 ]. Although its overexpression has already been identified in several literatures on ATAAD [ 45 , 54 , 55 ], molecular mechanisms of how HIF-1α promotes dissection progression remain still little known. Here, we have demonstrated a novel mechanism of VSMC ferroptosis through HIF-1α/HO-1 axis in response to oxidative stress.…”

NINJ1 regulates ferroptosis via xCT antiporter interaction and CoA modulation

“…2I), pitstop2 (42) (clathrin-mediated endocytosis inhibitor, Supplemental Fig. 2J), EML425 and C646 (43) (p300/CBP Inhibitor, Supplemental Fig. 2K-L), brequinar (44) (dihydroorotate dehydrogenase (DHODH) inhibitors, Supplemental Fig.…”

α-Ketoglutarate improves cardiac insufficiency through NAD+-SIRT1 signaling-mediated mitophagy and ferroptosis in pressure overload-induced mice

“…Studies have shown that bilirubin or biliverdin and carbon monoxide, which are metabolites of HO1, can reduce oxidative stress and inhibit ferroptosis (Ryter et al 2006 ; Sugimoto et al 2012 ). However, ferrous ions produced during HO1 metabolism can promote ferroptosis (Papanikolaou and Pantopoulos 2005 ; Shi et al 2023 ). In our study, the change in HO1 expression was synchronized with SIRT1 and GPX4, and ferroptosis was inhibited.…”