Up-regulation of HDACs, a harbinger of uraemic endothelial dysfunction, is prevented by defibrotide.

Marta Palomo,Sergi Torramadé‐Moix,Susana Martin,Aleix Cases,Ana Belen Moreno,Maribel Díaz‐Ricart,Ginés Escolar,Manel Vera,Enric Carreras,Julia Martinez‐Sanchez

doi:10.1111/jcmm.14865

Abstract

Endothelial dysfunction is an earlier contributor to the development of atherosclerosis in chronic kidney disease (CKD), in which the role of epigenetic triggers cannot be ruled out. Endothelial protective strategies, such as defibrotide (DF), may be useful in this scenario. We evaluated changes induced by CKD on endothelial cell proteome and explored the effect of DF and the mechanisms involved. Human umbilical cord vein endothelial cells were exposed to sera from healthy donors (n = 20) and patients with end‐stage renal disease on haemodialysis (n = 20). Differential protein expression was investigated by using a proteomic approach, Western blot and immunofluorescence. HDAC1 and HDAC2 overexpression was detected. Increased HDAC1 expression occurred at both cytoplasm and nucleus. These effects were dose‐dependently inhibited by DF. Both the HDACs inhibitor trichostatin A and DF prevented the up‐regulation of the endothelial dysfunction markers induced by the uraemic milieu: intercellular adhesion molecule‐1, surface Toll‐like receptor‐4, von Willebrand Factor and reactive oxygen species. Moreover, DF down‐regulated HDACs expression through the PI3/AKT signalling pathway. HDACs appear as key modulators of the CKD‐induced endothelial dysfunction as specific blockade by trichostatin A or by DF prevents endothelial dysfunction responses to the CKD insult. Moreover, DF exerts its endothelial protective effect by inhibiting HDAC up‐regulation likely through PI3K/AKT.

Highlights

Chronic kidney disease (CKD) is a major public health concern due to its increased prevalence, high morbidity and mortality associated, high costs, poorer quality of life and reduced life expectancy.[1]
Our present study explored the protein signature of the endothelium exposed to chronic kidney disease (CKD) sera in the presence and absence of DF, and pointed out to HDACs as key molecules that mediate the endothelial response to the CKD milieu
Both trichostatin A (TSA) and DF prevented the endothelium from developing its pro-inflammatory, prooxidant, prothrombotic and activated innate immunity phenotype induced by the CKD sera

Summary

| INTRODUCTION

Chronic kidney disease (CKD) is a major public health concern due to its increased prevalence, high morbidity and mortality associated, high costs, poorer quality of life and reduced life expectancy.[1]. Our previous in vitro results suggest that the CKD setting induces activation of specific genes in the endothelium that promotes a prooxidant, pro-inflammatory, prothrombotic and proproliferative state In this regard, HDACs enzymes are important epigenetic factors that regulate pro-inflammatory gene expression, in ECs and in other tissues, and deacetylate non-histone proteins that regulate inflammatory signalling.[16] they appear to be promising therapeutic targets for the treatment of atherosclerosis and other cardiovascular diseases.[17,18] HDAC activity is linked to a number of cardio-renal pathologies, including heart failure,[19] hypertension,[20] and diabetes or diabetic kidney disease.[21,22] the dysfunctional endothelium and its epigenetic regulation arise as an attractive target for interventions designed to reduce the risk and burden of CVD in CKD patients.[23,24] Defibrotide (Defitelio®) (DF) is a drug composed by a complex single-stranded oligodeoxyribonucleotides derived from porcine intestinal mucosal DNA that has demonstrated profibrinolytic, antithrombotic-thrombolytic, antiischaemic, antishock, antiatherosclerotic, antirejection and anti-angiogenic effects.[25] The endothelial protective activity of this drug has been demonstrated in different clinical settings.[26,27,28] This drug interacts with the membrane of ECs where it displays its anti-inflammatory and anti-oxidant effects.[29]. We aimed at identifying the mechanisms involved in the protective effect exerted by DF in this setting

| METHODS

Findings

| DISCUSSION