Di Pietrantonio Nadia, Sánchez-Ceinos Julia, Shumliakivska Mariana, Rakow Alexander, Mandatori Domitilla, Di Tomo Pamela, Formoso Gloria, Bonfini Tiziana, Baldassarre Maria Pompea Antonia, Sennström Maria, Almahmeed Wael, Pandolfi Assunta, Cosentino Francesco
Cardiology Unit, Department of Medicine Solna, Karolinska Institutet, Stockholm 171 76, Sweden.
Department of Medical, Oral and Biotechnological Sciences, Center for Advanced Studies and Technology-CAST, University G. D'Annunzio of Chieti-Pescara, Chieti 66100, Italy.
Eur Heart J. 2024 Dec 23;45(48):5171-5185. doi: 10.1093/eurheartj/ehae688.
Hyperglycaemia during gestational diabetes (GD) predisposes women and their offspring to later cardiometabolic disease. The hyperglycaemia-mediated epigenetic changes remain to be elucidated. Methyltransferase MLL1-induced trimethylation of histone 3 at lysine 4 (H3K4me3) activates inflammatory and oxidative phenotype. This epigenetic mark in GD women and its transmission to the offspring were investigated.
Peripheral blood mononuclear cells (PBMC) were collected from GD and control (C) women and also from adolescents born to women of both groups. Endothelial human umbilical vein endothelial cells (HUVEC) and cord blood mononuclear cells (CBMC) were from umbilical cords. The NF-κBp65 and NOX4 expressions were investigated by reverse transcription quantitative polymerase chain reaction and immunofluorescence (IF). MLL1 and H3K4me3 were investigated by immunoblotting and IF. H3K4me3 on NF-κBp65 and NOX4 promoters was studied by chromatin immunoprecipitation. Superoxide anion generation was measured by electron spin resonance spectroscopy. Plasma cytokines were measured by enzyme-linked immunosorbent assay. To investigate the role of MLL1, HUVEC were exposed to inhibitor MM102 or siRNA transfection.
PBMC, CBMC, and HUVEC showed an increase of NF-κBp65, IL-6, ICAM-1, MCP-1, and VCAM-1 mRNAs. These findings were associated with H3K4me3 enrichment in the promoter of NF-κBp65. Elevated H3K4me3 and cytokine levels were observed in GD adolescents. MLL1 drives H3K4me3 not only on NF-kB p65, but also on NOX4 promoter. Inhibition of MLL1 blunted NF-κBp65 and NOX4 by modulating inflammatory and oxidative phenotype.
Such proof-of-concept study shows persistence of MLL1-dependent H3K4me3 in offspring born to GD women, suggesting an epigenetic-driven transmission of maternal phenotype. These findings may pave the way for pharmacological reprogramming of adverse histone modifications to mitigate abnormal phenotypes underlying early ASCVD.
妊娠期糖尿病（GD）期间的高血糖会使女性及其后代日后易患心脏代谢疾病。高血糖介导的表观遗传变化仍有待阐明。甲基转移酶MLL1诱导的组蛋白3赖氨酸4位点三甲基化（H3K4me3）会激活炎症和氧化表型。本研究对GD女性中的这种表观遗传标记及其向后代的传递情况进行了调查。
从GD女性和对照（C）女性以及两组女性所生青少年中采集外周血单个核细胞（PBMC）。人脐静脉内皮细胞（HUVEC）和脐血单个核细胞（CBMC）取自脐带。通过逆转录定量聚合酶链反应和免疫荧光（IF）检测NF-κBp65和NOX4的表达。通过免疫印迹和IF检测MLL1和H3K4me3。通过染色质免疫沉淀研究NF-κBp65和NOX4启动子上的H3K4me3。通过电子自旋共振光谱法测量超氧阴离子的产生。通过酶联免疫吸附测定法测量血浆细胞因子。为了研究MLL1的作用，将HUVEC暴露于抑制剂MM102或进行小干扰RNA转染。
PBMC、CBMC和HUVEC中NF-κBp65、IL-6、ICAM-1、MCP-1和VCAM-1的mRNA水平升高。这些发现与NF-κBp65启动子中H3K4me3富集有关。在GD青少年中观察到H3K4me3和细胞因子水平升高。MLL1不仅在NF-κB p65上驱动H3K4me3，而且在NOX4启动子上也有此作用。抑制MLL1通过调节炎症和氧化表型减弱了NF-κBp65和NOX4。
此类概念验证研究表明，MLL1依赖的H3K4me3在GD女性所生后代中持续存在，提示母代表型存在表观遗传驱动的传递。这些发现可能为不良组蛋白修饰的药理学重编程铺平道路，以减轻早期动脉粥样硬化性心血管疾病（ASCVD）潜在的异常表型。
