NSD2驱动TM4SF1-MUC13-ASCT2复合物促进谷氨酰胺代谢激化肺动脉高压的表观遗传机制

代谢重编程促使肺动脉内皮细胞增殖引发的肺血管恶性重构是肺动脉高压(PAH)主要原因。基于NSD2上调TM4SF1、MUC13、ASCT2促进谷氨酰胺(Gln)分解加重PAH的前期研究，我们假设NSD2驱动TM4SF1-MUC13-ASCT2复合物促进Gln代谢激化PAH。拟用野百合碱腹腔注射及低氧孵育建立PAH体内外模型，基于Warburg效应及线粒体功能明确肺动脉内皮细胞Gln成瘾性，阐述ASCT2增强Gln代谢促发PAH之效应；以H3K36me2为介导探索NSD2驱动TM4SF1、MUC13表达的机制，确定TM4SF1、MUC13共表达促进Gln代谢对PAH的触发作用；联合Co-IP、免疫荧光、FRET技术探讨TM4SF1-MUC13-ASCT2复合物之形成，探索以其为介导NSD2抑制ASCT2降解之机理，揭示NSD2促进Gln代谢激化PAH的表观遗传机制，为研发PAH靶向药物奠定基础

Pulmonary vascular malignant remodeling induced by metabolic reprogramming motivated pulmonary artery endothelial cell proliferation is the main cause of pulmonary arterial hypertension (PAH). According to the preliminary studies of H3K36me2-mediated NSD2 upregulation of TM4SF1, MUC13 and ASCT2 expression to promote glutamine catabolism and aggravate PAH, we hypothesize that NSD2 enhances glutamine metabolism via epigenetically driving TM4SF1-MUC13-ASCT2 complex, which inhibits ASCT2 degradation and then intensifies PAH through promoting the malignant proliferation of pulmonary artery endothelial cells (PAECs). With the monocrotaline intraperitoneal injection and hypoxic incubation to establishing PAH in vitro and in vivo models, we intend to clarify the glutamine addiction of PAECs around Warburg effect and mitochondrial function in metabolic reprogramming, the histological analysis and angiography were used to expounded the stimulating PAH effect of ASCT2-enhanced glutamine metabolism. Using H3K36me2 as a medium to explore the mechanism of NSD2-driven TM4SF1 and MUC13, which will beneficial to determine the triggering effect of TM4SF1, MUC13 co-expression on PAH by promoting Gln metabolism. Combining Co-IP, immunofluorescence and FRET technologies to explore the formation of TM4SF1-MUC13-ASCT2 complex, which mediates the mechanism of NSD2 inhibition of ASCT2 degradation. All of these will conduce to reveal the epigenetic mechanism that NSD2 stimulates PAH through promoting Gln metabolism, which hope to provide theoretical basis for the development of molecular targeted drugs to PAH.