Vergani Elisabetta, Beretta Giovanni L, Aloisi Mariachiara, Costantino Matteo, Corno Cristina, Frigerio Simona, Tinelli Stella, Dugo Matteo, Accattatis Felice Maria, Granata Agnese, Arnaboldi Lorenzo, Rodolfo Monica, Perego Paola, Gatti Laura
Unit of Immunotherapy of Human Tumors, Department of Research, Fondazione IRCCS Istituto Nazionale dei Tumori di Milano, Milan, Italy.
Unit of Molecular Pharmacology, Department of Applied Research and Technological Development, Fondazione IRCCS Istituto Nazionale dei Tumori, Milan, Italy.
Front Cell Dev Biol. 2022 Jul 13;10:927118. doi: 10.3389/fcell.2022.927118. eCollection 2022.
Drug resistance limits the achievement of persistent cures for the treatment of melanoma, in spite of the efficacy of the available drugs. The aim of the present study was to explore the involvement of lipid metabolism in melanoma resistance and assess the effects of its targeting in cellular models of melanoma with acquired resistance to the BRAF-inhibitor PLX4032/Vemurafenib. Since transcriptional profiles pointed to decreased cholesterol and fatty acids synthesis in resistant cells as compared to their parental counterparts, we examined lipid composition profiles of resistant cells, studied cell growth dependence on extracellular lipids, assessed the modulation of enzymes controlling the main nodes in lipid biosynthesis, and evaluated the effects of targeting Acetyl-CoA Acetyltransferase 2 (ACAT2), the first enzyme in the cholesterol synthesis pathway, and Acyl-CoA Cholesterol Acyl Transferase (ACAT/SOAT), which catalyzes the intracellular esterification of cholesterol and the formation of cholesteryl esters. We found a different lipid composition in the resistant cells, which displayed reduced saturated fatty acids (SFA), increased monounsaturated (MUFA) and polyunsaturated (PUFA), and reduced cholesteryl esters (CE) and triglycerides (TG), along with modulated expression of enzymes regulating biosynthetic nodes of the lipid metabolism. The effect of tackling lipid metabolism pathways in resistant cells was evidenced by lipid starvation, which reduced cell growth, increased sensitivity to the BRAF-inhibitor PLX4032, and induced the expression of enzymes involved in fatty acid and cholesterol metabolism. Molecular targeting of ACAT2 or pharmacological inhibition of SOAT by avasimibe showed antiproliferative effects in melanoma cell lines and a synergistic drug interaction with PLX4032, an effect associated to increased ferroptosis. Overall, our findings reveal that lipid metabolism affects melanoma sensitivity to BRAF inhibitors and that extracellular lipid availability may influence tumor cell response to treatment, a relevant finding in the frame of personalized therapy. In addition, our results indicate new candidate targets for drug combination treatments.
尽管现有药物有效,但耐药性限制了黑色素瘤治疗中持久治愈的实现。本研究的目的是探讨脂质代谢在黑色素瘤耐药中的作用,并评估在对BRAF抑制剂PLX4032/维莫非尼获得性耐药的黑色素瘤细胞模型中靶向脂质代谢的效果。由于转录谱显示,与亲代细胞相比,耐药细胞中胆固醇和脂肪酸合成减少,我们检测了耐药细胞的脂质组成谱,研究了细胞生长对细胞外脂质的依赖性,评估了控制脂质生物合成主要节点的酶的调节情况,并评估了靶向胆固醇合成途径中的首个酶乙酰辅酶A乙酰转移酶2(ACAT2)以及催化胆固醇细胞内酯化和胆固醇酯形成的酰基辅酶A胆固醇酰基转移酶(ACAT/SOAT)的效果。我们发现耐药细胞中的脂质组成不同,其饱和脂肪酸(SFA)减少,单不饱和脂肪酸(MUFA)和多不饱和脂肪酸(PUFA)增加,胆固醇酯(CE)和甘油三酯(TG)减少,同时调节脂质代谢生物合成节点的酶的表达也发生了变化。脂质饥饿证明了在耐药细胞中处理脂质代谢途径的效果,脂质饥饿可减少细胞生长,增加对BRAF抑制剂PLX4032的敏感性,并诱导参与脂肪酸和胆固醇代谢的酶的表达。对ACAT2进行分子靶向或用阿伐斯汀对SOAT进行药理抑制,在黑色素瘤细胞系中显示出抗增殖作用,并与PLX4032产生协同药物相互作用,这种作用与铁死亡增加有关。总体而言,我们的研究结果表明脂质代谢会影响黑色素瘤对BRAF抑制剂的敏感性,细胞外脂质的可用性可能会影响肿瘤细胞对治疗的反应,这在个性化治疗框架中是一个相关发现。此外,我们的结果指出了联合药物治疗的新候选靶点。