Fatty Acid Synthase and lipophagy as therapeutic targets of glioblastoma

脂肪酸合酶和脂肪吞噬作为胶质母细胞瘤的治疗靶点

• 批准号: 528480730
• 负责人: Professor Dr. Donat Kögel
• 金额: --
• 依托单位: Institut für Klinische Pharmakologie
• 依托单位国家: 德国
• 项目类别: Research Grants
• 资助国家: 德国
• 项目状态: 未结题
• 来源: https://gepris.dfg.de/gepris/projekt/528480730?language=en
• 关键词: Fatty; Acid; Synthase; lipophagy; therapeutic

Blocking de novo fatty acid synthesis has been considered as an anti-cancer strategy since the discovery of orlistat-mediated effects. Recent work shows that brain tumours including GBM and brain metastases heavily rely on de novo fatty acid synthesis via fatty acid synthase (FASN). A novel FASN inhibitor, TVB-2640 (Denifanstat) has recently entered a phase-3 clinical trial for glioblastoma (GBM). It is now commercially available for preclinical research and has overcome previous pharmacokinetic and toxicity limitations. But tumour cells may escape from lipid shortage via reutilization from lipid droplets. Therefore, the present project shall investigate anti-GBM efficacy of TVB-2640 in vitro and in vivo alone and in combination with chloroquine and define the underlying molecular determinants of GBM susceptibilities. The molecular studies address lipidomic and metabolomic patterns, mitochondrial respiration, cell viability and autophagy pathways in high-to-low sensitive GBM cells. Our preliminary data have revealed strong synergistic effects of TVB-2640 and the autophagy inhibitor, cholorquine (CQ). Hence, the project shall specifically address the ability of the tumour cells to reutilize lipids from lipid droplets (LD) as putative escape mechanism. To address lipophagy, knockdown/knockout cell lines of candidate LD targeting proteins (e.g. spartin) and of autophagy proteins (ATG5/7), and co-therapy with TVB-2640 + CQ shall be tested in GBM cell lines and in organotypic brain slice cultures. Furthermore, we investigate the in vivo therapeutic efficacy of TVB-2640 monotherapy and TVB-2640+CQ co-therapy (versus CQ mono, versus placebo, 4 groups) in orthotopic GBM xenograft and syngeneic models in mice. Readouts shall be survival, tumour growth, histology, cognitive behaviour and final lipidomic and metabolomic patterns. FASN inhibition causes a state of tumour starvation. To reveal how lipid shortage affects clinical GBM, lipidomic and metabolomic data and histology of auto/lipophagy shall be obtained from syngeneic and xenotransplant mouse GBM and from clinical GBM samples of patients who participated in the ERGO3 trial (collaboration with Dept. of Neurooncology, FFM) that addresses the impact of fasting to enhance anti-GBM treatments. We plan to establish mass spectrometry imaging (MSI) for visualization of lipid gradients across GBM sections and surrounding brain tissue. Together, the studies shall unravel the putative usage of FASN inhibition for GBM treatment.

自从奥利司他介导的作用被发现以来，阻断从头合成脂肪酸一直被认为是一种抗癌策略。最近的研究表明，包括基底膜和脑转移瘤在内的脑肿瘤严重依赖脂肪酸合成酶(FASN)的从头合成。一种新型的FASN抑制剂TVB-2640(Denifanstat)最近进入了治疗胶质母细胞瘤(GBM)的3期临床试验。它现在可以用于临床前研究，并克服了以前的药代动力学和毒性限制。但是，肿瘤细胞可以通过重新利用脂滴来逃脱脂类短缺。因此，本项目将研究TVB-2640在体外和体内单独以及与氯喹联合使用时的抗GBM效果，并确定GBM易感性的潜在分子决定因素。分子研究涉及脂类和代谢组模式、线粒体呼吸、细胞活力和自噬途径。我们的初步数据显示，TVB-2640和自噬抑制剂氯喹(CQ)具有很强的协同作用。因此，该项目将具体解决肿瘤细胞重新利用脂滴(LD)中的脂类作为假定的逃逸机制的能力。为了解决脂噬作用，应在GBM细胞系和器官型脑片培养中测试候选LD靶向蛋白(如Sartin)和自噬蛋白(ATG5/7)的敲除/敲除细胞系，以及与TVB-2640+CQ的联合治疗。此外，我们研究了TVB-2640单独治疗和TVB-2640+CQ联合治疗(CQ单独治疗和安慰剂治疗，4组)在小鼠原位GBM异种移植和同基因移植模型中的体内治疗效果。读数应为存活率、肿瘤生长、组织学、认知行为以及最终的脂类和代谢组型。FASN抑制会导致肿瘤饥饿状态。为了揭示脂肪短缺如何影响临床GBM，应从同基因和异种移植小鼠GBM以及参与ERG03试验的患者的临床GBM样本中获得自体/脂噬的脂组和代谢组学数据和组织学(与DEP.神经肿瘤学，FFM)，解决禁食的影响，以加强抗GBM治疗。我们计划建立质谱学成像(MSI)，以可视化跨GBM切片和周围脑组织的脂质梯度。总之，这些研究将揭开FASN抑制剂在GBM治疗中的推定用途。