基于靶向代谢组学的肺腺癌干细胞葡萄糖代谢重编程通路分配定量研究

Metabolic reprogramming has revived to be thought as one of the hallmark of cancer. Modern Metabolic reprogramming study has evolved from previous descriptive analysis into the exact quantitative analysis of the metabolic pathway allocation and the metabolites. Glucose metabolism stands at the center of the substance and energy metabolism of the organism and the lung cancer stem cell is a source for lung cancer metastasis, resistance to drugs, and relapse. However, so far, there is very limited study directly focusing on the lung cancer stem cells for the quantitative analysis of glucose metabolic flux allocation into the various pathways of glycolysis, pentose phosphate, et al in the glucose metabolism, which is one of the bottleneck for the deeper understanding of the metabolic reprogramming. Based on our previous findings, the project will carry out the non-labeled and the stable isotope-labeled quantitative targeted metabolomics study on the glucose metabolic pathways of lung adenocarcinoma stem cells by using LC-MS/MS analytical platform established by our group. After drawing a blueprint of the profile and quantitative allocation of the metabolic flux into the pathways and selectively interfering the candidate key metabolites and key nodes/enzymes for controlling the metabolic networks, the project will identify and determine the key points and the tunable points for the control of the phenotypes of glucose metabolism and cancer stem cell biology of the lung cancer stem cells. This will provide theoretical and experimental foundation for the pathway mechanism of the metabolic reprogramming in lung cancer stem cells and for the discovery of the potential metabolic pathway key molecule-based drug targets and will offer and optimize the new techniques of pathway-targeted metabolomics for basic study and translational medicine.

近年重新认识到代谢重编程对肿瘤的关键作用。现今代谢研究已逐渐从定性描述进展到对代谢通量分配和代谢物分子的精确定量研究层面。葡萄糖代谢通路分配重编程及其代谢物分子异常变化对包括肺癌在内的肿瘤起重要作用。肺癌干细胞是肺癌发生、转移、耐药和复发的重要源泉，然而迄今对其葡萄糖代谢动态流向各通路分配的定量研究极为有限，成为阻碍精确认识肺癌代谢重编程通路机制的重要瓶颈之一。项目拟在前期工作的基础上，利用自主建立的色谱串联质谱联用检测分析体系，对肺腺癌干细胞的葡萄糖代谢通路进行非标记和稳定性同位素标记定量靶向代谢组学研究，结合生物学和药理学干预手段，揭示肺腺癌干细胞葡萄糖代谢的通路流向定量分配模式并确定该代谢网络中控制干细胞代谢表型和生物学表型的关键节点，为肺癌代谢重编程通路机制研究和基于该机制的潜在新药靶的发现提供理论和实验依据，同时为肿瘤研究提供及优化通路靶向代谢组学研究体系新技术手段。

近年重新认识到代谢重编程对肿瘤的关键作用。现今代谢研究已逐渐从定性描述进展到对代谢通量分配和代谢物分子的定量研究层面。葡萄糖代谢通路分配重编程及其代谢物分子异常变化对包括肺癌在内的肿瘤起重要作用。肺癌干细胞是肺癌发生、转移、耐药和复发的重要源泉，然而迄今对其葡萄糖代谢动态流向各通路分配的定量研究极为有限，成为阻碍认识其代谢重编程通路机制的重要瓶颈之一。本项目建立了基于高效液相色谱串联四级杆飞行时间质谱（Ultraperformance liquid chromatography tandem quadrupole time-of-flight mass spectrometry, UPLC-QTOF/MS）的非靶向代谢组学方法和基于高效液相色谱串联三重四级杆质谱（Ultraperformance liquid chromatography tandem triple-quadrupole mass spectrometry, UPLC-QQQ/MS）的葡萄糖靶向代谢组学方法，应用非标记靶向代谢组学研究肺腺癌干细胞与对照细胞差异代谢组中与葡萄糖代谢通路相关的代谢物，应用基于稳定性同位素标记示踪技术的靶向代谢组学方法定量葡萄糖代谢动态流向及各通路分配情况，确定和绘制肺腺癌干细胞特征性葡萄糖代谢动态流向及通路分配图谱，确定肺腺癌干细胞葡萄糖代谢重编程通路的关键节点和可干预节点。研究结果发现肺癌干细发生代谢重编程，其中糖代谢偏好流向酵解及磷酸戊糖途径，而流向三羧酸循环途径与非干细胞未见明显差异。针对该发现，筛选和确定对肺癌和肺癌耐药具有决定作用的代谢通路关键分子磷酸甘油酸变位酶和醛脱氢酶。靶向磷酸甘油酸变位酶和醛脱氢酶明显对抗肺癌生长转移，并且可以克服耐药。本项目为肺癌干细胞代谢重编程通路机制研究和基于该机制的潜在新药靶的发现提供理论和实验依据；同时，实验中优化的稳定同位素（无放射性）示踪通路靶向代谢组学技术体系还可扩展应用于多种内源性分子和药物代谢的研究，为肿瘤基础科学和转化医/药学提供和优化强有力的研究体系新技术手段。