靶向LDH-A能量代谢对T细胞急性淋巴细胞白血病的抗白血病效应及机制研究

项目名称： 靶向LDH-A能量代谢对T细胞急性淋巴细胞白血病的抗白血病效应及机制研究

项目编号： No.81200368

项目类型： 青年科学基金项目

立项/批准年度： 2013

项目学科： 医学一处

项目作者： 彭宏凌

作者单位： 中南大学

项目金额： 23万元

中文摘要： LDH－A是糖酵解途径的关键限速酶，在急性T细胞淋巴细胞白血病（T－ALL）患者，血清LDH常显著增高，并为影响ALL预后的独立危险因素。已知LDH－A的表达受NOTCH1、MYC调节。本项目以NOTCH1突变及野生型细胞株（DU528、MOLT4）为研究对象，观察低氧刺激对细胞株LDH活性、细胞增殖及线粒体凋亡的影响和差异，进一步采用LDH-A抑制剂干预细胞株和特异性敲除LDH-A基因，观察白血病细胞增殖活力和凋亡状态等细胞生物学行为，确立LDH-A的促白血病细胞增殖和诱导凋亡耐受活力。同时以MYC诱导的T-ALL转基因斑马鱼模型为研究模型，构建强力霉素调控性I-SceI-Rag2-rtTA-M2-I-SceI，I-SceI-LDH-A-TetR-EGFP-I-SceI载体，联合注入单细胞期受精卵，建立可调控性LDH-A高表达的T-ALL转基因模型，观察Tet on 状态下LDH-A基因

中文关键词： 急性T淋巴细胞白血病；乳酸脱氢酶；靶向治疗；CRISPR Cas9；斑马鱼

英文摘要： Lactate dehydrogenase A（LDH-A）is a key enzyme of the glycolytic pathway, and is regulated by NOTCH1 and MYC.Our study will investigate NOTCH1 mutant and wild type cell lines（DU528,MOLT4）through treatment with LDHA inhibitor and LDHA knocked down by means of LDHA short hairpin RNAs.We will observe the affections of LDHA suppression on proliferation and apoptosis.And genetically inhibit LDHA to investigate the affections of attenuated LDHA expression increase GSI sensitive.Using MYC-induced T-ALL transgenic zebrafish model, we are going to construct tetracycline（Tet）-On/off system in zebra?sh.The plasmids: I-SceI-Rag2-rtTA-M2-I-SceI, I-SceI-LDHA-TetR-EGFP-I-SceI modulated conveniently by doxycycline（Dox）, co-microinjection into one-cell stage zebrafish embryos, generation Dox induction LDHA high expression in T-ALL transgenic zebrafish.We will investigate the process that LDHA promotes the T-ALL progression upon Tet on and delays the disease upon Tet off； We will also treat on two groups fishes with LDH-A inhibitor, and observe the anti tumor affection of LDHA.By exploring the LDHA as energy metabolism target in T-ALL, the study will provide theoretical proofs for new targeted therapy and the decreasing of T-ALL resistant rate.

英文关键词： T cell llymphoblastic leukemia；Lactate dehydrogenase A；target therapy；CRISPR Cas9；zebrafish