Altered Glycolytic Pathway in in vitro Models of leukemia developed in hypoxia

体外糖酵解途径的改变在缺氧条件下开发白血病模型

• 批准号: 8100621
• 负责人: Min Hee Kang
• 金额: $ 44.44万
• 依托单位: TEXAS TECH UNIVERSITY HEALTH SCIS CENTER
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-03-01 至 2014-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8100621
• 关键词: Acute Lymphocytic Leukemia; Adolescent; Aerobic; Affect; Air; Apoptosis; Biochemical Pathway; Biological Assay; Biological Preservation; Bone Marrow; Cancer Biology; Cancer cell line; Cell Culture Techniques; Cell Line; Cell Proliferation; Cells; Child; Childhood; Childhood Acute Lymphocytic Leukemia; Consumption; Cultured Cells; Data; Dexamethasone; Diagnosis; Drug Combinations; Drug Delivery Systems; Enzymes; Flow Cytometry; Gene Expression; Gene Proteins; Genes; Glucocorticoids; Glucose; Glycolysis; Goals; Growth; Hypoxia; Immunocompromised Host; Immunosuppression; In Vitro; Incidence; Laboratories; Malignant Neoplasms; Malignant lymphoid neoplasm; Metabolic Pathway; Metabolism; Mitochondria; Molecular Profiling; Mus; NOD/SCID mouse; Non-Malignant; Oncogenic; Oxidative Phosphorylation; Oxygen; Oxygen measurement, partial pressure, arterial; Pathway interactions; Patients; Pharmaceutical Preparations; Physiological; Preclinical Testing; Production; Proliferating; RNA; Regulation; Regulatory Pathway; Relapse; Research Personnel; Sampling; Short Tandem Repeat; Signal Pathway; Sirolimus; Study models; Surface Antigens; Testing; Therapeutic; Time; United States; Warburg Effect; Western Blotting; Xenograft Model; Xenograft procedure; aerobic glycolysis; asparaginase; base; cancer cell; cytotoxicity; density; drug sensitivity; established cell line; genome-wide; glucose metabolism; human FRAP1 protein; in vitro Model; in vitro testing; in vivo; inhibitor/antagonist; leukemia; mTOR Inhibitor; novel; pre-clinical; programs; repository; research study

DESCRIPTION (provided by applicant): The majority of cancers employ an elevated level of glycolysis, even under mildly aerobic conditions. This bias towards aerobic glycolysis, known as the Warburg effect, is a hallmark of cancer that regulates various cellular attributes including drug sensitivity. Many studies show that the major pathway regulating glycolysis is PI3K/Akt/mTOR pathway. We hypothesize that cancer cell lines established in physiological hypoxia can more accurately mimic leukemia in vivo, and that in vitro studies in physiological O2 of agents targeting the PI3K/Akt/mTOR Pathway in ALL will more closely resemble in vivo results. Our goals are 1) to show that the glycolytic pathway genes are affected by oxygen conditions in acute lymphoblastic leukemia (ALL) established in different oxygen conditions and therefore, the more glucose consumption and more lactate production are seen in cell lines established in hypoxia; and 2) using the cell lines established in hypoxia and direct xenografts we will evaluate drugs targeting the glycolytic pathway. By comparing new childhood acute lymphoblastic leukemia cell lines in atmospheric (20% O2) and physiologic bone marrow (5% O2) oxygen concentrations, we will demonstrate that atmospheric oxygen tension inhibits glycolysis-related gene expression, glucose consumption, lactate production, cell proliferation, and alters dug sensitivities. The effects of physiological and non-physiological (i.e. "standard" culture conditions) oxygen concentrations on glycolysis are not readily reversible by switching cell cultures from one oxygen condition to another, indicating the need to establish cell lines in physiological oxygen conditions in order to provide accurate in vitro models for studying cancer cell metabolism. Our preliminary observations suggest that preservation of the Warburg effect in leukemia requires culturing cells at physiologic oxygen concentrations from the time of their initial establishment. By establishing and evaluating a large panel of leukemia cell lines in two different oxygen conditions, and also for comparison as direct xenografts in immunocompromised mice, we will provide an understanding on the importance of culture conditions on metabolic networks in cancer in vitro models. In addition, we will evaluate whether glycolytic pathway could provide feasible targets for ALL treatment by evaluating new PI3K inhibitor with less immunosuppression in combination with current therapy of ALL (e.g. dexamethasone and L-asparaginase, another metabolism inhibitor). This project will also make available for other investigators via the COG repository (www.COGcell.org) a unique set of well characterized and validated laboratory models for studying cancer biology and preclinical therapeutics. PUBLIC HEALTH RELEVANCE: Lymphoid malignancies are the most common cancer in children and adolescents in the United States, and the most prevalent of these is acute lymphoblastic leukemia (ALL) with the incidence being gradually increasing over the last 25 years. Significant improvements in primary therapy for childhood ALL have led to an overall cure rate of approximately 80 %. However, of the 20% of patients who relapse, the majority die. The ultimate goals of our proposal are to identify how leukemia cells make energy to grow and proliferate, and to find drugs that can attack the energy generating mechanisms of leukemia cells.

描述（由申请人提供）：即使在轻度有氧条件下，大多数癌症也采用升高水平的糖酵解。这种有氧糖酵解的偏向被称为瓦伯格效应，是癌症的一个标志，它调节包括药物敏感性在内的各种细胞属性。许多研究表明调节糖酵解的主要途径是PI3K/Akt/mTOR途径。我们假设在生理性缺氧中建立的癌细胞系可以更准确地模拟体内白血病，并且针对 ALL 中 PI3K/Akt/mTOR 通路的药物的生理性 O2 体外研究将更接近体内结果。我们的目标是 1) 证明在不同氧气条件下建立的急性淋巴细胞白血病 (ALL) 中，糖酵解途径基因受到氧气条件的影响，因此，在缺氧条件下建立的细胞系中，葡萄糖消耗更多，乳酸产生更多； 2）使用在缺氧和直接异种移植中建立的细胞系，我们将评估针对糖酵解途径的药物。通过比较大气（20% O2）和生理骨髓（5% O2）氧浓度下的新儿童急性淋巴细胞白血病细胞系，我们将证明大气氧张力抑制糖酵解相关基因表达、葡萄糖消耗、乳酸产生、细胞增殖，并改变挖掘敏感性。通过将细胞培养物从一种氧条件切换到另一种氧条件，生理和非生理（即“标准”培养条件）氧浓度对糖酵解的影响不容易逆转，这表明需要在生理氧条件下建立细胞系，以便为研究癌细胞代谢提供准确的体外模型。我们的初步观察表明，在白血病中保留瓦尔堡效应需要从细胞最初建立时起就在生理氧浓度下进行培养。通过在两种不同的氧气条件下建立和评估大量白血病细胞系，并与免疫受损小鼠中的直接异种移植物进行比较，我们将了解培养条件对癌症体外模型中代谢网络的重要性。此外，我们将通过评估免疫抑制较少的新型PI3K抑制剂与目前的ALL治疗（例如地塞米松和另一种代谢抑制剂L-天冬酰胺酶）相结合，来评估糖酵解途径是否可以为ALL治疗提供可行的靶点。该项目还将通过 COG 存储库 (www.COGcell.org) 为其他研究人员提供一套独特的、经过充分表征和验证的实验室模型，用于研究癌症生物学和临床前治疗。 公共卫生相关性：淋巴恶性肿瘤是美国儿童和青少年中最常见的癌症，其中最常见的是急性淋巴细胞白血病 (ALL)，其发病率在过去 25 年中逐渐增加。儿童 ALL 初级治疗的显着改善使总体治愈率达到约 80%。然而，在 20% 的复发患者中，大多数死亡。我们提案的最终目标是确定白血病细胞如何产生生长和增殖的能量，并找到可以攻击白血病细胞能量产生机制的药物。

项目成果

Synergistic activity of rapamycin and dexamethasone in vitro and in vivo in acute lymphoblastic leukemia via cell-cycle arrest and apoptosis.

• DOI: 10.1016/j.leukres.2011.10.022
• 发表时间: 2012-03
• 期刊: Leukemia research
• 影响因子: 2.7
• 作者: Zhang C;Ryu YK;Chen TZ;Hall CP;Webster DR;Kang MH
• 通讯作者: Kang MH

Modulation of Glucocorticoid Resistance in Pediatric T-cell Acute Lymphoblastic Leukemia by Increasing BIM Expression with the PI3K/mTOR Inhibitor BEZ235.

• DOI: 10.1158/1078-0432.ccr-15-0114
• 发表时间: 2016-02-01
• 期刊: Clinical cancer research : an official journal of the American Association for Cancer Research
• 作者: Hall CP;Reynolds CP;Kang MH
• 通讯作者: Kang MH

Preservation of high glycolytic phenotype by establishing new acute lymphoblastic leukemia cell lines at physiologic oxygen concentration.

通过在生理氧浓度下建立新的急性淋巴细胞白血病细胞系来保留高糖酵解表型。

• DOI: 10.1016/j.yexcr.2015.03.024
• 发表时间: 2015
• 期刊: Experimental cell research
• 影响因子: 3.7
• 作者: Sheard,MichaelA;Ghent,MatthewV;Cabral,DanielJ;Lee,JoanneC;Khankaldyyan,Vazgen;Ji,Lingyun;Wu,SamuelQ;Kang,MinH;Sposto,Richard;Asgharzadeh,Shahab;Reynolds,CPatrick
• 通讯作者: Reynolds,CPatrick