Exploiting Metabolic Changes in HCC for Cancer Therapy

利用 HCC 的代谢变化进行癌症治疗

基本信息

批准号：
10447400
负责人：
Alexander Richard Terry
金额：
$ 4万
依托单位：
STATE UNIVERSITY NEW YORK STONY BROOK
依托单位国家：
美国
项目类别：
财政年份：
2018
资助国家：
美国
起止时间：
2018-02-16 至 2022-11-15
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10447400
关键词：
Adult Affinity Age-Months Alanine Anabolism Antioxidants BAY 54-9085 Binding Branched-Chain Amino Acids CRISPR/Cas technology Cancer Etiology Cell Proliferation Cell Survival Cells Cessation of life Cirrhosis Citric Acid Cycle Complex Data Dependence Dependovirus Development Doxycycline Drug Targeting Electron Transport Electrons Enterobacteria phage P1 Cre recombinase Enzymes Equilibrium Etiology Excretory function FDA approved Fatty Acids Financial compensation Glucokinase Glucose Glutaminase Glutamine Glycolysis Goals Grant Hepatocyte Hexokinase 2 Hexosamines Human In Vitro Injections Ion Channel Knock-out Label Liver Malates Malignant - descriptor Malignant Epithelial Cell Malignant Neoplasms Malignant neoplasm of liver Metabolic Metabolism Metformin Microarray Analysis Mitochondria Modeling Molecular Mus NADP Natural regeneration Non-Insulin-Dependent Diabetes Mellitus Nude Mice Oxidation-Reduction Oxygen Pathologic Pathway interactions Patients Pentosephosphate Pathway Pharmaceutical Preparations Pharmacotherapy Primary carcinoma of the liver cells Production Protein Isoforms Pyruvate Reactive Oxygen Species Research Project Grants Role Sampling Second Primary Cancers Serine Severities Source Subcutaneous Injections Survival Rate Therapeutic Therapeutic Intervention Thyroglobulin Tissue Microarray United States Warburg Effect cancer cell cancer therapy cell growth glucose metabolism hepatocellular carcinoma cell line hexokinase implantation in vivo inhibitor/antagonist interest knock-down malic enzyme mouse model mutant novel therapeutics prevent promoter side effect small hairpin RNA subcutaneous synergism tumor tumor growth tumor initiation tumor progression tumorigenesis voltage

项目摘要

Project Summary: Hepatocellular carcinoma (HCC) is the 10th most common cancer but the fourth leading cause of cancer-related death in the United States. To date, the only FDA-approved molecular therapy, sorafenib, has not been effective. This grant is aimed at determining the role of hexokinase 2 (HK2) in HCC development and progression, and whether or not HK2 is a viable drug target for HCC treatment. Normal healthy adult hepatocytes express glucokinase, a low affinity hexokinase. However, during pathological transition to cirrhosis and HCC, transformed hepatocytes silence glucokinase and induce HK2. We have shown this isoform transition in both human HCC cell lines and in a tumor microarray of 312 samples from 153 patients. This transition also represents a more general metabolic alteration whereby cancer cells increase glucose utilization and lactate secretion despite ample oxygen being present, also known as the Warburg Effect. HK2 is a “mitochondrial” hexokinase, meaning it is capable of binding to the voltage dependent ion channel on the mitochondria, and we have data suggesting this interaction is essential for both tumorigenesis and efficient glucose utilization. Part of this grant will determine by what mechanism the mitochondrial interaction of HK2 promotes tumorigenesis and more generally why mitochondrial binding is necessary for full glycolytic capacity. We have shown that knockdown of HK2 beyond 70% is lethal to HCC cells, and indeed another group has shown that having either HK1 or HK2 is essential for cancer cell viability using a CRISPR/Cas9 knockout screen. Since HCC cells only express HK2, we believe HK2 is an ideal drug target for HCC. We will determine the feasibility of this therapeutic strategy using established mouse models of HCC to determine the cell autonomous necessity of HK2 in HCC initiation and progression. We will emulate drug therapy by using a doxycycline-inducible shRNA targeting HK2 in human HCC cells following subcutaneous implantation in nude mice. We will also determine if targeting mitochondrial glutamine metabolism synergizes with HK2 inhibition. Preliminary data suggests that production of NADPH, an important electron source for fatty acid synthesis and redox balance in cancer cells, is completely dependent on malic enzyme 1 and glutamine metabolism in HCC cells. We will exploit this dependency with a glutaminase-1 inhibitor, BPTES, or a complex-1 inhibitor, metformin in concert with HK2 inhibition. HK2 deletion should prevent compensatory activation of the pentose phosphate pathway and compensatory flux of glycolytic pyruvate into the mitochondria for TCA-flux, creating synergism with BPTES or metformin.

项目概要：肝细胞癌 (HCC) 是美国第十大常见癌症，也是癌症相关死亡的第四大原因。迄今为止，FDA 批准的唯一分子疗法索拉非尼尚未有效。己糖激酶 2 (HK2) 在 HCC 发生和进展中的作用，以及 HK2 是否是 HCC 治疗的可行药物靶点正常健康成人肝细胞表达低亲和力的葡萄糖激酶。然而，在向肝硬化和 HCC 的病理转变过程中，转化的肝细胞沉默葡萄糖激酶并诱导 HK2。我们已经在人类 HCC 细胞系和来自 153 名患者的 312 个样本的肿瘤微阵列中显示了这种同工型转变。尽管存在充足的氧气，癌细胞仍会增加葡萄糖利用和乳酸分泌，这种一般代谢改变也称为“瓦伯格效应”。 “线粒体”己糖激酶，意味着它能够与线粒体上的电压依赖性离子通道结合，我们有数据表明这种相互作用对于肿瘤发生和有效的葡萄糖利用至关重要，这笔资助的一部分将确定线粒体相互作用的机制。 HK2 促进肿瘤发生，更一般地说，为什么线粒体结合对于完整的糖酵解能力是必需的。我们已经证明，HK2 的敲低超过 70% 对于 HCC 细胞来说是致命的，事实上，另一组已经表明，具有完整的糖酵解能力。 HK1 或 HK2 对于使用 CRISPR/Cas9 敲除筛选的癌细胞活力至关重要，由于 HCC 细胞仅表达 HK2，因此我们相信 HK2 是 HCC 的理想药物靶点，我们将使用已建立的小鼠模型来确定这种治疗策略的可行性。 HCC 以确定 HK2 在 HCC 发生和进展中的细胞自主必要性我们将在皮下植入后在人 HCC 细胞中使用靶向 HK2 的多西环素诱导型 shRNA 来模拟药物治疗。我们还将确定靶向线粒体谷氨酰胺代谢是否与 HK2 抑制协同作用。初步数据表明，NADPH（癌细胞中脂肪酸合成和氧化还原平衡的重要电子源）的产生完全依赖于苹果酸酶 1 和谷氨酰胺。我们将利用谷氨酰胺酶 1 抑制剂 BPTES 或复合物 1 抑制剂二甲双胍与 HK2 抑制相结合来利用这种依赖性。防止磷酸戊糖途径的代偿性激活和糖酵解丙酮酸进入线粒体以实现 TCA 通量的代偿性流动，从而与 BPTES 或二甲双胍产生协同作用。