Exploiting Metabolic Changes in HCC for Cancer Therapy

利用 HCC 的代谢变化进行癌症治疗

• 批准号: 10328967
• 负责人: Alexander Richard Terry
• 金额: $ 3.98万
• 依托单位: STATE UNIVERSITY NEW YORK STONY BROOK
• 依托单位国家: 美国
• 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/10328967
• 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; interest; knock-down; liver cancer model; 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

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）是美国第十大最常见的癌症，但却是癌症相关死亡的第四大原因。到目前为止，FDA批准的唯一分子疗法索拉非尼尚未有效。该基金旨在确定己糖激酶2（HK 2）在HCC发展和进展中的作用，以及HK 2是否是HCC治疗的可行药物靶点。正常健康成人肝细胞表达葡萄糖激酶，一种低亲和力己糖激酶。然而，在向肝硬化和HCC的病理转变期间，转化的肝细胞沉默葡萄糖激酶并诱导HK 2。我们已经在人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亚型转变。这种转变也代表了更普遍的代谢改变，由此癌细胞增加葡萄糖利用和乳酸分泌，尽管存在充足的氧气，也称为瓦尔堡效应。HK 2是一种“线粒体”己糖激酶，这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都是必不可少的。这项资助的一部分将确定HK 2的线粒体相互作用促进肿瘤发生的机制，以及更普遍地为什么线粒体结合对于完全糖酵解能力是必要的。我们已经证明，HK 2的敲除超过70%对HCC细胞是致命的，事实上，另一个小组已经证明，使用CRISPR/Cas9敲除筛选，具有HK 1或HK 2对于癌细胞活力是必不可少的。由于肝癌细胞仅表达HK 2，我们认为HK 2是肝癌的理想药物靶点。我们将使用已建立的HCC小鼠模型来确定这种治疗策略的可行性，以确定HK 2在HCC起始和进展中的细胞自主必要性。 我们将通过在裸鼠皮下植入人肝癌细胞后使用多西环素诱导的靶向HK 2的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HK 2抑制协同作用。初步数据表明，NADPH的生产，在癌细胞中脂肪酸合成和氧化还原平衡的重要电子源，是完全依赖于苹果酸酶1和谷氨酰胺在肝癌细胞中的代谢。我们将利用这种依赖性与转氨酶-1抑制剂，BPTES，或复合物-1抑制剂，二甲双胍与HK 2抑制。HK 2缺失应防止戊糖磷酸途径的补偿性激活和糖酵解丙酮酸进入线粒体的TCA通量的补偿性通量，从而与BPTES或二甲双胍产生协同作用。

项目成果

Sequential Activation of Guide RNAs to Enable Successive CRISPR-Cas9 Activities.

• DOI: 10.1016/j.molcel.2020.12.003
• 发表时间: 2021-01-21
• 期刊: MOLECULAR CELL
• 影响因子: 16
• 作者: Clarke, Ryan;Terry, Alexander R.;Pennington, Hannah;Hasty, Cody;MacDougall, Matthew S.;Regan, Maureen;Merrill, Bradley J.
• 通讯作者: Merrill, Bradley J.

The calcium-binding protein S100B reduces IL6 production in malignant melanoma via inhibition of RSK cellular signaling.

• DOI: 10.1371/journal.pone.0256238
• 发表时间: 2021
• 期刊: PloS one
• 影响因子: 3.7
• 作者: Alasady MJ;Terry AR;Pierce AD;Cavalier MC;Blaha CS;Adipietro KA;Wilder PT;Weber DJ;Hay N
• 通讯作者: Hay N