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.

项目摘要：肝细胞癌（HCC）是美国第十个最常见的癌症，但在美国与癌症有关的第四个主要原因。迄今为止，唯一经FDA批准的分子疗法索拉非尼尚未有效。该赠款旨在确定己糖酶2（HK2）在HCC开发和进展中的作用，以及HK2是否是HCC治疗的可行药物靶标。正常健康的成年肝细胞表达葡萄糖酶，一种低亲和力己糖酶。但是，在患者向肝硬化和HCC过渡过程中，转化的肝细胞静音葡萄糖酶并诱导HK2。我们已经在人类HCC细胞系和来自153例患者的312个样品的肿瘤微阵列中表明了这种同工型转变。这种过渡也代表了一种更一般的代谢改变，癌细胞增加了葡萄糖利用率，并使分泌量足够充分的氧气，也称为Warburg效应。 HK2是一种“线粒体”六动物酶，这意味着它能够与线粒体上的电压依赖离子通道结合，并且我们有数据表明这种相互作用对于肿瘤发生和有效的糖酵解利用都是必不可少的。该赠款的一部分将通过HK2的线粒体相互作用来确定哪种机制会促进肿瘤发生，以及更笼统地为什么要全糖酵解能力必不可少的线粒体结合。我们已经表明，HK2超过70％的敲低对HCC细胞是致命的，实际上另一组表明，使用CRISPR/CAS9敲除屏幕具有HK1或HK2对于癌细胞的生存能力至关重要。由于HCC细胞仅表达HK2，因此我们认为HK2是HCC的理想药物靶标。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HK2在HCC启动和进展中必需的细胞自主性。我们将通过在皮下植入裸鼠后使用强力霉素诱导的shRNA靶向HK2的shRNA来模仿药物治疗。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的产生是癌细胞中脂肪酸合成和氧化还原平衡的重要电子源，完全取决于HCC细胞中苹果酶1和谷氨酰胺代谢。我们将使用谷氨酰胺-1抑制剂，BPTE或复合-1抑制剂二甲双胍与HK2抑制作用来利用这种依赖性。 HK2缺失应防止磷酸戊糖途径的补偿性激活和糖酵解丙酮酸的补偿通量进入线粒体，以进行TCA-升力，从而与BPTES或二甲双胍产生协同作用。