Genetic and pharmacological manipulation of system xc in pancreatic cancer

胰腺癌中 xc 系统的遗传和药理学操作

基本信息

批准号：
8598355
负责人：
Michael Alexander Badgley
金额：
$ 4.22万
依托单位：
COLUMBIA UNIVERSITY HEALTH SCIENCES
依托单位国家：
美国
项目类别：
财政年份：
2013
资助国家：
美国
起止时间：
2013-12-01 至 2017-11-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8598355
关键词：
Ablation Accounting Affinity Amino Acid Transporter Amino Acids Antioxidants Apoptosis Autophagocytosis Binding Biological Cancer Etiology Cancer Patient Cell Death Cells Cessation of life Chemicals Clinical Complication Cysteine Cystine Dependency Development Diagnosis Disease Disulfides Drug Metabolic Detoxication Enzymes Essential Amino Acids Family Floods Free Radical Scavengers Genes Genetic Glutamates Glutathione Goals Growth Guanosine Triphosphate Human Human Genome In Vitro Intervention Iron KRAS2 gene Link MEKs Maintenance Malignant Neoplasms Malignant neoplasm of pancreas Metabolic Methionine Missense Mutation Modeling Modification Molecular Genetics Mus Mutate Mutation Names Necrosis Neurons Oncogenes Oncogenic Oxidation-Reduction Pancreatic Ductal Adenocarcinoma Pathway interactions Patients Pharmaceutical Preparations Phenotype Piperazines Pre-Clinical Model Production Property Proteins Radiation Reactive Oxygen Species Research Resistance Role Signal Transduction Solubility Survival Rate System Techniques Technology Testing Therapeutic Therapeutic Intervention Toxic effect Transgenic Mice Transgenic Organisms United States Viral Viral Vector Woman Work activating transcription factor analog antiporter base cell transformation cellular engineering chemotherapy clinically relevant effective therapy erastin fluoromethyl 2,2-difluoro-1-(trifluoromethyl)vinyl ether genetic manipulation improved in vivo inhibitor/antagonist insight killings men mouse model mutant neoplastic cell novel novel therapeutic intervention pancreatic cancer cells pancreatic neoplasm pre-clinical preclinical efficacy programs protein expression public health relevance research study small hairpin RNA tool tumor

项目摘要

DESCRIPTION (provided by applicant): RAS is among the most potent oncogenes in the human genome and is mutated in over one in four of all human cancers [1]. In pancreatic cancer specifically, KRAS is mutated at an alarmingly high rate, activated in over 95% of all cases [2]. While extinguishing both RAS signaling and mutant RAS protein expression in a transgenic mouse model of pancreatic cancer completely ablates tumors [3], developing specific RAS-inhibitors or other useful therapies for pancreatic cancer patients has proven near impossible. In keeping with this lack of progress, pancreatic ductal adenocarcinoma (PDA) remains one of the most lethal cancers, with a 5-year survival rate of just 5%. It is clear that there is a critical ned to develop novel therapeutic approaches in the field of pancreatic cancer in order to identify more effective and less toxic treatments. Current efforts have been made to identify pathways whose manipulation could prove lethal specifically in the context of cells containing RAS mutations. Such "synthetic lethal" combinations could provide great utility in a clinical setting, limiting the toxicities commonly associated with both radiation and chemotherapeutic approaches, and targeting RAS-driven tumors in a specific, effective way. In 2003, a synthetic lethal chemical screen carried out by Dr. Stockwell identified a specific killer of Ras-mutant cells [4]. This compound, called "erastin," causes an iron-dependent form of oxidative cell death, termed "ferroptosis," through the inhibition of a cystine-glutamate antiporter named system xc- [5]. However, the utility of this approach in vivo and the precise role of system xc- in Ras-driven tumors remains unknown. Given the role of mutant RAS in the development and maintenance of PDA, the overall goal of the proposed research is to test the hypothesis that system xc- function is required for the development and maintenance of KRAS mutant pancreatic cancers. Specifically, we aim to interrogate the consequences of genetic and pharmacological manipulation of system xc- on the proliferation and survival of pancreatic cancer cells. Leveraging expertise in molecular genetics, mouse modeling, and preclinical therapeutics, we will interrogate the role of system xc- in the growth and maintenance of PDA and evaluate the clinical viability of targeting this pathway as a means of therapeutic intervention for pancreatic cancer patients. Specifically, we will use shRNA and viral technologies to modulate system xc- in vitro. To study the effects of genetic ablation of system xc- in vivo, we will cross mice deficient for system xc- function to our preclinical model of PDA, the KPC mouse. Finally, we will use a compound, derived from erastin, to evaluate the effect of pharmacological inhibition of system xc- on pancreatic tumor cells both in vitro and in the KPC model. In summary, the experiments proposed herein will aid our understanding of the precise role of redox state maintenance in cancer growth and survival as well as validate the use of system xc- inhibitors in the effective treatment of pancreatic cancer.

描述（由申请人提供）：RAS 是人类基因组中最有效的癌基因之一，超过四分之一的人类癌症都会发生突变 [1]。特别是在胰腺癌中，KRAS 的突变率高得惊人，在超过 95% 的病例中被激活[2]。虽然在胰腺癌转基因小鼠模型中消除 RAS 信号传导和突变 RAS 蛋白表达可以完全消除肿瘤 [3]，但事实证明，为胰腺癌患者开发特异性 RAS 抑制剂或其他有用的疗法几乎是不可能的。由于缺乏进展，胰腺导管腺癌 (PDA) 仍然是最致命的癌症之一，5 年生存率仅为 5%。显然，在胰腺癌领域迫切需要开发新的治疗方法，以确定更有效和毒性更低的治疗方法。目前，人们正在努力寻找在含有 RAS 突变的细胞中，对其进行操作可能会致命的途径。这种“合成致死”组合可以在临床环境中提供巨大的实用性，限制通常与放射和化疗方法相关的毒性，并以特定、有效的方式针对 RAS 驱动的肿瘤。 2003年，博士进行了合成致命化学筛选。 Stockwell 发现了 Ras 突变细胞的特异性杀手 [4]。这种化合物称为“erastin”，通过抑制名为系统 xc- 的胱氨酸-谷氨酸逆向转运蛋白，导致铁依赖性形式的氧化细胞死亡，称为“铁死亡”[5]。然而，这种方法在体内的实用性以及系统 xc- 在 Ras 驱动的肿瘤中的确切作用仍然未知。鉴于突变 RAS 在 PDA 发生和维持中的作用，本研究的总体目标是检验系统 xc 功能对于 KRAS 突变胰腺癌的发生和维持所必需的假设。具体来说，我们的目的是探讨 xc- 系统的遗传和药理学操作对胰腺癌细胞增殖和存活的影响。利用分子遗传学、小鼠建模和临床前治疗方面的专业知识，我们将探讨系统 xc- 在 PDA 生长和维持中的作用，并评估针对该途径作为胰腺癌患者治疗干预手段的临床可行性。具体来说，我们将使用shRNA和病毒技术来体外调节系统xc-。为了研究体内 xc- 系统基因消融的影响，我们将缺乏系统 xc- 功能的小鼠与我们的 PDA 临床前模型（KPC 小鼠）杂交。最后，我们将使用一种源自erastin的化合物，在体外和KPC模型中评估系统xc-的药理抑制作用对胰腺肿瘤细胞的影响。总之，本文提出的实验将有助于我们理解氧化还原状态维持在癌症生长和存活中的精确作用，并验证系统xc-抑制剂在胰腺癌的有效治疗中的使用。