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：
8975162
负责人：
Michael Alexander Badgley
金额：
$ 4.31万
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/8975162
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 Health 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 genetic manipulation improved in vivo inducible gene expression inhibitor/antagonist insight killings men mouse model mutant neoplastic cell novel novel therapeutic intervention novel therapeutics pancreatic cancer cells pancreatic neoplasm pre-clinical preclinical efficacy programs protein expression 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肿瘤中之一，并且在四分之一的人类癌症中被突变[1]。特别是在胰腺癌中，Kras以惊人的高率突变，在所有情况下超过95％都激活了[2]。尽管在胰腺癌的转基因小鼠模型中熄灭了RAS信号传导和突变的RAS蛋白表达完全消融了肿瘤[3]，但事实证明，开发特定的RAS抑制剂或其他有用的胰腺癌患者疗法。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年的存活率仅为5％。显然，在胰腺癌领域开发新的治疗方法是为了确定更有效和毒性更少的治疗方法。当前的努力是为了确定在含有RAS突变的细胞的背景下，其操纵可能会被证明是致命的途径。这种“合成致命”组合可以在临床环境中提供极大的实用性，从而限制了与辐射和化学治疗方法相关的毒性，并以特定的，有效的方式靶向由RAS驱动的肿瘤。 2003年，Dr. Stockwell鉴定了RAS突变细胞的特定杀手[4]。这种称为“ erastin”的化合物通过抑制了名为System XC- [5]的胱氨酸 - 谷氨酸抗毒剂的氧化细胞死亡形式，称为“铁毒性”。但是，这种方法在体内的实用性以及系统XC-在RAS驱动肿瘤中的确切作用仍然未知。鉴于突变体RA在PDA的开发和维持中的作用，拟议研究的总体目标是检验以下假设：System XC-功能是开发和维持KRAS突变体胰腺癌所必需的。具体而言，我们旨在询问系统XC-遗传和药理学操纵对胰腺癌细胞增殖和存活的后果。利用分子遗传学，小鼠建模和临床前疗法方面的专业知识，我们将询问系统XC-在PDA生长和维持PDA中的作用，并评估针对该途径作为胰腺癌患者治疗干预的手段的临床生存能力。具体而言，我们将使用SHRNA和病毒技术在体外调节系统XC-。为了研究系统XC-体内遗传消融的影响，我们将跨小鼠XC功能缺乏的小鼠与我们的PDA临床前模型KPC小鼠。最后，我们将使用源自Erastin的化合物来评估系统XC-在体外和KPC模型中的药理抑制对胰腺肿瘤细胞的影响。总而言之，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生存中的确切作用，并验证系统XC-抑制剂在有效治疗胰腺癌中的使用。