Targeting Malic Enzyme 3 as a Synthetic Lethality Target in Pancreatic Cancer

将苹果酸酶 3 作为胰腺癌的合成致死靶点

基本信息

批准号：
10241331
负责人：
Prasenjit Dey
金额：
$ 24.9万
依托单位：
ROSWELL PARK CANCER INSTITUTE CORP
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-09-01 至 2022-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10241331
关键词：
3-Dimensional 5&apos -AMP-activated protein kinase Address Affect Allosteric Site Attention Binding Biological Assay Biological Models Branched-Chain Amino Acids Bypass CRISPR/Cas technology Catabolism Cell Death Cell Survival Cells Collection Computer Models Data Databases Dimerization Drug Design Enzymes Event Extinction (Psychology)Future Genes Genetic Transcription Genome Genomics Glutamates Glutamine Goals Grant Housekeeping Housekeeping Gene In Vitro Individual Investigation Lead MADH4 gene Maintenance Malignant Neoplasms Malignant neoplasm of pancreas Mediating Metabolic Mitochondria Modeling Molecular Molecular Target NADP Natural regeneration Neoplasm Metastasis Nitrogen Nucleotide Biosynthesis Oncogenes Outcome Pancreatic Ductal Adenocarcinoma Patients Pharmaceutical Preparations Play Production Protein Isoforms Proteins Regulation Resistance Role Route SRE-1 binding protein Site Site-Directed Mutagenesis Specificity Structure Survival Rate System Testing Tetanus Helper Peptide Therapeutic Tumor Burden Tumor Suppressor Genes Validation alpha ketoglutarate amino group base branched-chain-amino-acid transaminase cancer survival computer cluster experimental study gain of function mutation genomic locus high end computer high throughput screening in silico in vivo in vivo Model inhibitor/antagonist insight malic enzyme metabolic abnormality assessment metabolomics molecular targeted therapies mouse model mutant nucleobase pancreatic ductal adenocarcinoma cell precision medicine small molecule therapeutic target therapy resistant tumor tumor progression uptake virtual screening

项目摘要

Project Summary/Abstract Pancreatic ductal adenocarcinoma (PDAC) is the most common cancer of the pancreas and 5-year survival rate for PDAC patients is a dreadful ~6%. PDAC genome sustains frequent deletion of tumor suppressor gene loci, most notably SMAD4, which is homozygously deleted in approximately 30% of cases. As loss of neighboring housekeeping genes can confer collateral lethality, I sought to determine whether loss of the metabolic gene malic enzyme (ME) 2 in the SMAD4 locus would create a cancer-specific metabolic vulnerability upon targeting its paralogous isoform ME3. Using in vitro and in vivo model system, I demonstrated that ME3 depletion in ME2 null PDAC cells causes cell death. Mechanistically, integrated metabolomic and molecular investigation of mitochondrial ME-deficient cells revealed diminished NADPH production and consequent high ROS which activates AMP activated protein kinase (AMPK), and which in turn suppresses BCAT2 (Branched chain amino acid transaminase 2) gene via sterol regulatory element-binding protein 1 (SREBP1)-directed transcription. BCAT2 catalyzes the transfer of the amino group from branched chain amino acids (BCAA) to alpha-ketoglutarate (αKG) thereby regenerating glutamate, which functions in part to support de novo nucleobase synthesis. In summary, this data shows that mitochondrial ME is required for pancreatic cancer survival and can be exploited as vulnerability for treatment. The goal of the project is to determine the druggability of ME in PDAC. In order to achieve this goal, I propose the following: 1) Systematic validation of Malic Enzyme as a therapeutic target in pancreatic cancer, 2) Mechanistic understanding of metabolic deregulation upon ME3 extinction; 3) Identification of selective ME3 inhibitors. By addressing the above aim, I will gain further insights into the structural features of ME3 and generate a blueprint for rational drug design to target ME3. Moreover, I will also identify the potential resistance or metabolic bypass mechanism of ME3 treatment by tracing metabolites such as BCAA and glutamine. Moreover, my unique understanding of the BCAA regulation will identify new avenues for targeting surrogate target of ME3. Finally, will undertake structure-function analysis of ME3 and validate the top hits identified by in-silico analysis.

项目摘要/摘要胰腺导管腺癌（PDAC）是胰腺最常见的癌症和5年生存 PDAC患者的比率是可怕的6％。 PDAC基因组自杀经常删除肿瘤抑制基因基因座，最著名的是SMAD4，在大约30％的病例中被纯合删除。作为损失邻近的家政基因可以赋予附带致死性，我感觉到是否丢失了 SMAD4基因座中的代谢基因MALIC酶（ME）2将产生癌症特异性代谢瞄准其寄生虫同工型ME3时的脆弱性。使用体外和体内模型系统，I 证明ME3在ME2 NULL PDAC细胞中的部署会导致细胞死亡。机械上，集成线粒体ME缺陷细胞的代谢组学和分子研究显示NADPH减少生产和随之而来的高ROS激活AMP激活蛋白激酶（AMPK），然后又通过固醇调节元件结合抑制BCAT2（分支链氨基酸转氨酶2）基因蛋白1（SREBP1）指导转录。 BCAT2催化了分支的氨基群的转移链氨基酸（BCAA）至α-酮戊二酸（αkg），从而再生谷氨酸支持从头核酶合成的一部分。总而言之，这些数据表明需要线粒体我对于胰腺癌的生存，可以探索为治疗脆弱性。该项目的目的是确定我在PDAC中的可药用性。为了实现此目标，我提出以下建议：1）系统将苹果酶验证为胰腺癌的治疗靶点，2）机械理解对ME3扩展的代谢放松管制； 3）识别选择性ME3抑制剂。通过解决上述目标，我将进一步了解ME3的结构特征，并产生一个理性的蓝图靶向ME3的药物设计。此外，我还将确定潜在的抗药性或代谢旁路通过追踪代谢物（例如BCAA和谷氨酰胺）来处理ME3治疗的机理。而且，我的独特之处对BCAA法规的了解将确定针对ME3替代目标的新途径。最后，将对ME3进行结构 - 功能分析，并通过内部分析验证最高命中。