HTS for inhibitors of IDH1m & synthetic-lethal in tumor cells producing 2HG

IDH1m 抑制剂的 HTS

• 批准号: 8070296
• 负责人: Lenny Dang
• 金额: $ 3.38万
• 依托单位: AGIOS PHARMACEUTICALS, INC.
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-02-01 至 2012-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8070296
• 关键词: 2q33; Active Sites; Acute Myelocytic Leukemia; Agonist; Amino Acids; Arginine; Biochemical; Biological Assay; Carbon Dioxide; Cell Line; Cell Proliferation; Cells; Chemicals; Chromosomes; Citric Acid Cycle; Codon Nucleotides; Decarboxylation; Detection; Down-Regulation; Engineering; Enzymes; Genes; Genome; Genomics; Glioblastoma; Glioma; Goals; Isocitrate Dehydrogenase; Isocitrates; Lead; Malignant Neoplasms; Measures; Mutate; Mutation; NADP; Nature; Neoplasms; Oncogenes; Oncogenic; Pathogenesis; Pathway interactions; Patients; Point Mutation; Production; Protein Isoforms; Proteins; Recurrence; Reporting; Role; Screening procedure; Somatic Mutation; Specificity; Therapeutic Intervention; Translating; Tumor Biology; Validation; base; cancer cell; cancer type; cellular engineering; cofactor; dihydrolipoamide dehydrogenase; fibrosarcoma; gain of function; inhibitor/antagonist; insight; isocitrate; loss of function; mutant; neoplastic cell; novel; novel therapeutics; protein structure; resazurin; resorufin; stable cell line; tool; tumor

DESCRIPTION (provided by applicant): The goal of this project is to identify and optimize inhibitors that are specific for the isocitrate dehydrogenase 1 (IDH1). Unbiased genomic sequencing for 22 glioma genomes found recurrent mutation of IDH1 on chromosome 2q33-a gene encoding the cytosolic isoform of IIDH1 associated with the tricarboxylic acid cycle (TCA) that catalyzes the oxidative decarboxylation of isocitrate yielding 1-ketoglutarate and CO2 via NADP+ to NADPH conversion. Subsequent studies confirmed the recurrent IDH mutations in up to 70% of secondary gliomas and in 10% of AML cases. We have found that the somatic mutation of cancer-associated IDH1 is a point mutation resulting in various amino-acid substituent's at Arginine132 (IDH1 R132)-a key residue found in the enzyme's active site that when mutated results in the loss-of-function in metabolizing isocitrate but confers a gain-of-function to produce the oncometabolite 2-hydroxyglutarate (2HG). This in effect defines IDH1 as an oncogene and provides an extraordinary opportunity to discover chemical probes against mutant IDH1 that may translate into much needed new therapies for glioma and AML patients. The proposal has two specific aims: (1) Discovery of novel chemical probes against mutant IDH1 using a validated HTS assay for IDH1 R132H; (2) Performing a synthetic-lethal screen for chemical probes specific for 2HG-producing tumor cells using matched pair cell lines (parental U87MG glioblastoma, and a U87MG stable cell-line that constitutively expresses IDH1R132H mutant). PUBLIC HEALTH RELEVANCE: The aim of this proposal is to screen for chemical inhibitors that are specific for the mutant form of isocitrate dehydrogenase 1 (IDH1). Recently, 70% of grade II-IV gliomas were found to harbor recurrent IDH mutations. Somatic point mutations of IDH1-associated gliomas resulted in various substitution at Arginine 132 (IDH1 R132)-a residue found in the IDH1 active site that confers a gain-of-function when mutated, resulting in the neomorphic production of the oncometabolite 2- hydroxyglutarate (2HG). The assays described in this proposal provide a unique opportunity to identify chemical probes aimed at study the role of mutant IDH1 and 2HG in gliomas.

描述（由申请人提供）：该项目的目的是识别和优化针对异氯酸酯脱氢酶1（IDH1）的抑制剂。 22个神经胶质瘤基因组的公正基因组测序发现IDH1在染色体2q33-A基因上复发突变，编码与三羧酸周期（TCA）相关的IIDH1的胞质同工型，可催化同二羧酸脱羧酸脱羧酸含量的氧化二羧酸含量，并与1-固定物的氧化羧酸脱羧酸含量，并促成1-酮2的coctypation+ coptrogation+ coction+ coptroutaRation and coptrad croctyprate+ coptrad awardarate and croctyprate and croctyprate and croct croctyp aid a后。随后的研究证实了多达70％的继发性神经胶质瘤和10％的AML病例的复发性IDH突变。我们发现，与癌症相关的IDH1的体躯体突变是一个点突变，导致精氨酸132（IDH1 R132）的各种氨基酸取代基 - 在酶的活性部位中发现的一个关键残基，当突变时，突变导致了造成损失的损失，从而产生了代理化的同含量，从而产生了构造的同含量，从而产生了构造的同含量。 2-羟基戊二酸（2HG）。实际上，这将IDH1定义为一种癌基因，并提供了一个非凡的机会，可以发现针对突变体IDH1的化学探针，该探针可能转化为胶质瘤和AML患者的急需的新疗法。该提案具有两个具体的目的：（1）使用经过验证的HTS测定法对IDH1 R132H发现了针对突变体IDH1的新型化学探针； （2）使用匹配的成对细胞系（父母U87mg胶质母细胞瘤和U87MG稳定的细胞系），对具有2HG产生的肿瘤细胞的化学探针进行合成杀伤性筛选，并构成表达IDH1R132H突变体）。 公共卫生相关性：该提案的目的是筛选针对异急塞脱氢酶1（IDH1）突变形式的化学抑制剂。最近，发现70％的II-IV胶质瘤有复发性IDH突变。 IDH1相关的神经膜瘤的体细胞突变导致精氨酸132（IDH1 R132） - 在IDH1活跃部位发现的残基，在突变时赋予了功能良好，从而赋予了功能的收益，从而导致Neomorphic of Neomorthic产生oncometabolite 2- hydroxy-hydroxy-glutarate（2HG）。该提案中描述的分析提供了一个独特的机会，可以识别旨在研究突变体IDH1和2HG在神经胶质瘤中的作用的化学探针。