Rational design of anti-cancer therapeutics harnessing the synthetic lethality of methionine metabolism and arginine methyltransferases

利用蛋氨酸代谢和精氨酸甲基转移酶的合成杀伤力合理设计抗癌疗法

• 批准号: 10536888
• 负责人: Gabriel T Bedard
• 金额: $ 5.18万
• 依托单位: ALBERT EINSTEIN COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-07-01 至 2026-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10536888
• 关键词: Accounting; Adenosine; Affect; Animals; ApcMin/+ mice; Apoptosis; Architecture; Arginine; Biochemical; Body Weight; Cancer Model; Cancer cell line; Cells; Clinical Trials; Colon Carcinoma; Colonic Neoplasms; Colorectal Cancer; Combined Modality Therapy; Development; Dose; Enzymes; Familial Adenomatous Polyposis Syndrome; Foundations; Gene Expression; Genetic; Genotype; Glioblastoma; Goals; Growth; Heart; Histone H4; Histones; Human; Immunohistochemistry; In Vitro; Individual; Intestinal Neoplasms; Isotopes; Kinetics; Laboratories; Large Intestine Carcinoma; Lead; Link; Malignant Neoplasms; Mass Spectrum Analysis; Measurement; Measures; Metabolic; Metabolism; Methionine; Methionine Metabolism Pathway; Methylation; Methyltransferase; Molecular; Monitor; Mus; Oral; Patients; Phase I Clinical Trials; Phase I/II Clinical Trial; Phenotype; Phosphorylases; Post-Translational Protein Processing; Primary Neoplasm; Protein Chemistry; Protein Inhibition; Protein-Arginine N-Methyltransferase; Proteins; Proteomics; RNA Splicing; Radiolabeled; Reaction; Recycling; Reporter; Route; S-Adenosylhomocysteine; Safety; Schedule; Specificity; Structure; Tail; Techniques; Testing; Therapeutic; Tissues; Toxic effect; Transferase; Treatment Efficacy; Wild Type Mouse; Work; analog; anti-cancer; anti-cancer therapeutic; anticancer activity; arginine methyltransferase; base; body system; cancer survival; cell growth; colon cancer patients; colorectal cancer treatment; design; driver mutation; drug candidate; experimental study; histone methylation; in vivo; indexing; inhibitor; intestinal adenoma; metabolic rate; metabolomics; methionine adenosyltransferase; mouse model; neoplastic cell; novel; novel anticancer drug; novel drug combination; patient derived xenograft model; pharmacokinetics and pharmacodynamics; protein biomarkers; rational design; safety testing; single-cell RNA sequencing; small molecule inhibitor; therapy design; tool; tumor; tumor growth

Proposal Abstract Methionine adenosyltransferase 2 alpha (MAT2A) and protein arginine methyltransferase 5 (PRMT5) are cancer targets that are synthetically lethal with MTAP deletions and have several drug candidates in clinical trials targeting MTAP-/- cancers. MTAP is deleted in ~15% of human cancers and encodes the metabolic enzyme 5’-methylthioadenosine phosphorylase, the sole enzyme in humans responsible for recycling of methylthioadenosine (MTA) to methionine. MAT2A synthesizes S-adenosyl methionine (SAM), the methyl donor substrate for methyltransferase reactions. PRMT5 utilizes SAM as a substrate and is inhibited by MTA, and MTAP-/- cells in culture demonstrate elevated MTA levels. In vivo observations of glioblastoma tumors suggest however, that MTAP-/- does not always lead to increased tumoral MTA levels due to MTA efflux into matrix MTAP-competent cells. Additionally, MTAP deletions are a rare (~2%) occurrence in colorectal cancers (CRCs), precluding MAT2A and PRMT5 inhibitors’ use for most CRCs. The Schramm laboratory has previously solved the transition state (TS) structure of MTAP and synthesized a potent small molecule inhibitor methylthio-DADMe-immucillin-A (MTDIA) that recapitulates the in vitro effects of MTA accumulation within tissues. MTDIA has been shown to inhibit tumor growth in several cancer models, including CRC, and is linked to a decrease in PRMT5 activity through elevation of MTA levels. We propose that MTDIA be used in combination with MAT2A inhibitor AG-270, currently in Phase I clinical trials, to harness their synthetic lethality by targeting PRMT5. We will test the safety, target engagement, and anti-cancer efficacy of MTDIA in combination with AG-270 in ApcMin/+ and CRC patient-derived xenograft (PDX) mice. To determine mechanisms of anti-cancer effects, we will probe the upstream and downstream effects related to PRMT5 activity. We will perform tumor metabolomic quantification of relevant metabolites and histone and protein- arginine methylation characterization using immunohistochemistry and proteomic techniques. We will also profile the gene expression changes using single-cell RNA sequencing to determine how combination therapy alters tumor architecture and growth. Finally, we will solve the transition state structure of PRMT5 with the goal of laying the foundations for development of novel transition state analogue inhibitors. This work will expand upon the use of MAT2A and PRMT5 inhibitors beyond the ~15% of MTAP-deleted cancers and provide avenues for MTDIA to be used in clinical trials.

提案摘要 蛋氨酸腺苷转移酶2α（MAT2A）和蛋白精氨酸甲基转移酶5（PRMT5）为 与MTAP缺失合成致死的癌症靶标，并在临床上有几个候选药物 针对MTAP - / - 癌症的试验。 MTAP在约15％的人类癌症中删除并编码代谢 酶5'-甲基硫代腺苷磷酸化酶，人类的唯一酶，负责回收 MAT2A合成S-腺苷甲基二氨氨酸（SAM），甲基 甲基转移酶反应的供体底物。 PRMT5利用SAM作为底物，并被MTA抑制， 培养物中的MTAP - / - 细胞表现出MTA水平升高。体内观察胶质母细胞瘤肿瘤 但是，建议MTAP - / - 并不总是导致MTA外排入的肿瘤MTA水平增加 基质MTAP竞争细胞。此外，MTAP缺失是结直肠癌发生的罕见（约2％） （CRC），阻止MAT2A和PRMT5抑制剂用于大多数CRC。 Schramm实验室有 先前求解了MTAP的过渡态（TS）结构并合成潜在的小分子抑制剂 甲基硫硫代dadme-immucillin-A（mTDIA）概括了MTA在体外积累的体外效应 组织。 MTDIA已显示可抑制包括CRC在内的几种癌症模型的肿瘤生长，并连接 通过MTA水平升高，PRMT5活性的降低。我们建议将mtdia用于 与目前正在I期临床试验中的MAT2A抑制剂AG-270结合使用，以利用其合成杀伤力 通过靶向PRMT5。我们将测试mtdia的安全性，目标参与和抗癌效率 与APCMIN/+和CRC患者衍生的Xenographotic（PDX）小鼠中的AG-270结合。确定 抗癌作用的机制，我们将探测与PRMT5相关的上游和下游效应 活动。我们将对相关代谢产物以及组蛋白和蛋白质进行肿瘤代谢组学定量 精氨酸甲基化使用免疫组织化学和蛋白质组学技术。我们也会 介绍了使用单细胞RNA测序的基因表达变化，以确定联合治疗的方式 改变肿瘤结构和生长。最后，我们将解决PRMT5的过渡状态结构 为开发新型过渡状态模拟抑制剂的基础。这项工作将扩大 使用MAT2A和PRMT5抑制剂后，超出了约15％的MTAP骨骼癌症并提供 MTDIA的途径用于临床试验。