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: 10664872
• 负责人: Gabriel T Bedard
• 金额: $ 5.27万
• 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/10664872
• 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; Pathogenesis; Patients; Phase I Clinical Trials; Phase I/II Clinical Trial; Phenotype; Phosphorylases; Post-Translational Protein Processing; Primary Neoplasm; Proliferating; Protein Chemistry; Protein Inhibition; Protein-Arginine N-Methyltransferase; Proteins; Proteomics; RNA Splicing; Radiolabeled; Reaction; Recycling; Reporter; Route; S-Adenosylhomocysteine; S-Adenosylmethionine; 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; body system; cancer survival; cell growth; design; driver mutation; drug candidate; experimental study; histone methylation; in vivo; indexing; inhibitor; intestinal adenoma; 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 2 α (MAT2A) 和蛋白精氨酸甲基转移酶 5 (PRMT5) MTAP 缺失具有综合致死性的癌症靶点，并且有多种候选药物处于临床阶段 针对 MTAP-/- 癌症的试验。 MTAP 在大约 15% 的人类癌症中被删除，并编码代谢 5'-甲硫腺苷磷酸化酶，人体中唯一负责循环利用的酶 甲硫腺苷 (MTA) 转化为蛋氨酸。 MAT2A 合成 S-腺苷甲硫氨酸 (SAM)，即甲基 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， 培养中的 MTAP-/- 细胞表现出升高的 MTA 水平。胶质母细胞瘤的体内观察 然而，表明 MTAP-/- 并不总是导致肿瘤 MTA 水平升高，因为 MTA 外流到 基质 MTAP 感受态细胞。此外，MTAP 缺失在结直肠癌中罕见（~2%） (CRC)，排除了 MAT2A 和 PRMT5 抑制剂对大多数 CRC 的使用。施拉姆实验室拥有 先前解决了MTAP的过渡态（TS）结构并合成了一种有效的小分子抑制剂 甲硫基-DADMe-immucillin-A (MTDIA) 概括了 MTA 积累的体外影响 组织。 MTDIA 已被证明可以抑制多种癌症模型（包括结直肠癌）中的肿瘤生长，并且与 通过升高 MTA 水平来降低 PRMT5 活性。我们建议 MTDIA 用于 与目前处于 I 期临床试验的 MAT2A 抑制剂 AG-270 组合，以利用其合成致死率 通过针对 PRMT5。我们将测试 MTDIA 的安全性、靶点参与度和抗癌功效 与 AG-270 联合用于 ApcMin/+ 和 CRC 患者来源的异种移植 (PDX) 小鼠。确定 抗癌作用机制，我们将探讨PRMT5相关的上下游效应 活动。我们将对相关代谢物以及组蛋白和蛋白质进行肿瘤代谢组学定量- 使用免疫组织化学和蛋白质组技术表征精氨酸甲基化。我们还将 使用单细胞 RNA 测序分析基因表达变化，以确定联合治疗的效果 改变肿瘤结构和生长。最后，我们将解决 PR​​MT5 的过渡状态结构，目标是 为开发新型过渡态类似物抑制剂奠定基础。这项工作将扩大 使用 MAT2A 和 PRMT5 抑制剂后，超过 15% 的 MTAP 缺失癌症并提供 MTDIA 用于临床试验的途径。