Targeting Menin Protein Stability In AML

靶向 AML 中的 Menin 蛋白稳定性

• 批准号: 10644645
• 负责人: Jevon Cutler
• 金额: $ 12.96万
• 依托单位: DANA-FARBER CANCER INST
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-07-18 至 2025-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10644645
• 关键词: Acute Myelocytic Leukemia; Acute leukemia; Advisory Committees; Automobile Driving; Award; Binding; Biochemical; Biological Assay; Biology; CRISPR library; Cell Line; Chimeric Proteins; Chromatin; Clinical Trials; Co-Immunoprecipitations; Complex; Data; Development; Drug Targeting; Environment; Funding; Gastrointestinal Stromal Tumors; Gene Expression; Genetic; Genetic Transcription; Goals; Grant; Ikaros protein; Imides; In Vitro; Intention; Lead; Leadership; Leukemic Cell; Lysine; MLL gene; Malignant Neoplasms; Manuscripts; Mass Spectrum Analysis; Mediating; Menin; Mentors; Mentorship; Methods; Methyltransferase; Multiple Endocrine Neoplasia Type 1; Mutate; NPM1 gene; NUP98 gene; Oncogenic; Outcome; Patient-Focused Outcomes; Pharmaceutical Preparations; Play; Pre-Clinical Model; Process; Protac; Proteins; RUNX1 gene; Regulation; Reporter; Reporting; Research; Research Personnel; Resistance; Resources; Role; System; Therapeutic; Therapeutic Effect; Training; Transcriptional Regulation; Ubiquitin; Work; Writing; acute myeloid leukemia cell; design; drug sensitivity; experience; experimental study; high risk; immunoregulation; improved; improved outcome; in silico; in vivo; inhibitor; interest; leukemia; multicatalytic endopeptidase complex; novel; novel strategies; novel therapeutic intervention; prevent; programs; protein degradation; protein function; recognins; response; skills; small molecule; success; targeted treatment; tool; transcription factor; transcriptome sequencing; tumorigenic; ubiquitin-protein ligase

PROJECT SUMMARY Multiple Endocrine Neoplasia Type 1 (MEN1/MENIN) is a transcriptional regulator known to interact with Lysine Methyltransferase 2A (KMT2A/MLL1) driving aggressive subsets of leukemia including MLL1 rearranged (MLL-r), NPM1 mutated and NUP98 rearranged acute myeloid leukemia (AML) as well as other cancers including gastrointestinal stromal tumors. Early success has been reported in clinical trials using small molecule MENIN inhibitors (MI) that disrupt the MLL1/MENIN interaction to treat AML causing MENIN to emerge as an important drug target. As a result, efforts to understand MENIN function are paramount for the continued success of targeting this protein. I, as a research fellow working under the mentorship of Drs. Scott Armstrong and Eric Fischer, have recently characterized an exciting aspect of the mechanism driving the therapeutic effect of MI treatments. We have observed that protein stability changes to MENIN and other proteins is an essential feature of MI sensitivity. Our preliminary examination of this mechanism has uncovered that the loss of a specific E3 ligase can mediate decreased MI sensitivities via MENIN protein stabilization. These findings have inspired a new approach of targeting MENIN protein stability with the simple idea that more MENIN protein decreases drug sensitivity while less MENIN protein inhibits MLL-r transcriptional programs. The experiments outline in this proposal aim to characterize MENIN protein stability in AML by studying how this specific E3 ligase regulates MENIN contributing to MI sensitivity and investigate newly established rapid MENIN degradation systems as a proof-of-principle for improving both MENIN targeted therapies as well as the means to study MENIN function. We have recently established a novel set of tools to study MENIN protein function including (i) MLL-r leukemia cell lines with edited MEN1 loci inserting different protein tags enabling high throughput MENIN protein stability reporter systems and dTAG based rapid MENIN degradation systems, (ii) specific CRISPR libraries targeting MENIN protein function and (iii) early candidate small molecule MENIN proteolysis targeting chimeras (PROTAC). This proposal is also designed to provide additional training experiences in advanced methods of transcriptional analysis, biochemical binding assays, small molecule PROTAC design and mass spectrometry-based co- immunoprecipitation as well as mentoring in manuscript/grant writing and leadership skills. Anchored by the guidance of two well-resourced and established mentors, an excellent scientific advisory committee composed of experts in the fields of leukemia and chromatin biology and a highly dynamic research environment at DCFI, funding for this proposal will enable me to transition to an independent investigator.

项目摘要 多种内分泌肿瘤1型（Men1/Menin）是已知与之相互作用的转录调节剂 赖氨酸甲基转移酶2A（KMT2A/MLL1）驱动白血病的侵略性子集，包括MLL1 重新排列（MLL-R），NPM1突变和NUP98重新排列的急性髓样白血病（AML）以及其他 包括胃肠道肿瘤在内的癌症。在使用 小分子梅宁抑制剂（MI）破坏MLL1/MENIN相互作用以治疗AML引起MENIN 成为重要的药物靶标。结果，了解Menin功能至关重要 靶向该蛋白质的持续成功。我是在Drs的指导下工作的研究研究员。 斯科特·阿姆斯特朗（Scott Armstrong）和埃里克·菲舍尔（Eric Fischer）最近表征了机制驾驶的令人兴奋的方面 MI治疗的治疗作用。我们已经观察到蛋白质的稳定性变化了Menin和其他 蛋白质是MI敏感性的重要特征。我们对这种机制的初步检查 发现特定的E3连接酶的损失可以通过Menin蛋白介导MI敏感性降低 稳定。这些发现激发了一种针对梅宁蛋白稳定性的新方法 简单的想法是，更多的梅宁蛋白会降低药物敏感性，而较少的梅宁蛋白抑制了MLL-R 转录程序。该提案中的实验概述旨在表征Menin蛋白 通过研究该特定的E3连接酶如何调节Menin有助于MI敏感性和 调查新建立的快速梅宁降解系统，作为改善两者的原则证明 Menin靶向疗法以及研究Menin功能的手段。我们最近建立了一个 研究Menin蛋白功能的新型工具，包括（i）具有编辑MEN1的MLL-R白血病细胞系 基因座插入不同的蛋白质标签，使高吞吐量Menin蛋白稳定性报告基因系统和 基于DTAG的快速MENIN降解系统，（ii）针对Menin蛋白的特定CRISPR库 功能和（iii）早期候选小分子梅宁蛋白水解靶向嵌合体（Protac）。这 建议还旨在提供高级转录方法的额外培训经验 分析，生化结合测定，小分子Protac设计和基于质谱的共同 免疫沉淀以及手稿/赠款写作和领导能力的指导。由 两个资源和成熟的导师的指导，这是一个出色的科学咨询委员会 由白血病和染色质生物学领域的专家组成以及高度动态的研究 DCFI的环境，该提案的资金将使我能够过渡到独立的调查员。