Dissecting the Mechanism of SETDB1 and its K867 Monoubiquitination in Lung Cancer Progression

剖析 SETDB1 及其 K867 单泛素化在肺癌进展中的机制

• 批准号: 10674509
• 负责人: Jia Fang
• 金额: $ 39.4万
• 依托单位: ROSWELL PARK CANCER INSTITUTE CORP
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-08-05 至 2027-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10674509
• 关键词: Alleles; Attenuated; BRAF gene; Biochemical; CDX2 gene; Cancer Etiology; Cell physiology; Cells; Clinical; Common Neoplasm; Complex; Data; Development; Drug Targeting; Endogenous Retroviruses; Enzymes; Epidermal Growth Factor Receptor; Epigenetic Process; Extracellular Matrix; Family; Gene Silencing; Genes; Genetically Engineered Mouse; Goals; Heterochromatin; Histones; Human; In Vitro; Invaded; KRAS oncogenesis; KRAS2 gene; KRASG12D; Link; Lung; Lung Adenocarcinoma; Lysine; Malignant - descriptor; Malignant Neoplasms; Malignant neoplasm of lung; Mediating; Methylation; Methyltransferase; Modeling; Modification; Molecular; Monoubiquitination; Mus; Mutation; Neoplasm Metastasis; Non-Small-Cell Lung Carcinoma; Oncogenic; Pathway interactions; Patients; Phenotype; Play; Process; Prognosis; Proteins; Proteomics; Refractory; Regulation; Repression; Role; SETDB1 gene; Signal Transduction; System; Therapeutic; Transcriptional Activation; Tumor Suppressor Genes; Ubiquitination; Up-Regulation; Xenograft Model; biomarker development; biomarker driven; cell motility; cohort; driver mutation; gain of function; gene repression; genetic signature; histone modification; human disease; in vivo; innovation; insight; loss of function; lung cancer cell; migration; molecular targeted therapies; mortality; mouse model; mutant; neoplastic cell; novel; novel therapeutics; restraint; subcutaneous; targeted treatment; therapeutically effective; transcription factor; transcriptional reprogramming; tumor; tumor growth; tumor progression

Project Summary Dissecting the Mechanism of SETDB1 and its K867 Monoubiquitination in Lung Cancer Progression Non-small cell lung cancer (NSCLC) is one of the leading causes of cancer-related mortality worldwide and associated with high mutation burdens. During the last decade, biomarker-driven targeted therapies have achieved clear clinical benefits in patients with several oncogenic driver mutations such as EGFR, BRAF and ALK. However, the treatments for tumors with the most prevalent KRAS mutations remain challenging despite of recent breakthrough in targeting KRAS-G12C mutation. In KRAS mutant NSCLC, the constitutively activated downstream signaling ultimately leads to transcription reprogramming to sustain tumor growth and progression. This process is orchestrated by a cohort of crucial transcription factors, whose temporal and spatial expression is associated with distinct epigenetic changes. Thus, dissecting epigenetic mechanisms in this process could open new avenues for targeting strategies. SETDB1 is a principal methyltransferase catalyzing histone H3K9 methylation, a major repressive epigenetic modification. The focal amplification and upregulation of SETDB1 have been observed in a wide range of cancers including NSCLC, suggesting it has pro-oncogenic function. However, the mechanisms of action of SETDB1 in NSCLC remain elusive. In preliminary study, we uncovered that SETDB1 upregulation in lung adenocarcinoma (ADC) correlates with poorer prognosis and significantly co-occurs with KRAS gene alternations. In NSCLC cells harboring oncogenic RAS, SETDB1 loss attenuates migration and invasion, invadopodia formation and ECM degradation. These phenotypic changes correlate with reactivation of transcription factors FOXA2 and CDX2 which function as key regulatory nodes to impede metastatic progression of KRAS mutant lung ADC. More importantly, Setdb1 loss in tumor cells derived from metastatic KrasG12D;p53fl/fl lung ADC abolished their spontaneous ability to metastasize from the subcutaneous tumor to the lungs. These findings argue for a novel concept that SETDB1-mediated epigenetic mechanisms are critical for oncogenic KRAS-induced transcription reprogramming during lung ADC progression. Moreover, we have demonstrated that SETDB1 is monoubiquitinated at lysine-867 by the UBE2E family of E2 enzymes independent of E3s and this monoubiquitination is essential for SETDB1-mediated H3K9 methylation and ERV silencing. Accordingly, one objective of this proposal is to delineate SETDB1's pro-metastatic activity in KRAS mutant lung ADC using various genetically engineered mouse models and xenograft models. Another objective is to dissect the mechanisms of lysine-867 monoubiquitination in SETDB1's pro-metastatic activity in vivo and in vitro. Our central hypothesis is that SETDB1 plays important roles in KRAS mutant lung ADC progression through epigenetic mechanisms that require its monoubiquitination dependent enzymatic activity. This project is significant because it will characterize SETDB1 as an actionable drug target in KRAS mutant lung cancer and identify novel molecular interfaces for targeting. Our long-term goal is to develop better and more effective therapeutic strategies by delineating epigenetic mechanisms in lung cancer.

项目摘要 SETDB 1及其K867单泛素化在肺癌进展中的作用机制研究 非小细胞肺癌（Non-small cell lung cancer，NSCLC）是世界范围内癌症相关死亡的主要原因之一 并与高突变负担相关。在过去的十年中，生物标志物驱动的靶向治疗已经 在携带几种致癌驱动突变（如EGFR、BRAF和 ALK。然而，尽管KRAS基因突变的发生率很高，但对具有最普遍KRAS突变的肿瘤的治疗仍然具有挑战性。 最近在针对KRAS-G12 C突变方面取得了突破。在KRAS突变型NSCLC中， 下游信号传导最终导致转录重编程以维持肿瘤生长和进展。 这一过程是由一组重要的转录因子协调的，这些转录因子的时空表达 与明显的表观遗传变化有关因此，在这一过程中解剖表观遗传机制， 为目标战略开辟了新的途径。SETDB 1是催化组蛋白H3 K9的主要甲基转移酶 甲基化是一种主要的抑制性表观遗传修饰。SETDB 1的局部扩增和上调 已在包括NSCLC在内的多种癌症中观察到，表明其具有促癌功能。 然而，SETDB 1在NSCLC中的作用机制仍然难以捉摸。在初步研究中，我们发现 肺腺癌（ADC）中SETDB 1的上调与较差的预后相关， 与KRAS基因突变同时发生。在携带致癌RAS的NSCLC细胞中，SETDB 1丢失减弱 迁移和入侵，侵入伪足形成和ECM降解。这些表型变化与 转录因子FOXA 2和CDX 2的重新激活，它们作为关键的调节节点， KRAS突变型肺ADC的转移进展。更重要的是，Setdb 1在来源于 转移性KrasG 12 D; p53 fl/fl肺ADC消除了它们从皮下转移的自发能力 肿瘤转移到肺部这些发现为SETDB 1介导的表观遗传机制提出了新的概念 在肺ADC进展过程中对致癌KRAS诱导的转录重编程至关重要。此外，委员会认为， 我们已经证明SETDB 1被E2酶的UBE 2 E家族在赖氨酸-867处单泛素化 独立于E3，这种单泛素化对于SETDB 1介导的H3 K9甲基化和ERV是必需的 沉默因此，该建议的一个目的是描述SETDB 1在KRAS中的促转移活性 使用各种基因工程小鼠模型和异种移植物模型对突变型肺ADC进行了研究。另一个目的 是剖析赖氨酸-867单泛素化在SETDB 1体内促转移活性中的机制， 体外我们的中心假设是SETDB 1在KRAS突变型肺ADC进展中起重要作用 通过需要其单泛素化依赖性酶活性的表观遗传机制。这个项目 是重要的，因为它将把SETDB 1表征为KRAS突变型肺癌中的可操作的药物靶点 并鉴定用于靶向的新型分子界面。我们的长远目标是发展得更好、更有成效 通过描绘肺癌的表观遗传机制来制定治疗策略。