Identification of MSKs as critical integrators of chromatin and transcription factor activation in melanoma

鉴定 MSK 作为黑色素瘤染色质和转录因子激活的关键整合因子

• 批准号: 9519543
• 负责人: Douglas Barrows
• 金额: $ 5.9万
• 依托单位: ROCKEFELLER UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-03-01 至 2020-02-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9519543
• 关键词: Address; Affect; BRAF gene; Biochemical; Biological; Biological Assay; Biology; CD58 Antigens; CREB1 gene; CRISPR/Cas technology; Cancer Control; Cancer Model; Cells; Chromatin; Chromatin Fiber; Collaborations; Colorectal Cancer; Complex; Cues; Data; Dependence; Development; Disease; Drug resistance; EP300 gene; Epigenetic Process; Foundations; Future; Gene Activation; Genes; Genetic; Genetic Transcription; Goals; Growth; Histone H3; Histones; In Vitro; Lead; Link; Literature; MEKs; Maintenance; Malignant Neoplasms; Malignant neoplasm of lung; Malignant neoplasm of pancreas; Mass Spectrum Analysis; Mediating; Melanoma Cell; Mitogen-Activated Protein Kinases; Mitogens; Modeling; Mutation; Oncogenes; Oncogenic; Output; Pathway interactions; Patients; Pharmacology; Phosphorylation; Polycomb; Positioning Attribute; Post-Translational Protein Processing; Protein Kinase; Proteins; Proteome; RPS6KA5 gene; Regulation; Repression; Resistance; Role; Signal Pathway; Signal Transduction; System; Testing; Therapeutic; Therapeutic Intervention; Training; Transcriptional Activation; activating transcription factor; advanced disease; cancer genome; cancer type; career; combat; epigenomics; experimental study; genome sequencing; inhibitor/antagonist; insight; melanoma; new therapeutic target; novel; p300/CBP-Associated Factor; recruit; response; stress activated protein kinase; targeted treatment; training opportunity; transcription factor; treatment strategy; tumor; tumor growth; tumorigenesis

Project Summary Cancer genome sequencing has revealed high rates of mutations in chromatin-modifying proteins, highlighting the importance of chromatin regulation in cancer. However, mechanistic links between chromatin and the signaling pathways that are known to promote cancer development remain poorly understood. In melanoma, in addition to other cancers, activation of the MAPK signaling pathway is a driver of tumorigenesis. Current therapeutic strategies used to treat melanoma involve reducing the output of MAPK signaling through pharmacologic inhibition of upstream protein kinases. Unfortunately, resistance rapidly occurs to these therapies, highlighting the need for a more in-depth understanding of downstream signaling. There is an emerging appreciation that chromatin may be a key target for post-translational modifications downstream of MAPK signaling. Preliminary data from the Allis lab and others has established a novel understanding of the role of mitogen and stress activated protein kinases (MSKs) as critical intermediates in MAPK signaling that regulate histone H3 Ser28 phosphorylation (H3S28ph). H3S28ph has a potent role in signal-induced activation of transcription, potentially through the recruitment of p300 acetyltransferase and displacement of Polycomb complexes. MSKs also phosphorylate and activate transcription factors, including NF-κB and CREB, both of which have established roles in melanoma. The ability to dually regulate transcription factors and chromatin puts MSKs in a position to have a major impact on transcription in the context of MAPK pathway activation. Here we will specifically address the hypothesis that constitutive activation of MSKs in melanoma and other cancers controls the transcription of genes that are critical for the growth and maintenance of the tumor. Two Specific Aims are proposed to test this working hypothesis: 1) use a CRISPR-Cas9 system and complementary pharmacologic approaches to determine the requirements of MSK1 and MSK2 for oncogenic transcription in melanoma and 2) use biochemical assays in order to identify new MSK substrates/interacting proteins and understand the mechanisms by which MSKs regulate chromatin and transcription. Importantly, insights resulting from the completion of these aims will potentially lead to novel opportunities for therapeutic intervention in this deadly disease. In addition, BRAF-driven melanoma will be used here as a model cancer type to study the role of MSKs, but MAPK signaling is often dysregulated in cancer, and these findings will serve as the foundation to study these signaling paradigms in various tumor types, such as pancreatic, colorectal, and lung cancer.

项目摘要 癌症基因组测序揭示了染色质修饰蛋白的高突变率， 染色质调节在癌症中的重要性。然而，染色质和细胞质之间的机械联系， 已知促进癌症发展的信号传导途径仍然知之甚少。在黑色素瘤中， 除了其他癌症之外，MAPK信号通路的激活是肿瘤发生的驱动因素。电流 用于治疗黑色素瘤的治疗策略包括通过以下途径减少MAPK信号传导的输出： 上游蛋白激酶的药理学抑制。不幸的是，这些疗法很快就会产生耐药性， 突出了对下游信号传导的更深入理解的需要。正在形成 认识到染色质可能是MAPK下游翻译后修饰的关键靶点 发信号。来自Allis实验室和其他实验室的初步数据已经建立了一个新的理解， 丝裂原和应激活化蛋白激酶（MSKs）是MAPK信号传导的关键中间体， 组蛋白H3 Ser 28磷酸化（H3 S28 ph）。H3 S28 ph在信号诱导的细胞活化中具有有效作用， 转录，可能通过招募p300乙酰转移酶和置换Polycomb 配合物MSKs还磷酸化和激活转录因子，包括NF-κB和CREB，两者都是 它们在黑色素瘤中起着重要作用。双重调节转录因子和染色质的能力， 在MAPK通路激活的背景下，MSK能够对转录产生重大影响。这里我们 将具体阐述在黑色素瘤和其他癌症中MSK的组成性激活的假设， 控制对肿瘤生长和维持至关重要的基因的转录。两个具体 提出了测试该工作假设的目的：1）使用CRISPR-Cas9系统和互补的 药理学方法来确定MSK 1和MSK 2的致癌转录的要求， 黑色素瘤和2）使用生化测定来鉴定新的MK底物/相互作用蛋白， 了解MSK调节染色质和转录的机制。重要的是， 从这些目标的完成将可能导致新的机会，治疗干预， 致命的疾病此外，BRAF驱动的黑色素瘤将在这里作为一种模型癌症类型来研究的作用， 但是MAPK信号在癌症中经常失调，这些发现将作为研究的基础。 研究各种肿瘤类型中的这些信号模式，如胰腺癌、结肠直肠癌和肺癌。