KDM6A mutation as an epigenetic driver of multiple myeloma

KDM6A 突变作为多发性骨髓瘤的表观遗传驱动因素

• 批准号: 10229675
• 负责人: Jonathan D. Licht
• 金额: $ 5.7万
• 依托单位: UNIVERSITY OF FLORIDA
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-09-01 至 2021-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10229675
• 关键词: Adhesives; Affect; B-Lymphocytes; Binding; Biological; Biology; Bone Marrow; CREBBP gene; Cell Line; Cell Maturation; Cells; ChIP-seq; Chemicals; Chromatin; Chromatin Structure; Clustered Regularly Interspaced Short Palindromic Repeats; DNA; Data; Engineering; Enhancers; Epigenetic Process; Fellowship; Female; Gene Expression; Gene Expression Profile; Gene Mutation; Genes; Genetic; Grant; Growth; Growth and Development function; Homeostasis; Immune system; Interleukin 6 Receptor; Lead; Lymphocyte; Malignant Neoplasms; Maps; Modification; Multiple Myeloma; Mus; Mutate; Mutation; Pathway interactions; Patients; Property; Recurrence; Regulator Genes; Role; Tumor Biology; Tumor Suppressor Proteins; cell growth; chromatin modification; chromatin remodeling; histone acetyltransferase; histone demethylase; histone methyltransferase; male; mouse model; next generation sequencing; protein complex; recruit; relapse patients; response; therapeutic target; tool development; tumor; Adhesives; Affect; B-Lymphocytes; Binding; Biological; Biology; Bone Marrow; CREBBP gene; Cell Line; Cell Maturation; Cells; ChIP-seq; Chemicals; Chromatin; Chromatin Structure; Clustered Regularly Interspaced Short Palindromic Repeats; DNA; Data; Engineering; Enhancers; Epigenetic Process; Fellowship; Female; Gene Expression; Gene Expression Profile; Gene Mutation; Genes; Genetic; Grant; Growth; Growth and Development function; Homeostasis; Immune system; Interleukin 6 Receptor; Lead; Lymphocyte; Malignant Neoplasms; Maps; Modification; Multiple Myeloma; Mus; Mutate; Mutation; Pathway interactions; Patients; Property; Recurrence; Regulator Genes; Role; Tumor Biology; Tumor Suppressor Proteins; cell growth; chromatin modification; chromatin remodeling; histone acetyltransferase; histone demethylase; histone methyltransferase; male; mouse model; next generation sequencing; protein complex; recruit; relapse patients; response; therapeutic target; tool development; tumor

PROJECT SUMMARY Background: Next generation sequencing has identified recurrently mutated genes in MM. Together anomalies of epigenetic regulator genes are present in 25% MM patients [4]. Among these affected genes are those encoding the histone demethylase KDM6A, the histone methyltransferase KMT2C/D, the histone acetyltransferase CREBBP and the chromatin remodeling subunit ARID1A, which are known to interact together to activate enhancers. KMT2C mutations are more frequent in relapsed patients and this correlates with shorter therapy response duration. KDM6A gene copy loss occurs in about 50% of male and female patients, and mutations of this gene are associated with poor survival. These data emphasize the importance of enhancer deregulation in the biology of MM but yet no functional studies have explored this in detail. We have shown Loss of KDM6A stimulates growth, clonogenicity and adhesive properties of MM. Re-expression of KDM6A or a demethylase inactive form both suppresses cell growth. Hypothesis: KDM6A deletion causes loss of enhancer activity and gene expression favoring uncontrolled proliferation. The recruitment of activators and not demethylating activity of KDM6A is most critical for this effect. Specific Aims: 1a. Determine KDM6A's direct targets. KDM6A will be mapped by ChIP-Seq in MM cell lines CRISPR engineered to express V5-tagged and degradation-inducible KDM6A. 1b. Define the role of KDM6A demethylase activity in enhancer function. KMT2C/D, P300/CBP binding and H3K27me, H3K27Ac modifications will be mapped by ChIP-seq in engineered cells expressing endogenous KDM6A with no demethylase activity. 2. Explore biological effect of KDM6A loss in a mouse model of MM. MM will be generated in KDM6Aflox/floxCD19-Cre mice by transduction of bone marrow with activated IL6 receptor and the effects on tumor biology determined. The aims proposed in this LLS Special Fellowship are related to a grant R01CA180475 aiming to understand the importance of KMT2C and UTX/KDM6A in the control of chromatin structure and homeostasis of MM by identifying their genetic target and how they affect B-cell maturation. This fellowship is expanding the understanding of KDM6A as a tumor suppressor in multiple myeloma, and is facilitating the development of tool necessary to study the role of KMT2C in MM as well.

项目摘要 背景：下一代测序已鉴定出MM中反复突变的基因。在一起异常 25％MM患者存在表观遗传调节基因的基因[4]。其中这些受影响的基因是 编码组蛋白脱甲基酶KDM6A，组蛋白甲基转移酶KMT2C/D，组蛋白 乙酰基转移酶Crebbp和染色质重塑亚基ARID1A，已知它们相互作用 激活增强剂。 KMT2C突变在复发患者中更常见，这与较短 治疗反应持续时间。 KDM6A基因复制损失发生在约50％的男性和女性患者中，并且 该基因的突变与存活不良有关。这些数据强调了增强器的重要性 在MM的生物学中放松管制，但没有功能研究详细探讨了这一点。我们已经表现出损失 KDM6A的刺激刺激MM的生长，克隆性和粘合特性。重新表达KDM6A或 脱甲基酶非活性形式都抑制细胞生长。 假设：KDM6A缺失会导致增强子活性的丧失和基因表达有利于控制的基因表达 增殖。激活剂的募集而不是KDM6A的脱甲基活性对于这种效果至关重要。 具体目的： 1a。确定KDM6A的直接目标。 KDM6A将由MM Cell Lines中的芯片序列绘制 设计为表达V5标签和降解诱导的KDM6A。 1B。定义KDM6A脱甲基活性在增强子功能中的作用。 KMT2C/D，P300/CBP结合和 H3K27ME，H3K27AC修改将由表达内源性的工程细胞中的芯片序列映射 KDM6A无脱甲基活性。 2。探索MM小鼠模型中KDM6A损失的生物学效应。 MM将在 Kdm6aflox/floxcd19-cre小鼠通过活化的IL6受体转导骨髓和对肿瘤的影响 生物学确定。 该法学学士学位特别奖学金中提出的目的与授予R01CA180475有关 KMT2C和UTX/KDM6A在控制MM的染色质结构和稳态中的重要性 识别其遗传靶标以及它们如何影响B细胞的成熟。这个奖学金正在扩大 将KDM6A作为多发性骨髓瘤中的肿瘤抑制剂的理解，并促进工具的发展 还需要研究KMT2C在MM中的作用。