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

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.

项目总结 背景：下一代测序已经在MM中发现了反复突变的基因。 25%的多发性骨髓瘤患者中存在表观遗传调节基因[4]。在这些受影响的基因中有 编码组蛋白去甲基酶KDM6A、组蛋白甲基转移酶KMT2C/D、组蛋白 已知相互作用的乙酰基转移酶CREBBP和染色质重塑亚基ARID1A 来激活增强剂。KMT2C突变在复发患者中更常见，这与较短的时间相关 治疗反应持续时间。KDM6A基因拷贝丢失发生在约50%的男性和女性患者中，并且 这种基因的突变与较差的存活率有关。这些数据强调了增强剂的重要性。 多发性骨髓瘤生物学上的放松调控，但目前还没有功能研究详细地探讨这一点。我们已经表现出了损失 KDM6A对MM生长、克隆形成和黏附特性的刺激作用 去甲基酶非活性形式都会抑制细胞生长。 假设：KDM6A缺失导致增强子活性丧失和基因表达不受控制 扩散。KDM6A的激活剂募集和非去甲基化活性对这一作用最为关键。 具体目标： 1A.确定KDM6A的直接目标。KDM6A将被CHIP-SEQ映射到MM细胞系CRISPR中 设计表达V5标签和降解诱导的KDM6A。 1B.确定KDM6A去甲基酶活性在增强子功能中的作用。KMT2C/D、P300/CBP结合和 H3K27Me、H3K27Ac修饰将被CHIP-SEQ定位在表达内源基因的工程细胞中 KDM6A无脱甲基酶活性。 2.探讨KDM6A缺失对MM小鼠模型的生物学效应 活化IL-6受体骨髓转导KDM6Aflox/FloxCD19-Cre小鼠及其对肿瘤的影响 生物学决定的。 LLS特别奖学金中提出的目标与旨在理解以下内容的赠款R01CA180475有关 KMT2C和UTX/KDM6A在MM染色质结构和动态平衡调控中的作用 确定他们的遗传目标以及他们如何影响B细胞的成熟。这一奖学金正在扩大 了解KDM6A在多发性骨髓瘤中作为肿瘤抑制因子的作用，并正在促进工具的开发 有必要研究KMT2C在多发性骨髓瘤中的作用。

项目成果

Chromatin activation as a unifying principle underlying pathogenic mechanisms in multiple myeloma.

• DOI: 10.1101/gr.265520.120
• 发表时间: 2020-09
• 期刊: Genome research
• 影响因子: 7
• 作者: Ordoñez R;Kulis M;Russiñol N;Chapaprieta V;Carrasco-Leon A;García-Torre B;Charalampopoulou S;Clot G;Beekman R;Meydan C;Duran-Ferrer M;Verdaguer-Dot N;Vilarrasa-Blasi R;Soler-Vila P;Garate L;Miranda E;San José-Enériz E;Rodriguez-Madoz JR;Ezponda T;Martínez-Turrilas R;Vilas-Zornoza A;Lara-Astiaso D;Dupéré-Richer D;Martens JHA;El-Omri H;Taha RY;Calasanz MJ;Paiva B;San Miguel J;Flicek P;Gut I;Melnick A;Mitsiades CS;Licht JD;Campo E;Stunnenberg HG;Agirre X;Prosper F;Martin-Subero JI
• 通讯作者: Martin-Subero JI

Leveraging epigenetics to enhance the efficacy of immunotherapy.

• DOI: 10.1186/s13148-021-01100-x
• 发表时间: 2021-05-17
• 期刊: Clinical epigenetics
• 影响因子: 5.7
• 作者: Licht JD;Bennett RL
• 通讯作者: Bennett RL

H3K27 Methylation: A Focal Point of Epigenetic Deregulation in Cancer.

• DOI: 10.1016/bs.acr.2016.05.001
• 发表时间: 2016
• 期刊: Advances in cancer research
• 作者: Nichol JN;Dupéré-Richer D;Ezponda T;Licht JD;Miller WH Jr
• 通讯作者: Miller WH Jr

DISORDERED HISTONE METHYLATION IN HEMATOLOGICAL MALIGNANCIES THE CASE OF UTX/KDM6A.

血液系统恶性肿瘤中组蛋白甲基化紊乱（以 UTX/KDM6A 为例）。

• 发表时间: 2018
• 期刊: Transactions of the American Clinical and Climatological Association
• 作者: Licht,JonathanD

Acquired Resistance to EZH2 Inhibitor GSK343 Promotes the Differentiation of Human DLBCL Cell Lines toward an ABC-Like Phenotype.

• DOI: 10.1158/1535-7163.mct-21-0216
• 发表时间: 2022-04-01
• 期刊: Molecular cancer therapeutics
• 影响因子: 5.7
• 作者: Preston SEJ;Emond A;Pettersson F;Dupéré-Richer D;Abraham MJ;Riva A;Kinal M;Rys RN;Johnson NA;Mann KK;Del Rincón SV;Licht JD;Miller WH
• 通讯作者: Miller WH