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.

项目概要 背景：下一代测序已鉴定出 MM 中反复突变的基因。一起异常 25% 的多发性骨髓瘤患者中存在 25% 的表观遗传调节基因 [4]。这些受影响的基因包括 编码组蛋白去甲基化酶 KDM6A、组蛋白甲基转移酶 KMT2C/D、组蛋白 乙酰转移酶 CREBBP 和染色质重塑亚基 ARID1A，已知它们会相互作用 来激活增强子。 KMT2C 突变在复发患者中更为常见，这与较短的 治疗反应持续时间。 KDM6A 基因拷贝丢失发生在大约 50% 的男性和女性患者中，并且 该基因的突变与较差的生存率有关。这些数据强调了增强子的重要性 MM 生物学中的放松管制，但尚未有功能研究详细探讨这一点。我们已经显示了损失 KDM6A 刺激 MM 的生长、克隆形成和粘附特性。 KDM6A 或 a 的重新表达 去甲基酶失活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失控 增殖。 KDM6A 激活剂的募集而非去甲基化活性对于这种效果最为关键。 具体目标： 1a.确定KDM6A的直接目标。 KDM6A 将通过 ChIP-Seq 在 MM 细胞系 CRISPR 中定位 旨在表达 V5 标记和降解诱导型 KDM6A。 1b.定义 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 修饰将通过 ChIP-seq 在表达内源性的工程细胞中进行定位 KDM6A 没有脱甲基酶活性。 2. 探讨 KDM6A 缺失对 MM 小鼠模型的生物学影响。 MM 将生成于 激活IL6受体转导骨髓的KDM6Aflox/floxCD19-Cre小鼠及其对肿瘤的影响 生物学决定的。 此 LLS 特别奖学金中提出的目标与旨在了解的 R01CA180475 拨款相关 KMT2C 和 UTX/KDM6A 在控制 MM 染色质结构和稳态中的重要性 确定它们的遗传目标以及它们如何影响 B 细胞成熟。该奖学金正在扩大 了解 KDM6A 作为多发性骨髓瘤的肿瘤抑制因子，并正在促进工具的开发 研究 KMT2C 在 MM 中的作用也是必要的。

项目成果

Leveraging epigenetics to enhance the efficacy of immunotherapy.

利用表观遗传学增强免疫治疗的功效。

• 发表时间: 2021
• 影响因子: 5.7
• 作者: Licht, Jonathan D;Bennett, Richard L
• 通讯作者: Bennett, Richard L

Targeting histone acetylation dynamics and oncogenic transcription by catalytic P300/CBP inhibition.

通过催化 P300/CBP 抑制来靶向组蛋白乙酰化动力学和致癌转录。

• DOI: 10.1016/j.molcel.2021.04.015
• 发表时间: 2021-05-20
• 期刊: Molecular cell
• 影响因子: 16
• 作者: Hogg SJ;Motorna O;Cluse LA;Johanson TM;Coughlan HD;Raviram R;Myers RM;Costacurta M;Todorovski I;Pijpers L;Bjelosevic S;Williams T;Huskins SN;Kearney CJ;Devlin JR;Fan Z;Jabbari JS;Martin BP;Fareh M;Kelly MJ;Dupéré-Richer D;Sandow JJ;Feran B;Knight D;Khong T;Spencer A;Harrison SJ;Gregory G;Wickramasinghe VO;Webb AI;Taberlay PC;Bromberg KD;Lai A;Papenfuss AT;Smyth GK;Allan RS;Licht JD;Landau DA;Abdel-Wahab O;Shortt J;Vervoort SJ;Johnstone RW
• 通讯作者: Johnstone RW

Epigenetic regulatory mutations and epigenetic therapy for multiple myeloma.

表观遗传调控突变和多发性骨髓瘤的表观遗传治疗。

• 发表时间: 2017-07
• 影响因子: 3.2
• 作者: Dupéré;Licht, Jonathan D
• 通讯作者: Licht, Jonathan D

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

染色质激活是多发性骨髓瘤致病机制的统一原则。

• 发表时间: 2020
• 影响因子: 7
• 作者: Ordoñez, Raquel;Kulis, Marta;Russiñol, Nuria;Chapaprieta, Vicente;Carrasco;García;Charalampopoulou, Stella;Clot, Guillem;Beekman, Renée;Meydan, Cem;Duran;Verdaguer;Vilarrasa;S
• 通讯作者: S

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
• 作者: Licht; Jonathan D
• 通讯作者: Jonathan D