The role of epigenetic plasticity in acute myeloid leukemia cell persistence

项目总结 急性髓系白血病(AML)患者的预后很差，这主要是因为目前的治疗方法未能 根除所有AML细胞，留下促使复发的持久性癌细胞。急性髓系白血病细胞的存活能力 治疗与干细胞特性有关1，2。这些原始的AML细胞需要被根除才能实现 持久的减刑。在约翰·迪克博士的博士培训期间，我研究了造血干细胞1，3，其中 启发我专门研究急性髓细胞白血病干细胞状态的表观遗传调控。对于我的博士后培训，我 加入了表观遗传学专家伯恩斯坦博士的实验室，并开发了一种创新技术来绘制地图 稀有细胞中的组蛋白修饰4。利用这项技术，我以前所未有的精确度描述了干细胞是如何 基因被组蛋白3赖氨酸27三甲基化(H3K27me3)沉默，这是一种抑制性的表观遗传修饰， 被PRC2催化，被KDM6拮抗。PRC2在急性髓细胞白血病中经常受损，导致增强 干细胞特性，但H3K27去甲基酶KDM6的作用尚不清楚。我发现KDM6促进了 AML细胞适合性和在IDH1/2(IDHmut)突变的AML细胞中上调，表明对 IDHmut AML细胞对KDM6的作用。本提案的目的是(1)研究KDM6的功能 使用不同的人类AML细胞模型，(2)评估KDM6作为IDHmut AML细胞的治疗靶点，以及(3) 描述KDM6通过识别其下游靶点促进AML细胞适合性的机制。这些 研究将揭示驱动AML的一种新的表观遗传学机制，并为其发展奠定基础 KDM6抑制剂的应用，以促进AML细胞的有效根除，包括那些在当前 治疗。 伯恩斯坦博士是一位杰出的导师，他的受训人员曾在 学术界。他是表观基因组学、癌症和发展领域备受国际尊敬的领导者 在马萨诸塞州综合医院(MGH)、哈佛医学院、美国癌症学会 和布罗德学院。这项研究将在MGH进行，MGH是一家著名的研究机构和医疗机构 该中心是一个充满活力的社区的一部分，其中包括哈佛医学院、达纳-法伯癌症研究所、 布里格姆妇女医院和布罗德研究所，一个促进合作和 智力交流。由世界级内科科学家组成的研究咨询委员会将提供建议 指导：大卫·斯卡登博士、乔恩·阿斯特和安德鲁·莱恩。研究的关键方面将是 通过与安德鲁·莱恩、大卫·温斯托克和查尔斯·爱泼斯坦博士的合作完成。 K99/R00奖将为我实现职业目标提供最好的机会。详细的 培训计划包括一个研究咨询委员会以及领导和指导技能的发展， 对于成功过渡到独立将是无价的。颁奖的R00阶段将允许我 确保在顶级癌症研究所获得一个独立的职位，并建立一个雄心勃勃的研究计划。 1.P.van Galen等人，自然。510,268-272(2014)。 2.P.van Galen等人，《科学》杂志正在审查中(见附录)。 3.P.van Galen等人，细胞干细胞。14、94-106(2014)。 4.P.van Galen等人，Mol.牢房。61,170-180(2016)。

PROJECT SUMMARY Acute myeloid leukemia (AML) patients have poor outcomes, largely because current therapies fail to eradicate all AML cells, leaving persistent cancer cells that drive relapse. The ability of AML cells to survive therapy is associated with stem cell properties1,2. These primitive AML cells need to be eradicated to achieve durable remissions. During my Ph.D. training with Dr. John Dick, I studied hematopoietic stem cells1,3 which inspired me to specialize in epigenetic regulation of the stem cell state in AML. For my postdoctoral training, I joined the laboratory of Dr. Bernstein, an expert in epigenetics, and developed an innovative technology to map histone modifications in rare cells4. Using this technology, I describe with unprecedented precision how stem cell genes are silenced by histone 3 lysine 27 trimethylation (H3K27me3), a repressive epigenetic modification that is catalyzed by PRC2 and antagonized by KDM6. PRC2 is frequently impaired in AML, leading to enhanced stem cell properties, but the role of the H3K27 demethylase KDM6 is unknown. I discovered that KDM6 promotes AML cell fitness and is upregulated in AML cells with mutations in IDH1/2 (IDHmut), indicating a dependency of IDHmut AML cells on KDM6 function. The objectives of this proposal are to (1) investigate the function of KDM6 using various human AML cell models, (2) evaluate KDM6 as a therapeutic target in IDHmut AML cells, and (3) delineate the mechanism by which KDM6 promotes AML cell fitness by identifying its downstream targets. These studies will uncover a novel epigenetic mechanism that drives AML and lay the foundation for the development of KDM6 inhibitors to facilitate efficient eradication of AML cells, including those that persist through current therapies. Dr. Bernstein is an outstanding mentor with a history of trainees that obtained group leader positions in academia. He is an internationally respected leader in the fields of epigenomics, cancer and development with appointments at the Massachusetts General Hospital (MGH), Harvard Medical School, American Cancer Society and the Broad Institute. The research will be carried out at MGH, a prestigious research institute and medical center that is part of a vibrant community that includes Harvard Medical School, Dana-Farber Cancer Institute, Brigham and Women's Hospital and the Broad Institute, an environment that fosters collaborations and intellectual exchange. A Research Advisory Committee of world-class physician-scientists will provide advice and guidance: Drs. David Scadden, Jon Aster and Andrew Lane. Critical aspects of the research will be completed through collaborations with Drs. Andrew Lane, David Weinstock and Charles Epstein. The K99/R00 award will provide me with the best opportunity to succeed in my career goals. The detailed training plan includes a Research Advisory Committee and development of leadership and mentoring skills that will be invaluable for a successful transition to independence. The R00 phase of the award will allow me to secure an independent position at a top cancer institute and set up an ambitious research program. 1. P. van Galen et al., Nature. 510, 268–272 (2014). 2. P. van Galen et al., under review at Science (see appendix). 3. P. van Galen et al., Cell Stem Cell. 14, 94–106 (2014). 4. P. van Galen et al., Mol. Cell. 61, 170–180 (2016).