Defining the epigenetic landscape and therapeutic vulnerabilities of Richter's syndrome in CRISPR-based mouse models

项目总结/摘要 里希特综合征（RS）是高达10%的慢性淋巴细胞白血病（CLL）患者的严重并发症， 其发展为扩散性大B细胞组织学的侵袭性转化，并且大多数是难治的。 现有疗法。RS的发病机制在很大程度上仍然未知，细胞和小鼠模型的分子 研究有限。为了应对这一挑战，滕·哈肯博士开发了一种新型的人类遗传学， 通过CRISPR-Cas9多重B细胞编辑的小鼠模型，重现CLL转化为RS。 在她的初步研究中，滕·哈肯博士证明了选择性的突变共同发生是如何 促进疾病转化，并与不同的转录变化和治疗相关。 目前接近完成的工作。腾·哈肯博士现在正计划推出一套新的 参与B细胞表观遗传编程的基因突变，被鉴定为推定的RS驱动因子 最近的人类基因组分析。在目标1中，ten Hacken博士将在小鼠中引入表观遗传驱动因素，以评估 所选改变（及其组合）对CLL转化的影响。作为这个目标的一部分，博士。 ten Hacken还将评估这些模型对人类疾病的转录和遗传忠诚度。在 目标2，博士ten Hacken将功能性地描述模型化的基因突变，同时解剖 CLL转化为RS的表观遗传景观。表观遗传依赖性通过 这些研究将与人类RS数据集进行交叉比较，并在人类原始样本中进行验证， 相似的基因组成在目标3中，ten Hacken博士将对一个面板进行体外和体内临床前测试。 药物（化疗和新型靶向药物的综合），以设计突变（或共同- 突变）特异性治疗策略。为了完成这项工作，腾·哈肯博士 合作者是计算生物学、系统免疫学、小鼠病理学、分子生物学、 药理学、生物统计学、功能基因组学和表观遗传学。滕·哈肯博士概述了他三年的职业生涯 发展计划，这将使她能够促进她的个人专业发展（包括领导，赠款 写作、谈判和沟通技能），并获得生物信息学方面的额外科学培训， 生物统计学博士十哈肯的独立研究计划将集中在翻译研究， 血液恶性肿瘤，长期目标是开展临床相关研究， 其他淋巴和骨髓恶性肿瘤的功能基因组分析。通过她提出的工作，博士十 Hacken预计将在奖项期限内贡献2篇高影响力的手稿。她将每年在 国际会议，并将准备好她的第一个R 01提交对年底2。拟议 这些研究有望为里希特综合征的生物学和自然史提供重要的见解， ten Hacken博士正在开发的小鼠模型将成为剖析致病机制的有用工具， 为这种基本上无法治愈的恶性肿瘤测试新的治疗策略。

Project Summary/Abstract Richter’s syndrome (RS) is a critical complication of up to 10% of chronic lymphocytic leukemia (CLL) patients, which develops as an aggressive transformation into a diffuse large B cell histology, and is mostly refractory to existing therapies. RS pathogenesis remains largely unknown and cellular and mouse models for molecular studies are limited. To address this challenge, Dr. ten Hacken has developed novel human-genetics inspired mouse models through CRISPR-Cas9 multiplexed B-cell editing, recapitulating CLL transformation into RS. Already through her preliminary studies, Dr. ten Hacken demonstrated how selected mutational co-occurrences facilitate disease transformation, and are associated to distinct transcriptional changes and therapeutic vulnerabilities—work that is presently near completion. Dr. ten Hacken is now planning to introduce a new set of mutations in genes involved in epigenetic programming of B cells, which were identified as putative RS drivers in recent human genomic analyses. In Aim 1, Dr. ten Hacken will introduce epigenetic drivers in mice to assess the impact of the selected alterations (and their combinations) on CLL transformation. As part of this Aim, Dr. ten Hacken will also assess the transcriptional and genetic faithfulness of these models to human disease. In Aim 2, Dr. ten Hacken will functionally characterize the modeled gene mutations, while dissecting changes in the epigenetic landscape underlying transformation of CLL into RS. Epigenetic dependencies identified through these studies will be cross-compared with human RS datasets and validated in human primary samples with similar genetic make-ups. In Aim 3, Dr. ten Hacken will perform in vitro and in vivo preclinical testing of a panel of agents (comprehensive of chemotherapy and novel targeted agents) in order to design mutation (or co- mutation) specific treatment strategies. To carry out the proposed work, Dr. ten Hacken has enlisted collaborators who are experts in computational biology, systems immunology, mouse pathology, molecular pharmacology, biostatistics, functional genomics and epigenetics. Dr. ten Hacken has outlined a 3-year career development plan that will allow her to foster her personal professional development (including leadership, grant writing, negotiation and communication skills), and to gain additional scientific training in bioinformatics and biostatistics. Dr. ten Hacken’s independent research program will be focused on translational research in hematological malignancies, with the longer-term objective of undertaking clinical correlative research and functional genomic analyses of other lymphoid and myeloid malignancies. Through her proposed work, Dr. ten Hacken anticipates to contribute 2 high-impact manuscripts within the award term. She will present yearly at international conferences, and will be ready for her first R01 submission towards the end of Year 2. The proposed studies are expected to provide critical insight into the biology and natural history of Richter’s syndrome, and the mouse models Dr. ten Hacken is developing will represent useful tools to dissect pathogenic mechanisms and test novel treatment strategies for this largely incurable malignancy.