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

项目摘要/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， 它发展为一种侵袭性转化为弥漫性大B细胞组织学，并且大多对 现有的治疗方法。RS的发病机制在很大程度上仍不清楚，细胞和小鼠的分子模型 研究是有限的。为了应对这一挑战，滕·哈肯博士开发了一种新的人类遗传学启发 小鼠模型通过CRISPR-Cas9多路B细胞编辑，概括CLL转化为RS。 通过她的初步研究，Ten Hacken博士已经展示了如何选择突变共存 促进疾病转化，并与不同的转录变化和治疗相关 漏洞--目前即将完成的工作。Ten Hacken博士现在计划推出一套新的 参与B细胞表观遗传编程的基因突变，被确认为假定的RS驱动因素 在最近的人类基因组分析中。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 所选更改(及其组合)对CLL转换的影响。作为这一目标的一部分，Dr。 Ten Hacken还将评估这些模型对人类疾病的转录和遗传忠实性。在……里面 目标2，Ten Hacken博士将从功能上描述模拟的基因突变，同时剖析 CLL向RS转化的表观遗传景观。表观遗传依赖性通过以下方式确定 这些研究将与人类遥感数据集进行交叉比较，并在人类初级样本中进行验证 相似的基因构成。在目标3中，Ten Hacken博士将在体外和体内进行一个小组的临床前测试 药物(化疗和新型靶向药物的综合)，以设计突变(或联合 突变)特定的治疗策略。为了执行这项拟议的工作，滕·哈肯博士招募了 合作者是计算生物学、系统免疫学、小鼠病理学、分子生物学方面的专家 药理学、生物统计学、功能基因组学和表观遗传学。Ten Hacken博士勾勒出了他三年的职业生涯 使她能够促进个人职业发展的发展计划(包括领导力、赠款 写作、谈判和沟通技能)，并获得额外的生物信息学和 生物统计学。Ten Hacken博士的独立研究计划将专注于翻译研究 恶性血液病，长期目标是进行临床相关研究和 其他淋巴系和髓系恶性肿瘤的功能基因组分析。通过她提议的工作，Ten博士 Hacken预计在获奖期限内贡献2篇高影响力的手稿。她将在每年一度的 国际会议，并将在第二年年底准备好提交她的第一份R01报告。 预计研究将为里希特综合征的生物学和自然病史提供关键的见解，以及 Ten Hacken博士正在开发的小鼠模型将成为剖析致病机制和 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.