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

在基于 CRISPR 的小鼠模型中定义里氏综合症的表观遗传景观和治疗脆弱性

基本信息

批准号：
10425662
负责人：
Elisa ten Hacken
金额：
$ 19.6万
依托单位：
WEILL MEDICAL COLL OF CORNELL UNIV
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-05-19 至 2026-04-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10425662
关键词：
Acceleration Address Agammaglobulinaemia tyrosine kinase Apoptotic Award B-Lymphocytes BCL2 gene Bioinformatics Biological Biological Assay Biology Biometry Bromodomain CD19 gene CDKN2A gene CRISPR/Cas technology Cell model ChIP-seq Characteristics Chronic Lymphocytic Leukemia Clinical Clinical Management Clonal Evolution Clustered Regularly Interspaced Short Palindromic Repeats Communication Complication Computational Biology DNMT3a Data Data Set Dependence Development Development Plans Diffuse Disease Disease model EP300 gene EZH2 gene Engineering Enhancers Epigenetic Process Exhibits Family Flow Cytometry Fostering Gene Mutation Generations Genes Genetic Genetic Engineering Genetic Transcription Genetically Engineered Mouse Genome Genomics Genotype Grant Hematologic Neoplasms Hematopoietic stem cells Histology Human Human Genetics Immunology In Vitro Individual International Knock-in Leadership Lesion Link Malignant Neoplasms Malignant lymphoid neoplasm Manuscripts Mediation Methylation Modeling Molecular Mus Mutation Myeloproliferative disease NOTCH1 gene Natural History Oncogenic Pathogenesis Pathogenicity Pathology Pathway interactions Patients Pharmaceutical Preparations Pharmacology Phenotype Postdoctoral Fellow Preclinical Testing Proteins Recurrence Refractory Reporting Research Richter&apos s Syndrome Sampling Signal Transduction System TP53 gene Testing Therapeutic Therapeutic Intervention Training Translational Research Transplantation Tumor Suppressor Proteins Tyrosine Kinase Inhibitor Up-Regulation Validation Work Writing XCL1 gene Yin cancer cell career development cell type chemotherapy chromosome 13q loss cohort design efficacy evaluation functional genomics genetic architecture genetic makeup genomic locus human disease improved in vivo in vivo evaluation inhibitor insight interest large cell Diffuse non-Hodgkin&apos s lymphoma leukemic transformation loss of function loss of function mutation molecular modeling mouse model novel pre-clinical preclinical study programs skills symposium targeted agent tool transcriptome transcriptome sequencing treatment strategy

项目摘要

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.

项目概要/摘要里氏综合征 (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变为弥漫性大 B 细胞组织学，并且大多难以抵抗现有的治疗方法仍然很大程度上未知，并且分子细胞和小鼠模型。为了应对这一挑战，十哈肯博士开发了一种受人类遗传学启发的新型药物。通过 CRISPR-Cas9 多重 B 细胞编辑的小鼠模型，重现了 CLL 向 RS 的转化。十号博士已经通过她的初步研究证明了如何选择突变共现促进疾病转化，并与独特的转录变化和治疗相关漏洞——目前已接近完成的工作，十哈肯博士现在正计划推出一套新的漏洞。参与 B 细胞表观遗传编程的基因突变，被确定为假定的 RS 驱动因素在最近的人类基因组分析中，十哈肯博士将在小鼠中引入表观遗传驱动因素来评估。作为该目标的一部分，选定的改变（及其组合）对 CLL 转化的影响。十哈肯还将评估这些模型对人类疾病的转录和遗传忠实度。目标 2，十哈肯博士将从功能上表征建模的基因突变，同时剖析基因突变的变化通过表观遗传依赖性确定 CLL 转化为 RS 的表观遗传景观。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在目标 3 中，十号 Hacken 博士将对一组进行体外和体内临床前测试。药物（综合化疗和新型靶向药物）以设计突变（或共同为了开展拟议的工作，十哈肯博士已经入伍。合作者是计算生物学、系统免疫学、小鼠病理学、分子生物学等领域的专家十哈肯博士概述了药理学、生物统计学、功能基因组学和表观遗传学的 3 年职业生涯。发展计划将使她能够促进个人职业发展（包括领导力、资助写作、谈判和沟通技巧），并获得生物信息学和十哈肯博士的独立研究项目将侧重于生物统计学的转化研究。血液系统恶性肿瘤，长期目标是开展临床相关研究和通过她提出的工作，十博士对其他淋巴和骨髓恶性肿瘤进行了功能基因组分析。哈肯预计将在获奖期间贡献两篇高影响力的手稿，她将每年在会上发表演讲。国际会议，并将在第二年年底为她的第一份 R01 提交做好准备。研究预计将为里氏综合症的生物学和自然史提供重要的见解，以及十哈肯博士正在开发的小鼠模型将成为剖析致病机制和测试针对这种基本上无法治愈的恶性肿瘤的新治疗策略。