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

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

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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.

项目摘要/摘要里希特综合征（RS）是多达10％的慢性淋巴细胞白血病（CLL）患者的关键并发症，它作为侵略性转化发展为弥漫性大B细胞组织学，并且大多是难治性的现有疗法。 RS发病机制在很大程度上仍然未知，分子的细胞和小鼠模型仍然研究是有限的。为了应对这一挑战，Ten Hacken博士开发了新颖的人类基因学启发通过CRISPR-CAS9多路复用B细胞编辑的小鼠模型，将CLL转换概括为Rs。 Ten Hacken博士已经通过她的初步研究证明了如何选择突变共发生促进疾病转化，与独特的转录变化和治疗有关。漏洞 - 即将完成的工作。 Ten Hacken博士现在计划介绍一套新的 B细胞表观遗传编程的基因突变，被鉴定为假定的RS驱动因素在最近的人类基因组分析中。在AIM 1中，Ten Hacken博士将在小鼠中引入表观遗传驱动器来评估所选变化（及其组合）对CLL转化的影响。作为此目标的一部分，博士十个哈肯还将评估这些模型对人类疾病的转录和遗传忠诚。在 AIM 2，Ten Hacken博士将在功能上表征建模的基因突变，同时剖析变化 CLL转化为Rs的表观遗传景观。通过这些研究将与人类RS数据集交叉相比，并在人类主要样本中进行验证类似的基因组成。在AIM 3中，Ten Hacken博士将在面板上进行体外和体内临床前测试为了设计突变（或共同）突变）特定的治疗策略。为了进行拟议的工作，十Hacken博士已入伍是计算生物学，系统免疫学，小鼠病理学，分子专家的合作者药理学，生物统计学，功能基因组学和表观遗传学。 Ten Hacken博士概述了3年的职业生涯制定计划将使她能够促进自己的个人专业发展（包括领导力，格兰特写作，谈判和沟通技巧），并获得生物信息学和生物统计学。 Ten Hacken博士的独立研究计划将重点介绍血液学恶性肿瘤，其长期目标是进行临床相关研究和其他淋巴机和髓样恶性肿瘤的功能基因组分析。通过她提出的工作，十博士 Hacken预计将在奖励期内贡献2个高影响力的手稿。她将每年在国际会议，并将为她在第二年底的第一个R01提交准备。期望研究将对Richter综合征的生物学和自然历史以及鼠标模型Ten Hacken正在开发中，将代表有用的工具来剖析病原机制和测试这种基本无法治愈的恶性肿瘤的新型治疗策略。