(PQ8) Genetically faithful murine models for studying disease progression in chronic lymphocytic leukemia

(PQ8) 用于研究慢性淋巴细胞白血病疾病进展的遗传忠实小鼠模型

• 批准号: 10161748
• 负责人: Lili Wang
• 金额: $ 53.9万
• 依托单位: DANA-FARBER CANCER INST
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-05-11 至 2022-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10161748
• 关键词: ATM Gene Mutation; Acceleration; Adoption; Affect; Age-Months; Aging; Animal Model; B lymphoid malignancy; B-Lymphocytes; Biological; Blood; CD19 gene; Cell Line; Chronic Lymphocytic Leukemia; Chronic Phase; Clinical; Clustered Regularly Interspaced Short Palindromic Repeats; DNA Damage; Data; Development; Disease; Disease Progression; Disease model; Engraftment; Evaluation; Event; Experimental Models; Foundations; Gene Mutation; Genes; Genetic; Genetic Diseases; Genetic Heterogeneity; Genome; Goals; Histology; Human; Human Genetics; In Vitro; Individual; Indolent; Investigation; Knock-in Mouse; Knowledge; Lead; Lesion; Lymphoma; Malignant Neoplasms; Marrow; Mature B-Lymphocyte; Medicine; Methylation; Modeling; Modernization; Molecular; Monitor; Mus; Mutate; Mutation; Oncogenic; Onset of illness; Patients; Penetrance; Positron-Emission Tomography; Preclinical Testing; Progressive Disease; RNA Splicing; Reporting; Resources; Role; Sampling; Spleen; Study models; Subgroup; Techniques; Testing; Therapeutic; Therapeutic Agents; Time; Vision; Work; aged; base; cancer cell; cancer gene expression; chronic lymphocytic leukemia cell; clinical practice; cohort; combinatorial; del(11q); driver mutation; exome; gain of function; gain of function mutation; genetic makeup; genetic manipulation; genome editing; genome sequencing; genomic data; human disease; in vivo; individual patient; insight; knock-down; lentiviral-mediated; leukemia; mouse development; mouse model; network models; novel; novel therapeutics; overexpression; patient variability; personalized therapeutic; precision medicine; pressure; prognostic; programs; response; stem cells; therapeutic evaluation; tool; transcriptome sequencing; whole genome

Project Summary A key goal of modern-day medicine is to use our knowledge of the unique genetic makeup of an individual patient to make personalized therapeutic decisions. For chronic lymphocytic leukemia (CLL), the advances in our knowledge of its disease genetics as well as the adoption of novel therapeutic agents in clinical practice have been rapid over recent years. Thus, the need now is ever more urgent to match patients with the appropriate therapeutic choice. Given the wealth of available human genetic data in CLL and our understanding of its genetic heterogeneity, our vision is that the path to precision medicine can be trail-blazed with CLL. Critical to this vision is the development of faithful animal models, since these would undoubtedly accelerate the preclinical testing of agents against genetically-defined subgroups. Herein, we propose leveraging the entirety of CLL genomics data, including genetic to methylation studies, to rationally create mouse models that provide the full range of genetic variability in order to recapitulate the clinical variability of patients. This goal is implementable because we recently demonstrated that the combined expression of 2 putative CLL driver events, identified from unbiased sequencing of patient samples, generates CLL-like disease that is highly faithful to the human disease. Specifically, co-expression of mutated Sf3b1 with Atm deletion (significantly associated together in patient samples) resulted in the development of clonal pathognomonic CD19+CD5+ B cells in blood, marrow and spleen at low penetrance in aged (18 months) mice, that can be propagated by in vivo passaging. With this work as a foundation, we now propose to investigate the hypothesis that distinct evolutionary paths are undertaken in CLL depending on the starting points of disease and that specific combinations of genetic events function to initiate malignancy, while others are critical for disease acceleration and even oncogenic transformation. To achieve this goal, we propose to leverage recently available facile genome-editing approaches, and a robust workflow we optimized to genetically manipulate mature B cells through engraftment of genome-edited B cell progenitor cells, in order to nimbly screen the functional impact of a variety of candidate driver mutations within a B cell context in vivo. We aim to identify the stepwise events required to initiate disease from normal B cells to a state of indolent malignancy (Aim 1), from indolent malignancy to more progressive disease (Aim 2), and even to aggressive lymphoma transformation (Aim 3). Generating such animal models is expected to provide an invaluable resource which will enable deep understanding of the functional impact of driver alterations, to accelerate disease prognostication, to facilitate rational evaluation of novel and combinatorial therapeutics, and to dissect the interaction of CLL cells with their in vivo microenvironment. Thus, we seek to create the same genetic heterogeneity in mice as in CLL patients so that we can faithfully model disease and provide a means to test therapeutics in advance—a stepping stone towards achieving precision medicine.

项目摘要 现代医学的一个关键目标是利用我们对个体独特基因构成的了解 让患者做出个性化的治疗决定。慢性淋巴细胞性白血病（CLL）， 我们对其疾病遗传学的了解以及在临床实践中采用新的治疗药物 近年来发展迅速。因此，现在更加迫切需要将患者与 适当的治疗选择。鉴于CLL中丰富的人类遗传数据和我们的研究， 了解其遗传异质性，我们的愿景是，精准医疗的道路可以 用CLL开辟了道路。这一愿景的关键是发展忠实的动物模型，因为这些模型将 毫无疑问，加速了针对基因定义的亚组的药物的临床前测试。在此我们 建议利用整个CLL基因组学数据，包括遗传甲基化研究， 创建小鼠模型，提供完整的遗传变异范围，以概括临床 患者的变异性。这个目标是可以实现的，因为我们最近证明， 从患者样品的无偏测序中鉴定的2个推定的CLL驱动事件的表达产生 CLL样疾病是高度忠实于人类疾病。具体地，突变的Sf 3b 1与 ATM缺失（在患者样本中显著相关）导致克隆性 老年人（18岁）外周血、骨髓和脾脏中特异性CD 19 + CD 5 + B细胞的低表达 月）小鼠，其可以通过体内传代繁殖。以这项工作为基础，我们现在提出 研究CLL中不同的进化路径取决于 疾病的起始点，以及遗传事件的特定组合， 恶性肿瘤，而另一些是疾病加速甚至致癌转化的关键。到 为了实现这一目标，我们建议利用最近可用的简易基因组编辑方法，以及一个强大的 我们优化了工作流程，以通过植入基因组编辑的B细胞来遗传操作成熟的B细胞 祖细胞，以便灵活地筛选细胞内多种候选驱动突变的功能影响。 体内B细胞环境。我们的目标是确定从正常B细胞开始疾病所需的逐步事件 惰性恶性肿瘤状态（目标1），从惰性恶性肿瘤到更进展的疾病（目标2），以及 甚至到侵袭性淋巴瘤转化（Aim 3）。建立这样的动物模型有望提供 这是一个宝贵的资源，将使人们能够深入了解驾驶员变更的功能影响， 加速疾病诊断，促进对新型和组合疗法的合理评价，以及 剖析CLL细胞与其体内微环境的相互作用。因此，我们寻求创造同样的 小鼠和CLL患者的遗传异质性，因此我们可以忠实地模拟疾病，并提供一个 提前测试治疗方法的手段-实现精确医学的垫脚石。

项目成果

Emerging Therapies in CLL in the Era of Precision Medicine.

• DOI: 10.3390/cancers15051583
• 发表时间: 2023-03-03
• 期刊: Cancers
• 影响因子: 5.2

SF3B1 mutation and ATM deletion codrive leukemogenesis via centromeric R-loop dysregulation.

• DOI: 10.1172/jci163325
• 发表时间: 2023-09-01
• 期刊: JOURNAL OF CLINICAL INVESTIGATION
• 影响因子: 15.9
• 作者: Cusan, Martina;Shen, Haifeng;Zhang, Bo;Liao, Aijun;Yang, Lu;Jin, Meiling;Fernandez, Mike;Iyer, Prajish;Wu, Yiming;Hart, Kevyn;Gutierrez, Catherine;Nik, Sara;Pruett-Miller, Shondra M.;Stark, Jeremy;Obeng, Esther A.;Bowman, Teresa, V;Wu, Catherine J.;Lin, Ren-Jang;Wang, Lili
• 通讯作者: Wang, Lili