Investigating High-Risk Epigenetic Modifying Alterations on JAK2VF Dependency and Fibrotic Progression in Myeloproliferative Neoplasms (MPNs)

研究骨髓增生性肿瘤 (MPN) 中 JAK2VF 依赖性和纤维化进展的高风险表观遗传修饰改变

• 批准号: 10723901
• 负责人: Andrew Jeffrey Dunbar
• 金额: $ 16.76万
• 依托单位: SLOAN-KETTERING INST CAN RESEARCH
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-08-20 至 2028-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10723901
• 关键词: Academic Medical Centers; Alleles; Bioinformatics; Biological Assay; Bone Marrow; Cell surface; Cells; Chromatin; Chromatin Remodeling Factor; Chronic; Clinical; Clonal Evolution; Complement; Coupled; DNA sequencing; Dedications; Dependence; Development; Development Plans; Disease; Disease Progression; EZH2 gene; Engraftment; Epigenetic Process; Evolution; Fibrosis; Frequencies; Funding; Gene Expression; Genetic Transcription; Genomics; Genotype; Germany; Glean; Goals; Hematologic Neoplasms; Hematopoietic; Hematopoietic stem cells; Human; Immunophenotyping; Individual; Inflammatory; JAK2 gene; Knock-in; Knock-out; Knockout Mice; Laboratories; Longitudinal Studies; Maintenance; Maps; Mediating; Memorial Sloan-Kettering Cancer Center; Mentors; Methods; Modeling; Molecular; Mus; Mutation; Myelofibrosis; Myelogenous; Myeloproliferative disease; Oncogenic; Output; Paper; Pathogenesis; Pathway interactions; Patients; Polycomb; Polycythemia Vera; Primary Myelofibrosis; Process; Production; Prognosis; Proliferating; Publishing; Reporter; Research; Research Personnel; Resistance; Risk; Sampling; Secondary to; Signal Pathway; Signal Transduction; Specimen; Technical Expertise; Techniques; Therapeutic; Therapeutic Studies; Time; Training; Translating; Transplant Recipients; University Hospitals; Work; burden of illness; career development; cell type; clinical translation; cohort; conditional knockout; cytokine; epigenetic regulation; epigenomics; fitness; gain of function mutation; high risk; improved; in vivo; inhibitor; inhibitor therapy; insight; leukemic transformation; loss of function; loss of function mutation; mesenchymal stromal cell; mouse model; mutant; mutant mouse model; novel; novel therapeutics; patient subsets; programs; protein profiling; recombinase; response; therapy resistant; treatment response

PROJECT SUMMARY/ABSTRACT Myeloproliferative Neoplasms (MPNs) are chronic, progressive hematopoietic disorders characterized by aberrant proliferation of myeloid lineage constituents, pro-inflammatory sequelae, progressive bone marrow fibrosis, and increased risk of leukemic transformation. Gain-of-function mutations of the JAK/STAT pathway, including JAK2VF, are present in the majority of MPN patients and are amenable to targeted inhibition; however, clinically-improved JAK2 inhibitors fail to reduce mutant allele burden, and response wanes over time. Somatic alterations in high-risk chromatin modifiers ASXL1 and EZH2 co-occur frequently with JAK2VF in MPN and confer adverse prognosis, myelofibrotic progression, and reduced response to JAK2 therapy. The mechanisms by which chromatin dysregulation secondary to ASXL1/EZH2 alterations enhance clonal evolution and pro-fibrotic inflammatory pathways to promote fibrosis progression and JAK2 inhibitor resistance have not been elucidated. Recently, we demonstrate an absolute requirement for mutant Jak2VF in MPN maintenance using a dual- recombinase knock-in/knock-out mouse model of Jak2VF. I hypothesize that Jak2VF cooperates with Asxl1 or Ezh2 loss to alter dependency on JAK/STAT signaling for disease maintenance and promote pro-inflammatory signaling pathways favoring fibrotic progression. In this proposal, I will investigate the requirement for oncogenic Jak2VF signaling and reversibility of epigenetic dysregulation and pro-inflammatory mediated fibrosis/niche remodeling in high-risk dual-mutant MPN using my dual-recombinase Jak2VF allele. I will complement this with single-cell DNA sequencing + immunophenotyping/cytokine analysis of clinical MPN specimens evaluating cytokine production, clonal evolution, and order of mutation acquisition in MF progression. Further understanding of the cooperative effects of chromatin dysregulation in fibrotic progression and therapeutic resistance in MPNs might lead to the identification of therapeutically tractable dependencies for this high-risk MPN patient subset. Andrew Dunbar, MD, an Assistant Attending at MSKCC, will conduct this project as part of a 5-year career development plan, dedicating >75% of his time to research with remainder on clinical work. Dr. Dunbar is mentored by Dr. Ross Levine, a world expert in the study of hematologic malignancies. Dr. Dunbar is advised by Drs. Raajit Rampal, Omar Abdel-Wahab, Richard Koche and Andriy Derkach at MSKCC, Dr. P. Brent Ferrell at Vanderbilt University Medical Center, and Dr. Rebekka Schneider at University Hospital RWTH Aachen, Germany. Andrew’s training will include gaining technical skills in performing and analyzing single-cell genotypic assays with formal courses in bioinformatics, modeling the bone marrow microenvironment, and methods to investigate the epigenetic regulation of hematopoietic stem cells. In the short term, the project goal is to publish two papers on the findings from this research. In the long term, the goal is for developing a research program and obtaining R01 funding to become an independent laboratory investigator in hematologic malignancies.

项目摘要/摘要 骨髓增生性肿瘤（MPN）是慢性进行性造血系统疾病，其特征在于： 骨髓谱系成分异常增殖，促炎后遗症，进行性骨髓 纤维化和白血病转化的风险增加。JAK/STAT途径的功能获得性突变， 包括JAK 2 VF，存在于大多数MPN患者中并且适于靶向抑制;然而， 临床上改进的JAK 2抑制剂不能减少突变等位基因负荷，并且应答随时间减弱。体细胞 高危染色质修饰物ASXL 1和EZH 2的改变经常与JAK 2 VF共同发生在MPN中， 不良预后、骨髓纤维化进展和对JAK 2治疗的反应降低。的机制 继发于ASXL 1/EZH 2改变的染色质失调增强了克隆进化和促纤维化 促进纤维化进展和JAK 2抑制剂抗性的炎性途径尚未阐明。 最近，我们证明了突变体Jak 2 VF在MPN维护中的绝对要求，使用双- 重组酶敲入/敲除Jak 2 VF小鼠模型。我假设Jak 2 VF与Asxl 1合作， Ezh 2损失改变对JAK/STAT信号传导的依赖性以维持疾病并促进促炎 促进纤维化进展的信号通路。在这个建议中，我将研究致癌的要求， Jak 2 VF信号传导和表观遗传失调的可逆性以及促炎介导的纤维化/生态位 使用我的双重重组酶Jak 2 VF等位基因在高危双重突变型MPN中重塑。我将补充说， 临床MPN标本的单细胞DNA测序+免疫表型/细胞因子分析， 细胞因子产生、克隆进化和MF进展中突变获得的顺序。进一步理解 染色质失调在MPN纤维化进展和治疗耐药性中的协同作用 可能导致识别治疗上易处理的依赖性，这种高风险的MPN患者子集。 安德鲁邓巴，医学博士，助理出席MSKCC，将进行这个项目的一部分，5年的职业生涯 发展计划，将超过75%的时间用于研究，其余时间用于临床工作。邓巴医生 由Ross Levine博士指导，他是血液系统恶性肿瘤研究的世界专家。邓巴医生建议 作者：MSKCC的Raajit Rampal、Omar Abdel-Wahab、Richard Koche和Andriy Derkach博士，P.布伦特费雷尔博士 在范德比尔特大学医学中心，以及亚琛RWTH大学医院的Rebekka Schneider博士， 德国。安德鲁的培训将包括获得技术技能，在执行和分析单细胞基因型 生物信息学的正式课程，骨髓微环境建模，以及 研究造血干细胞的表观遗传调控。在短期内，该项目的目标是发布 两篇关于这项研究结果的论文。从长远来看，我们的目标是制定一个研究计划， 并获得R 01资金，成为血液恶性肿瘤的独立实验室研究者。