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，进行性造血性疾病，其特征是 髓样谱系一致的异常增殖，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途径的功能收益突变， 包括JAK2VF在内，大多数MPN患者都存在，并且可以受到靶向抑制作用；然而， 临床改良的JAK2抑制剂无法减轻突变等位基因负担，并且随着时间的推移反应减弱。躯体 高危染色质修饰符ASXL1和EZH2的变化经常与MPN和会议中的JAK2VF共同发生 不良预后，骨髓纤维化进展以及对JAK2治疗的反应降低。这些机制 继发于ASXL1/EZH2改变的染色质失调增强了克隆进化和促纤维化 尚未阐明促进纤维化进展和JAK2抑制剂耐药性的炎症途径。 最近，我们证明了使用双重的MPN维护中突变的JAK2VF的绝对要求 JAK2VF的重组酶敲入/敲除小鼠模型。我假设JAK2VF与ASXL1或 EZH2损失改变对疾病维持的JAK/STAT信号的依赖并促进促炎性 信号通路有利于纤维化进展。在此提案中，我将调查对致癌的要求 表观遗传失调和促炎性介导的纤维化/利基的JAK2VF信号传导和可逆性 使用我的双重组酶JAK2VF等位基因重塑高风险双突变MPN。我会用 临床MPN标本评估的单细胞DNA测序 +免疫表型/细胞因子分析 MF进展中的细胞因子产生，克隆进化和突变获取的顺序。进一步的理解 染色质失调在MPN中纤维化进展和热耐药性的合作作用 可能导致对该高风险MPN患者子集的历史可依赖依赖性鉴定。 在MSKCC参加助理的医学博士Andrew Dunbar将作为5年职业生涯的一部分进行此项目 发展计划，奉献精神> 75％的时间研究临床工作。邓巴博士是 由罗斯·莱文（Ross Levine）博士指导，他是血液学恶性肿瘤研究的世界专家。建议邓巴博士 由Drs。 MSKCC的Raajit Rampal，Omar Abdel-Wahab，Richard Koche和Andriy Derkach博士P. Brent Ferrell博士 在范德比尔特大学医学中心和丽贝卡·施耐德博士的大学医院RWTH AACHEN， 德国。安德鲁的培训将包括在表演和分析单细胞基因型方面获得技术技能 具有生物信息学的正式课程的测定，对骨髓微环境进行建模和方法 研究造血干细胞的表观遗传调节。在短期内，项目目标是发布 有关这项研究结果的两篇论文。从长远来看，目标是制定研究计划 并获得R01资金成为血液系统恶性肿瘤的独立实验室研究者。