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Community Outreach and Education Core

社区外展和教育核心

基本信息

批准号：
8376863
负责人：
Kelly A. Edwards
金额：
$ 19.13万
依托单位：
UNIVERSITY OF WASHINGTON
依托单位国家：
美国
项目类别：
财政年份：
资助国家：
美国
起止时间：
至
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/8376863
关键词：
AML1-ETO fusion protein Alleles CCAAT-Enhancer-Binding Protein-alpha Cathepsin G Cells Clinical Trials Combined Modality Therapy Community Health Education Community Outreach Complement Data Development Disease Disease model Environmental Health FLT3 gene FLT3 inhibitor Flow Cytometry Funding Generations Hematopoietic Histone Deacetylase Inhibitor Human JAK2 gene Knock-in Mouse Light Lymphoid Mediating Modeling Multipotent Stem Cells Mus Mutate Mutation Myelogenous Myeloproliferative disease PML-RARalpha protein Pathogenesis Penetrance Phase Phase I/II Trial Phenotype Relative (related person)Role STAT5A gene Signal Transduction Signal Transduction Inhibitor Site Speed Staging Testing Therapy Clinical Trials Tretinoin Validation Work dosage in vivo in vivo Model inhibitor/antagonist leukemia leukemogenesis loss of function mutant novel pre-clinical progenitor retroviral transduction success

项目摘要

Significant progress has been made in the assessment of the role of FLT3 mutations in AML. These have included analysis of FLT3-ITD expression alone, and in combination with cooperating alleles such as PML-RARa. Working with Project 1, Project 2 has been instrumental in preclinical development of FLT3 inhibitors by showing efficacy in murine models of disease, generating data from therapeutic trials in murine models of disease. In Specific Aim 1, we will explore the in vivo activity of FLT3-ITD and activation loop alleles, and try to understand the relative predilection of FLT3-ITD for myeloid lineage disease and of the FLT3 actiavtion loopalleles for lymphoid disease. We will use multiparameter flow cytometry to test the hypothesis that these alleleshave differential effect on cell fate determination at the multipotent progenitor stage (MPP or LMPP) where FLT3 is highly expressed during hematopoietic development. We will try to understand the mechanism whereby FLT3ITD, in contrast with FLT3 WT, is a potent activator of STAT5 using mutations that abrogate this activity. In Specific Aim 2, we will explore cooperating effects of these accurate genotypic models of FLT3-ITD mediated disease, working with Projects 3 and 4. These in turn will serve as useful in vivo models for testing novel combination therapies that are developed in Project 1. In Specific Aim 3, we will develop murine models of myeloproliferative disease mediated by the JAK2V617F allele, use these models to understand phenotypicpleiotropy of disease in humans, and as a platform for testing novel JAK2 inhibitors for development of clinical trials in Project 5. Overall, this is a highly interactive Project that will build on a proven track record of success and preclinical development of novel therapies for myeloid malignancies. SA 1. Generation and characterization of accurate models of leukemia mediated by mutated FLT3 using knock-in strategies. We will characterize the phenotype of each of these alleles. a. Generation and characterization of a FLT3-ITD conditional knock-in allele b. Generation and characterization of FLT3 D835Y and I836del conditional knock-in alleles c. Generation of FLT3-ITD Y589F/Y598F conditional knock-in allele that is defective in signaling to STATS SA 2. Characterize the cooperative effects of these accurate genotypic models of FLT3-ITD mediated disease with crosses to other germline alleles a. FLT3-ITD KI crossed with Cathepsin G PML-RARalpha and C/EBPalpha knock-in alleles (Interaction with Tenen Project 3) b. FLT3-ITD I836del KI crossed with MIL fusion alleles (interaction with Armstrong, Project 4) c. FLT3-ITD KI crossed with AML1-ETO conditional KI allele Use these models to test combination therapy delineated in Project 1 that could include combination signal transduction inhibitors, ATRA, HDAC inhibitors, or HSP inhibitors SA 3. Develop accurate murine models of JAK2V617F mediated MPD a. Develop and characterize a retroviral transduction model of JAK2V617F disease b. Generate and characterize JAK2V617F conditional knock-in allele c. Characterize novel potentiating mutations of JAK2V617F disease including MPL d. Characterize novel JAK2 inhibitors in murine models as developed in Project 1

在评估FLT3突变在AML中的作用方面取得了重大进展。这些包括单独分析FLT3-ITD表达，以及与合作等位基因如PML-RAR α组合分析。与项目1合作，项目2通过在小鼠疾病模型中显示疗效，从小鼠疾病模型的治疗试验中生成数据，在FLT3抑制剂的临床前开发中发挥了重要作用。在具体目标1中，我们将探索FLT3-ITD和激活环等位基因的体内活性，并试图了解FLT3-ITD对髓系疾病和FLT3激活环等位基因对淋巴疾病的相对偏好。我们将使用多参数流式细胞术来检验这一假设，即这些等位基因对多能祖细胞阶段（MPP或LMPP）的细胞命运决定具有不同的影响，在该阶段，FLT3在造血发育过程中高度表达。我们将尝试理解FLT3ITD与FLT3 WT相比是STAT5的有效激活剂的机制，使用消除这种活性的突变。在具体目标2中，我们将与项目3和4合作，探索这些FLT3-ITD介导疾病的精确基因型模型的协同作用。这些反过来将作为有用的体内模型，用于测试项目1中开发的新型联合疗法。在具体目标3中，我们将开发由JAK2V617F等位基因介导的骨髓增生性疾病的小鼠模型，使用这些模型来了解人类疾病的表型多效性，并作为测试新型JAK2抑制剂的平台，用于项目5中的临床试验。总的来说，这是一个高度互动的项目，将建立在成功和临床前开发的新型骨髓治疗的良好记录之上。恶性肿瘤 SA 1.使用敲入策略生成和表征突变FLT3介导的白血病的精确模型。我们将描述这些等位基因中每一个的表型。 a. FLT3-ITD条件性敲入等位基因的产生和表征 B. FLT3 D835Y和I836del条件性敲入等位基因的产生和表征 C.产生FLT3-ITD Y589F/Y598F条件性敲入等位基因，其在向STATS的信号传导中有缺陷 SA 2.表征FLT3-ITD介导疾病的这些精确基因型模型与其他生殖系等位基因杂交的协同效应 a. FLT3-ITD KI与组织蛋白酶G PML-RAR α和C/EBPalpha敲入等位基因交叉（与Tenen项目3相互作用） B. FLT3-ITD I836del KI与MIL融合等位基因杂交（与Armstrong相互作用，项目4） C. FLT3-ITD KI与AML 1-ETO条件性KI等位基因交叉使用这些模型测试项目1中描述的联合治疗，可能包括联合信号转导抑制剂、ATRA、HDAC抑制剂或HSP抑制剂 SA 3.开发JAK2V617F介导的MPD的准确鼠模型 a. JAK2V617F疾病的逆转录病毒转导模型的开发和表征 B.产生并表征JAK2V617F条件性敲入等位基因 C.表征JAK2V617F疾病（包括MPL）的新型增强突变 D.在项目1中开发的鼠模型中表征新型JAK 2抑制剂