MURINE MODELS OF MYELOID MALIGNANCIES

在前一个供资期间，我们在评估FLT3的作用方面取得了重大进展 急性心肌梗死的突变，以及测试Flt3抑制剂的小鼠模型的发展。这些都有 包括单独分析Flt3-ITD的表达，并结合协同等位基因 作为PML-RARAlpha。与项目1合作，项目2在临床前工作中发挥了重要作用 Flt3抑制剂在小鼠疾病模型中显示疗效的研究进展 项目5中两种不同的Flt3抑制剂的I期和II期试验。我们计划继续 使用条件性敲入等位基因进一步研究Flt3-ITD在白血病发生中的作用。在……里面 具体目的1，我们将分析Flt3-ITD和激活环等位基因的体内活性，并尝试 了解Flt3-ITD对髓系疾病和Flt3激活的相对偏好 淋巴疾病的环状等位基因。我们将使用高速多参数流式细胞术来检测 假设这些等位基因在多能性中对细胞命运决定有不同的影响 造血祖细胞阶段(LMPP/MPP)，在造血发育过程中，Flt3高表达。 我们将研究与Flt3 WT相比，Flt3-ITD是一种有效的激活剂的机制。 在Flt3的上下文中使用突变来废除这一活动的统计数据。在具体目标2中，我们将 探索这些精确的Flt3-ITD介导的疾病基因模型的协同作用 通过与包括C/EBPalpha和MLL融合在内的几个互补等位基因的杂交， 与项目3和4一起工作。这些反过来将作为测试新小说的有用的活体模型 在项目1中开发的联合疗法。我们还将继续研究最新的发现 提示逆转录病毒Flt3-ITD逆转录病毒整合位点可能参与了病毒性肝炎的发病机制。 AML在组织蛋白酶G-PML-RARpha模型中的协同作用。在具体目标3中，我们将 JAK2V617F等位基因介导的小鼠骨髓增生性疾病(MPD)模型的建立 我们和其他人最近确认了..最后，我们将使用MPD的小鼠模型作为平台 在项目5中测试用于临床试验开发的新型JAK2抑制剂。总体而言，这是一个高度 互动项目，将建立在已证实的成功记录和临床前开发的基础上 治疗髓系恶性肿瘤的新疗法。

We have made significant progress during the prior funding period in assessing the role of FLT3 mutations in AMI, and the development of murine models to test FLT3 inhibitors. These have included analysis of FLT3-ITD expression alone, and in combination with cooperating alleles such as PML-RARalpha. Working with Project 1, Project 2 has been instrumental in preclinical development of FLT3 inhibitors by showing efficacy in murine models of disease, resulting in Phase I and Phase II trials of two different FLT3 inhibitors in Project 5. We plan to move forward with additional studies of the role of FLT3-ITD in leukemogenesis using conditional knock-in alleles. In Specific Aim 1, we will analyze 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 FLT3 activation loop alleles for lymphoid disease. We will use high speed multiparameter flow cytometry to test the hypothesis that these alleles have differential effect on cell fate determination at the multipotent progenitor stage (LMPP/MPP) where FLT3 is highly expressed during hematopoietic development. We will study the mechanism whereby FLT3-ITD, in contrast with FLT3 WT, is a potent activator of STATS using mutations in the context of FLT3 that abrogate this activity. In Specific Aim 2, we will explore cooperating effects of these accurate genotypic models of FLT3-ITD mediated disease through crosses with several complementing alleles including, C/EBPalpha, and MLL fusions, 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. We .will also pursue recent findings suggesting that retroviral FLT3-ITD retroviral integration sites may contribute to pathogenesis of AML in the Cathepsin G - PML-RARalpha model of cooperativity. In Specific Aim 3, we will develop murine models of myeloproliferative disease (MPD) mediated by the JAK2V617F allele that we and others have recently identified.. Finally, we will use murine models of MPD 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.