Role of Nuclear Transport in Acute Leukemia

描述(申请人提供)：应聘者是一名内科科学家 最近被任命为该学院终身教职助理教授 西北大学医学院的病理学专业。他的主要关注点(80 百分比)将是对核运输及其在 白血病；其余的致力于临床血液病理学和教学。 核孔蛋白(核孔蛋白)Nup98基因是一种常见的 急性髓系白血病(AML)的染色体重排 以髓系分化失败为特征的。这些重新安排 产生嵌合蛋白，最具特性的是Nup98-HOXA9。 Nup98嵌合体导致白血病的机制尚不清楚。 核孔素和核粘附素(重要蛋白)是这一机制的一部分 调节核质运输的物质。髓系分化是 由需要核转运的转录因子(TF)调控 在原子核内到达目的地的机械。在这些TF中，AML1 和RAR-α在正常髓系分化和 AML。有大量证据表明，下降和/或异常 AML1和RAR-α在核内的分布有助于 急性髓系白血病的发病机制。我们的假设是Nup98基因重排导致 通过干扰介导转录因子的核进口而导致急性髓细胞白血病 髓系分化。关注Nup98-HOXA9对核的影响 AML1和RAR-α的运输与髓系分化，我们将：(I) Nup98和核粘附素在AML1和AML1核进口中的作用 RAR-阿尔法。(Ii)确定Nup98-HOXA9是否通过以下方式中断这一进程 监测AML1和RAR-的核进口、本地化和活性 稳定地导入可诱导的Nup98-HOXA9构建体的细胞中。 我们的假设预测AML1和RAR-α和/或核进口减少 它们的核内分布和转录活性的变化 在Nup98-HOXA9诱导后。(Iii)确定Nup98-HOXA9对 HL60髓系分化过程中髓系特异性基因转录的研究 细胞。髓系分化伴随着转录增强。 一些基因，如CD11b和CD18，以及其他基因的转录减少 即髓过氧化物酶(MPO)。我们的假设预测Nup98-HOXA9将 消除这些倾向。我们将监测Nup98-HOXA9在 内源性和瞬时转染型CD11b、CD18和MPO的分化 推动者。查看整个推广者将提供对 Nup98-HOXA9对髓系转录的影响

DESCRIPTION (provided by applicant): The candidate is a physician-scientist recently appointed as a tenure-track Assistant Professor in the Department of Pathology at Northwestern University Medical School. His major focus (80 percent) will be basic research into nuclear transport and its role in leukemia; with the rest dedicated to clinical hematopathology and teaching. The nuclear pore protein (nucleoporin) Nup98 gene is a frequent target of chromosomal rearrangements in acute myelogenous leukemia (AML), a disease characterized by failure of myeloid differentiation. These rearrangements produce chimeric proteins, the best characterized of which is Nup98-HOXA9. T h e mechanism by which Nup98 chimeras cause leukemia is not known. Nucleoporins, along with karyopherins (importins), are part of the machinery that mediates nucleocytoplasmic transport. Myeloid differentiation is regulated by transcription factors (TFs) that need the nuclear transport machinery to reach their destinations inside the nucleus. Of these TFs, AML1 and RAR-alpha play a central role in both normal myeloid differentiation and AML. There is substantial evidence that a decrease and/or abnormal distribution of AML1 and RAR-alpha within the nucleus contribute to the pathogenesis of AML. Our hypothesis is that Nup98 gene rearrangements cause AML by disrupting the nuclear import of the transcription factors that mediate myeloid differentiation. Focusing on the effect of Nup98-HOXA9 on the nuclear trafficking of AML1 and RAR-alpha and myeloid differentiation, we will: (I) Delineate the role of Nup98 and karyopherins in the nuclear import of AML1 and RAR-alpha. (ii) Determine whether Nup98-HOXA9 disrupts this process by monitoring the nuclear import, localization, and activity of AML1 and RAR- alpha in cells stably transfected with an inducible Nup98-HOXA9 construct. Our hypothesis predicts reduced nuclear import of AML1 and RAR-alpha and/or alteration in their intranuclear distribution and transcriptional activity upon induction of Nup98-HOXA9. (iii) Determine the effect of Nup98-HOXA9 on myeloid-specific gene transcription during myeloid differentiation of HL60 cells. Myeloid differentiation is accompanied by increased transcription of some genes such as CD11b and CD18 and decreased transcription of others such as myeloperoxidase (MPO). Our hypothesis predicts that Nup98-HOXA9 would counteract these tendencies. We will monitor the effect of Nup98-HOXA9 during differentiation on endogenous and transiently transfected CD11b, CD18, and MPO promoters. Looking at entire promoters will provide a global assessment of the effects of Nup98-HOXA9 on myeloid transcription.