Mechanisms and effects of NF-E2 and PRV-1 overexpression in PV: Role of Jak2V617F

转录因子NF-E2和PRV-1编码基因在慢性支气管炎患者中的过度表达 真性红细胞增多症(PV)。最近对PV患者JAK2V617F突变的描述提出了一个问题 该等位基因对恶性克隆的作用部分是通过诱导核因子-E2的表达来实现的。核因子-E2 是PV病理生理学领域很有前途的候选者，原因如下： 核因子-E2在干细胞中以及在PV中受影响的所有三种细胞系中都过表达。在小鼠细胞NFE2中 过度表达导致EPO非依赖的生长和分化。因此，核因子-E2可能起到关键作用。 在引起PV的红细胞增多症方面。然而，导致核因子-E2的分子机制 目前尚不清楚该基因的过度表达及其对人类造血的影响。此外，PRV-1的起因 过度表达仍不清楚。因此，本项目的目的是研究核因子-E2的原因 PV中PRV-1过表达及造血细胞中NF-E2过表达的影响。 因此，基于以下假设，本项目的具体目标是： 1.假设：核因子-E2是PV细胞EPO非依赖性生长或其过度表达所必需的 调节造血分化。特定目的：通过siRNA敲除来调节核因子-E2的表达 和逆转录病毒或慢病毒转导，并检查对EPO依赖和非依赖的后果 在体外生长。 2.假说：PV中NF-E_2和PRV-1的过度表达是由JAK2V617F等位基因特异的目的介导的： 体内、外引入JAK2wt和V617F等位基因并检测其对NF-E2和PRV-1的影响 在不同的模型中表达。 3.假设：核因子-E2和PRV-1的过度表达是转录激活异常的结果特定目的： PV和健康对照细胞中NF-E2和PRV-1启动子的蛋白质/DNA相互作用特征 以确定异常激活的转录因子。 对公共卫生的影响：通过更好地理解导致 开发光伏，这个项目将指出治疗新药可以针对的分子 发展起来的。

The transcription factor NF-E2 and the gene encoding PRV-1 are overexpressed in patients with polycythemia vera (PV). The recent description of a Jak2V617F mutation in PV patients raises the question whether this allele exerts its effect on the malignant clone in part through inducing NF-E2 expression. NF-E2 is a promising candidate in the pathophysiology of PV for several reasons: NF-E2 is overexpressed in stem cells as well as in all three cell lineages affected in PV. In murine cells NFE2 overexpression leads to Epo-independent growth and differentiation. NF-E2 may thus play a pivotal role in causing the erythrocytosis of PV. However, both the molecular mechanism leading to NF-E2 overexpression and its effect on human hematopoiesis are not known. In addition, the cause of PRV-1 overexpression remains unclear. Therefore, it is the aim of this project to investigate the cause of NF-E2 and PRV-1 overexpression in PV and the effect of NF-E2 overexpression in hematopoietic cells. Based on the following hypotheses, the specific aims of this project are therefore: 1. Hypothesis: NF-E2 is required for the Epo-independent growth of PV cells or its overexpression modulates hematopoietic differentiation. Specific Aim: To modulate NF-E2 expression via siRNA knock down and retroviral or lentiviral transduction and examine the consequences on Epo-dependent and -independent growth in vitro. 2. Hypothesis: NF-E2 and PRV-1 overexpression in PV are mediated by the Jak2V617F allele Specific Aim: To introduce Jak2 wt and V617F alleles in vivo and in vitro and examine the effects on NF-E2 and PRV-1 expression in various models. 3. Hypothesis: NF-E2 and PRV-1 overexpression result from aberrant transcriptional activation Specific Aim: To characterize protein/DNA interaction on the NF-E2 and PRV-1 promoters in PV and healthy control cells to determine aberrantly activated transcription factors. Public Health Implications: By leading to a better understanding of the molecular changes that lead to the development of PV, this project will point out molecules against which new drugs for treatment can be developed.