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

转录因子NF-E2和编码PRV-1的基因在患有高脂血症的患者中过表达。 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， 该等位基因是否部分通过诱导NF-E2表达对恶性克隆发挥作用。NF-E2 是PV病理生理学中有前途的候选药物，原因如下： NF-E2在干细胞以及PV中受影响的所有三种细胞谱系中过表达。在鼠细胞中，NFE 2 过表达导致Epo-independent生长和分化。因此，NF-E2可能发挥关键作用 在引起PV的红细胞增多症方面。然而，导致NF-E2的分子机制 过表达及其对人造血的影响尚不清楚。此外，PRV-1的病因 过度表达仍不清楚。因此，本项目的目的是调查NF-E2的原因 PV中PRV-1过表达和造血细胞中NF-E2过表达的影响。 基于以下假设，该项目的具体目标是： 1.假设：NF-E2是PV细胞的Epo非依赖性生长或其过表达所必需的 调节造血分化。具体目的：通过siRNA敲低调节NF-E2表达 和逆转录病毒或慢病毒转导，并检查Epo依赖性和非依赖性 体外生长 2.假设：PV中的NF-E2和PRV-1过表达由Jak2V617F等位基因介导。 在体内和体外引入Jak2 wt和V617F等位基因，并检查对NF-E2和PRV-1的影响 在不同的模型中表达。 3.假设：NF-E2和PRV-1过表达由异常转录激活引起。 表征PV和健康对照细胞中NF-E2和PRV-1启动子上的蛋白质/DNA相互作用 以确定异常激活的转录因子。 公共卫生影响：通过更好地了解导致疾病的分子变化， PV的发展，该项目将指出分子对新的治疗药物可以 开发

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.