Investigating the role of AP-1 in the transcriptional regulation of hypoxic cell adaptation

大多数低氧研究都集中在HIF1α和HIF2α，它们是在低氧中稳定的转录因子(TF)。数据表明，发生的转录变化受到许多转录因子和表观遗传调节因子的调控。我们的证据表明，AP-1转录因子(c-fos、c-jun和ATF蛋白的异源二聚体)是低氧分子适应的关键调节因子。我们计划验证AP-1转录因子的新下游靶点在低氧反应中被激活的假设，并调节分子对低氧适应的关键方面。我们将：1.基于内部RNA-SEQ数据和CHIP-SEQ2发现AP-1在缺氧中的新靶点。确认缺氧时靶点的激活，并确认AP-1.3直接激活。确定新的AP-1靶点在分子低氧适应中的作用。研究新的AP-1通路在癌细胞中的表型效应。使用CRISPR产生的基因敲除系识别新的AP-1靶标的HIF非依赖的低氧调节。除了在实验室轮换中发展的技能之外，学生将使用慢病毒shRNA/CRISPR来分析对癌细胞的表型影响，包括干细胞能力、血管生成、增殖、三维培养和免疫组织化学。学生将在支持下对内部RNA-SEQ数据和来自癌症基因组图谱的公开数据进行生物信息学分析。

The majority of hypoxic research has focused on HIF1alpha and HIF2alpha, transcription factors (TFs) which are stabilised in hypoxia. Data suggests that the transcriptomic changes that occur are regulated by a number of TFs and epigenetic regulators. Our evidence suggests that the AP-1 transcription factors (a heterodimer of proteins from c-FOS, c-JUN and ATF proteins) are key regulators of hypoxic molecular adaptation.We plan to test the hypothesis that novel downstream targets of the AP-1 transcription factor are activated in response to hypoxia and regulate key aspects of molecular adaptation to hypoxia. We will:1. Identify novel targets of AP-1 in hypoxia based on in-house RNA-SEQ data and ChIP-SEQ.2. Validate the activation of targets in hypoxia and confirm direct activation by AP-1.3. Identify the role of novel AP-1 targetas in molecular adaptation to hypoxia.4. Investigate phenotypic effects of novel AP-1 pathways in cancer cells.5. Identify HIF-independent hypoxic regulation of novel AP-1 targets using CRISPR-generated knockout lines.Further to the skills developed in the lab rotation the student will utilise lentiviral-shRNA/CRISPR to analyse the phenotypic effects on cancer cells including stem cell capacity angiogenesis, proliferation, 3 Dimensional culture and immunohistochemistry. The student will with support undertake bioinformatics analyses of in house RNA-SEQ data and publically available data from The Cancer Genome Atlas.