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

研究 AP-1 在缺氧细胞适应转录调控中的作用

• 批准号: 2074487
• 金额: --
• 依托单位: University of Nottingham
• 依托单位国家: 英国
• 项目类别: Studentship
• 财政年份: 2018
• 资助国家: 英国
• 起止时间: 2018 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=studentship-2074487
• 关键词: Investigating; role; AP; transcriptional; regulation

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.

大多数低氧研究都集中在HIF 1 α和HIF 2 α上，它们是在低氧中稳定的转录因子（TF）。数据表明，发生的转录组学变化是由一些TF和表观遗传调节因子调节的。我们的证据表明，AP-1转录因子（来自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应答中被激活并调节低氧分子适应的关键方面的假设。我们将：1.基于内部RNA-SEQ数据和ChIP-SEQ.2鉴定缺氧中AP-1的新靶点。在缺氧条件下检测靶点的激活，并确认AP-1的直接激活。确定新的AP-1靶点在低氧分子适应中的作用.研究新型AP-1途径在癌细胞中的表型效应。5.利用CRISPR产生的敲除细胞系鉴定新型AP-1靶点的HIF非依赖性缺氧调节。除了在实验室轮换中培养的技能外，学生还将利用慢病毒-shRNA/CRISPR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数据和癌症基因组图谱中的可用数据进行生物信息学分析。