Finding specific treatments for cancer patients with mutations in the ribosome production/p53 pathway

寻找针对核糖体生成/p53 通路突变的癌症患者的具体治疗方法

• 批准号: 1960427
• 金额: --
• 依托单位: Newcastle University
• 依托单位国家: 英国
• 项目类别: Studentship
• 财政年份: 2017
• 资助国家: 英国
• 起止时间: 2017 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=studentship-1960427
• 关键词: Finding; specific; treatments; cancer; patients

Keywords: Molecular datasets and disease; Genetic influence on Health; Bioinformatics and personalised medicineSummary:BACKGROUNDBroad activity chemotherapeutics are used to treat multiple types of cancer. However, due to the complex and diverse genetic background underlying cancer types and individual tumours, this does not always work and a "personalised medicine" approach is now seen as a way forward.The TP53 gene, which encodes the tumour suppressor p53, is mutated in more than half of all cancers. Many anti-cancer drugs function through activating p53, leading to cell cycle arrest, apoptosis or senescence.The majority of anti-cancer chemotherapeutics, including commonly used drugs such as 5FU, are known to block ribosome production (Burger et al., JBC 2010). Sequencing of 12,000 cancers and 1001 cancer cell lines revealed that ribosomal protein (RP) genes are "cancer genes" in 12 of the 30 cancer types studied (including leukaemia, breast, lung and prostate cancers (Iorio et al., Cell 2016).The primary supervisor's lab demonstrated that defects in ribosome production (e.g. RP gene mutation) leads to the activation of p53 (Sloan et al., Cell Reports 2013). It is, however, currently unclear how p53 activation caused by ribosome production defects would lead to cancer in patients carrying specific mutations in RP genes. Here, we therefore plan to characterise a selection of cancer cell lines, which faithfully recapitulate oncogenic alterations in tumours and carry known mutations in RP genes (Iorio et al., Cell 2016). This approach will allow us to investigate the effect of the mutations on p53 signalling and to determine the sensitivity of these cell lines to a range of chemotherapeutics.PROJECTWe have selected a series of cancer cell lines containing stop codons/frame-shift mutations early in the ribosomal protein coding sequence to ensure a functional defect. The student will use a wide variety of approaches, including high-throughput screening, to address the following aims:1) Use a combination of cellular, molecular biology and RNA analysis approaches to determine the impact of RP gene mutations on ribosome production, p53 signalling and cellular growth in the cancer cell lines (primary and secondary supervisors).2) Use CRISPR/Cas9 editing (established in lab of secondary supervisor) to generate select RP gene cancer mutations in a control cell line (e.g. U2OS) to validate that this mutation, and not other variations in the cancer cell line, is responsible for changes in ribosome production and p53 signaling (secondary and primary supervisors).3) Use high-throughput screening to test the hypothesis that mutations in RP genes will affect the response of these tumours to anti-cancer chemotherapeutics that impact ribosome production (primary and secondary supervisors and the High-Throughput Screening Facility).4) Use chemicals (e.g. mTOR activators), to increase general RP production, and long non-coding RNAs (Sineups), to increase expression from the non-mutated RP gene, in cancer cell lines to determine whether this is a valid approach to treat tumours with RP gene mutations (primary supervisor).This project will form the basis for future collaborations with the NICR including patient-associated material (e.g. Christine Harrison for leukaemia's) and the patient-derived xenograft (PDX) system.

关键词：分子数据集和疾病；遗传对健康的影响；生物信息学和个性化医疗摘要：背景广泛活性的化疗药物用于治疗多种类型的癌症。然而，由于癌症类型和个体肿瘤的遗传背景复杂多样，这种方法并不总是有效，“个性化医疗”方法现在被视为一种前进的方向。编码肿瘤抑制因子 p53 的 TP53 基因在超过一半的癌症中发生突变。许多抗癌药物通过激活 p53 发挥作用，导致细胞周期停滞、细胞凋亡或衰老。大多数抗癌化疗药物，包括 5FU 等常用药物，已知会阻断核糖体产生（Burger 等人，JBC 2010）。对 12,000 个癌症和 1001 个癌细胞系的测序表明，核糖体蛋白 (RP) 基因在所研究的 30 种癌症类型中的 12 种中是“癌症基因”（包括白血病、乳腺癌、肺癌和前列腺癌（Iorio 等人，Cell 2016）。主要主管实验室证明核糖体生产缺陷（例如 RP 基因突变）会导致 p53 的激活 （Sloan 等人，Cell Reports 2013）。然而，目前尚不清楚核糖体产生缺陷引起的 p53 激活如何导致携带 RP 基因特定突变的患者罹患癌症。因此，在这里，我们计划表征一系列癌细胞系，这些细胞系忠实地再现了肿瘤中的致癌改变，并携带已知的 RP 基因突变（Iorio 等人，Cell 2016）。这种方法将使我们能够研究 p53 信号传导突变，并确定这些细胞系对一系列化疗药物的敏感性。项目我们选择了一系列在核糖体蛋白编码序列早期含有终止密码子/移码突变的癌细胞系，以确保功能缺陷。学生将使用多种方法，包括高通量筛选，来实现以下目标：1) 结合使用细胞、分子生物学和 RNA 分析方法可确定 RP 基因突变对癌细胞系（初级和二级主管）中核糖体产生、p53 信号传导和细胞生长的影响。2) 使用 CRISPR/Cas9 编辑（在二级主管实验室建立）在对照细胞系（例如 U2OS）中生成选定的 RP 基因癌症突变，以验证该突变（而不是癌细胞系中的其他变异）导致了 核糖体产生和 p53 信号转导（二级和一级监管器）。3) 使用高通量筛选来检验以下假设：RP 基因突变将影响这些肿瘤对影响核糖体产生的抗癌化疗药物的反应（一级和二级监管器以及高通量筛选设施）。4) 使用化学品（例如 mTOR 激活剂）来增加一般 RP 产量，并长期 非编码RNA（Sineups），以增加癌细胞系中非突变RP基因的表达，以确定这是否是治疗RP基因突变肿瘤的有效方法（主要主管）。该项目将构成未来与NICR合作的基础，包括患者相关材料（例如用于白血病的Christine Harrison）和患者来源的异种移植（PDX）系统。