Understanding and targeting oncogenic biomolecular condensates of ALK kinase

了解和靶向 ALK 激酶的致癌生物​​分子缩合物

• 批准号: MR/X008673/1
• 负责人: Richard Bayliss
• 金额: $ 101.02万
• 依托单位: University of Leeds
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2023
• 资助国家: 英国
• 起止时间: 2023 至 无数据
• 项目状态: 未结题
• 来源: https://gtr.ukri.org/projects?ref=MR%2FX008673%2F1
• 关键词: Understanding; targeting; oncogenic; biomolecular; condensates

Lung cancer is the 3rd most common cancer in the UK, with almost 50,000 new cases diagnosed annually. Lung cancer is classified as small cell or the more common non-small cell (80% of cases). Approximately 5% of these patients have a specific mutation in the DNA of their cancer cells, resulting in a gene fusion called EML4-ALK. EML4-ALK patients are on average much younger than most lung cancer patients. They are treated with ALK tyrosine kinase inhibitors, and 5-year survival for these patients has improved to 50%-60%, much more effective than standard chemotherapy. There is still room for improvement, and so we need to decipher the molecular mechanisms through which EML4-ALK proteins promote cancer cell survival and proliferation and to determine how the cancers become resistant to ALK inhibitor therapy. This will allow us to develop new and optimised treatments.In this project, we will focus on the molecular interactions of EML4-ALK proteins, the cellular structures they form, and how these respond to cancer therapies. In healthy cells, EML4 regulates the formation of cellular structures called microtubules. The ALK protein normally functions on the surface of cells in the developing brain where, in response to a signalling molecule from outside the cell, it instructs the cell to proliferate. In lung cancer, parts of these proteins are fused together, and interactions between EML4-ALK protein molecules allows them to instruct lung cancer cells to proliferate out of the control of the rest of the body. These interactions also allow EML4-ALK proteins to form compartments within cells that are enriched in other proteins required to promote cell proliferation and survival. We previously discovered that some ALK inhibitors used in cancer treatment destabilise the compartments, whereas one commonly used ALK inhibitor stabilises the compartments. Based on this and other observations, we think that the compartments are formed from interactions between the ALK parts of the EML4-ALK protein molecules, as well as interactions between the EML4 parts. If we could block these interactions, this might lead to a new way to treat EML4-ALK lung cancers. We will focus on the ALK interactions because we think this is more feasible, and because uncontrolled ALK activity is the cause of many other cancers, whereas EML4 dysfunction is rarely found in other cancer types. Our project will begin with the production of molecular tools to disrupt the ALK interactions. We have already made a set of eight synthetic protein molecules called nanobodies that bind to EML4-ALK in cells. We will develop these molecular tools further by characterising precisely how they affect ALK interactions and activity. In parallel, we will determine precisely how ALK molecules interact with each other. Both strands of work will use experimental structural biology methods and computational methods such as artificial intelligence. Also in parallel, we will find out which other proteins interact with EML4-ALK and determine which of them are present in the cellular compartments. The organisation of these proteins within the compartments, and the movement of molecules in and out of the compartments will be studied using state-of-the-art cell imaging methods. This part of the project will tell us how the compartments are built, and how molecules are passed from the compartment to the rest of the cell with the instructions to proliferate. The final step of the project will be to use this new information and our new molecular tools to ask whether disrupting or stabilising the compartments can effectively block the cell proliferation and survival signals from EML4-ALK. We will test this in cells that were cultured from lung cancer patients, and cells that have become resistant to ALK inhibitors used in cancer therapy . If successful, we will initiate a follow-up project towards improving the treatment of lung cancer based on our findings.

肺癌是英国第三大常见癌症，每年诊断出近 50,000 例新病例。肺癌分为小细胞或更常见的非小细胞（占病例的 80%）。这些患者中大约 5% 的癌细胞 DNA 发生特定突变，从而产生称为 EML4-ALK 的基因融合。 EML4-ALK 患者平均比大多数肺癌患者年轻得多。他们接受ALK酪氨酸激酶抑制剂治疗，这些患者的5年生存率提高到50%-60%，比标准化疗有效得多。仍有改进的空间，因此我们需要破译 EML4-ALK 蛋白促进癌细胞存活和增殖的分子机制，并确定癌症如何对 ALK 抑制剂治疗产生耐药性。这将使我们能够开发新的和优化的治疗方法。在这个项目中，我们将重点研究 EML4-ALK 蛋白的分子相互作用、它们形成的细胞结构，以及它们如何响应癌症治疗。在健康细胞中，EML4 调节称为微管的细胞结构的形成。 ALK 蛋白通常在发育中的大脑细胞表面发挥作用，响应细胞外的信号分子，指示细胞增殖。在肺癌中，这些蛋白质的部分融合在一起，EML4-ALK 蛋白质分子之间的相互作用使它们能够指示肺癌细胞在不受身体其他部分控制的情况下增殖。这些相互作用还允许 EML4-ALK 蛋白在细胞内形成区室，这些区室富含促进细胞增殖和存活所需的其他蛋白质。我们之前发现，一些用于癌症治疗的 ALK 抑制剂会破坏区室的稳定性，而一种常用的 ALK 抑制剂则可以稳定区室。基于这一观察和其他观察，我们认为区室是由 EML4-ALK 蛋白分子的 ALK 部分之间的相互作用以及 EML4 部分之间的相互作用形成的。如果我们能够阻断这些相互作用，这可能会带来一种治疗 EML4-ALK 肺癌的新方法。我们将重点关注 ALK 相互作用，因为我们认为这更可行，并且因为不受控制的 ALK 活性是许多其他癌症的原因，而 EML4 功能障碍在其他癌症类型中很少发现。我们的项目将从生产分子工具来破坏 ALK 相互作用开始。我们已经制造了一组八种合成蛋白质分子，称为纳米抗体，可以与细胞中的 EML4-ALK 结合。我们将通过精确表征它们如何影响 ALK 相互作用和活性来进一步开发这些分子工具。与此同时，我们将精确确定 ALK 分子如何相互作用。这两项工作都将使用实验结构生物学方法和人工智能等计算方法。同时，我们还将找出哪些其他蛋白质与 EML4-ALK 相互作用，并确定其中哪些蛋白质存在于细胞区室中。将使用最先进的细胞成像方法研究这些蛋白质在区室内的组织以及分子进出区室的运动。该项目的这一部分将告诉我们隔室是如何构建的，以及分子如何按照增殖指令从隔室传递到细胞的其余部分。该项目的最后一步将是利用这些新信息和我们的新分子工具来探究破坏或稳定隔室是否可以有效阻断来自 EML4-ALK 的细胞增殖和存活信号。我们将在肺癌患者培养的细胞以及对癌症治疗中使用的 ALK 抑制剂产生耐药性的细胞中对此进行测试。如果成功，我们将根据我们的发现启动一个后续项目，以改善肺癌的治疗。

项目成果

ALKing the flames of lung cancer immunosensitivity.

• DOI: 10.1002/1878-0261.13533
• 发表时间: 2023-11
• 期刊: MOLECULAR ONCOLOGY
• 影响因子: 6.6
• 作者: Bayliss, Richard;Sarnowska, Elzbieta;Yeoh, Sharon;Sampson, Josephina
• 通讯作者: Sampson, Josephina

EML4-ALK biology and drug resistance in non-small cell lung cancer: a new phase of discoveries.

• DOI: 10.1002/1878-0261.13446
• 发表时间: 2023-06
• 期刊: Molecular oncology
• 影响因子: 6.6