Targeting Integrin-Linked (pseudo) Kinase in glioblastoma

靶向胶质母细胞瘤中的整合素连接（伪）激酶

• 批准号: MR/X001180/1
• 负责人: Margaret Frame
• 金额: $ 95.79万
• 依托单位: University of Edinburgh
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2022
• 资助国家: 英国
• 起止时间: 2022 至 无数据
• 项目状态: 未结题
• 来源: https://gtr.ukri.org/projects?ref=MR%2FX001180%2F1
• 关键词: Targeting; Integrin; Linked; pseudo; Kinase

ILK (Integrin-Linked Kinase) is a protein found at sites where cells adhere to their environment. For many years ILK was considered to be a classical enzyme whose activity could be inhibited, but it is now known that instead, it works as a scaffold to bind other proteins in complexes that control how cells behave. In examining several important proteins at adhesion sites in mouse brain progenitor cells that harbour cancer-causing mutations, so as to model the most aggressive brain cancer known as glioblastoma (GBM), we have found that loss of ILK protein causes profound effects associated with reduced malignancy. In brief, cells and tumours can no longer grow properly, and the cells lose their ability to invade surrounding material. Also, the cells lose their 'stem-like' state, often associated with problematic cells in tumours that do not respond the treatment. Other researchers have used unbiased genetic screens and also found that ILK is a promising protein to change the cancerous behaviour of human GBM cells.In this project, we propose to use the most up to date form of gene editing to remove the ILK gene, and so protein, from human GBM cells that were recently derived from patients in Edinburgh. The idea here is to perform proof-of-principle experiments that will tell us whether, and if so how, the ILK protein plays important roles in human GBM cells. In beginning of our experiments in human GBM cells, we made an intriguing finding, namely that one of the proteins that binds to ILK is perturbed. We will investigate whether or not this is an important change, and might predict if the cells have adopted new ways to function. As well as addressing ILK function in human GBM cells in depth, we will also try to find out how best to combine loss of ILK with drugs that enhance the biological effects, and the duration of effects. For this, we will use state-of-the-art drug discovery methods that "paint" cells and study their shape using high-throughput microscopy. We have used this before to successfully identify combinations between deletion of other adhesion proteins and drugs in a different cancer type. A series of well-understood chemical (drug) libraries that we have built for such purposes in our Institute will be used. Agents that work together with ILK loss to enhance advantageous biological effects will be studied in more detail and the strongest hits will be prioritised for further study to understand how they are working.Finally, we propose to take the bold approach of generating a novel kind of drug that binds to ILK and cause its protein complexes to disintegrate. The strategy will ensure that biological effects are guiding the synthesis of chemical series (so selecting for agents that get into GBM cells and have good effects ('selecting the winners') early in the drug discovery process. These will be optimised by further chemical modification and then testing in GBM cells. We think this strategy will work because we already have agents that bind to the region of ILK that is needed to maintain the levels of its protein partners, and these can be further chemically modified to improve them. As proof this can work, we have recently used this strategy to develop drug candidates licensed to Pharma. Our vision is to deepen understanding of GBM cell biology and the role of one important adhesion protein, namely ILK; this is much-needed because no new treatments for this dismal disease have been forthcoming over the past two decades. ILK is not a new target, but we have substantial new information in more physiologically relevant systems to study GBM cells than used previously, implying that ILK may be an excellent target for therapy. New experiments to fully understand its role in GBM, and bold new approaches to: a) determine how best to combine ILK-deficiency with other drugs, and b) make our own small molecule inhibitors for onward translation, will further our aim of ultimately providing benefit to patients.

ILK（整合素连接激酶）是在细胞粘附于其环境的位点处发现的蛋白质。多年来，ILK被认为是一种经典的酶，其活性可以被抑制，但现在已知的是，它作为一种支架结合其他蛋白质的复合物，控制细胞的行为。在检查小鼠脑祖细胞中的粘附位点处的几种重要蛋白质时，这些蛋白质具有致癌突变，以便模拟称为胶质母细胞瘤（GBM）的最具侵袭性的脑癌，我们发现ILK蛋白的缺失引起与恶性程度降低相关的深远影响。简而言之，细胞和肿瘤不再能正常生长，细胞失去了侵入周围物质的能力。此外，细胞失去了它们的“干细胞样”状态，通常与肿瘤中对治疗没有反应的问题细胞有关。其他研究人员使用无偏见的遗传筛选，也发现ILK是一种有希望改变人类GBM细胞癌行为的蛋白质。在这个项目中，我们建议使用最新形式的基因编辑来删除ILK基因，因此蛋白质，从最近来自爱丁堡患者的人类GBM细胞中。这里的想法是进行原理验证实验，告诉我们ILK蛋白是否在人类GBM细胞中起重要作用，如果是的话，如何起重要作用。在我们在人类GBM细胞中的实验开始时，我们发现了一个有趣的发现，即与ILK结合的蛋白质之一受到干扰。我们将研究这是否是一个重要的变化，并可能预测细胞是否采用了新的功能方式。除了深入探讨ILK在人GBM细胞中的功能外，我们还将尝试找出如何最好地将ILK的联合收割机损失与增强生物效应和效应持续时间的药物相结合。为此，我们将使用最先进的药物发现方法，“油漆”细胞，并使用高通量显微镜研究它们的形状。我们以前曾使用这种方法成功地鉴定了不同癌症类型中其他粘附蛋白缺失和药物之间的组合。我们将使用我们在研究所为此目的建立的一系列众所周知的化学（药物）图书馆。将更详细地研究与ILK损失一起工作以增强有利的生物效应的药剂，并且将优先考虑最强的命中以进行进一步研究以了解它们是如何工作的。最后，我们建议采取大胆的方法来产生一种新型药物，该药物与ILK结合并导致其蛋白质复合物分解。该策略将确保生物效应指导化学系列的合成（因此在药物发现过程的早期选择进入GBM细胞并具有良好效果的药物（“选择赢家”））。这些将通过进一步的化学修饰进行优化，然后在GBM细胞中进行测试。我们认为这种策略会起作用，因为我们已经有了与ILK区域结合的试剂，该区域是维持其蛋白质伴侣水平所必需的，并且这些试剂可以进一步化学修饰以改善它们。为了证明这是可行的，我们最近使用这一策略开发了授权给制药公司的候选药物。我们的愿景是加深对GBM细胞生物学和一种重要的粘附蛋白（即ILK）的作用的理解;这是非常必要的，因为在过去的二十年里，这种令人沮丧的疾病没有新的治疗方法。ILK不是一个新的靶点，但我们在更生理相关的系统中获得了大量新的信息来研究GBM细胞，这意味着ILK可能是一个很好的治疗靶点。新的实验，以充分了解其在GBM中的作用，和大胆的新方法：a）确定如何最好地结合联合收割机ILK缺陷与其他药物，和B）使我们自己的小分子抑制剂向前翻译，将进一步我们的目标，最终为患者提供好处。