MICA: Liverpool Imaging Partnership: Molecular physiology and drug response

在多细胞生物中，细胞暴露于分子信号，如炎症分子和应激信号。细胞必须解释这些信号以适应并做出适当的反应。为了理解导致特定反应（如细胞死亡、细胞生长、细胞迁移等）的分子机制，生物学家必须测量蛋白质的水平、定位和修饰，以及随后对特定基因的调节。人们越来越认识到，在细胞群中，每个细胞都是不同的，因此测量平均行为可能会导致错误的结论。活细胞成像加上荧光和发光标签和报告器的使用，使生物学家能够实时监测单个细胞中的动态事件。纳米技术为单个分子的可视化提供了额外的工具。我们项目的目的是建立我们在细胞成像中心现有的成像能力，以及我们在纳米颗粒合成和成像方面的世界领先的专业知识，以改进最新显微镜的设备。我们还将开发新的技术、软件和图像分析方法，使我们能够提高图像的分辨率及其解释（以便更好地看到细胞内部的详细结构）。我们将确保这些设备在回答重要的生物学和生物医学问题上发挥最大的作用。为此，我们将在英国利物浦及其他地区选择最好的项目，我们将为用户提供从实验概念到结果分析和解释的支持。因此，CCI平台将允许来自广泛研究领域的研究人员通过在分子水平上实时研究细胞事件来解决21世纪生物医学研究中的重要挑战。为了说明该平台将为科学服务的广度，我们简要介绍以下三个范例项目，其中许多项目将从改进的设施中受益。在纳米尺度上理解急性胰腺炎。胰腺细胞产生消化肠道内食物的酶。在急性胰腺炎中，酶不是分泌到肠道，而是在胰腺细胞中变得活跃，损害胰腺，导致剧烈疼痛、住院治疗，在某些情况下甚至死亡。这些酶的激活涉及到动态结构的形成，这是很难通过传统显微镜研究的，因为它们比传统显微镜的分辨率小。当胰腺细胞暴露于可能诱发或预防急性胰腺炎的分子中时，新的平台将使我们能够独特地研究这些结构的反应，并最终开发出防止这种情况发生的新方法。成像氧化应激在衰老中的微妙作用；50多年来，氧化应激导致氧化损伤一直与衰老过程有关，但最近的数据表明，与其说氧化应激损伤的总体变化，不如说是还原氧化反应调节的更微妙的变化，在衰老过程中起着根本作用。新平台将使我们能够控制和测量氧气水平和氧化应激，为衰老机制提供新的见解。纳米药物肠道吸收机制的探讨药物活性成分的不溶性及其导致的生物利用度的缺乏是新药开发的一个主要问题。基于纳米技术的方法已经被证明是成功的，但是关于其机制基础所知甚少。目前的应用将使我们能够研究纳米药物与传统溶解药物在肠通和毒性方面的差异。

In multicellular organisms, cells are exposed to molecular signals such as inflammatory molecules and stress signals. Cells have to interpret these signals to adapt and respond appropriately. To understand the molecular mechanisms leading to a particular response (e.g. cell death, cell growth, cell migration, etc), biologists have to measure the levels, localisation and modifications of proteins, and the subsequent regulation of specific genes. There is an increasing recognition that in a population of cells, each cell is different and therefore measuring the average behaviour can lead to erroneous conclusions. Live cell imaging coupled with the use of fluorescent and luminescent labels and reporters enables biologists to monitor dynamic events in individual cells in real time. Nanotechnology provides additional tools to visualise single molecules.The aim of our project is to build on our existing imaging capabilities in the Centre for Cell Imaging and our world-leading expertise in the synthesis and imaging of nanoparticles to improve the equipment available with the most up-to-date microscopes. We will also develop new technologies, software and image analysis methods that will allow us to increase the resolution of the images and their interpretation (to better see the detailed structures inside the cells). We will make sure that the equipment is used to the best of its capabilities to answer important biological and biomedical questions. For this purpose, we will select the best projects, in Liverpool, the UK and beyond, and we will support users from the conception of their experiments to the analysis and interpretation of their results.The CCI platform will therefore permit researchers from a broad range of research fields to tackle important 21st century challenges in biomedical research, by investigating in real-time cellular events at the molecular level. To illustrate the breadth of science that will be served by the platform, we briefly present below three exemplar projects that, among many others, will benefit of the improved facility. UNDERSTANDING ACUTE PANCREATITIS ON THE NANOMETRE SCALE. Cells of the pancreas produce the enzymes that digest our food inside the intestine. In acute pancreatitis, instead of being secreted to the intestine, enzymes become active in the pancreas cells themselves and damage the pancreas leading to severe pain, hospitalisation and in some cases, death. The activation of these enzymes involves the formation of dynamic structures that are very difficult to study by conventional microscopy because they are smaller than the resolution of conventional microscopes. The new platform will uniquely allow us to study the response of those structures when pancreatic cells are exposed to molecules that may induce or prevent acute pancreatitis and ultimately develop novel approaches to prevent this from happening.IMAGING THE SUBTLE ROLE OF OXIDATIVE STRESS IN AGEING; Oxidative stress leading to oxidative damage has been implicated in the processes underlying ageing for over 50 years, but recent data demonstrated that rather than gross changes in oxidative stress damage, it is the more subtle changes in regulation of reduction-oxidation reactions that play a fundamental role in ageing processes. The new platform will uniquely allow us to control and measure oxygen level and oxidative stress providing new insights into mechanisms of ageing.PROBING THE MECHANISM FOR INTESTINAL ABSORPTION OF NANOMEDICINES; Insolubility of active pharmaceutical ingredients and the resulting lack of bioavailability is a major concern for the development of new drugs. Approaches based on nanotechnology have proven successful, but very little is known regarding the mechanistic basis. The present application will enable us to study differences in trans-intestinal passage and toxicity of nanomedicines compared to conventional dissolved drugs.