Cellular homeostasis in response to transition metals associated with shipping emissions

空气中的颗粒物（PM）每年与全球近900万人过早死亡有关。PM由其在空气中的浓度来调节，但这忽略了其成分，其成分因来源而异，并且被认为是其健康影响的重要决定因素。来自船舶的PM是港口和沿海地区空气污染的一部分，但很少有人研究。船舶经常燃烧重油作为燃料，产生PM，我已经证明，与其他来源的PM相比，PM中钒，镍和钴的浓度增加。这些金属尤其存在于超细颗粒（UFPM）中，直径小于100纳米，小到足以到达肺部深处的气囊（肺泡）。肺泡允许吸入的氧气进入血液，因此保持健康的肺泡至关重要。肺泡衬里细胞，称为上皮细胞，可以对损伤做出反应，而引起损伤的物质，如细菌或颗粒，可以被称为巨噬细胞的细胞包裹和消化。然而，人们对这些细胞在这些不同金属存在下是否正常工作知之甚少。PM暴露是我们一生中健康状况不佳的主要危险因素，因此了解我们的肺部如何保持健康是非常重要的。我将开始研究钒、镍和钴如何影响上皮细胞和巨噬细胞执行其功能的能力，并将其效果与在忙碌航运区附近收集的UFPM进行比较。然后，我将研究对PM已知反应的影响，包括激活对金属的防御和释放化学信息以协调细胞内和细胞之间的反应。我将使用一种称为转录组学的技术，它可以提供有关细胞内所有信号的信息，这些信号在对这些金属的反应中发生了变化，这意味着可以发现以前未记录的反应。因此，该项目的第一部分将重点关注暴露的电池如何受到金属的影响。接下来，我将重点介绍细胞如何处理金属，目的是保持最佳功能状态，这一过程称为稳态。我将使用一种称为电感耦合等离子体质谱（ICP-MS）的技术来研究这些金属进入并保留在暴露细胞内的程度，以及细胞对控制这些金属浓度的反应是否也会影响其他重要金属的浓度，包括铁，铜和锌。我将使用荧光染料来观察细胞内这些金属浓度的变化，然后研究细胞产生蛋白质的速度，这些蛋白质使金属进入，保留和排泄，以了解细胞如何调节它们所含的这些金属的量。接下来，我将使用巨噬细胞和上皮细胞以及血管细胞的培养物，代表整个肺泡，然后是整个肺组织的薄片，暴露于这些金属，以研究单个细胞的转录组学变化，显示细胞与细胞之间的反应差异。然后，我将使用最先进的分析化学技术来研究单个细胞中的金属浓度，看看一些与船舶相关的金属是否被不同的细胞吸收得更多，以及这是否会影响对细胞正常功能至关重要的其他金属的吸收。最后，我将把这两组信息结合起来，以了解细胞内一系列金属的浓度与同一细胞内激活和失活过程之间的关系。这个项目将提高对空气中PM中这些金属如何影响我们肺部的理解。反过来，这将表明是否应该进一步研究其中的一些金属，以更好地为保护公众健康的政策提供信息。这可能会导致限制这些金属在燃料中的浓度，或对船舶在港口使用发动机的规定。

Airborne particulate matter (PM), is associated with almost 9 million premature deaths per year worldwide. PM is regulated by its concentration in the air, but this neglects its composition, which varies depending on source, and is thought to be an important determinant of its health effects. PM from ships contributes to air pollution in port and coastal areas but is rarely studied. Ships often burn heavy oil as fuel, producing PM which I have shown to contain increased concentrations of vanadium, nickel, and cobalt, compared to PM from other sources. These metals are found especially in ultrafine PM (UFPM), less than 100 nanometres in diameter and small enough to reach the air sacs (alveoli) deep in the lungs. Alveoli allow inhaled oxygen to enter the blood, so maintaining healthy alveoli is critical. Alveolar lining cells, called epithelial cells, can heal in response to damage, while damage-causing agents such as bacteria or particles can be enveloped and digested by cells called macrophages. However, little is known about whether these cells function normally in the presence of these different metals. PM exposure is a leading risk factor for ill health over our lives, so understanding how our lungs can stay healthy is of great importance.I will begin by studying how vanadium, nickel, and cobalt affect the ability of the epithelial cells and macrophages to perform their functions, comparing effects to UFPM collected near a busy shipping area. I will then examine effects on known responses to PM, including activation of defences against metals and release of chemical messages to coordinate responses within and between cells. I will use a technique called transcriptomics, which allows gives information about all the signals within a cell which are altered in response to these metals, meaning previously undocumented responses can be discovered. Thus, these first parts of the project will focus on how exposed cells are affected by the metals. Next I will focus on how the metals are dealt with by the cells, which aim to maintain a state of optimal functioning, a process called homeostasis. I will use a technique called inductively coupled plasma mass spectrometry (ICP-MS) to study the extent to which these metals enter, and are retained within, exposed cells, and whether responses of cells to keep concentrations of these metals under control might also affect concentrations of other vital metals, including iron, copper, and zinc. I will use fluorescent dyes to visualise the changes in concentrations of these metals within cells, and then study the rate at which the cell produces proteins which enable entry, retention, and excretion of the metals, to understand how cells regulate the amount of these metals which they contain.I will next use cultures of macrophages and epithelial cells together with blood vessel cells, representing the whole alveolus, and then thin slices of whole lung tissue, exposed to these metals, to study transcriptomic changes in individual cells, showing cell-to-cell variation in responses. I will then use state-of-the-art analytical chemistry techniques to study concentrations of metals in individual cells, to see whether some of the ship-associated metals are taken up more by different cells, and whether this affects uptake of other metals critical to the normal functioning of the cells. Finally, I will match the two sets of information together, to understand the relationship between concentrations of a range of metals within cells and the processes activated and deactivated within the same cells.This project will improve understanding of how these metals in airborne PM might affect our lungs. In turn, this will suggest whether some of these metals should be further studied, to better inform policy to protect the health of the public. This might lead to restrictions in the concentrations of these metals in fuels, or regulations regarding ships' use of engines in port.