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受其在空气中的浓度控制，但这忽略了其组成，其组成因来源而异，被认为是其对健康影响的重要决定因素。船舶PM会造成港口和沿海地区的空气污染，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含有更高浓度的钒、镍和钴。这些金属尤其存在于超细颗粒物(UFPM)中，直径不到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是至关重要的。肺泡衬里细胞，称为上皮细胞，可以在损伤后愈合，而导致损伤的因素，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，关于这些细胞在这些不同金属存在下是否正常工作，人们知之甚少。PM暴露是我们一生中健康不佳的主要风险因素，因此了解我们的肺部如何保持健康非常重要。我将首先研究钒、镍和钴如何影响上皮细胞和巨噬细胞执行其功能的能力，并将其与在繁忙航运地区收集的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。我将使用一种名为转录组学的技术，它可以给出细胞内所有信号的信息，这些信号会因这些金属而改变，这意味着以前没有记录的反应可以被发现。因此，该项目的这些第一部分将集中在暴露的电池如何受到金属的影响。接下来，我将重点介绍细胞是如何处理金属的，细胞的目标是保持最佳功能状态，这一过程被称为体内平衡。我将使用一种名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。我将使用荧光染料将这些金属在细胞内浓度的变化可视化，然后研究细胞产生使金属进入、保留和排泄的蛋白质的速率，以了解细胞如何调节它们所含金属的数量。接下来，我将使用代表整个肺泡的巨噬细胞和上皮细胞以及血管细胞的培养，然后使用整个肺组织的薄片，暴露于这些金属，研究单个细胞的转录变化，显示细胞对细胞的反应差异。然后，我将使用最先进的分析化学技术来研究单个细胞中金属的浓度，看看与船舶相关的一些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运作至关重要的其他金属的吸收。最后，我将把这两组信息配对，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的过程之间的关系。这个项目将提高对空气中PM中这些金属可能如何影响我们肺部的理解。反过来，这将表明是否应该对其中一些金属进行进一步研究，以更好地为保护公众健康的政策提供信息。这可能会导致对燃料中这些金属浓度的限制，或者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.