Epithelial stress sensors: novel roles for cytochrome P450s and organellar calcium in integrated stress and immune responses.

上皮应激传感器：细胞色素 P450 和细胞器钙在综合应激和免疫反应中的新作用。

• 批准号: BB/G020620/1
• 负责人: Shireen Davies
• 金额: $ 77.29万
• 依托单位: University of Glasgow
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2009
• 资助国家: 英国
• 起止时间: 2009 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=BB%2FG020620%2F1
• 关键词: Epithelial; stress; sensors; novel; roles

All cells produce reactive oxygen species (ROS) due to chemical reactions, which also occur in the organelles contained within cells. For example, the production of energy from the mitochondria and chemical reactions in the peroxisomes generate ROS. Cells and tissues deal with the production of ROS using different enzymes and processes, which 'neutralise' ROS. When there is an imbalance between the production of ROS and the ability of the cells or tissues in question to deal with the charged ions, 'oxidative stress' results. Cellular responses to oxidative stress are regulated by signal transduction mechanisms which utilise small signalling molecules including calcium ions (Ca2+). Oxidative stress affects cellular proteins, lipids and DNA, and in humans is associated with neurodegenrative disease, cardiovascular disease including stroke and heart attack, and with the ageing process. However, ROS production can also closely linked to immunity, as ROS can be used in defence against infection. It is increasingly known that mechanisms of oxidative stress and immune function are linked, and these are important areas of research for the basic fundamentals of biological mechanisms, and for human health and ageing. Invertebrate model organisms eg., Drosophila melanogaster, are central for investigating the impact of gene function in stress responses in the organism. Furthermore, studies in Drosophila have also have provided the clearest links between manipulation of stress-associated genes and physiological output. The importance of the tissue- or cell-specific context of stress-response genes in oragnismal responses to stress, makes Drosophila particularly useful, as cell- and tissue-specific targeting of genes in Drosophila is well-established. The Drosophila epithelial Malpighian (renal) tubule is equivalent to vertebrate kidney and liver and plays critical roles in detoxification, fluid and ion transport. Tubules are packed with mitochondria and peroxisomes, and produce ROS as part of their normal function. We have shown that tubules have specific adaptations to counter the high mitochondrial activity and production of ROS, including being enriched for 'anti-oxidant' genes which are also conserved in humans. We have also shown that de-regulation of tubule mitochondrial calcium signalling by excess ROS leads to tubule cell death (apoptosis) and death of the whole fly. Thus, this epithelial tissue has a central role in stress-sensing for the whole organism. The tubule is also a stand-alone immune-sensing tissue which uses 'innate' immune mechanisms to sense bacterial or fungal challenge. Such innate immune mechanisms in epithelia are also conserved in vertebrates, and constitute an increasingly important area of study, as it is now known that diseases including Crohns' Disease and asthma are due to deregulation of innate immune mechanisms in these fluid-transporting epithelial tissues (intestine, and lungs, respectively). Our recent data from survival studies in response to oxidative stress and immune challenge, work on calcium signals in mitochondria and analysis of the ~13500 genes of the fly in response to stress shows that key novel tubule-specific genes involved with the stress/immune responses the fly. These genes encode 'detoxification' proteins-the cytcochrome P450s, normally associated with mitochondria or peroxisomes and here, we have shown involvement with the stress response. We will investigate these genes in the context of calcium signals in the peroxisome and mitochondria, and assess the impact of each of these genes/proteins on survival of the whole organism using functional assays for novel genes, genetics/transgenics with calcium reporters targeted to mitochondria and peroxisomes in the intact tissue. This will allow us to uncover new ways of regulating stress responses in epithelial cells, which may impact on human biology.

所有的细胞都会产生活性氧（ROS），这是由于化学反应，也发生在细胞内的细胞器。例如，从线粒体产生能量和过氧化物酶体中的化学反应产生ROS。细胞和组织使用不同的酶和过程来处理ROS的产生，这些酶和过程“中和”ROS。当ROS的产生与细胞或组织处理带电离子的能力之间不平衡时，就会产生“氧化应激”。细胞对氧化应激的反应由信号转导机制调节，该机制利用包括钙离子（Ca 2+）的小信号分子。氧化应激影响细胞蛋白质、脂质和DNA，并且在人类中与神经变性疾病、心血管疾病（包括中风和心脏病发作）以及衰老过程相关。然而，ROS的产生也与免疫力密切相关，因为ROS可用于防御感染。人们越来越多地认识到，氧化应激和免疫功能的机制是相互关联的，这些都是生物机制的基本原理以及人类健康和衰老的重要研究领域。无脊椎动物模式生物，果蝇是研究基因功能对生物体应激反应影响的核心。此外，对果蝇的研究也提供了操纵压力相关基因和生理输出之间最清晰的联系。组织或细胞特异性环境的压力反应基因在oragnismal应激反应的重要性，使果蝇特别有用，作为细胞和组织特异性靶向基因在果蝇是公认的。果蝇上皮Malpighian（肾）小管相当于脊椎动物的肾脏和肝脏，在解毒，液体和离子运输中起着关键作用。小管中充满了线粒体和过氧化物酶体，并产生ROS作为其正常功能的一部分。我们已经表明，肾小管具有特定的适应性，以对抗高线粒体活性和ROS的产生，包括富含在人类中也是保守的“抗氧化剂”基因。我们还表明，过量的ROS对小管线粒体钙信号的失调导致小管细胞死亡（凋亡）和整个果蝇的死亡。因此，这种上皮组织在整个生物体的压力传感中起着核心作用。肾小管也是一个独立的免疫感应组织，它使用“先天”免疫机制来感知细菌或真菌的挑战。上皮细胞中的这种先天免疫机制在脊椎动物中也是保守的，并且构成了越来越重要的研究领域，因为现在已知包括克罗恩病和哮喘在内的疾病是由于这些流体运输上皮组织（分别为肠和肺）中先天免疫机制的失调。我们最近的数据来自对氧化应激和免疫挑战的生存研究，线粒体中钙信号的研究以及对果蝇对应激反应的~13500个基因的分析表明，关键的新的微管特异性基因参与了果蝇的应激/免疫反应。这些基因编码“解毒”蛋白-细胞色素P450，通常与线粒体或过氧化物酶体相关，在这里，我们已经表明参与应激反应。我们将研究这些基因的背景下的钙信号的过氧化物酶体和线粒体，并评估这些基因/蛋白质的每一个对整个生物体的生存使用功能检测新的基因，遗传学/转基因与钙报告靶向线粒体和过氧化物酶体在完整的组织。这将使我们能够发现调节上皮细胞应激反应的新方法，这可能会影响人类生物学。

项目成果

A biogenic amine and a neuropeptide act identically: tyramine signals through calcium in Drosophila tubule stellate cells.

• DOI: 10.1098/rspb.2012.2943
• 发表时间: 2013-04-22
• 期刊: Proceedings. Biological sciences
• 作者: Cabrero P;Richmond L;Nitabach M;Davies SA;Dow JA
• 通讯作者: Dow JA

The nitric oxide-cyclic GMP pathway regulates FoxO and alters dopaminergic neuron survival in Drosophila.

• DOI: 10.1371/journal.pone.0030958
• 发表时间: 2012
• 期刊: PloS one
• 影响因子: 3.7
• 作者: Kanao T;Sawada T;Davies SA;Ichinose H;Hasegawa K;Takahashi R;Hattori N;Imai Y
• 通讯作者: Imai Y

Modulation of epithelial innate immunity by autocrine production of nitric oxide

• DOI: 10.1016/j.ygcen.2008.09.012
• 发表时间: 2009-05-15
• 期刊: GENERAL AND COMPARATIVE ENDOCRINOLOGY
• 影响因子: 2.7
• 作者: Davies, Shireen-Anne;Dow, Julian A. T.
• 通讯作者: Dow, Julian A. T.