Defining mechanisms of mucosal innate defence using the X. tropicalis tadpole

使用热带蝌蚪定义粘膜先天防御机制

• 批准号: BB/M021688/1
• 负责人: David Thornton
• 金额: $ 55.21万
• 依托单位: University of Manchester
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2015
• 资助国家: 英国
• 起止时间: 2015 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=BB%2FM021688%2F1
• 关键词: Defining; mechanisms; mucosal; innate; defence

Pathogens are disease causing microorganisms and viruses, which represent a severe challenge to the health and survival of animals. When animals succumb to infectious disease, it is because pathogens have breached the body's natural defence mechanisms. Broadly, these defences fall into two types, innate and adaptive immune defences. Adaptive immunity is a specific defence, where antibodies generated against a particular pathogen are used to rapidly respond to infection. However, it is advantageous for animals if pathogens never have the opportunity to infect the body in the first place. Innate immunity (non-specific immunity) is the first line of defence against infection and includes the generation of physical barriers to entry and the production of molecules that actively destroy pathogens. One such physical barrier used by exposed tissues in the body (e.g. lungs and gut) is the production of a mucus layer. Mucus traps pathogens preventing them from accessing the underlying cells and also contains other innate defence molecules such as antibacterial agents. It is important to study mucus to understand how it functions and what happens when it becomes dysfunctional, and is breached. Severe diseases can arise due to lack of a properly functioning mucus barrier; these include inflammatory bowel disease, stomach ulcers, and asthma. Mammalian models, including mice, are often used to study mucus in the lungs and gut. However, simple non-mammalian organisms can also provide valuable insight into evolutionarily conserved mechanisms of mucus function. In this proposal, we introduce the frog, Xenopus tropicalis, and specifically the mucus on the surface of the tadpole stage, as a model to study innate defence.X. tropicalis is a model organism commonly used in developmental biology to understand basic concepts in how embryos develop and the molecular processes that occur. In recent years, we have been studying how the embryonic skin changes as the embryo develops into a tadpole, prior to becoming a frog. We have identified a number of cell types in the tadpole skin, including two types of secretory cells that together secrete molecules that form a mucus layer over the surface of the skin. We are proposing that this simple model can be used to study aspects of mucosal innate defence that will be applicable to other organisms, including humans. Indeed, we have identified a number of innate defence molecules in the tadpole mucus that have known homologues in humans and yet their functions are largely unexplored. In this grant proposal, we intend to interrogate their functions through a number of means. The X. tropicalis tadpole model has some distinct advantages in that the skin directly faces the environment and so conditions can be altered to observe the effects. This is not the case with mammalian mucus barriers, which are usually found on tissues within the body and are thus more difficult to access. In addition, X. tropicalis tadpoles have yet to acquire adaptive immunity (which happens later in their development) so any observed effects will be due to innate defence mechanisms, rather than a more complicated, combined adaptive response to infection. We intend to genetically alter the expression of the innate defence molecules in the tadpoles skin mucus layer and then challenge the tadpoles with a potential pathogen found in their native environment (the bacterium, Aeromonas hydrophila), in order to understand the importance of each molecule. We will look at their structural roles in the physical barrier and their potential functional roles as anti-microbial agents. We aim to advance the understanding of how the tadpole defends itself against infection and ultimately how evolutionarily conserved innate defence mechanisms function in mucus barriers. This could potentially lead to new targets for treating disease.

病原体是引起疾病的微生物和病毒，对动物的健康和生存构成严重挑战。当动物死于传染病时，这是因为病原体破坏了身体的自然防御机制。广义上，这些防御分为两种类型，先天性和适应性免疫防御。适应性免疫是一种特异性防御，其中针对特定病原体产生的抗体用于快速响应感染。然而，如果病原体从一开始就没有机会感染身体，这对动物是有利的。先天免疫（非特异性免疫）是抵抗感染的第一道防线，包括产生进入的物理屏障和产生积极破坏病原体的分子。身体中暴露的组织（例如肺和肠道）使用的一种这样的物理屏障是粘液层的产生。粘液捕获病原体，防止它们进入底层细胞，还含有其他先天防御分子，如抗菌剂。重要的是要研究粘液，以了解它是如何发挥作用的，以及当它变得功能失调和被破坏时会发生什么。严重的疾病可能会出现由于缺乏正常运作的粘液屏障;这些包括炎症性肠病，胃溃疡和哮喘。包括小鼠在内的哺乳动物模型通常用于研究肺和肠道中的粘液。然而，简单的非哺乳动物生物体也可以提供有价值的见解粘液功能的进化保守机制。在这个提议中，我们引入了青蛙，热带爪蟾，特别是蝌蚪阶段表面的粘液，作为研究先天防御的模型。tropicalis是发育生物学中常用的模式生物，用于了解胚胎发育的基本概念和发生的分子过程。近年来，我们一直在研究胚胎皮肤如何在胚胎发育成蝌蚪时发生变化，然后才成为青蛙。我们已经在蝌蚪皮肤中鉴定了许多细胞类型，包括两种类型的分泌细胞，它们共同分泌分子，在皮肤表面形成粘液层。我们建议，这个简单的模型可用于研究粘膜先天防御方面，将适用于其他生物，包括人类。事实上，我们已经在蝌蚪粘液中发现了一些先天防御分子，这些分子在人类中有已知的同源物，但它们的功能在很大程度上尚未被探索。在这项拨款建议中，我们打算通过一些手段来询问它们的功能。副艇长Tropicalis蝌蚪模型具有一些明显的优点，因为皮肤直接面对环境，因此可以改变条件以观察效果。哺乳动物的粘液屏障则不是这种情况，它们通常存在于体内的组织上，因此更难以进入。此外，X. tropicalis蝌蚪尚未获得适应性免疫（这发生在它们发育的后期），因此任何观察到的影响都将是由于先天防御机制，而不是对感染的更复杂的组合适应性反应。我们打算从基因上改变蝌蚪皮肤粘液层中先天防御分子的表达，然后用在其原生环境中发现的潜在病原体（细菌，Aeromonaserma）挑战蝌蚪，以了解每个分子的重要性。我们将研究它们在物理屏障中的结构作用以及它们作为抗微生物剂的潜在功能作用。我们的目标是促进了解蝌蚪如何防御感染，并最终如何进化保守的先天防御机制在粘液屏障的功能。这可能会导致治疗疾病的新靶点。

项目成果

Functional characterization of the mucus barrier on the Xenopus tropicalis skin surface.

• DOI: 10.1073/pnas.1713539115
• 发表时间: 2018-01-23
• 期刊: Proceedings of the National Academy of Sciences of the United States of America
• 影响因子: 11.1
• 作者: Dubaissi E;Rousseau K;Hughes GW;Ridley C;Grencis RK;Roberts IS;Thornton DJ
• 通讯作者: Thornton DJ