Opportunities to modulate extracellular matrix secretion and assembly for long term health

胶原蛋白是人体内含量最丰富的结构蛋白，占我们体重的三分之一。它形成厘米长的纤维。这种组织赋予富含胶原蛋白的组织不同的特性(例如，可伸展皮肤上的扁平胶合板格子和绳状肌腱中的平行束)。胶原蛋白的变化是我们与衰老相关的许多变化的基础，例如皮肤弹性丧失、伤口愈合不良、纤维化、骨折和骨关节炎的易感性。由于胶原蛋白的维护失败，大多数人的生活质量都会下降。然而，尽管它的基本重要性，我们仍然不完全了解如何合成前体前胶原，从细胞输出，组装和维持胶原蛋白网络。该计划汇集了曼彻斯特大学和布里斯托尔大学的研究人员，他们在胶原蛋白生物学的关键方面具有互补的专业知识。我们已经发现了胶原蛋白分泌的新机制，表明胶原蛋白基质的分泌和组装受到昼夜节律(我们组织中的内部时钟对昼夜活动和光线模式的反应)的控制，并定义了免疫系统如何调节创伤时胶原蛋白基质的修复。现在，我们希望利用我们的多学科技能，包括细胞和组织生物学的基本方面，使用体外和体内模型的综合实验，昼夜生物学，数学建模，以及新型合成支架来回答基质生物学中的主要问题。共同努力，共享工具、人员和专业知识，我们将能够产生比单独行动更大的影响。我们有5个具体目标：1.使用细胞和斑马鱼来确定被称为前胶原的胶原前体如何通过细胞。我们将确定高尔基体(细胞分泌的一切东西都通过的中央分拣站)中关键蛋白质机械的作用。2.了解新合成的胶原蛋白的运输是如何在空间和时间上协调的。我们将确定昼夜节律如何调节胶原蛋白的形成、保持模式和输出。3.推导出一个数学框架，将胶原蛋白的合成信息与其在体内的使用情况联系起来。这将使我们能够预测制造和组装胶原蛋白的途径的任何部分的变化如何影响其他成分，我们可以在细胞或动物身上进行测试。4.测试胶原蛋白网络对损伤和老化的反应，并测试昼夜节律和我们的免疫系统对此有何影响。我们将对半透明的斑马鱼进行轻微损伤，并使用荧光标记的胶原蛋白来观察细胞如何反应以及胶原蛋白网络是如何重建的。5.我们将生产3D支架，模拟老组织和年轻组织的表现。我们知道，随着年龄的增长，组织会变得更加坚硬；使用定制的支架，我们可以测试年轻细胞对类似旧组织的支架的反应，反之亦然。该项目还将培训下一代科学家，让职业早期的研究人员接触到最先进的技术和设备，并接受对他们的职业生涯有利的量身定制的培训。合作提供了与业界、临床医生和更广泛的公众接触的更多机会，以确保工作产生最大影响。除了进一步了解胶原蛋白基质是如何组装和调节的，该计划还将产生重要的新工具，使更广泛的学术界受益。这些包括能够可视化胶原如何在细胞中移动的新试剂，定义皮肤、肌腱和骨骼中的胶原在发育过程中和受伤后如何重塑的新工具，以及可在工业或临床上用于帮助修复主要皮肤伤口或受伤后肌腱和韧带修复的新合成支架。

Collagen is the most abundant structural protein in the body, making up 1/3 of our mass. It is formed into centimetre-long fibrils. This organisation gives collagen-rich tissues their differing properties (e.g. flat plywood lattices in stretchable skin and parallel bundles in rope-like tendons). Changes to collagen underpin many of the changes we associate with ageing, such as loss of skin elasticity, poor wound healing, fibrosis, susceptibility to fracture and osteoarthritis. Most people will experience reduced quality of life due to a failure of collagen maintenance. Yet, despite its fundamental importance, we still do not fully understand how synthesis of the precursor procollagen, export from the cell, assembly and maintenance of the collagen network are regulated. This programme brings together researchers from the Universities of Manchester and Bristol, with complementary expertise in key aspects of collagen biology. We have discovered new mechanisms of collagen secretion, shown that secretion and assembly of the collagen matrix is controlled by the circadian rhythm (the internal clocks in our tissues that cycle in response to day and night patterns of activity and light), and defined how the immune system modulates the repair of a collagen matrix on wounding. Now, we wish to exploit our multidisciplinary skills that include fundamental aspects of cell and tissue biology, integrated experiments using in vitro and in vivo models, circadian biology, mathematical modelling, and novel synthetic scaffolds to answer major questions in matrix biology. Working together, sharing tools, personnel, and expertise we will be able to make more impact than we could individually. We have 5 specific aims: 1. Use cells and zebrafish to determine how the precursor of collagen, called procollagen, passes through the cell. We will define the role of key protein machineries in the Golgi apparatus (the central sorting station through which everything that is secreted by cells passes). 2. Understand how transport of newly-made collagen is coordinated in space and time. We will determine how the circadian rhythm regulates the formation, holding pattern, and export of collagen. 3. Derive a mathematical framework that links information on how collagen is made to how it is used in the body. This will allow us to predict how changes to any part of the pathway that makes and assembles collagen affects other components, which we can test in cells or animals. 4. Test how the collagen network responds to damage, in injury and ageing, and test how day/night rhythm and our immune system influence this. We will make minor injuries into translucent zebrafish and use fluorescently-labelled collagen to watch how cells respond and how the collagen network is rebuilt. 5. We will produce 3D scaffolds that mimic how old and young tissues perform. We know that tissues become stiffer as we age; using custom built scaffolds we can test how young cells respond to scaffolds that resemble old tissue and vice versa.This project will also train the next generation of scientists, exposing the early-career researchers to state-of-the-art technology and equipment and to tailored training that will benefit them in their careers. Working together offers enhanced opportunities to engage with industry, clinicians and the wider public to ensure the work has the maximum impact. As well as furthering our understanding of how the collagen matrix is assembled and regulated, the programme will generate significant new tools that will benefit the wider academic community. These include new reagents that will enable visualisation of how collagen moves through the cell, new tools to define how remodelling of collagen in skin, tendon and bone occurs during development and following injury, and new synthetic scaffolds that could be used industrially or clinically to help in repair of major skin wounds, or tendon and ligament repair following injury.