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

胶原蛋白是人体中含量最多的结构蛋白，占人体质量的三分之一。它形成厘米长的原纤维。这种组织使富含胶原蛋白的组织具有不同的特性（例如，可拉伸皮肤中的扁平胶合板晶格和绳状肌腱中的平行束）。胶原蛋白的变化是许多与衰老相关的变化的基础，比如皮肤失去弹性、伤口愈合不良、纤维化、易骨折和骨关节炎。由于胶原蛋白维持失败，大多数人的生活质量都会下降。然而，尽管它具有基本的重要性，我们仍然不完全了解前体前胶原的合成，从细胞输出，胶原网络的组装和维护是如何被调节的。该项目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是由昼夜节律控制的（我们组织中的内部时钟，根据昼夜活动和光线的模式进行循环），并定义了免疫系统如何调节胶原蛋白基质在受伤时的修复。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，昼夜节律生物学，数学建模和新型合成支架来回答基质生物学中的主要问题。通过共同努力，共享工具、人员和专业知识，我们将比单打独斗产生更大的影响。我们有5个具体目标：使用细胞和斑马鱼来确定胶原蛋白的前体，称为前胶原蛋白，是如何通过细胞的。我们将定义高尔基体（细胞分泌的一切物质通过的中央分选站）中关键蛋白质机制的作用。2. 了解新生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，将胶原蛋白是如何产生的信息与它在体内的使用方式联系起来。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中进行测试。4. 测试胶原蛋白网络对损伤、损伤和衰老的反应，以及昼夜节律和免疫系统对其的影响。我们将在半透明的斑马鱼身上制造轻微的损伤，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如何重建。5. 我们将制作3D支架来模拟年老和年轻组织的表现。我们知道，随着年龄的增长，组织会变得更硬；使用定制的支架，我们可以测试年轻细胞对类似于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，使早期职业研究人员接触到最先进的技术和设备，并接受量身定制的培训，这将使他们在职业生涯中受益。合作为与行业、临床医生和更广泛的公众接触提供了更多的机会，以确保工作产生最大的影响。除了进一步加深我们对胶原基质如何组装和调节的理解外，该计划还将产生重要的新工具，这将使更广泛的学术界受益。其中包括能够可视化胶原蛋白如何在细胞中移动的新试剂，用于定义皮肤、肌腱和骨骼中胶原蛋白在发育期间和损伤后如何重构的新工具，以及可用于工业或临床的新型合成支架，以帮助修复主要皮肤伤口，或损伤后的肌腱和韧带修复。

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.