Biomechanical characterisation of joints in osteoarthritis mutant zebrafish; studying interactions between genotype and biomechanics in osteoarthritis

骨关节炎突变斑马鱼关节的生物力学特征；

• 批准号: MR/L002566/1
• 负责人: Chrissy Hammond
• 金额: $ 51.83万
• 依托单位: University of Bristol
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2014
• 资助国家: 英国
• 起止时间: 2014 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=MR%2FL002566%2F1
• 关键词: Biomechanical; characterisation; joints; osteoarthritis; mutant

The degenerative joint condition osteoarthritis (OA) affects tens of millions of people worldwide. Although a number of genes have recently been identified that increase susceptibility to osteoarthritis, it currently remains unclear how many of these genes lead to pathogenic changes to the joint. We do, however, understand that joint shape and the subsequent effects of shape on loading and distribution of strain in the joint affects the pathogenesis of OA. Our group has identified a zebrafish line which carries a mutation in a gene identified as increasing susceptibility to OA in humans (CHST11). We have preliminary evidence showing that zebrafish carrying this mutant gene have altered joint shape; the joints have a more flattened profile that doesn't form the 'ball and cup' shape associated with a normal joint. This change to the shape appears to be progressive, ultimately this shape change leads to joint failure and death of the fish at around 2 weeks of age as they can no longer open their jaws to feed.This project uses experiments in zebrafish, combined with computational modelling to establish how the developing cartilages of the jaw respond to strains, allowing us to study the relationship between OA genes, joint shape and joint function. We will use state of the art high resolution microscopy with materials testing to determine the differences in stiffness between different parts of the jaw tissues, and whether there are material property differences between the normal and mutant fish. We will use microscopy to visualise the zebrafish jaw musculoskeletal system, which is comprised of muscle, cartilage, bone and connective tissue. From the images that we generate we will develop 3-dimensional computational models to visualise which parts of the jaw cartilages are under stress and strain in normal (wild type) fish, compared to 'mutant' fish carrying the OA (CHST11) gene. We predict that the change in shape of the joints in mutant fish changes how the developing cartilages experience stress and strain. We will also determine the morphology of jaw tissues from mutant fish that have their jaw muscles immobilised, this will allow us to test how muscle activity influences explore the influence of muscle loads on determinin joint shape. We will then, using this biomechanical data, study the effect of changes to the biomechanical environment on the cartilage cells (called chondrocytes) testing whether changes to strain can predict the changes in behaviour that these cells exhibit. For example, are cells under higher strains more or less likely to divide, to mature or to undergo death by a process called apoptosis? Using lines of zebrafish that express fluorescent proteins when various collagen genes are switched on, we will test whether the cells under the highest strain change the types of cartilage matrix which they secrete. Finally, we will use the models along with data that we have accumulated about the activity of a major signalling pathway, known as the wnt pathway which controls cartilage cell behaviour. We will use this to predict how wnt-signalling is involved in mediating the cell's response to the change in joint shape, which we can then test in the mutant fish. This will help us start to understand how the changes in mechanical strain are interpreted by the cell in a way that leads to a change in cell behaviour. This research is highly interdisciplinary in nature, therefore researchers from a variety of disciplines will benefit including anatomists, biomechanists, evolutionary and developmental biologists, cell biologists and biomedical engineers. The results will be of particular relevance to the study of OA at all levels, from the genetics underpinning the disease to the development of orthopaedic implants and replacement joints. There will be benefits to the UK science base through multidisciplinary training of young scientists and through international collaborations.

退行性关节疾病骨关节炎（OA）影响着全世界数千万人。虽然最近已经确定了一些基因增加骨关节炎的易感性，但目前仍不清楚这些基因中有多少导致关节的致病性变化。然而，我们确实了解关节形状以及形状对关节负荷和应变分布的后续影响会影响OA的发病机制。我们的研究小组已经确定了一种斑马鱼品系，该品系携带一种基因突变，该基因被确定为人类对OA的易感性增加（CHST 11）。我们有初步证据表明，携带这种突变基因的斑马鱼改变了关节形状;关节具有更扁平的轮廓，不会形成与正常关节相关的“球和杯”形状。这种形状的变化似乎是渐进的，最终这种形状的变化导致关节衰竭，并在大约2周龄时死亡，因为它们不再能张开下巴进食。该项目使用斑马鱼的实验，结合计算模型来建立发育中的颌骨软骨如何对应变做出反应，使我们能够研究OA基因之间的关系，关节形态和关节功能。我们将使用最先进的高分辨率显微镜和材料测试来确定颌组织不同部分之间的刚度差异，以及正常鱼和突变鱼之间是否存在材料特性差异。我们将使用显微镜来观察斑马鱼的下颌肌肉骨骼系统，该系统由肌肉、软骨、骨骼和结缔组织组成。从我们生成的图像中，我们将开发三维计算模型，以可视化正常（野生型）鱼与携带OA（CHST 11）基因的“突变”鱼相比，颌软骨的哪些部分处于应力和应变下。我们预测，突变鱼类关节形状的变化会改变发育中软骨的应力和应变。我们还将确定突变鱼的颌骨组织的形态，这些鱼的颌骨肌肉固定不动，这将使我们能够测试肌肉活动如何影响关节形状的确定。然后，我们将使用这些生物力学数据，研究生物力学环境变化对软骨细胞（称为软骨细胞）的影响，测试应变的变化是否可以预测这些细胞表现出的行为变化。例如，在较高的压力下，细胞是否更有可能分裂、成熟或通过称为细胞凋亡的过程而死亡？使用当各种胶原蛋白基因被打开时表达荧光蛋白的斑马鱼品系，我们将测试最高应变下的细胞是否改变它们分泌的软骨基质的类型。最后，我们将使用沿着数据的模型，我们已经积累了关于一个主要的信号通路的活动，被称为wnt通路控制软骨细胞的行为。我们将利用这一点来预测wnt信号是如何参与调节细胞对关节形状变化的反应的，然后我们可以在突变鱼中进行测试。这将帮助我们开始了解细胞如何解释机械应变的变化，从而导致细胞行为的变化。这项研究本质上是高度跨学科的，因此来自不同学科的研究人员将受益，包括解剖学家，生物力学家，进化和发育生物学家，细胞生物学家和生物医学工程师。这些结果将与OA的各个层面的研究特别相关，从支持疾病的遗传学到骨科植入物和关节置换的发展。通过对年轻科学家的多学科培训和国际合作，英国的科学基础将从中受益。

项目成果

The mechanical impact of col11a2 loss on joints; col11a2 mutant zebrafish show changes to joint development and function, which leads to early onset osteoarthritis

col11a2损失对关节的机械影响；

• DOI: 10.1101/302307
• 发表时间: 2018
• 作者: Lawrence E

Finite element modelling predicts changes in joint shape and cell behaviour due to loss of muscle strain in jaw development.

• DOI: 10.1016/j.jbiomech.2015.07.017
• 发表时间: 2015-09-18
• 期刊: Journal of biomechanics
• 影响因子: 2.4
• 作者: Brunt LH;Norton JL;Bright JA;Rayfield EJ;Hammond CL
• 通讯作者: Hammond CL

A zebrafish model of developmental joint dysplasia: Manipulating the larval mechanical environment to drive the malformation and recovery of joint shape

• DOI: 10.1101/155911
• 发表时间: 2017-06
• 期刊: bioRxiv
• 作者: K. Roddy;Roderick E. H. Skinner;Lucy H. Brunt;E. Kague;Stephen J. Cross;E. Rayfield;C. Hammond

Differential effects of altered patterns of movement and strain on joint cell behaviour and skeletal morphogenesis.

• DOI: 10.1016/j.joca.2016.06.015
• 发表时间: 2016-11
• 期刊: OSTEOARTHRITIS AND CARTILAGE
• 影响因子: 7
• 作者: Brunt, L. H.;Skinner, R. E. H.;Roddy, K. A.;Araujo, N. M.;Rayfield, E. J.;Hammond, C. L.
• 通讯作者: Hammond, C. L.

The role of muscle activity on zebrafish jaw joint shape and cell behaviour

肌肉活动对斑马鱼颌关节形状和细胞行为的作用

• 发表时间: 2015
• 作者: Brunt LH