Development of human ex vivo bone-tumour niche for the study of cancer bone disease

骨骼是肿瘤转移的首选部位，通常影响年轻人的原发性骨肿瘤（如骨肉瘤）的发病率和管理在十多年内保持不变。此外，肿瘤扩散到骨骼往往代表疾病的最后阶段。因此，有一个大的研究领域，旨在减少骨骼内肿瘤的扩散和发展，并涉及广泛的动物模型。这些模型包括将肿瘤注射到远处的部位（例如心脏）并使其扩散到骨骼，或者将肿瘤细胞直接植入骨骼部位（例如胫骨）。此外，人类肿瘤细胞通常用于这种小鼠模型中，并且其中使用缺乏有效免疫系统的动物，并且其不完全屈服于在患有癌症-骨疾病的患者中观察到的人骨-人肿瘤微环境。该提案旨在解决这一问题，并通过开发一种新的离体模型来减少/取代将动物用作此类研究的一线工具，在该模型中，在无菌、伦理和人体组织法批准的程序下分离人骨，并在培养物中与人肿瘤细胞一起定殖。将评估骨破坏和肿瘤生长的程度，如用于从癌症-骨病小鼠模型解剖的骨。这将包括使用计算机生成的骨骼和组织学的3D图像，并存储组织培养基以供将来分析或骨和肿瘤衍生因子。此外，该提案将重复使用我们组内先前进行的体内研究的数据，以直接与使用该离体试验生成的数据进行比较，其中包括抗癌疗法的检测。

The skeleton is the preferential site for tumour metastases and the incidence and management of primary bone tumours (e.g. osteosarcoma) typically affecting young adults, have remained unchanged for over a decade. Moreover, the spread of tumours to the skeleton often represents the final stages of disease. Consequently, there is a large field of research aimed at reducing the spread and development of tumours within the skeleton, and which involve a wide range of animal models. These include models where tumours are injected into distant sits (e.g. heart) and allowed to spread to bone, or where tumour cells are implanted directly into skeletal sites (e.g. tibia). In addition, human tumour cells are often used in such mouse models, and where animals lacking an efficient immune system are used, and which do not fully capitulate the human bone-human tumour microenvironment seen in patients with cancer-bone disease. This proposal aims to address this issue and reduce/replace the use of animals as a first line tool in such studies, through the development of a new ex vivo model where human bone is isolated under sterile, ethical- and human tissue act-approved procedures, and colonised in culture with human tumour cells. The extent of bone destruction and tumour growth will be assessed as is used for bones dissected from mouse models of cancer-bone disease. This will include using computer-generated 3D images of the bone and histology, with tissue culture media stored for future analysis or bone- and tumour-derived factors. In addition, this proposal will re-use data from previous in vivo studies conducted within our group to compare directly against data generated using this ex vivo assay, and which will include the testing of anti-cancer therapies.