Three-dimensional analysis of the patterns of growth and mechanisms of evading therapy of pancreatic cancer in whole organs and thick slabs of tissue using active and passive SHANEL technology

胰腺癌是最致命的癌症之一。为了改善胰腺癌的治疗，研究必须基于最新的可用方法。这是改善患者长期预后的唯一途径。在本申请中，将使用称为SHANEL的新型显微镜方法研究胰腺癌的三维生长模式。SHANEL代表“小胶束介导的人体器官有效清除和标记”。与早期的方法不同，SHANEL用于检查整体肿瘤。这具有可以同时分析肿瘤的几个区域的优点。通过这种方式，可以评估肿瘤细胞与其微环境的关系。这是第一次，现在可以检查肿瘤生长到血管并导致转移的部分。此外，临床上经常观察到的化疗耐药性的潜在机制也可以进行研究。约翰霍普金斯大学的Wood博士和Hruban博士的研究小组计划进行这项研究，他们已经对较小的肿瘤部分进行了三维显微镜分析。在该小组现有专业知识的帮助下，SHANEL研究现在将扩展到胰腺癌。因此，该研究项目基于三个主要目标。在第一步中，SHANEL技术将适应非常坚硬和结缔组织丰富的胰腺癌的特殊要求。第二个主要目标是分析人类胰腺癌细胞在组织中相对于其他结构（如神经、血管和胰管）的3D生长。主要的重点是评估是否有一个定向的，非随机的增长与无定向的，随机的增长。背景是对伍德博士和赫鲁班博士周围的群体的观察，这些群体假设有定向增长。第三个主要目的是研究接受新辅助化疗（即手术前化疗）的胰腺癌的形态学变化。用新辅助化疗治疗的胰腺癌的三维显微镜将显示存活的癌细胞的位置。将研究是否存在保护肿瘤细胞的解剖学小生境。最近的数据表明，癌细胞已经侵入正常的胰腺导管系统的化疗反应不如那些位于stroma.In总结，目前的研究项目提出，以确定整个胰腺肿瘤的高分辨率三维显微镜的转移和耐药的治疗机制。从长远来看，这一结果可以提高对胰腺癌的认识。

Pancreatic cancer is one of the most deadly cancers of all. In order to improve the treatment of pancreatic cancer, research must be based on the latest available methods. This is the only way to improve the prognosis of affected patients in the long term. In the present application, the three-dimensional growth pattern of pancreatic cancer is to be investigated using a novel microscopic method called SHANEL. SHANEL stands for "small-micelle-mediated human organ efficient clearing and labelling". In contrast to earlier methods, SHANEL is used to examine whole tumours in one piece. This has the advantage that several regions of the tumour can be analysed simultaneously. In this way, the relationship of the tumor cells to their microenvironment can be assessed. For the first time, it is now possible to examine parts of the tumour that grow into vessels and lead to metastases. In addition, mechanisms underlying the clinically frequently observed resistance to chemotherapy can be investigated.The proposed investigations are planned in the group of Dr. Wood and Dr. Hruban at Johns-Hopkins University, who have already carried out three-dimensional microscopic analyses of smaller tumor parts. With the help of the existing expertise of this group, the SHANEL investigation will now be extended to pancreatic cancer.The research project is therefore based on three main objectives. In the first step, the SHANEL technology will be adapted to the special requirements of the very hard and connective tissue-rich pancreatic cancer. The second main goal is the analysis of the 3D growth of human pancreatic cancer cells in tissue in relation to other structures such as nerves, vessels and pancreatic ducts. The main focus is to evaluate whether there is a directed, non-random growth versus an undirected, random growth. Background are observations of the group around Dr. Wood and Dr. Hruban, which assume a directed growth. The third main objective is to investigate the morphological changes in pancreatic carcinomas that have undergone neoadjuvant chemotherapy, i.e. chemotherapy that precedes surgery. Three-dimensional microscopy of pancreatic carcinomas treated with neoadjuvant chemotherapy will show where the surviving cancer cells are located. It will be investigated whether there are anatomical niches that protect tumour cells. Recent data from the group suggest that cancer cells that have invaded the normal pancreatic duct system respond less well to chemotherapy than those located in the stroma.In summary, the present research project proposes to identify mechanisms of metastasis and resistance to therapy by high-resolution three-dimensional microscopy of whole pancreatic tumors. The results could improve the understanding of pancreatic carcinoma in the long term.