Generation and initial characterization of novel tumour organoid models to study human pancreatic cancer-induced cachexia

Background The majority of patients with pancreatic cancer develops cachexia. The mechanisms underlying cancer cachexia development and progression remain elusive, although tumour-derived factors are considered to play a major role. Pancreatic tumour organoids arein vitrothree-dimensional organ-like structures that retain many pathophysiological characteristics of thein vivotumour. We aimed to establish a pancreatic tumour organoid biobank from well-phenotyped cachectic and non-cachectic patients to enable identification of tumour-derived factors driving cancer cachexia. Methods Organoids were generated from tumour tissue of eight pancreatic cancer patients. A comprehensive pre-operative patient assessment of cachexia-related parameters including nutritional status, physical performance, body composition, and inflammation was performed. Tumour-related and cachexia-related characteristics of the organoids were analysed using histological stainings, targeted sequencing, and real-time-quantitative PCR. Cachexia-related factors present in the circulation of the patients and in the tumour organoid secretome were analysed by enzyme-linked immunosorbent assay. Results The established human pancreatic tumour organoids presented typical features of malignancy corresponding to the primary tumour (i.e. nuclear enlargement, multiple nucleoli, mitosis, apoptosis, and mutatedKRASand/orTP53). These tumour organoids also expressed variable levels of many known cachexia-related genes including interleukin-6 (IL-6),TNF-alpha,IL-8,IL-1 alpha,IL-1 beta,Mcp-1,GDF15, andLIF. mRNA expression ofIL-1 alpha andIL-1 beta was significantly reduced in organoids from cachectic vs. non-cachectic patients (IL-1 alpha: -3.8-fold,P = 0.009, andIL-1 beta: -4.7-fold,P = 0.004).LIF,IL-8, andGDF15mRNA expression levels were significantly higher in organoids from cachectic vs. non-cachectic patients (LIF: 1.6-fold,P = 0.003;IL-8: 1.4-fold,P = 0.01;GDF15: 2.3-fold,P <0.001). In line with theGDF15andIL-8mRNA expression levels, tumour organoids from cachectic patients secreted more GDF15 and IL-8 compared with organoids from non-cachectic patients (5.4 vs. 1.5 ng/mL,P = 0.01, and 7.4 vs. 1.3 ng/mL,P = 0.07, respectively). Conclusions This novel human pancreatic tumour organoid biobank provides a valuable tool to increase our understanding of the mechanisms driving cancer cachexia. Our preliminary characterization of the secretome of these organoids supports their application in functional studies including conditioned medium approaches andin vivotransplantation models.