Endocrine Abstracts

Gonadotroph tumours (GoTs) are frequent intracranial neoplasms that represent 30% of all pituitary tumours (PiTs). While GoTs are responsible of an important morbidity, their tumorigenesis is not yet understood, and their treatment is limited to surgical resection and radiotherapy. The absence of identified driver-genes combined with their heterogeneity and silent behaviour (i.e. lack of hormone hypersecretion) limit the development of medical treatment. The current lack of relevant preclinical animal-models and patient-derived cell-lines has further slowed the study of the biological and molecular mechanism underlying gonadotroph tumorigenesis. Here we questioned whether we could scale down the culture and the functional analysis of tumour-cells derived from resected gonadotroph tumour to improve our knowledge of GoTs behaviour and intra-tumoral heterogeneity. We developed a series of approaches combining custom-made agarose-based micro-culture inserts and computerized-analyses of 3D-images to: i) characterize GoT intra-tumoral heterogeneity and ii) screen molecules to develop new therapeutics. The feasibility of these approaches was first addressed through the analysis of pituispheres obtained from rodent-pituitary cell-lines. Subsequently pituispheres obtained from GoT-patients were analyzed. Parameters such a size and percentage of tumor cells (CHGA+) vs non tumor-cells (i.e. Microenvironment, TME) and response to candidate therapeutic molecule were addresses in a 3D semi-high throughput manner (30-50 spheroids acquired through confocal scanning). Magnetic sort based on surface expressing-markers was also performed on surgically-resected gonadotroph tumours to compare the growing capabilities of different tumour-cell-populations isolated from single patients. In conclusion, we have built and validated custom-made micro-culture inserts for growing, imaging and analyzing pituitary rodent cell-lines and patient-derived cells. This work confirms that scaling down the analysis of patient-derived pituispheres could overcome the lack of patient-derived cell lines and could serve the screening of novel therapeutics for personalized medicine.