ECE2023 Poster Presentations Adrenal and Cardiovascular Endocrinology (72 abstracts)
Characterisation of a Three-Dimensional (3D) Cell Culture Model of Adrenocortical Carcinoma
Sarah Feely 1 , Padraig Donlon 1 , Nathan Mullen 1 , Anna Sorushanova 1 , David P Finn 2 , Brendan Hernan 2 , Oliver Carroll 3 , Peter Owens 4 , Abhay Pandit 3 , Constanze Hantel 5 & Michael C Dennedy 1
1Lambe Institute, Discipline of Pharmacology & Therapeutics, School of Medicine, Galway, Ireland; 2Galway Neuroscience Centre and Centre for Pain Research, NCBES, Discipline of Pharmacology & Therapeutics, Galway, Ireland; 3Science Foundation Ireland (SFI) Centre for Research in Medical Devices (CURAM), Biomedical Science Building, Galway, Ireland; 4University of Galway, Centre for Microscopy and Imaging, Galway, Ireland; 5Department of Medicine IV,, University Hospital, Munich, Germany
Adrenocortical carcinoma (ACC) is a rare malignancy associated with a poor prognosis (1). Current treatments are limited with surgical resection the only option for a complete cure (2). The development of translational therapies is limited by pre-clinical disease models. Three-dimensional (3D) cell culture models can accurately reflect the tumour micro-environment but are lacking in ACC (3). In the current study, we developed and characterised novel 3D models of MUC-1, HAC15 and H295R within a type-1 Collagen matrix. Viability of each model was assessed by Sytox Blue staining. Live/dead imaging using confocal microscopy was carried out to determine the distribution of live and dead cells within the 3D models. Metabolic activity was assessed via AlamarBlue staining with Ki67 detecting proliferation. Steroidogenic capacity was determined using Liquid chromatography tandem mass spectrometry and real time- polymerase chain reaction (RT-qPCR). The morphology of the cells was imaged using confocal microscopy. All cells were successfully cultured in a type 1 collagen matrix. H295R and MUC-1 models show optimum viability up until day 7 but a decrease compared to when the corresponding cell line was cultured in monolayer. HAC15 model maintains a constant level of viability over 21 days in culture. All three models increase their metabolic and proliferative activity over time. All were steroidogenic, with HAC15 and H295R cells showing an increased expression of aldosterone when stimulated with angiotensin II when cultured in 3D compared to monolayer. Decreased levels of cortisol production were reported in all 3D models compared to when cultured in monolayer. In the current study, we successfully developed 3D cell culture models for ACC. Each cell line in a 3D model showed behaviour reflective of the 3D tumour micro-environment. Each cell line in a 3D model showed: (i) lower overall viability compared to monolayer- reflective of cell turnover and necrosis in the 3D tumour microenvironment, and (ii) an increase in metabolic activity and proliferation reflective of cell turnover in the 3D microenvironment. Similar findings have been shown in the literature for other 3D cancer cell culture models. All were steroidogenic in nature. This model will be used in the future studies to test novel and traditional therapeutics to test the translational relevance of this model to ACC, as a reliable, animal sparing model.
References: 1. Fassnacht et al., Cancer. 115(2): 243-50, 2009. 2. Porpiglia et al., Eur Endocrinol 14: 62-66, 2018 3. Terzolo et al., N Engl J Med 356:2372- 2380, 2007
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Endocrine Abstracts
ISSN 1470-3947 (print) | ISSN 1479-6848 (online)
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