From operating room to laboratory: a pathologist’s perspective on tubo-ovarian patient-derived organoids

Tubo-ovarian carcinoma remains a leading cause of gynaecological-related mortality and continues to lack robust preclinical models that accurately reflect its biological heterogeneity. Through a joint effort between Gulbenkian Institute of Molecular Medicine and CUF Descobertas Hospital, the research team established patient-derived organoids from high- and low-grade serous carcinoma, as well as normal fallopian tube. Despite the limited cohort, the organoids preserved the morphological and immunophenotypical features of the original tissues. These findings support the potential of patient-derived organoids as experimental models for investigating tumour biology and evaluating therapeutic strategies. The study also underscores the pivotal role of the pathologist in sample collection and organoid characterization, in addition to highlighting the importance of a multidisciplinary collaboration in developing models with translational relevance.

Authors and Affiliations:

Catarina Alves-Vale^1,2,3,4*, Beatriz Galvão^2*, Ana Rita Silvestre¹, José Silva Pereira⁴, Li Bei⁴, João Paulo Fernandes⁴, Paula Borralho^1,3, Maria Carmo-Fonseca ^2,3, Noélia Custódio^2,3

1 – Department of Pathology, CUF Oncologia, Lisboa, Portugal

2 – Gulbenkian Institute for Molecular Medicine, Lisboa, Portugal

3 – Faculdade de Medicina, Universidade de Lisboa, Lisboa, Portugal

4 – Gynaecological Oncology Unit, CUF Oncologia, Lisboa, Portugal

*These authors equally contributed to this paper.

Abstract:

Tubo-ovarian carcinoma, a leading cause of gynaecological-related mortality, holds substantial biological and clinical heterogeneity. Despite advancements in drug development, predicting therapeutic efficacy remains challenging, partly due to the limited availability of in vitro models that accurately replicate tumour behaviour. We present a concise overview of the intrahospital workflow for establishing patient-derived organoids and analyse the morphological and immunophenotypical features of high-grade serous carcinoma (HGSC), serous borderline tumour (SBT)/low-grade serous carcinoma (LGSC), and normal fallopian tube (FT) organoids.

Samples were collected from patients undergoing surgery or paracentesis. Tissue underwent mechanical and enzymatical digestion. Resulting cell suspensions were resuspended in an extracellular matrix substitute for subsequent culture. Despite the low efficacy in establishing HGSC organoids (n = 1/7, 14%; 96 days, 11 passages), we successfully established two organoid lines of SBT/LGSC (n = 2/2, 100%; 65 days, 7 passages; 134 days, 16 passages) and normal FT (n = 2/2, 100%; 73 days, 10 passages; 58 days, 8 passages). HGSC organoids exhibited limited growth and mostly irregular structures, while preserving the p53 immunostaining pattern of the original tumour. SBT/LGSC and FT organoids maintained features of architectural complexity and faithfully recapitulated the original immunoprofile.

This study highlights the need for a multidisciplinary collaboration in both clinical and research settings to establish patient-derived organoids. It emphasises the pivotal contribution of pathologists in meticulous sampling and organoid characterisation. The integration of diverse expertise is essential for maximising the potential of organoids as preclinical tools, advancing our understanding of tubo-ovarian carcinoma, and ultimately improving patient outcomes.

Journal: Journal of Ovarian Research

Link: https://ovarianresearch.biomedcentral.com/articles/10.1186/s13048-025-01766-4