Generation and Maintenance of Patient-Derived Endometrial Cancer Organoids

Abstract

Endometrial cancer (EC) is the leading cause of gynecologic cancer morbidity and mortality in the U.S. Despite advancements in cancer research, EC death rates are increasing, particularly high-grade endometrial cancers. The development of three-dimensional (3D) patient-derived organoid (PDO) models for EC is crucial, as they provide a more accurate representation of the biological and genetic complexity of a patient's tumor compared to traditional 2D cell lines. Here, we describe a protocol for cultivating PDO models from normal endometrium and EC across different EC subtypes. These EC PDO models can be expanded across multiple passages and facilitate the exploration of tumor behavior and drug responses, thereby advancing our understanding of the disease and potentially leading to more effective and individualized novel therapeutic strategies. Key features • Establishment of patient-derived EC and normal endometrium organoids. • Accurate replication of the various histologic and molecular subtypes of EC within the organoids. • Our approach provides a clinically relevant platform for studying EC development, metastasis, progression, and recurrence. • It offers potential for developing targeted therapeutic interventions tailored to specific EC subtypes. Graphical overview Schematic overview of endometrial cancer and normal organoid preparation from patient-derived samples.

Keywords: Endometrial cancer; Long-term expansion; Patient-derived organoids; Translational research; Tumor selectivity.

Figure 1.. Patient-derived organoids from normal endometrium and cancerous tissues.

A. Organoids from normal endometrium, FIGO1 (low grade), FIGO2 (intermediate grade), FIGO3 (high grade), serous cancer, and carcinosarcoma demonstrating that all histologic subtypes can be cultured and maintained through several passages. B. Serous cancer organoids demonstrating survival and maintenance over several passages in culture at passage 1 (P1), passage 7 (P7), and passage 14(P14). Scale bars: 100 μm (10×) and 200 μm (4×).