Abstract B024: Hypoxia at 3D organoid establishment selects essential subclone within heterogenous pancreatic cancer

Abstract

Abstract Hypoxia is a key microenvironment associated with malignant traits, including epithelial-mesenchymal transition (EMT) and chemotherapy resistance. Pancreatic ductal adenocarcinoma (PDAC) is extremely hypoxic due to hypovascularity and abundant stromal proliferation. This severe condition accelerates the emergence of hypoxia-adapted cells, which contaminate PDAC as a heterogeneous population. Thus, analyzing hypoxia-adapted cells should be prioritized in analyzing/developing PDAC therapeutic strategies. Recently, 3D organoid culture has been widely used to simulate various organ cancers, featuring precise control with respect to cellular selection through diverse culture conditions. However, as the technology of single-cell analysis has revealed more details of the heterogeneity, the current 3D organoid culture system was also known to have limitation that the system has some potential selection bias. Although several selection biases have been identified, very few reports have focused on oxygen concentrations as a selection bias, leaving room for intervention. To overcome the potential selection bias, we focused on oxygen concentration during the establishment of 3D organoids. We subjected identical PDAC surgical samples to normoxia (O2 20%) or hypoxia (O2 1%) from primary culture, yielding glandular and solid organoid morphology, respectively. Pancreatic cancer organoids established under hypoxia displayed higher expression of EMT-related proteins, a Moffitt basil-like subtype transcriptome, and higher 5-FU resistance in contrast to established organoids under normoxia. We suggest that hypoxia during organoid establishment efficiently selects for hypoxia-adapted cells possibly responsible for PDAC malignant traits, facilitating a fundamental source for elucidating and developing new treatment strategies against recalcitrant PDAC. Citation Format: Koichiro Kumano, Hiromitsu Nakahashi, Pakavarin Louphrasitthiphol, Yukihito Kuroda, Yoshihiro Miyazaki, Osamu Shimomura, Shinji Hashimoto, Yoshimasa Akashi, Bryan James Mathis, Jaejeong Kim, Yohei Owada, Tatsuya Oda. Hypoxia at 3D organoid establishment selects essential subclone within heterogenous pancreatic cancer [abstract]. In: Proceedings of the AACR Special Conference in Cancer Research: Pancreatic Cancer; 2023 Sep 27-30; Boston, Massachusetts. Philadelphia (PA): AACR; Cancer Res 2024;84(2 Suppl):Abstract nr B024.