Establishment and Molecular Characterization of Patient-Derived Organoids for Primary Central Nervous System Lymphoma

Abstract

Primary central nervous system lymphoma (PCNSL) exhibits substantial heterogeneity, both intra-tumoral and intertumoral, posing challenges in developing effective treatment methods. Existing in vitro models fail to simulate the inherent microenvironment and the cellular and mutational diversity of native tumors and require a prolonged generation time. To address this concern, we described an organoid culture method for patient-derived PCNSL organoids (CLOs) and evaluated them through extensive molecular characterization. These CLOs accurately mimicked the histological attributes, gene expression landscapes and mutational profiles of their original tumors through rigorous histopathological analysis, RNA sequencing and whole-exome sequencing. Notably, CLOs were generated within 2 weeks, demonstrating rapid development and reliability. Furthermore, therapeutic profiling was performed on three selected CLOs using four standard treatment drugs. High concordance was observed between the drug responses of patients and those observed in the CLOs, with specific sensitivity to ibrutinib and methotrexate and resistance to dexamethasone and rituximab. Taken together, these results emphasize that CLOs can effectively emulate the key characteristics of PCNSL, enhancing the understanding of the genetic landscape of this complex disease. CLOs provide a rapid and reliable platform for exploring individualized treatment strategies, potentially accelerating the transition of research findings to clinical practice.