Patient-Derived Meningioma Organoids: A Reliable Model for Studying Human Tumor Pathophysiology
Introduction: Meningiomas are the most common primary central nervous system tumors, constituting 39.7% of intracranial tumors. Although generally benign, some exhibit aggressive be-havior and risk of recurrence, necessitating adjuvant therapy and repeat surgical in-terventions. Molecular studies have identified tumor-driving mutations, leading to tar-geted therapies and clinical trials. However, translating preclinical findings into clinical success is often hindered by limitations in current meningioma tumor models. This study aims to develop and validate a standardized protocol for establishing patient-derived meningioma organoids (MEN-O) that faithfully replicate human disease. Methods: We obtained meningioma samples from neurosurgical resections and MEN-O were developed using an optimized culture protocol. Histological and immunohistochemical analyses were used to assess the resemblance of MEN-O to original tumor tissues. RNA sequencing compared transcriptional signatures between MEN-O and corresponding patient-resected tissues. Results: MEN-O were successfully established from patient-resected samples and maintained in culture for up to four weeks, showing stable growth and structural integrity. Histopathological analysis revealed that MEN-O preserved key architectural features, including cellular organization, nuclear mor-phology, and proliferation rates. Immunohistochemical staining for meningioma-specific markers, such as progesterone receptor, confirmed similar expression patterns to parental tumors. Transcriptomic profiling demonstrated that MEN-O retained the transcriptional signatures of original tissues, including genes associated with meningioma pathology (NF2, CDKN2A, TP53). Differential expression and deconvolution analyses showed that MEN-O contained diverse cell populations, including tumor and stromal cells, while preserving the immune microenvironment, as validated by histopathological and tran-scriptomic profiling. Conclusion: We established a robust, reproducible protocol for generating MEN-O, which faithfully replicates the histopathological, molecular, and cellular characteristics of original tumors. MEN-O provide a valuable model for studying meningioma biology and evaluating therapeutic strategies. Overall design: RNA-seq profiling of human resected meningiomas and correspondingly created tumor organoid cultures