Patient-Derived Meningioma Organoids: A Reliable Model for Studying Human Tumor Pathophysiology

Abstract

Introduction: Meningiomas are the most common primary central nervous system tumors, constituting 39.7% of intracranial tumors. Although generally benign, some exhibit aggressive behavior and risk of recurrence, necessitating adjuvant therapy and repeat surgical interventions. Molecular studies have identified tumor-driving mutations, leading to targeted 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-Os) that faithfully replicate human disease. Methods: MEN-Os were successfully established from 15 meningioma samples (11 grade 1, 4 grade 2) from neurosurgical resections using an optimized culture protocol. Histological and immunohistochemical analyses were used to assess the resemblance of MEN-Os to original tumor tissues. RNA sequencing compared transcriptional signatures between MEN-Os and corresponding patient-resected tissues. Results: MEN-Os 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-Os preserved key architectural features, including cellular organization, nuclear morphology, and proliferation rates. Immunohistochemical staining for meningioma-specific markers, such as the progesterone receptor, confirmed similar expression patterns to parental tumors. Transcriptomic profiling demonstrated that MEN-Os retained the transcriptional signatures of original tissues, including genes associated with meningioma pathology (NF2, CDKN2A, TP53). Differential expression and deconvolution analyses showed that MEN-Os contained diverse cell populations, including tumor and stromal cells, while preserving the immune microenvironment, as validated by histopathological and transcriptomic profiling. Conclusion: We established a robust, reproducible protocol for generating MEN-Os, which faithfully replicates the histopathological, molecular, and cellular characteristics of original tumors. MEN-Os provide a valuable model for studying meningioma biology and evaluating therapeutic strategies.

Keywords: meningioma; organoids.

Figure 1

Establishment of patient-derived meningioma organoids. (a) Meningioma tissues were obtained from patients undergoing neurosurgical tumor resection. (b) Tumor tissues were minced into 0.5–1 mm diameter pieces and washed with DPBS++, lysis buffer, then DMEM/F12. (c) Processed tumor pieces were then plated in untreated 6-well plates and cultured at 37 °C, 5% CO₂, and 90% humidity on an orbital shaker for up to 4 weeks. Bright-light pictures showing tumor pieces at day 0 (d) and 4 weeks in culture (e). (f) Dot plots showing the change in organoid circularity (left-panel) and 2D size normalized to timepoint zero (right-panel). Dots and error bars represent mean ± SEM. DPBS++, Dulbecco’s phosphate buffered saline with calcium and magnesium; RPM, rotations per minute. CO₂.

Figure 2

Histopathological analysis of patient-resected meningioma and corresponding meningioma organoid. (a) H&E staining of a patient resected meningioma (left panel) and corresponding established organoid (right panel) showing recapitulation of the tumor’s histological cytoarchitecture and cellular composition. (b) Organoid containing psammoma bodies, likely retained from the original tumor given the short incubation period. (c) Organoid displaying characteristic whorl pattern seen in meningiomas. (d) Strong SSTR2 immunostaining in tumor organoids (left panel) and CD163 immunostaining of tumor organoids highlighting tissue macrophages/histiocytes (right panel). MEN-O, meningioma organoid; SSTR2, somatostatin receptor 2. Scale bar, 100 µm.

Figure 3

Immunofluorescent analysis of patient-resected meningioma and corresponding meningioma organoid. (a) KI67 staining of original tumor tissues and corresponding tumor organoids to analyze and quantify cellular proliferation. (b) Progesterone receptor (PR) staining and quantification of the patient-resected meningioma and corresponding organoids. Scale bar, 50 µm. Error bars represent ±SEM.

Figure 4

Transcriptomic analysis of patient-resected meningioma and corresponding meningioma organoid. (a) Principal component analysis of the original tumor and corresponding established organoids at 4 weeks showing a degree of right cultural shift on PC1, yet little-to-no variation on PC2. (b) Heatmap showing Pearson’s correlation (r) between original tumor and corresponding organoids (p < 0.001). (c) Expression analysis of specific genes in original tumors and corresponding organoids at 4 weeks. Dots and error bars represent mean ± SEM. (d) Deconvolution analysis showing the percentage of cellular composition of the original tumors and tumor organoids. MEN-O, meningioma organoids; MSC, mesenchymal stem cells; DC, dendritic cells; GMP, granulocyte/macrophage progenitors; NK cell, natural-killer cell. P#, patient sample (original tumor); PO#, patient organoid (MEN-O).

Figure 5

Transcriptomic mutation analysis of original tumors and MEN-Os. (a) Percentage overlap of SNPs and indels between original tumors and their corresponding MEN-Os. Calculated as nonsignificant difference in mutation frequency using 2-tailed t-test at p < 0.05. (b) Mutant transcript fractions for specific SNPs/indels between original tumors and MEN-Os. P#, patient sample (original tumor); PO#, patient organoid (MEN-O).