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. RNA-seq profiling of human resected meningiomas and correspondingly created tumor organoid cultures

引言：脑膜瘤是最为常见的原发性中枢神经系统肿瘤，占颅内肿瘤总数的39.7%。尽管多数脑膜瘤属于良性病变，但部分亚型可表现出侵袭性行为与复发风险，因此需采取辅助治疗并实施重复手术干预。分子研究已明确驱动肿瘤发生的突变，由此催生了靶向治疗方案与临床试验。然而，当前脑膜瘤肿瘤模型存在诸多局限，阻碍了临床前研究成果向临床应用的转化。本研究旨在开发并验证一套标准化方案，用于构建可精准复现人类疾病特征的患者来源脑膜瘤类器官（Meningioma Organoids, MEN-O）。
方法：本研究从神经外科切除标本中获取脑膜瘤组织，采用优化后的培养方案构建MEN-O。通过组织学与免疫组织化学分析，评估MEN-O与原始肿瘤组织的相似性；利用RNA测序对比MEN-O与对应患者切除组织的转录特征。
结果：本研究成功从患者切除标本中构建出MEN-O，可在体外培养长达4周，且呈现稳定的生长状态与结构完整性。组织病理学分析显示，MEN-O保留了原始肿瘤的关键结构特征，包括细胞组织结构、细胞核形态与增殖速率。针对脑膜瘤特异性标志物（如孕激素受体）的免疫组织化学染色结果证实，MEN-O的标志物表达模式与亲本肿瘤一致。转录组分析表明，MEN-O保留了原始肿瘤组织的转录特征，包括与脑膜瘤病理相关的NF2、CDKN2A、TP53等基因。差异表达与反卷积分析显示，MEN-O包含肿瘤细胞与基质细胞等多种细胞群，且保留了免疫微环境，该结论经组织病理学与转录组分析验证。
结论：本研究建立了一套稳定且可重复的MEN-O构建方案，该方案可精准复现原始肿瘤的组织病理学、分子与细胞特征。MEN-O为研究脑膜瘤生物学特性及评估治疗策略提供了极具价值的模型。对人类切除的脑膜瘤及相应构建的肿瘤类器官培养物开展RNA测序分析。