Patch-seq of human patient-derived diffuse midline glioma xenografts. Patch-seq of human patient-derived diffuse midline glioma xenografts

Treatment-resistance of lethal high-grade brain tumors including H3K27M diffuse midline gliomas is thought to arise in part from transcriptional and electrophysiological heterogeneity. These phenotypes are readily studied in isolation using single-cell RNA-seq and whole-cell patch clamping, respectively, but their simultaneous capture is now possible by aspirating a cell's contents into a patch pipet after electrophysiology recordings using a method called 'patch-seq'. Here, we adapt this method to characterize the gene expression programs and functional responses of patient-derived glioma xenografts to neuronal firing at single-cell resolution. Overall design: Xenografted patient-derived diffuse midline glioma cells were analyzed by whole-cell patch clamp with neuronal stimulation and subsequently by single cell RNA-seq ***Raw data not available due to patient privacy concerns***

致死性高级别脑肿瘤（包括H3K27M弥漫性中线胶质瘤（H3K27M diffuse midline gliomas））的治疗抵抗，被认为部分源于转录组与电生理异质性。这两种表型可分别通过单细胞RNA测序（single-cell RNA-seq）与全细胞膜片钳（whole-cell patch clamping）技术独立开展研究，但如今借助名为“膜片测序（patch-seq）”的实验方法，可在电生理记录完成后将细胞内容物吸入膜片钳电极，从而实现两种表型的同步捕获。本研究对该方法进行适配优化，以单细胞分辨率表征患者来源的胶质瘤异种移植瘤（patient-derived glioma xenografts）在神经元放电刺激下的基因表达程序与功能反应。实验设计概述：对移植建模的患者来源弥漫性中线胶质瘤细胞施加神经元刺激并开展全细胞膜片钳记录，随后进行单细胞RNA测序。***原始数据因患者隐私保护无法公开***