Single-cell RNA-sequencing reveals enrichment of different macrophage subsets following radiation or anti-CD47 therapy in Diffuse Midline Glioma and Glioblastoma

Pediatric diffuse midline gliomas (DMGs), previously known as diffuse intrinsic pontine glioma (DIPG), is one of the most common malignant, fatal brain tumors of childhood arising in the ventral pons. Currently, radiation therapy (RT) is the mainstay treatment for DIPG. However, RT is not a curative treatment and provides only temporary relief in most patients. Therefore, new, and effective therapies are desperately needed for children with these tumors, but decades of clinical trials have so far failed to improve outcomes. Recent advances in immunotherapy have yielded some fantastic opportunities to effectively treat patients with high-grade pediatric brain tumors. The goal of the current study is to perform scRNA-seq on macrophages that either phagocytose-or not. Briefly, a DMG cell line, BT245 was mock-treated or exposed to fractionated RT and co-incubated with macrophages-derived from human peripheral blood mononuclear cells for 24 hrs in the presence of anti-CD47 mAb. Macrophages that either phagocytosed tumor cells- or not were sorted using flow cytometry and scRNA-seq was performed. Overall design: 10x Genomics single cell RNA sequencing of tumor-derived phagocytosing and non-phagocytosing macrophage populations.