Single-cell RNA sequencing  of orthotopic GBM tumors following oncolytic adenovirus treatment
Mice bearing intracranial GL261-CAR tumors were treated intratumorally with PBS (control), oAd, or oAd-mCD47/PD-L1-mIL-12 (1 × 109 pfu). Tumors were harvested three days after the second viral injection and dissociated into single-cell suspensions using the SeekMate Tissue Dissociation Reagent Kit A Pro (SeekGene, K01801-30).Single-cell RNA-seq libraries were constructed using the SeekOne Digital Droplet Single-Cell 3' Library Preparation Kit (SeekGene, #K00202).Single-cell transcriptomic data demonstrate that oAd-mCD47/PDL1-mIL-12 orchestrates a multi-faceted immune response: expanding cytotoxic and memory T-cell subsets, polarizing macrophages toward an antigen-presenting and chemokine-secreting phenotype, and enhancing reciprocal signaling between immune and cancer cells to promote tumor clearance. Overall design: To dissect the immune landscape and tumor-immune interactions following treatment, we performed single-cell RNA sequencing (scRNA-seq) on tumors harvested three days after intratumoral injection of PBS, oAd, or oAd-mCD47/PDL1-mIL-12.