Single-cell RNA-seq analysis of PDX (Patient-Derived Xenograft) tumors isolated from mouse spleens, treated with Empty CAR, anti-CD72 H24 nanoCAR or anti-CD72 NbD4.13 nanoCAR.

Single-cell RNA-seq analysis was performed on human mantle cell lymphoma PDX (Patient-Derived Xenograft) tumors isolated from mouse spleens to examine gene expression changes after treatment of Empty CAR, humanized anti-CD72 nanobody-based CAR-T (“H24 nanoCAR”) or NbD4.13-based anti-CD72 nanoCAR-T. Overall design: Human mantle cell lymphoma PDX tumor (DFBL44685) was implanted at 1 e6 cells/mouse, mice were then treated with Empty, H24 or NbD4.13 CAR at 5e6 CAR+ cells per mouse on day 10. Spleens were collected at day 70 after tumor implantation. Tumor cells were isolated from mouse spleens and sorted with gating on human CD45+ positive cells. The samples were processed using 10X chromium platform (Chromium Next GEM Immune Profiling 5'HT kit v2, 1000374). Illumina sequencing was performed on the PDX tumor samples using Illumina NovaSeq X 1.5B platform with paired-end 150 bp reads.