Exploring the immune environment of glioblastoma in humanized mouse models

Glioblastoma (GBM) is the deadliest primary brain tumor in adults, and progress in therapy development has been hindered by the lack of preclinical models that faithfully recapitulate interactions between human tumor cells and human immune cells, particularly in therapy-resistant disease. To address this limitation, we established a humanized mouse model of GBM using radiation-resistant patient-derived xenografts (PDXs) implanted into immunodeficient mice reconstituted with human hematopoietic stem progenitor cells derived from umbilical cord blood. For single-cell RNA sequencing, peripheral blood (PB), tumor-infiltrating immune cells (TILs), and tumor cells were collected from humanized NOG-EXL mice bearing EGFP-positive JX14P-RT tumors. Tumor cells from the same JX14P-RT model implanted into non-humanized naive NOG-EXL mice were also analyzed for comparison. Single-cell suspensions were prepared from tumor tissues, and CD45-positive immune cells and EGFP-positive tumor cells were isolated by cell sorting. Human CD45-positive cells in peripheral blood were purified separately. The dataset includes three TIL samples from humanized mice, three PB samples from humanized mice, three tumor-cell samples from humanized mice, and two tumor-cell samples from naive mice. Individual samples were labeled with hashtag oligonucleotide antibodies, pooled, and then analyzed separately by compartment as TIL, PB, and tumor groups. Libraries were generated using the 10x Genomics Chromium Single Cell 3' platform and sequenced on an Illumina NovaSeq 6000. The data show infiltration of diverse human immune populations, including T cells, natural killer cells, and myeloid lineage cells, and support the utility of this model for studying the immune microenvironment of recurrent and therapy-resistant GBM.