Fig. 1: A head and neck cancer in vivo CRISPR screen identifies epigenetic regulators of tumor immunity.
From: In vivo CRISPR screening in head and neck cancer reveals Uchl5 as an immunotherapy target
A Schematic for in vivo epigenome-wide CRISPR screening using the SCAR system and screen design, created in BioRender108. B Tumor volume over time for SCAR-vector-system-library-transduced MOC1-esc1 cells in NSG, WT untreated, and anti-PD-1- and anti-CTLA-4-treated C57BL/6 mice (n = 40 mice per group). C, D Volcano plots illustrating the comparison of ICB-treated wild-type and untreated (C) or NSG (D) mice with genes whose knockout (KO) can enhance (red) or inhibit (blue) sensitivity to combination of anti-PD-1 and anti-CTLA-4 treatment. E Histograms showing guide performance relative to library distribution for some top depleted or enriched genes in ICB-treated versus NSG animals. F Depletion (negative ratios) of targeted genes in ICB-treated wild-type versus NSG mice compared to ICB-treated versus untreated wild-type. HIRA, INO80, TFIID, SIN3, EP300, E3 ligase or Deubiquitinating enzymes and positive control genes highlighted. G Schematic of validation pool screen, figure created in BioRender109. H Calculated log2 fold change in the ratio of tumor cells with sgRNAs targeting selected genes in the validation pool versus control sgRNA within MOC1-esc1 tumors treated with anti-PD1 normalized to the ratio for tumors implanted in NSG mice, n = 5 mice each. Genes whose knockout (KO) can enhance (red) or inhibit (blue) sensitivity to anti–PD1 treatment. I Histograms showing guide performance relative to library distribution for Uchl5 gene in ICB-treated versus NSG animals across different tumor models. J Schematic for tumor growth competition, created in BioRender110. K Calculated log2 fold changes of Uchl5 versus control sgRNA barcode abundance is plotted on the y-axis, where negative values represent the depletion of Uchl5 sgRNAs, n = 6 tumors for NSG and PD1 treated groups, 10 tumors for untreated WT group, data are representative of four independent experiments. Data in B were calculated by two-way ANOVA represented as mean ± s.d. The -log P-values as in F were calculated by two-sided hypergeometric test. Data in H and K were analyzed by unpaired, two-sided Student’s t-test and are represented as mean ± s.d. Source data are provided as a Source Data file.
