Linda Fong

Differential expression analyses of CD8 T and macrophages populations of proliferating (p53 off) vs. senescent (p53 on) tumors by scRNA-seq.

Low (50 pg/ml) and high (1 ng/ml) dose of IFN-γ treatment in proliferating and senescent NSP cells.

Supplementary Video S1 from Senescence Rewires Microenvironment Sensing to Facilitate Antitumor Immunity

Supplementary Video S2 from Senescence Rewires Microenvironment Sensing to Facilitate Antitumor Immunity

Supplementary figures complement main figures to show that senescent cells have a rewired environmental signal sensing phenotype, exemplified by an enhanced IFN-g signaling, to facilitate anti-tumor immunity.

RNA-Seq data of proliferating (PRO) or senescent (SEN) NSP liver tumor cells, for both p53-restoration and drug-induced (trametinib+palbociclib) settings. PRO and SEN cells were also treated with the BET inhibitor JQ-1 (500 n, 48 h), to expose BRD4-mediated transcriptional output in each cellular state.

Cellular senescence involves a stable cell-cycle arrest coupled to a secretory program that, in some instances, stimulates the immune clearance of senescent cells. Using an immune-competent liver cancer model in which senescence triggers CD8 T cell–mediated tumor rejection, we show that senescence also remodels the cell-surface proteome to alter how tumor cells sense environmental factors, as exemplified by type II interferon (IFNγ). Compared with proliferating cells, senescent cells upregulate the IFNγ receptor, become hypersensitized to microenvironmental IFNγ, and more robustly induce the antigen-presenting machinery—effects also recapitulated in human tumor cells undergoing therapy-induced senescence. Disruption of IFNγ sensing in senescent cells blunts their immune-mediated clearance without disabling the senescence state or its characteristic secretory program. Our results demonstrate that senescent cells have an enhanced ability to both send and receive environmental signals and imply that each process is required for their effective immune surveillance.Significance:Our work uncovers an interplay between tissue remodeling and tissue-sensing programs that can be engaged by senescence in advanced cancers to render tumor cells more visible to the adaptive immune system. This new facet of senescence establishes reciprocal heterotypic signaling interactions that can be induced therapeutically to enhance antitumor immunity.See the interview with Direna Alonso-Curbelo, PhD, recipient of the inaugural Cancer Discovery Early Career Award: https://vimeo.com/992987447See related article by Marin et al., p. 410.This article is highlighted in the In This Issue feature, p. 247

A p53-restorable tumor model to study senescence immune surveillance. A, Generation of the p53-restorable, NRAS-driven mouse liver cancer model using the sleeping beauty transposon system delivered through HTVI. (Created with BioRender.com.) B, Representative ultrasonogram of HTVI and orthotopic injection liver cancer models at indicated time after p53 restoration. C, Survival analysis of mice in the HTVI model. D, Representative hematoxylin and eosin (H&E), immunofluorescence (IF), and senescence-associated β-galactosidase (SA-β-gal) staining of p53-suppressed (p53 off) and -restored (p53 on for 14 days) tumor sections generated from the HTVI model. Scale bars, 50 μm. E–G, Orthotopic injection of GFP-luciferase vector-transduced NSP tumor cells into the livers of immunocompetent and immunodeficient mouse strains. E, Tumor size change measured by ultrasound upon p53 restoration. R2G2, Rag2-Il2rg double-knockout mouse. Data are presented as mean ± SEM. n e 9 for each strain. F, Representative macroscopic pictures at 21 days of p53 on or endpoint p53 off tumor. G, Representative IHC staining of GFP-labeled tumor cells at day 21 upon p53 restoration. Scale bars, 100 μm. **, P < 0.01; ***, P < 0.001.

Differential expression analyses of CD8 T and macrophages populations of proliferating (p53 off) vs. senescent (p53 on) tumors by scRNA-seq.

Low (50 pg/ml) and high (1 ng/ml) dose of IFN-γ treatment in proliferating and senescent NSP cells.