Highlights d The majority of disseminating cells of colorectal cancer are Lgr5 À d Lgr5 À cancer cells are the main seeds of colorectal cancer metastatic lesions d Long-term metastatic growth from Lgr5 À cells requires appearance of Lgr5 + cells d Lgr5 À metastases have the intrinsic capacity to re-establish the cellular hierarchy

Highlights d Lineage tracing of partial and full EMT cells in breast cancer metastasis d Partial EMT cells cycle between hybrid EMT and epithelial stages d Partial, but not full, EMT cells are required for metastasis formation d Both partial and full EMT cells contribute to chemoresistance Authors Fabiana L€ uo ¨nd, Nami Sugiyama,

T-cell-secreted interferon (IFN)-γ can exert pleiotropic effects on tumor cells that include induction of immune checkpoints and antigen presentation machinery components, and inhibition of cell growth. Despite its role as a key effector molecule, little is known about the spatiotemporal spreading of IFN-γ secreted by activated CD8 + T cells within the tumor environment. Using multiday intravital imaging, we demonstrate that T cell recognition of a minor fraction of tumor cells leads to sensing of IFN-γ by a large part of the tumor mass. Furthermore, imaging of tumors in which antigen-positive and antigen-negative tumor cells are separated in space reveals spreading of the IFN-γ response, reaching distances of >800 µm. Notably, long-range sensing of IFN-γ can modify tumor behavior, as shown by both induction of PD-L1 expression and inhibition of tumor growth. Collectively, these data reveal how, through IFN-γ, CD8 + T cells modulate the behavior of remote tumor cells, including antigen-loss variants. Articles Nature CaNCerExtended Data Fig. 5 | analysis of T cell mediated loss of ag-presenting tumor cells over time. a, relative GFP + volume in tumors from imaging experiments described in Fig. 2 quantified over time. Mean and SEM are depicted for n=5 mice (n=5 mice for time 0, 16, 24, and 32 h; n=4 for 40, 48, and 72 h; n=3 for 120 h, from data obtained in all independent experiments. b, The distance between CFP + bystander tumor cells and the closest GFP + Ag + tumor cell was determined at indicated time points from tumors described in Fig. 2 for n=2 mice. Data are obtained from two independent experiments, boxplot presenting the minimum, 25 th percentile, median, 75 th percentile and the maximum For total sample size per timepoint see Source Data ED_Fig5_source table.