Fig. 2: thGPRs mobilize NK cells to cancer cell factors and spheroids and mark specific types of tumor-infiltrating cells.

a–c, Chemotaxis and spheroid infiltration CRISPRa screens in NK-92 cells. a, Experimental scheme. b,c, Screen results shown as the significance (MAGeCK P values combined via Fisher’s method with BH correction for multiple hypothesis testing, y axis) and fold change (x axis) of each target gene (dot). Positive and negative values denote enrichment and depletion, respectively, in the NK cell population that migrated to the breast cancer supernatant (b) and infiltrated into breast cancer spheroids (c) compared to the NK-92 cell library. d, Differential expression of top hits (rows) in different immune cell types (columns) when comparing tumor and blood samples of individuals with breast cancer based on scRNA-seq data¹⁵ (MAST¹⁷; Methods). e, Differential expression of top hits (rows) in NK cells when comparing tumor and blood samples in different cancer types (column) based on the pan-cancer NK single-cell atlas¹⁶ (edgeR¹⁸ with BH correction for multiple hypothesis testing; Methods). f, UMAP of NK cells in tumor and blood samples from individuals with breast cancer (matching Fig. 1c), with cells colored based on the expression of GPR183 and genes positively (IL7R and CCR7) and negatively (PRF1 and FCGR3A encoding CD16) correlated with GPR183, marking early and late stages of NK cell maturation, respectively. Expression values are shown as log1p -transformed transcripts per million (TPM). g, Expression levels of top hits and top GPR183 program genes across NK cells from breast cancer tumor and blood samples, stratified by NK cell subtype. h, GPR183 is positively and negatively associated with early and late stages of lymphocyte maturation from precursor/naive to cytotoxic effector cells both in NK and CD8⁺ T cells. Spearman correlation coefficients (r_s) of the expression of single genes (dots) with the expression of GPR183 in NK¹⁶ (x axis) and CD8⁺ T cells¹⁵ in breast cancer tumors are shown. GPR183 coexpression patterns in the two cell types were correlated (r_s = 0.29, P < 1 × 10⁻²⁰); cDC2, conventional type 2 dendritic cells; pDC, plasmacytoid DC; ALL, acute lymphoblastic leukemia; BRCA, breast cancer; CLL, chronic lymphocytic leukemia; CRC, colorectal cancer; ESCA, esophageal cancer; GC, gastric cancer; HCC, hepatocellular carcinoma; HNSCC, head and neck squamous cell carcinoma; LC, lung cancer; MELA, melanoma; MM, multiple myeloma; NB, neuroblastoma; NPC, nasopharyngeal cancer; OV, ovarian cancer; PACA, pancreatic cancer; PRAD, prostate cancer; RC, renal carcinoma; THCA, thyroid carcinoma; UCEC, uterine corpus endometrial carcinoma. Panel a created in BioRender; Jerby Lab https://biorender.com/c1gnk9g (2026).