Figure 5.

FAPCAR_ΔPD1_{ML CAR} UT cells efficiently target FAP⁺ML⁺ tumors and impede CAF⁺ tumor growth with no “off-target” toxicity in vivo

(A) Schematic of UCAR T cell treatment and analysis of bilateral subcutaneous tumors implanted in NSG mice. Data representative of two independently conducted studies, each with different T cell donors. (B) Bar graph representing quantitation of total number of hCD45⁺ cells per gram of tumors from mice treated with indicated UT cells 7 days post administration, as determined by flow cytometry. p values determined by Student t test (two-tailed, paired), n = 3 mice per cohort. ns, not significant, ∗p ≤ 0.05, ∗∗p ≤ 0.01. (C) Bar graph representing percentage of FAP CAR⁺ cells among total hCD45⁺ cells in tumors from mice treated with indicated UT cells 7 days post administration, as determined by flow cytometry. p values determined by Student t test (two-tailed, paired), n = 3 mice per cohort. ns, not significant, ∗p ≤ 0.05, ∗∗p ≤ 0.01. (D) Bar graph representing percentage of ML CAR⁺ cells among total hCD45⁺ cells in tumors from mice treated with indicated UT cells 7 days post administration, as determined by flow cytometry. p values determined by Student t test (two-tailed, paired), n = 3 mice per cohort. ns, not significant, ∗p ≤ 0.05. (E) Top panel: Graph representing growth kinetics of subcutaneous left flank NCI-H226 tumors of remaining mice treated as indicated over time until study endpoint. p values determined by Student t test (two-tailed, unpaired), n = 5 mice per cohort. ns, not significant. Bottom panel: Graph representing growth kinetics of subcutaneous right flank NCI-H226-FAP_40% tumors of remaining mice treated as indicated over time until study endpoint. p values determined by Student t test (two-tailed, unpaired), n = 5 mice per cohort. ns, not significant, ∗∗p ≤ 0.01. (F) Schematic of UT cell treatment and analysis of orthotopic TNBC tumors co-implanted with patient-derived CAFs in NSG mouse mammary fat pad. Data representative of two independently conducted studies, each with different T cell donors. (G) Graph representing growth kinetics of orthotopic TNBC tumors in mice treated as indicated over time. p values determined by Student t test (two-tailed, unpaired), n = 3–5 mice per cohort. ns, not significant, ∗p ≤ 0.05. (H) Kaplan-Meier curve for survival analysis of orthotopic TNBC tumor-implanted NSG mice treated as indicated (n = 3–5 per cohort). p values determined by log rank (Mantel-Cox) test. ns, not significant, ∗p ≤ 0.05.