Journal: Molecular Therapy

Article Title: TALEN-edited allogeneic inducible dual CAR T cells enable effective targeting of solid tumors while mitigating off-tumor toxicity

doi: 10.1016/j.ymthe.2024.08.018

Figure Lengend Snippet: Tumor localized FAP + CAFs inhibit CAR T cell intra-tumoral infiltration and anti-tumor cytotoxicity (A) Schematic illustrating T cell infiltration assay in tumor spheroids plated with or without CAFs. (B) Graph depicting flow cytometry quantitation of T cells infiltrated per tumor spheroids with or without CAFs, represented as percentage of T cell input. Bars show means ± SD, n = 3; p values determined by Student t test (two-tailed, unpaired). ns, not significant, ∗ p ≤ 0.05, ∗∗ p ≤ 0.01, ∗∗∗ p ≤ 0.001. (C) Schematic of TRAC KO ML CAR T cell cytotoxicity assay against tumor spheroids of TNBC cell line MDA-MB-231-Luc alone or co-cultured with TNBC-derived CAFs. (D) Bar graph representing percentage MDA-MB-231-Luc tumor cell survival post cytotoxicity assay outlined in (C), at Effector:Target ratio = 5:1. Bars show means ± SD, n = 3; p values determined by Student t test (two-tailed, unpaired). ns, not significant, ∗ p ≤ 0.05, ∗∗ p ≤ 0.01. (E) Immunohistochemistry for detection of human FAP protein in tissue microarray of patient samples from different cancers. (F) Heatmap depicting percentage positive area stained for human FAP protein in immunohistochemical analysis of healthy human tissue microarray.

Article Snippet: Human tumor tissue and healthy tissue microarray slides were obtained from Novus Biologicals (NBP2-30234, NBP2-78082, NBP2-30232, NBP2-30233, NBP2-30189).

Techniques: Flow Cytometry, Quantitation Assay, Two Tailed Test, Cytotoxicity Assay, Cell Culture, Derivative Assay, Immunohistochemistry, Microarray, Staining, Immunohistochemical staining