Fig. 2: On-target killing of prostate cancer organoids by LeY CAR T cells. | Nature Communications

Fig. 2: On-target killing of prostate cancer organoids by LeY CAR T cells.

From: Low-dose carboplatin modifies the tumor microenvironment to augment CAR T cell efficacy in human prostate cancer models

Fig. 2

a Representative immunohistochemical staining (n = 3 samples per PDX/organoid) of LeY in PDXs or organoids from PDXs (scale bars = 25 µm). b Representative images of LeY-positive organoids killed by infiltrating LeY CAR T cells compared to empty vector T (ev-T) cells (scale bars = 50 µm). c Propidium iodide (PI; red) uptake by organoids after 24-h culture with LeY CAR T cells, but not ev-T cells (scale bars = 50 µm). b, c Representative images of data presented in d. d PI staining of organoids with high or low-moderate LeY expression co-cultured with LeY CAR T cells (red) and ev-T cells (blue) for 48 h. Organoids were established from PDX-287R (n = 100 and 90 organoids co-cultured with CAR T and ev-T cells, respectively), PDX-224R-Cx (n = 42 and 15 organoids for CAR T and ev-T cells, respectively), PDX-167.2M-Cx (n = 50 organoids for CAR T or ev-T cells, respectively), PDX-472M (n = 53 and 46 organoids for CAR T and ev-T cells, respectively), PDX-435.1A-Cx (n = 50 organoids for CAR T or ev-T cells) and PDX-201.A-Cx (n = 33 and 31 organoids for CAR T and ev-T cells, respectively). Normalized percent lysis was calculated for CAR T or ev-T organoids as (MFI of organoid – minmean)/(maxmean – minmean) x 100, with MFI of Triton set as maximal lysis (max) and MFI of organoids alone as spontaneous lysis (min). The percentage membrane LeY-positive cells is shown for each organoid model. e Killing by LeY-positive CAR T cells is abrogated by granzyme and perforin inhibitors. Graphs show normalized percent lysis of organoids by CAR T cells over 48 h established from PDX-287R and treated with DMSO, perforin inhibitor (SN34960), granzyme B inhibitor (C20) and combined inhibition (n = 100 organoids per treatment), or established from PDX-472M and treated with DMSO (n = 54 organoids), SN34960 (n = 56 organoids), C20 (n = 60 organoids), and combined inhibition (n = 53 organoids). Data in d and e are shown as mean ± SEM. Significance was determined by two-tailed unpaired t test. Source data are provided as a Source Data file.

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