Additional file 1 of FAP-targeted CAR-T suppresses MDSCs recruitment to improve the antitumor efficacy of claudin18.2-targeted CAR-T against pancreatic cancer
Creators: name: Liu, Yifan
name: Sun, Yansha
name: Wang, Peng
name: Li, Songling
name: Dong, Yiwei
name: Zhou, Min
name: Shi, Bizhi
name: Jiang, Hua
name: Sun, Ruixin
name: Li, Zonghai
Publisher: name: figshare
Year: 2024
Subjects: subject: Biochemistry
subject: Molecular Biology
subject: Pharmacology
subject: Biotechnology
subject: Chemical Sciences not elsewhere classified
subject: Immunology
subject: FOS: Clinical medicine
subject: Biological Sciences not elsewhere classified
subject: Cancer

description: Additional file 1: Figure S1. (a) Box plot comparing fibroblast characteristics between high-FAP vs. low-FAP groups estimated by IOBR. (b) Heatmap showing fibroblast characteristics between high-FAP vs. low-FAP groups estimated by IOBR. (c) Pan-cancer analysis of FAP protein expression by IHC staining. Figure S2. (a) The transduction efficiency of 806-28Z CAR on splenic T cells derived from C57BL/6 was determined by flow cytometry. UTD cells served as negative controls. (b) The body weight of mice of each treatment group. All data are presented as the mean ± SEM of triplicate experiments. Figure S3. (a-d) In vivo experimental design. C57BL/6 mice were injected s.c. with PANC02-A2 cells and allowed to establish for 10 days. Mice were assigned to four experimental groups. Then, CAR-T cells were injected on day 10 (i.v.; n = 5 mice per group). Additional three mice in each group were used for harvesting tumor tissue during and after the CAR-T treatment. (b) The volume of tumors of each treatment group. (c) The body weight of mice of each treatment group. (d) CAR copy numbers in genomic DNA of residual tumors from each treatment group were measured by qRT-PCR (TaqMan probe). All data are presented as the mean ±SEM of triplicate experiments. *p &lt; 0.05. Figure S4. Hematoxylin and eosin (HE) staining of important organs. Heart, liver, spleen, lung, and kidney were analyzed by HE staining. After study termination, the specimens were harvested from PANC02-bearing mice. The images were obtained under 200×magnification. The scale bar was 100 μm. The data shown are representative of experiments with similar results. Figure S5. Hematoxylin and eosin (HE) staining of important organs. Heart, liver, spleen, lung, and kidney were analyzed by HE staining. After study termination, the specimens were harvested from KPC1199-bearing mice. The images were obtained under 200×magnification. The scale bar was 100 μm. The data shown are representative of experiments with similar results. Figure S6. (a-b) Analysis of the immune cell in tumor tissues of PANC02-A2 allografts. (a) Quantitation of tumor-infiltrating immune cells of each treatment group by flowcytometry. (b) Surface expression of exhaustion marker in T cells and PD-L1 in tumor cells of each treatment group determined by flow cytometry. (c-d) Analysis of the immune cell in tumor tissues of KPC1199 allografts. (c) Quantitation of tumor-infiltrating immune cells of each treatment group by flow cytometry. (d) Surface expression of exhaustion marker in T cells and PD-L1 in tumor cells of each treatment group determined by flow cytometry. (e) Correlation between FAP expression and MDSCs characteristics estimated by IOBR in TCGA-PDAC patients. (f) Correlation between CAFs and MDSCs characteristics estimated by IOBR in TCGA-PDAC patients. (g) Gating strategies for flow cytometry. All data are presented as the mean ± SEM of triplicate experiments.