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

Journal: Nature Communications

Article Title: Targeting of SLC25A22 boosts the immunotherapeutic response in KRAS-mutant colorectal cancer

doi: 10.1038/s41467-023-39571-6

Figure Lengend Snippet: a The construction and validation of PBMC humanized mice model. b SLC25A22 knockout significantly inhibited the growth of DLD1 xenografts in PBMC humanized mice (sgControl: n = 15; SLC-KO1: n = 10). Each dot represents an independent tumor. c SLC25A22 knockout inhibited tumor infiltration of human MDSC (HLA-DR - CD11b + CD33 + ) while increasing T-cell (hCD3 + CD4 + / hCD3 + CD8 + ) infiltration (sgControl: n = 15; SLC-KO1: n = 10). Each dot represents an independent tumor. d Co-immunofluorescence of SLC25A22 and CD33 in a tissue microarray CRC cohort ( n = 207). Positive correlation was revealed in only KRAS-mutant CRC ( upper ). Increased MDSC infiltration was found in SLC25A22-high CRC with mutant KRAS ( lower ). Each dot represents an independent patient. e In TCGA cohort ( n = 383), SLC25A22 mRNA expression is significantly higher in MDSC-high tumors compared to MDSC-low tumors in KRAS-mutant CRC. Each dot represents an independent patient. f Immunohistochemistry of CD8 and SLC25A22 in CRC tissue microarray ( n = 202). CD8 + T cells were reduced in KRAS-mutant CRC compared to wildtype CRC patients. g High SLC25A22 expression correlated with reduced CD8 + T-cell in KRAS-mutant CRC (All cases: n = 202; KRAS-mutant: n = 102). Each dot represents an independent patient. Data are shown as mean ± SD ( b , c , d , e , g ) and ± SEM for growth curve b . Two-tailed Student’s t test ( b , c , d , e , g ). Two-tailed two-way ANOVA for growth curves ( b ). Chi-Square test ( g ) and Pearson correlation test ( d ). Source data are provided as a Source Data file.

Article Snippet: Slides were incubated with primary antibodies overnight, including anti-CD4 (1:2000, ab183685, Abcam), anti-CD8 (1:2000, ab217344, Abcam), anti-S100A8 (1:800, 15792-1-AP, Proteintech), or anti-SLC25A22 (HPA014662; Sigma-Aldrich).

Techniques: Knock-Out, Immunofluorescence, Microarray, Mutagenesis, Expressing, Immunohistochemistry, Two Tailed Test

Journal: Molecular and Clinical Oncology

Article Title: Somatic BRCA1-associated protein 1 (BAP1) loss is an early and rare event in esophageal adenocarcinoma

doi: 10.3892/mco.2017.1286

Figure Lengend Snippet: Staining for BAP1 on the TMAs of 332 esophageal adenocarcinomas. (A) Nuclear staining for BAP1 (original magnification, ×50). (B) Loss of nuclear staining for BAP1 with positive internal control of fibroblasts (original magnification, ×50). TMA, tissue microarray; BAP1, BRCA1-associated protein.

Article Snippet: IHC analysis IHC was performed on TMA slides using the primary mouse anti-BAP1 monoclonal antibody (cat. no. SC-28383; dilution 1:100; Santa Cruz Biotechnology, Santa Cruz, CA, USA).

Techniques: Staining, Control, Microarray