Journal: Cell Reports Medicine

Article Title: CAR-T triggers TAM reeducation and adaptive anti-tumor response via TREM2 deficiency or CD40 agonist

doi: 10.1016/j.xcrm.2025.102539

Figure Lengend Snippet: TREM2 + TAMs hinder the therapeutic effectiveness of GPC3-CAR-T cells (A) UMAP visualization of scRNA-seq data from tumor biopsies following IL15-GPC3-CAR-T therapy (data from Steffin et al., Nature 2025; GEO: GSE253352 ). (B) The heatmap displayed the signature differentially expressed genes across distinct myeloid cell subpopulations. (C) The bar chart showed the proportions of myeloid cell subsets in the PD and PR groups. (D) The boxplot demonstrated the statistical differences among various myeloid subpopulations. (E) In vitro cytotoxicity measured by lactate dehydrogenase (LDH) release at indicated effector cell vs. target cell ratios (E:T ratios) using Hepa1-6 cells overexpressing human GPC3 or vector control were co-cultured with Ctrl-CAR-T (targeted CD19) or GPC3-CAR-T cells. (F) Subcutaneous tumor growth and survival in C57BL/6J mice ( n = 6/group) inoculated with GPC3-Hepa1-6 cells and treated with Ctrl- or GPC3-CAR-T cells (1 × 10 6 ) via tail vein. (G) Orthotopic liver tumor model: tumor weight at day 28 and survival in mice receiving intrahepatic GPC3-Hepa1-6 implants followed by CAR-T treatment. (H) Flow cytometric analysis of immune and stromal cell populations in tumors from subcutaneous and orthotopic models following GPC3-CAR-T treatment. (I) Expression of TAM-associated genes (Trem2, Spp1, Cd274, Arg1, Tgfb1, and Cd206) in F4/80 + TAMs sorted from tumors of GPC3-CAR-T-treated mice, measured by RT-qPCR. (J) Schematic of the orthotopic HCC model for CAR-T therapy in Trem2 -WT/KO mice. (K) Tumor representative photographs of different groups were shown. Dot plots showed liver weight, liver-to-body weight ratio, per liver, n = 8. (L) UMAP plots of individual groups displayed distinct intratumoral cell populations. n = 2 for all treatment groups on scRNA sequence. (M) Proportion of the distinct intratumoral cell populations. Data were represented by mean ± SEM. ns, no significance, ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001.

Article Snippet: Rabbit anti-mouse F4/80 antibody , CST , Cat#70076.

Techniques: In Vitro, Plasmid Preparation, Control, Cell Culture, Expressing, Quantitative RT-PCR, Sequencing

Journal: Nature Communications

Article Title: Living sensor display implanted on skin for long-term biomarker monitoring

doi: 10.1038/s41467-025-67384-2

Figure Lengend Snippet: a Bright-field images of the engrafted tissue-engineered skin before LPS stimulation, more than 2 months post-transplantation. b Fluorescence images of the engrafted tissue-engineered skin before LPS administration, c 2 days after the intraperitoneal administration of LPS, d 7 days after administration, e Fluorescence image of the engrafted tissue-engineered skin at 2 days after subcutaneous administration of LPS on the 9th day post-initial administration. f Time course of fluorescence intensity calculated from the image analysis of the tissue-engineered skin. The vertical dotted lines indicate the time points of fluorescence observation, and the downward‑pointing triangles indicate the timing of LPS administration. g Comparison of fluorescence intensities at 7 days and 9 days after administration. Data are presented as mean ± SD from n = 5 biologically independent mice. p = 0.00000056. The statistical analysis was performed using t-tests (two-tailed). *** p < 0.001. h Comparison of fluorescence intensities at 2 days and 7 days after administration. The statistical analysis was performed using t-tests (two-tailed). Data are presented as mean ± SD from n = 5 biologically independent mice. p = 0.3898 i Immunostaining image merged for human mitochondria and GFP from a skin section taken from around the engrafted skin at 9 days after LPS administration. Representative of three independent experiments with similar results. j Fluorescence image confirming the localisation of inflammatory cells positive for Ly6g and F4/80 in the magnified area. Representative of three independent experiments with similar results.

Article Snippet: Following the removal of the BSA solution, the tissue sections were incubated with the diluted primary antibody: Cytokeratin 14 Polyclonal antibody (Proteintech・10143-1-AP,CK14), Anti-Collagen IV antibody(abcam・ab6586, Col4),Vimentin (D21H3) XP Rabbit mAb (CST 5741S, Vimentin), Anti-GFP (Rat IgG2a) Monoclonal (GF090R) CC (nacalali,04404-84,GFP), Anti-GFP Antibody (abcam・ab290, GFP), Purified Rat Anti-Mouse CD31 (BD pharmingen・553370, CD31), Anti-Mitochondria antibody (Merck (Sigma)・MAB1273, Human Mitochondria), Rat Anti-Mouse F4/80 antibody (Bio-Rad・MCA497GA, F4/80), Anti-CD86 antibody (Cell Signalling Technology・ #19589S, CD86) Anti-CD163 antibody (abcam・ab182422, CD163), Anti-Ly6g antibody (abcam・ab25377, Ly6g), overnight at 4 °C within the moisture chamber.

Techniques: Transplantation Assay, Fluorescence, Comparison, Two Tailed Test, Immunostaining