Journal: Advanced Science

Article Title: Granzyme B PET Imaging Reveals Lgmn + Macrophage‐Mediated Immune Evasion and Guides Immunotherapy in EGFR‐TKI‐Resistant NSCLC

doi: 10.1002/advs.77837

Figure Lengend Snippet: Lgmn recruits Chip to ubiquitinate Eno1 to regulate the dysfunction of CD8 + T cells. (A) The expression of glycolysis‐related enzymes in CD8 + T cells treated with Lgmn fl/fl /EVs and Lgmn cko /EVs from proteomics analysis by LC‐MS/MS. (B) Cell lysates were immunoprecipitated using anti‐Lgmn antibodies, and Eno1 expression was detected by Western blotting. (C, D) Eno1 expression in CD8 + T cells treated with Lgmn fl/fl /EVs and Lgmn cko /EVs was detected by qRT‐PCR (C) and Western blotting (D). (E, F) Detection of Eno1 expression in CD8 + T cells overexpressing Lgmn by immunoblotting (E) and qRT‐PCR (F). (G, H) Detection of Eno1 protein stability in CD8 + T cells overexpressing Lgmn (G) or treated with EVs (H) by immunoblotting. (I) Detect Eno1 expression in CD8 + T cells overexpressing Lgmn or treated with MG132 using Western blotting. (J) Detect Eno1 expression in CD8 + T cells overexpressing Lgmn or treated with CQ using Western blotting. (K) Detect Eno1 ubiquitination in CD8 + T cells overexpressing Lgmn using Western blotting. (L) Cell lysates were immunoprecipitated using anti‐Lgmn or anti‐Eno1 antibodies, and the proteins interacting with Lgmn and Eno1 were detected by LC‐MS. (M) Co‐IP assays were conducted to observe the interaction between Lgmn, Chip, and Eno1. (N) Chip was knocked down, and Eno1 ubiquitination in CD8 + T cells was detected by Western blotting. (O) Chip was knocked down, and Lgmn was overexpressed; Eno1 expression and ubiquitination in CD8 + T cells were detected by Western blotting. (P) Cell lysates were immunoprecipitated using anti‐Eno1 antibodies, and Chip expression was detected by Western blotting. (Q, R) Chip was knocked down, and Lgmn was overexpressed; glucose uptake in CD8 + T cells was detected (Q), and TIM‐3 + and PD‐1 + CD8 + T cells were detected by flow cytometry (R). (S) Illustration of Lgmn enhancing the interaction between Chip and Eno1, and subsequently modulating CD8 + T cell exhaustion through metabolic reprogramming.

Article Snippet: Primary antibodies used are as follows: anti‐β‐actin (#4970, Cell Signaling Technology, 1:2000), anti‐Lgmn (A23776, ABclonal, 1:1000), anti‐Eno1 (A11448, ABclonal, 1:1000), anti‐Chip (#2080, Cell Signaling Technology, 1:1000), anti‐HA (AE105, ABclonal, 1:20000), anti‐FLAG (AE005, ABclonal, 1:10000), anti‐Ubiquitin (A19686, ABclonal, 1:1000).

Techniques: Expressing, Liquid Chromatography with Mass Spectroscopy, Immunoprecipitation, Western Blot, Quantitative RT-PCR, Ubiquitin Proteomics, Co-Immunoprecipitation Assay, Flow Cytometry

Journal: Cell Reports Medicine

Article Title: Ferroptotic tumor cells reprogram tumor-associated macrophage antigen presentation to enhance the efficacy of immune checkpoint blockade

doi: 10.1016/j.xcrm.2026.102774

Figure Lengend Snippet: Ferroptosis signature associates with better responses to immunotherapy (A) Volcano plot analysis of differentially expressed genes from RNA-seq data of RSL3- and vehicle-treated MC38-bearing mice. (B) Correlation between ferroptosis signature score and cytotoxic T cell (CTL) signature score and CD38 expression in patients across solid malignancies. The Spearman correlation coefficient and associated probability were determined. All the data were obtained from TCGA. (C) Ferroptosis signature is positively correlated with CTL signatures and CD38 expression in most clinical trials for immunotherapy. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001; Spearman correlation. (D) Ferroptosis signature score is higher in patients from the Riaz 2017 cohort evaluated as responders than in those evaluated as nonresponders. Data are presented as mean ± SD. ∗ p < 0.05; two-sided unpaired Student’s t test. (E) Ferroptosis signature score is higher in patients from Liu 2019 cohort evaluated as responders than in those evaluated as nonresponders. Data are presented as mean ± SD. ∗ p < 0.05; two-sided unpaired Student’s t test. (F) Patients with higher ferroptosis signature score have better progress-free survival in patients from the Liu 2019 cohort treated with anti-PD-1; log rank (Mantel-Cox) test. (G and H) Representative multiplex immunofluorescence images of TFRC, IDO1, ENO1, and ACSL4 (G) were shown from biopsy samples of responders ( n = 6) or non-responders ( n = 6) to immunotherapy in CRC, and the fluorescence signals were quantified (H). Scale bar: 50 μM. Data are presented as mean ± SD. ∗∗∗ p < 0.001; two-sided unpaired Student’s t test. (I) Representative multiplex immunofluorescence images of CD68 and MHC-II co-location from biopsy samples of responders ( n = 6) or non-responders ( n = 6) to immunotherapy in CRC. Scale bar: 50 μM. (J) Correlation between the number of MHC-II + CD68 + macrophages and ferroptosis signature score. Spearman correlation. (K) Representative positron emission tomography-computed tomography (PET-CT) of patients receiving anti-PD-1 antibody treatment. (L) Representative CT of patients receiving anti-PD-1 antibody treatment.

Article Snippet: Rabbit polyclonal anti-human/mouse ENO1 antibody , ABclonal , Cat# A1033; RRID: AB_2757874.

Techniques: RNA Sequencing, Expressing, Clinical Proteomics, Multiplex Assay, Immunofluorescence, Fluorescence, Positron Emission Tomography, Computed Tomography, Positron Emission Tomography-Computed Tomography