Journal: Cell Reports Medicine

Article Title: LIFUS-driven engineered bacteria reprogram immunosuppressive niches via mechano-NOTCH signaling

doi: 10.1016/j.xcrm.2026.102658

Figure Lengend Snippet: Mechano-regulation of CAF-CD8 + T cell NOTCH signaling interactions (A and B) Ultrasound elastography (A) shows a significant reduction in tumor stiffness with the combination group (12.95 ± 2.18 kPa) compared with controls (57.19 ± 15.02 kPa, ∗∗∗ p < 0.005) and VNP /ARG-GV monotherapy (46.82 ± 8.13 kPa, ∗∗ p < 0.01), LIFUS monotherapy group (36.93 ± 6.812 kPa), and the control group (57.19 ± 15.02 kPa, ∗ p < 0.05) ( n = 4) (B). (C) AFM further confirms reduced matrix elastic modulus ( n = 12). (D) Collagen I distribution secreted by fibroblasts in B supported the hypothesized changes in fibroblasts within the TME. Quantitative assessment (D) showed a significant reduction in the combined therapy group (42.24 ± 8.46 mm 2 ) compared with the control group (248.62 ± 9.19 mm 2 , ∗∗∗∗ p < 0.0001; n = 3). (E) The differential analysis visualizes changes in interaction numbers and strength, where red indicates enhanced and blue indicates reduced interactions in the combined group vs. control. The highlighted box reveals Cd8t_04 in the differential number of interactions and Cd8t_00 in differential interaction strength with reduced interactions with CAF. (F) Relative information flow of major stromal-immune signaling pathways. Relative contribution of fibroblast-derived signaling pathways to Cd8t_04 cells in control and combination treatment groups, normalized to a scale of 0–1. The dashed line at 0.5 indicates equal contribution between groups. NOTCH signaling dominates CAF–Cd8t_04 cell communication in control tumors and is selectively reduced following combination treatment, whereas CXCL and IFN-II pathways show a relative increase. (G–I) Notch1 and Jagged1 expression at the transcript (G and H) ( n = 6) and protein levels (I) are significantly reduced after combination therapy. Data are representative of three independent experiments. Data are mean ± SD. Statistical analysis was performed using one-way ANOVA (B–D) or two-tailed unpaired Student’s t test (G and H). See also .

Article Snippet: Anti-Notch1 antibody , Proteintech , Cat# 10062-2-AP; RRID: AB_2153338.

Techniques: Control, Protein-Protein interactions, Derivative Assay, Expressing, Two Tailed Test

Journal: Cell Reports Medicine

Article Title: LIFUS-driven engineered bacteria reprogram immunosuppressive niches via mechano-NOTCH signaling

doi: 10.1016/j.xcrm.2026.102658

Figure Lengend Snippet: Dual mechanotransductive regulation of CAFs-CD8 + T cell crosstalk via NOTCH signaling (A) The schematic diagram of co-culture systems of CAFs and CD8 + T cells with GVs responsive to LIFUS constructs in vitro . Before 60 s of LIFUS exposure, cell co-incubation with purified GVs allowed sufficient GV-cell contact. (B and C) Confocal imaging revealed a significant reduction in Notch1 receptor and Jagged1 ligand expression in CAFs exposed to LIFUS-driven GVs (GVs + LIFUS group) compared to unstimulated controls (GVs − LIFUS group). Scale bars, 100 μm. (D and E) In CD8 + T cells, Jagged1 expression was decreased following LIFUS-driven GV stimulation (GVs + LIFUS group), whereas Notch1 receptor levels remained unchanged, indicating asymmetric NOTCH pathway modulation between CAF and CD8 + T cell. Scale bars, 100 μm. (F) Experimental workflow for assessing CAFs’ functional changes following LIFUS-driven GV treatment. (G and H) Transwell migration assays revealed significantly reduced CAF motility in the LIFUS treatment group (56 ± 4.0 cells/field) compared with the control group (150.67 ± 15.04 cells/field, ∗∗∗∗ p < 0.0001) and CAFs exposed to treatment without LIFUS (101.67 ± 3.79 cells/field, ∗∗ p < 0.01; n = 3). Scale bars, 100 μm. (I and J) ELISA quantification of CAF-secreted factors showed decreased TGF-β (0.39 ± 0.07 vs. 0.68 ± 0.38 ng/mL in control, ∗∗∗ p < 0.005) ( n = 3) (I) and collagen I (0.27 ± 0.11 vs. 0.76 ± 0.06 ng/mL in control, ∗∗ p < 0.01) ( n = 3) (J) after LIFUS-driven GV stimulation, consistent with reduced CAF activation. (K) Schematic of co-culture assay evaluating CD8 + T cell (precondition with or without LIFUS-driven GVs) and 4T1 tumor cell interactions. (L and M) CD8 + T cells stimulated by LIFUS-driven GVs exhibited significantly enhanced adhesion to 4T1 tumor cells (51.89% ± 10.47% adherent cells/field) compared with control (3.25% ± 1.19%, ∗∗∗ p < 0.005) and GV-only groups (15.72% ± 7.53%, ∗∗ p < 0.01) ( n = 3). Scale bars, 100 μm. (N) Functional validation showed the cytotoxicity of CD8 + T cells to 4T1-Luc cells ( C); the luminescence intensity, which represented the apoptosis of 4T1-Luc tumor cells, showed significant luminescence change at 12 h between CD8 + T −LIFUS ((48 ± 6.3) × 10 5 ps −1 cm −2 sr −1 ) and CD8 + T +LIFUS ((17 ± 5.2) × 10 5 ps −1 cm −2 sr −1 , ∗∗ p < 0.01) group, and a stronger decrease at 36 h compared to control group (∗∗∗∗ p < 0.0001) ( n = 3). A representative image is shown in C. Data are representative of three independent experiments. Data are mean ± SD. Statistical analysis was performed using one-way ANOVA (H–J and M) or two-way ANOVA (N). Also see A–S5C.

Article Snippet: Anti-Notch1 antibody , Proteintech , Cat# 10062-2-AP; RRID: AB_2153338.

Techniques: Co-Culture Assay, Construct, In Vitro, Incubation, Purification, Imaging, Expressing, Functional Assay, Migration, Control, Enzyme-linked Immunosorbent Assay, Activation Assay, Co-culture Assay, Biomarker Discovery