Journal: Military Medical Research

Article Title: Octopus-inspired engineered bacteria with a plug-and-play surface display system achieves enhanced tumor-specific colonization and antitumor immunity

doi: 10.1016/j.mmr.2026.100030

Figure Lengend Snippet: Schematic of the construction and therapeutic mechanism of the octopus-inspired triple-engineered bacteria (OITE strain) . The antitumor attenuated Salmonella typhimurium AISI strain was triple-engineered. (1) ST/SC-RGD×4 “tentacles”-Surface modification: the ST sequence was expressed within the external third loop of OmpA in the AISI strain, resulting in the AISI-ST strain. The ST protein was coexpressed with OmpA (OmpA-ST fusion protein) localized to the bacterial outer membrane. An incubation with SC-RGD×4 protein (SC-RGD×4) led to the formation of AISI-ST/SC-RGD×4 strain (AISI-ST/SC-RGD×4) through ST-SC-mediated covalent conjugation. (2) Dynamic EPS “camouflage”-Immunoactivation engineering: the AISI-H-ST strain was created by introducing the quorum-sensing (QS) promoter pLuxI to control HtrA expression into AISI-ST strain, which specifically increased HtrA-mediated extracellular polysaccharide (EPS) production to amplify bacteria-mediated immune activation. (3) Anti-PD1nanobody (PD1nb) “secretion”-Checkpoint blockade engineering: further programming of AISI-H-ST strain enabled QS-triggered anti-PD1nb secretion, generating the AISI-HP-ST strain. The incubation of AISI-HP-ST strain with SC-RGD×4 protein finally produced the OITE strain (AISI-HP-ST/SC-RGD×4). The intravenously administered OITE strain highly selectively accumulates in tumors through RGD-αvβ3 integrin interactions, with subsequent bacterial proliferation initiating two therapeutic actions: HtrA-mediated immune activation via increasing immune cells infiltration and activation, and secreted PD1nb-based blockade of PD-1/PD-L1 immunosuppressive signaling. OITE. Octopus-inspired, triple-engineered bacterium; AISI. Attenuated Salmonella Δ htrA :: luxI - VNP20009 strains; ST. SpyTag; SC. SpyCatcherΔ; RGD. Arginine-glycine-aspartic acid; HtrA. High-temperature requirement A; PD1. Programmed cell death protein 1; H. HtrA; HP. HtrA and PD1nb; AHL. N-acyl homoserine lactone; MACS. Macrophages; Teffs. Effector T cells; Tregs. Regulatory T cells.

Article Snippet: To prepare the surfaces for the strain adhesion assays, the recombinant mouse integrin αvβ3 (TGAV & ITGB3) heterodimer protein (HY-P700761, MCE, USA) was first diluted to 10 μg/ml in 50 mmol/L carbonate buffer (pH 9.6).

Techniques: Bacteria, Modification, Sequencing, Membrane, Incubation, Conjugation Assay, Control, Expressing, Activation Assay, Produced

Journal: Molecular Medicine

Article Title: Integrin αVβ3 antagonist-c(RGDyk) peptide attenuates the progression of ossification of the posterior longitudinal ligament by inhibiting osteogenesis and angiogenesis

doi: 10.1186/s10020-024-00822-x

Figure Lengend Snippet: Integrin αVβ3 is upregulated in OPLL. a Typical preoperative CT and image of a patient with cervical OPLL (the red square at the left indicates OPLL of the cervical spine) and postoperative X-ray image indicating anterior en bloc resection of the cervical ossified posterior longitudinal ligament (the blue square at the right). b RNA-seq visualized by heatmap analysis. c GSEA plots illustrating upregulation of ECM receptor signaling pathway in LFs derived from patients with OPLL and non-OPLL. d Integrin subfamily molecule expression levels using the RNA-seq sequencing results visualized by heatmap analysis. e Relative mRNA expression levels of Integrin subfamily molecule. f The expression of integrin αVβ3 was detected by western blot. g HE staining, immunohistochemical analysis of OCN and integrin αVβ3, and immunofluorescence analysis of CD31 in ligament tissues from patients with OPLL and non-OPLL patients. h Quantitative analysis of OCN-positive cells in g. n = 3 per group. i Quantitative analysis of integrin αVβ3-positive cells in g. n = 3 per group. j Quantitative analysis of CD31-positive cells in g, n = 3 per group. k Quantitative analysis of integrin αVβ3 by western blot. n = 3 per group. All experimental data are presented as the mean value ± SD. Student’s t-test used in ( h ), ( i ), ( j ), and ( k ). * p < 0.05, ** p < 0.01, *** p < 0.001

Article Snippet: Integrin αVβ3 is involved in vascular lumen formation and vascular network construction by regulating the adhesion and migration of EC in angiogenesis(Rocha et al. ).

Techniques: RNA Sequencing, Derivative Assay, Expressing, Sequencing, Western Blot, Staining, Immunohistochemical staining, Immunofluorescence

Journal: Molecular Medicine

Article Title: Integrin αVβ3 antagonist-c(RGDyk) peptide attenuates the progression of ossification of the posterior longitudinal ligament by inhibiting osteogenesis and angiogenesis

doi: 10.1186/s10020-024-00822-x

Figure Lengend Snippet: The c(RGDyk) peptide effectively inhibits the expression of integrin αVβ3 in LFs and ECs. a Cell proliferation of LFs when cultured with different concentrations of C(RGDyk) peptide. b Immunofluorescence of phalloidin and integrin αVβ3 in LFs. Scale bar:100 μm. c Quantitative analysis of integrin αVβ3 fluorescence intensity in LFs. d Cell proliferation of ECs when cultured with different concentrations of c(RGDyk) peptide. e Immunofluorescence of phalloidin and integrin αVβ3 in ECs. Scale bar:100 μm. f Quantitative analysis of integrin αVβ3 fluorescence intensity in ECs. All experiments were repeated three times. All experimental data are presented as the mean value ± SD. One-way ANOVA for multiple comparisons is used in ( a ), ( c ), ( e ), and ( f ). * p < 0.05, ** p < 0.01, *** p < 0.001

Article Snippet: Integrin αVβ3 is involved in vascular lumen formation and vascular network construction by regulating the adhesion and migration of EC in angiogenesis(Rocha et al. ).

Techniques: Expressing, Cell Culture, Immunofluorescence, Fluorescence

Journal: Molecular Medicine

Article Title: Integrin αVβ3 antagonist-c(RGDyk) peptide attenuates the progression of ossification of the posterior longitudinal ligament by inhibiting osteogenesis and angiogenesis

doi: 10.1186/s10020-024-00822-x

Figure Lengend Snippet: The c(RGDyk) peptide inhibits osteogenesis and angiogenesis by targeting integrin αVβ3 and regulating the FAK/ERK1/2 pathway. a and b Western blotting and quantitative analysis of the protein expression of the FAK/ERK1/2 pathway in ECs, including integrin αVβ3, p-VEGFR2, p-FAK, FAK, p-ERK1/2, and ERK1/2. c and d Western blotting and quantitative analysis of the protein expression of the FAK/ERK1/2 pathway in LFs, including integrin αVβ3, p-FAK, FAK, p-ERK1/2, ERK1/2, and RUNX2. β-actin was selected as the internal control. All experiments were repeated three times. All experimental data are presented as the mean value ± SD. One-way ANOVA for multiple comparisons is used in ( b ) and ( d ). * p < 0.05, ** p < 0.01, *** p < 0.001

Article Snippet: Integrin αVβ3 is involved in vascular lumen formation and vascular network construction by regulating the adhesion and migration of EC in angiogenesis(Rocha et al. ).

Techniques: Western Blot, Expressing, Control

Journal: Molecular Medicine

Article Title: Integrin αVβ3 antagonist-c(RGDyk) peptide attenuates the progression of ossification of the posterior longitudinal ligament by inhibiting osteogenesis and angiogenesis

doi: 10.1186/s10020-024-00822-x

Figure Lengend Snippet: c(RGDyk) peptide inhibits heterotopic bone formation in vivo. a The procedures of the heterotopic bone formation assay in vivo. LFs were treated with c(RGDyk) peptide before implantation. b Representative 3D-reconstructed micro-CT images of a scaffold implanted in coculture with LFs after 8 weeks. c Micro-CT scan analysis of the BV/TV and BMD of scaffolds among different groups ( n = 3). d HE staining, immunohistochemical analysis of OCN and integrin αVβ3, and immunofluorescence analysis of CD31 among different implanted scaffolds. e Quantitative analysis of OCN-positive, integrin αVβ3-positive, and CD31-positive cells in d. n = 3 per group. All experimental data are presented as the mean value ± SD. One-way ANOVA for multiple comparisons is used in ( c ) and ( e ). * p < 0.05, ** p < 0.01, *** p < 0.001

Article Snippet: Integrin αVβ3 is involved in vascular lumen formation and vascular network construction by regulating the adhesion and migration of EC in angiogenesis(Rocha et al. ).

Techniques: In Vivo, Tube Formation Assay, Micro-CT, Staining, Immunohistochemical staining, Immunofluorescence

Journal: Molecular Medicine

Article Title: Integrin αVβ3 antagonist-c(RGDyk) peptide attenuates the progression of ossification of the posterior longitudinal ligament by inhibiting osteogenesis and angiogenesis

doi: 10.1186/s10020-024-00822-x

Figure Lengend Snippet: Schematic diagram of the mechanism by which c(RGDyk) suppressed integrin αVβ3 to inhibit OPLL

Article Snippet: Integrin αVβ3 is involved in vascular lumen formation and vascular network construction by regulating the adhesion and migration of EC in angiogenesis(Rocha et al. ).

Techniques: