Engineered E. coli OMVs Carrying the Membrane-Binding hGC33 Fragment Precisely Target Liver Cancer and Effectively Treat Tumor DOI Creative Commons
Yufei Deng,

Bangya Yang,

Zelan Yang

et al.

International Journal of Nanomedicine, Journal Year: 2025, Volume and Issue: Volume 20, P. 6573 - 6590

Published: May 1, 2025

Glypican-3 (GPC3), which is a membrane-associated antigen that overexpressed in hepatocellular carcinoma (HCC). hGC33, humanized anti-GPC3 antibody, has been validated as potential antibody drug with good antitumor activity by preclinical studies and the Phase II clinical trial. However, free usually lack tumor penetration. Outer membrane vesicles (OMVs) are secreted Escherichia coli function natural vectors for molecule delivery mediators of biological signals across tissues. Our study aimed to engineer E. use platform precisely deliver hGC33 single-chain variable fragment (hGC33-scFv) targeted treatment HCC. In this study, we utilized BL21(DE3) express Hbp-hGC33-scFv fusion protein generated hGC33-OMVs. After isolation characterization, assessed their chemotaxis toward HepG2 cells Transwell, coimmunoprecipitation (co-IP) confirm hGC33-GPC3 binding, immunofluorescence (IF) evaluate localization on OMV membranes. The vivo efficacy was BALB/c nude mice harboring cell-derived xenografts, targeting analyzed Cy7-labeled OMVs live imaging. Proliferation assays, cell cycle analysis, Wnt pathway expression analysis were performed elucidate underlying mechanisms. hGC33-OMVs exhibited spherical bilayered nanostructures displayed hGC33-scFv surface. preferentially accumulated tumors, significantly reducing volume compared controls downregulating proliferation markers Ki67 PCNA. Transwell assays revealed increased tropism cells, while Co-IP confirmed direct interaction between GPC3. Meanwhile, suppressed proliferation, induced G1-phase arrest, reduced Wnt3a, β-catenin, Cyclin D1, C-myc expression. Engineered effectively target HCC via interaction, inhibit growth suppressing signaling, demonstrate versatile delivery.

Language: Английский

Engineered E. coli OMVs Carrying the Membrane-Binding hGC33 Fragment Precisely Target Liver Cancer and Effectively Treat Tumor DOI Creative Commons
Yufei Deng,

Bangya Yang,

Zelan Yang

et al.

International Journal of Nanomedicine, Journal Year: 2025, Volume and Issue: Volume 20, P. 6573 - 6590

Published: May 1, 2025

Glypican-3 (GPC3), which is a membrane-associated antigen that overexpressed in hepatocellular carcinoma (HCC). hGC33, humanized anti-GPC3 antibody, has been validated as potential antibody drug with good antitumor activity by preclinical studies and the Phase II clinical trial. However, free usually lack tumor penetration. Outer membrane vesicles (OMVs) are secreted Escherichia coli function natural vectors for molecule delivery mediators of biological signals across tissues. Our study aimed to engineer E. use platform precisely deliver hGC33 single-chain variable fragment (hGC33-scFv) targeted treatment HCC. In this study, we utilized BL21(DE3) express Hbp-hGC33-scFv fusion protein generated hGC33-OMVs. After isolation characterization, assessed their chemotaxis toward HepG2 cells Transwell, coimmunoprecipitation (co-IP) confirm hGC33-GPC3 binding, immunofluorescence (IF) evaluate localization on OMV membranes. The vivo efficacy was BALB/c nude mice harboring cell-derived xenografts, targeting analyzed Cy7-labeled OMVs live imaging. Proliferation assays, cell cycle analysis, Wnt pathway expression analysis were performed elucidate underlying mechanisms. hGC33-OMVs exhibited spherical bilayered nanostructures displayed hGC33-scFv surface. preferentially accumulated tumors, significantly reducing volume compared controls downregulating proliferation markers Ki67 PCNA. Transwell assays revealed increased tropism cells, while Co-IP confirmed direct interaction between GPC3. Meanwhile, suppressed proliferation, induced G1-phase arrest, reduced Wnt3a, β-catenin, Cyclin D1, C-myc expression. Engineered effectively target HCC via interaction, inhibit growth suppressing signaling, demonstrate versatile delivery.

Language: Английский

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