Engineered hypoxia-responding Escherichia coli carrying cardiac peptide genes, suppresses tumor growth, angiogenesis and metastasis in vivo

Abstract

Development of engineered non-pathogenic bacteria, capable of expressing anti-cancer proteins under tumor-specific conditions, is an ideal approach for selectively eradicating proliferating cancer cells. Herein, using an engineered hypoxia responding nirB promoter, we developed an engineered Escherichia coli BW25133 strain capable of expressing cardiac peptides and GFP signaling protein under hypoxic condition for spatiotemporal targeting of mice mammary tumors. Following determination of the in vitro cytotoxicity profile of the engineered bacteria, selective accumulation of bacteria in tumor microenvironment was studied 48 h after tail vein injection of 108 cfu bacteria in animals. For in vivo evaluation of antitumoral activities, mice with establishment mammary tumors received 3 consecutive intravenous injections of transformed bacteria with 4-day intervals and alterations in expression of tumor growth, invasion and angiogenesis specific biomarkers (Ki-67, VEGFR, CD31and MMP9 respectively), as well as fold changes in concentration of proinflammatory cytokines were examined at the end of the 24-day study period. Intravenously injected bacteria could selectively accumulate in tumor site and temporally express GFP and cardiac peptides in response to hypoxia, enhancing survival rate of tumor bearing mice, suppressing tumor growth rate and expression of MMP-9, VEGFR2, CD31 and Ki67 biomarkers. Applied engineered bacteria could also significantly reduce concentrations of IL-1β, IL-6, GC-SF, IL-12 and TNF-α proinflammatory cytokines while increasing those of IL-10, IL-17A and INF-γ. Overall, administration of hypoxia-responding E. coli bacteria, carrying cardiac peptide expression construct could effectively suppress tumor growth, angiogenesis, invasion and metastasis and enhance overall survival of mice bearing mammary tumors.

Recombinant protein expression in proteome‐reduced cells under aerobic and oxygen‐limited regimes

“…This, together with lower ATP requirements (as explained above), may explain the greater formation of biomass and FbFP in strain PR1 under oxygen limitation. The performance of PR1 strain in oxygen‐limited regimes makes it an attractive host for the production of vaccines and antimicrobial peptides under microaerobic conditions, which can be preferred over aerobic regimes, to enhance the product activity (Abdolmohammadi Khiav & Zahmatkesh, 2021; Christmann et al, 2023; Samadi et al, 2021).…”

Anti-cancer therapeutic strategies based on HGF/MET, EpCAM, and tumor-stromal cross talk

“…One of the problems with using antibodies is their large size; thus, researchers are trying to increase the tumor permeability by replacing antibodies with smaller peptides and aptamers [143,207]. Using specific peptides instead of antibodies could solve this limitation (large size) and enhance the tumor permeability.…”

Bacteria-Based Nanoprobes for Cancer Therapy

“…186 Many molecules could activate apoptosis of cancer cells, which could be further investigated. 187 Aside from apoptosis induction, disruption of tumor angiogenesis is also promising in retarding tumor growth. Basic fibroblast growth factor (BFGF), as a compensative molecule of VEGF, can cause resistance to anti-VEGF therapy and is regarded as another potential antitumor target.…”