ArticleMicrobial cell factories2018
Secretion of tumoricidal human tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) by recombinant Lactococcus lactis: optimization of in vitro synthesis conditions.
Article in Microbial cell factories, 2018. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 11 papers.
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Who cites it
11 citing papers in PubMed, 20 citations in OpenAlex.
- Potential applications of engineered bacteria in disease diagnosis and treatment.Microbiome research reports · 2025Review
- Effects ofAnnals of medicine and surgery (2012) · 2024Review
- EV-T synergizes with AZD5582 to overcome TRAIL resistance through concomitant suppression of cFLIP, MCL-1, and IAPs in hepatocarcinoma.Journal of molecular medicine (Berlin, Germany) · 2022Article
- A De Novo Optimized Cell-Free System for the Expression of Soluble and Active Human Tumor Necrosis Factor-Alpha.Biology · 2022Article
- Engineered Bacteria-Based Living Materials for Biotherapeutic Applications.Frontiers in bioengineering and biotechnology · 2022Review
- Recent advancements in the exploitation of the gut microbiome in the diagnosis and treatment of colorectal cancer.Bioscience reports · 2021Review
- The Anti-Tumor Effect ofCancers · 2021Article
- Plasmid Replicons for the Production of Pharmaceutical-Grade pDNA, Proteins and Antigens byInternational journal of molecular sciences · 2021Review
- Generation of the tumor-suppressive secretome from tumor cells.Theranostics · 2021Article
- Construction of Genetically Modified Lactococcus lactis Producing Anti-human-CTLA-4 Single-Chain Fragment Variable.Molecular biotechnology · 2020Article
- Identification and characterization of a moonlighting protein-enolase for surface display in Streptococcus thermophilus.Microbial cell factories · 2020Article
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Authors and funding
7 authors at 1 institution in 1 country.
Funding
Abstract
backgroundTumor necrosis factor-related apoptosis-inducing ligand (TRAIL) selectively eliminates tumor cells. However, the short biological half-life of this molecule limits its potential use in the clinic. Our aim was to construct a recombinant strain of nonpathogenic Lactococcus lactis bacteria as a vector for effective and prolonged human TRAIL production. Herein, we examined the expression and secretion conditions leading to the production of biologically active protein in vitro.
resultsThe human soluble TRAIL-cDNA (hsTRAIL-cDNA) with optimized codons was designed to fit the codon usage pattern (codon bias) of the L. lactis host. This cDNA construct was synthesized and cloned in lactococcal plasmid secretion vector pNZ8124 under the control of the nisin-induced PnisA promoter. The pNZ8124-hsTRAIL plasmid vector was transformed into the L. lactis NZ9000 host strain cells by electroporation. Secretion of the protein occurred at the neutral pH during induction, with optimized concentration of the inducer and presence of serine proteases inhibitor. Using Western blotting and amino acid sequencing method we found that TRAIL was secreted in two forms, as visualized by the presence of two distinct molecular size bands, both deprived of the usp45 protein, the bacterial signal peptide. By the use of MTS assay we were able to prove that hsTRAIL present in supernatant from L. lactis (hsTRAIL+) broth culture was cytotoxic to human HCT116 colon cancer cells but not to normal human fibroblasts. Flow cytometry analysis revealed TRAIL-induced apoptosis of cancer cells.
conclusionsWe designed recombinant L. lactis bacteria, which efficiently produce biologically active, anti-tumorigenic human TRAIL in vitro. Further studies in tumor-bearing NOD-SCID mice will reveal whether the TRAIL-secreting L. lactis bacteria can be used as a safe carrier of this protein, capable of inducing effective elimination of human colon cancer cells in vivo.
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