Inhibition of neovascularisation in human endothelial cells using anti NRP-1 nanobody fused to truncated form of diphtheria toxin as a novel immunotoxin.
Shamsi NaderiReyhaneh RoshanMahdi BehdaniFatemeh Kazemi-LomedashtPublished in: Immunopharmacology and immunotoxicology (2021)
Neuropilin-1 (NRP-1) regulates a range of physiological and pathological processes, including angiogenesis. Targeting of NRP1 is considered a significant approach in cancer therapy. In the present study, a novel antiNRP1 immunotoxin (αNRP1 IT) was developed by genetic fusion of a single domain (VHH) anti-NRP-1 antibody fragment to a truncated diphtheria toxin. The αNRP1 IT was expressed into bacterial cells as an inclusion body (IB). Expression of αNRP1 IT was confirmed by SDS-PAGE and western blotting. Recombinant αNRP1 IT was purified using nickel affinity chromatography. Toxicity and antiangiogenesis effect of αNRP1 IT was investigated both in vitro and in vivo. Results showed that αNRP1 IT significantly reduced the viability of human umbilical vein endothelial cell line (HUVEC) (p < .05). The αNRP1 IT significantly inhibited tube formation of HUVEC cells (p < .001). Furthermore, αNRP1 IT inhibited angiogenesis in Chick Chorioallantoic Membrane (CAM) Assay. These data suggest the potential of αNRP1 IT as a novel therapeutic in targeted cancer therapy.
Keyphrases
- endothelial cells
- cancer therapy
- escherichia coli
- induced apoptosis
- vascular endothelial growth factor
- south africa
- poor prognosis
- machine learning
- climate change
- artificial intelligence
- cell cycle arrest
- dna methylation
- deep learning
- cell death
- endoplasmic reticulum stress
- liquid chromatography
- electronic health record
- reduced graphene oxide
- carbon nanotubes
- pi k akt