Anti-c-MET Fab-Grb2-Gab1 Fusion Protein-Mediated Interference of c-MET Signaling Pathway Induces Methuosis in Tumor Cells

Int J Mol Sci. 2022 Oct 10;23(19):12018. doi: 10.3390/ijms231912018.

Abstract

Bio-macromolecules have potential applications in cancer treatment due to their high selectivity and efficiency in hitting therapeutic targets. However, poor cell membrane permeability has limited their broad-spectrum application in cancer treatment. The current study developed highly internalizable anti-c-MET antibody Fab fusion proteins with intracellular epitope peptide chimera to achieve the dual intervention from the extracellular to intracellular targets in tumor therapy. In vitro experiments demonstrated that the fusion proteins could interfere with the disease-associated intracellular signaling pathways and inhibit the uncontrolled proliferation of tumor cells. Importantly, investigation of the underlying mechanism revealed that these protein chimeras could induce vacuolation in treated cells, thus interfering with the normal extension and arrangement of microtubules as well as the mitosis, leading to the induction of methuosis-mediated cell death. Furthermore, in vivo tumor models indicated that certain doses of fusion proteins could inhibit the A549 xenograft tumors in NOD SCID mice. This study thus provides new ideas for the intracellular delivery of bio-macromolecules and the dual intervention against tumor cell signaling pathways.

Keywords: Fab fusion proteins; bio-macromolecules; dual intervention; intracellular delivery; methuosis; signaling pathways.

MeSH terms

  • Adaptor Proteins, Signal Transducing / metabolism
  • Animals
  • Antibodies / metabolism
  • Epitopes
  • GRB2 Adaptor Protein / metabolism
  • Humans
  • Mice
  • Mice, SCID
  • Peptides / chemistry
  • Proto-Oncogene Proteins c-met* / genetics
  • Proto-Oncogene Proteins c-met* / metabolism
  • Signal Transduction*

Substances

  • Adaptor Proteins, Signal Transducing
  • Antibodies
  • Epitopes
  • GAB1 protein, human
  • GRB2 Adaptor Protein
  • GRB2 protein, human
  • Gab1 protein, mouse
  • Peptides
  • Proto-Oncogene Proteins c-met

Grants and funding

This research received no external funding.