Intracellular Protein Delivery System Using a Target-Specific Repebody and Translocation Domain of Bacterial Exotoxin

ACS Chem Biol. 2017 Nov 17;12(11):2891-2897. doi: 10.1021/acschembio.7b00562. Epub 2017 Oct 25.

Abstract

With the high efficacy of protein-based therapeutics and plenty of intracellular drug targets, cytosolic protein delivery in a cell-specific manner has attracted considerable attention in the field of precision medicine. Herein, we present an intracellular protein delivery system based on a target-specific repebody and the translocation domain of Pseudomonas aeruginosa exotoxin A. The delivery platform was constructed by genetically fusing an EGFR-specific repebody as a targeting moiety to the translocation domain, while a protein cargo was fused to the C-terminal end of the delivery platform. The delivery platform was revealed to efficiently translocate a protein cargo to the cytosol in a target-specific manner. We demonstrate the utility and potential of the delivery platform by showing a remarkable tumor regression with negligible toxicity in a xenograft mice model when gelonin was used as the cytotoxic protein cargo. The present platform can find wide applications to the cell-selective cytosolic delivery of diverse proteins in many areas.

MeSH terms

  • ADP Ribose Transferases / chemistry*
  • Animals
  • Antineoplastic Agents, Phytogenic / administration & dosage*
  • Antineoplastic Agents, Phytogenic / therapeutic use
  • Bacterial Toxins / chemistry*
  • Cell Line, Tumor
  • Drug Carriers / chemistry*
  • Drug Delivery Systems
  • Exotoxins / chemistry*
  • Humans
  • Male
  • Mice
  • Mice, Inbred BALB C
  • Models, Molecular
  • Neoplasms / drug therapy*
  • Protein Domains
  • Pseudomonas aeruginosa Exotoxin A
  • Ribosome Inactivating Proteins, Type 1 / administration & dosage*
  • Ribosome Inactivating Proteins, Type 1 / therapeutic use
  • Virulence Factors / chemistry*

Substances

  • Antineoplastic Agents, Phytogenic
  • Bacterial Toxins
  • Drug Carriers
  • Exotoxins
  • Ribosome Inactivating Proteins, Type 1
  • Virulence Factors
  • GEL protein, Gelonium multiflorum
  • ADP Ribose Transferases