Tuned Density of Anti-Tissue Factor Antibody Fragment onto siRNA-Loaded Polyion Complex Micelles for Optimizing Targetability into Pancreatic Cancer Cells

Biomacromolecules. 2018 Jun 11;19(6):2320-2329. doi: 10.1021/acs.biomac.8b00507. Epub 2018 May 22.

Abstract

Antibody fragment (Fab')-installed polyion complex (PIC) micelles were constructed to improve targetability of small interfering RNA (siRNA) delivery to pancreatic cancer cells. To this end, we synthesized a block copolymer of azide-functionalized poly(ethylene glycol) and poly(l-lysine) and prepared PIC micelles with siRNA. Then, a dibenzylcyclooctyne (DBCO)-modified antihuman tissue factor (TF) Fab' was conjugated to azido groups on the micellar surface. A fluorescence correlation spectroscopic analysis revealed that 1, 2, or 3 molecule(s) of Fab'(s) were installed onto one micellar nanoparticle according to the feeding ratio of Fab' (or DBCO) to micelle (or azide). The resulting micelles exhibited ∼40 nm in hydrodynamic diameter, similar to that of the parent micelles before Fab' conjugation. Flow cytometric analysis showed that three molecules of Fab'-installed PIC micelles (3(Fab')-micelles) had the highest binding affinity to cultured pancreatic cancer BxPC3 cells, which are known to overexpress TF on their surface. The 3(Fab')-micelles also exhibited the most efficient gene silencing activity against polo-like kinase 1 mRNA in the cultured cancer cells. Furthermore, the 3(Fab')-micelles exhibited high penetrability and the highest cellular internalization amounts in BxPC3 spheroids compared with one or two molecule(s) of Fab'-installed PIC micelles. These results demonstrate the potential of anti-TF Fab'-installed PIC micelles for active targeting of stroma-rich pancreatic tumors.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antibodies, Neoplasm* / chemistry
  • Antibodies, Neoplasm* / pharmacology
  • Cell Cycle Proteins / antagonists & inhibitors*
  • Cell Cycle Proteins / biosynthesis
  • Cell Cycle Proteins / genetics
  • Cell Line, Tumor
  • Drug Delivery Systems*
  • Gene Silencing*
  • Humans
  • Immunoglobulin Fab Fragments* / chemistry
  • Immunoglobulin Fab Fragments* / pharmacology
  • Micelles*
  • Pancreatic Neoplasms / drug therapy*
  • Pancreatic Neoplasms / enzymology
  • Pancreatic Neoplasms / genetics
  • Pancreatic Neoplasms / pathology
  • Polo-Like Kinase 1
  • Polyethylene Glycols / chemistry
  • Polyethylene Glycols / pharmacology
  • Polylysine / chemistry
  • Polylysine / pharmacology
  • Protein Serine-Threonine Kinases / antagonists & inhibitors*
  • Protein Serine-Threonine Kinases / biosynthesis
  • Protein Serine-Threonine Kinases / genetics
  • Proto-Oncogene Proteins / antagonists & inhibitors*
  • Proto-Oncogene Proteins / biosynthesis
  • Proto-Oncogene Proteins / genetics
  • RNA, Small Interfering* / chemistry
  • RNA, Small Interfering* / genetics
  • RNA, Small Interfering* / pharmacology
  • Thromboplastin / antagonists & inhibitors*
  • Thromboplastin / metabolism

Substances

  • Antibodies, Neoplasm
  • Cell Cycle Proteins
  • Immunoglobulin Fab Fragments
  • Micelles
  • Proto-Oncogene Proteins
  • RNA, Small Interfering
  • Polylysine
  • Polyethylene Glycols
  • Thromboplastin
  • Protein Serine-Threonine Kinases