Student award winner in the Ph.D. category for the 2013 society for biomaterials annual meeting and exposition, april 10-13, 2013, Boston, Massachusetts : biomaterial-mediated cancer-specific DNA delivery to liver cell cultures using synthetic poly(beta-amino ester)s

J Biomed Mater Res A. 2013 Jul;101(7):1837-45. doi: 10.1002/jbm.a.34616. Epub 2013 Apr 5.

Abstract

Liver cancer is a leading cause of cancer death. Most patients are treated by arterial injection of chemoembolizing agents, providing a convenient avenue for local treatment by novel therapies, including gene therapy. Poly(beta-amino ester)s (PBAEs) were synthesized and used to form nanoparticles for non-viral transfection of buffalo rat hepatoma (MCA-RH7777) and hepatocyte (BRL-3A) lines with eGFP and luciferase DNA. Hepatoma cells were transfected with up to (98 ± 0.4)% efficacy with no measurable cytotoxicity. Hepatocytes were transfected with as high as (73 ± 0.4)% efficacy with (10 ± 4)% non-specific cytotoxicity. In contrast, positive controls (branched polyethylenimine, Lipofectamine™ 2000, and X-tremeGENE(®) DNA HP) caused 30-90% toxicity in BRL-3A cells at doses required for >50% transfection. Of the 21 optimized PBAE-DNA formulations tested, 12 showed significant specificity for hepatoma cells over hepatocytes in monoculture (p < 0.05 for both percentage transfected and eGFP expression intensity). Top polymers from eGFP studies also delivered luciferase DNA with 220 ± 30-fold and 470 ± 30-fold greater specificity for hepatoma cells than hepatocytes. Transfections of co-cultures of hepatoma and hepatocytes with eGFP DNA also showed high specificity (1.9 ± 0.1- to 5.8± 1.4-fold more transfected hepatoma cells than hepatocytes, measured by percentage transfected and flow cytometry). By eGFP intensity, up to 530 ±60-fold higher average expression per cell was measured in hepatoma cells. One top formulation caused (95 ± 0.2)% transfection in hepatoma cells and (27 ± 0.2)% in hepatocytes [(96 ± 9)% relative hepatocyte viability]. PBAE-based nanoparticles are a viable strategy for liver cancer treatment, delivering genes to nearly 100% of cancer cells while maintaining high biomaterial-mediated specificity to prevent toxic side-effects on healthy hepatocytes.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Animals
  • Awards and Prizes
  • Biocompatible Materials*
  • Carcinoma, Hepatocellular / therapy
  • Cell Line, Tumor
  • Cell Survival
  • Cells, Cultured
  • Coculture Techniques
  • Drug Delivery Systems*
  • Genetic Therapy / methods*
  • Green Fluorescent Proteins / genetics
  • Liver / cytology*
  • Liver Neoplasms / therapy
  • Luciferases / genetics
  • Nanoparticles
  • Neoplasms / therapy*
  • Polyesters / chemical synthesis
  • Polyesters / chemistry*
  • Rats
  • Transfection

Substances

  • Biocompatible Materials
  • Polyesters
  • Green Fluorescent Proteins
  • Luciferases