Systemic delivery of modified mRNA encoding herpes simplex virus 1 thymidine kinase for targeted cancer gene therapy

Mol Ther. 2013 Feb;21(2):358-67. doi: 10.1038/mt.2012.250. Epub 2012 Dec 11.

Abstract

Failure of clinical trials of nonviral vector-mediated gene therapy arises primarily from either an insufficient transgene expression level or immunostimulation concerns caused by the genetic information carrier (e.g., bacteria-generated, double-stranded DNA (dsDNA)). Neither of these issues could be addressed through engineering-sophisticated gene delivery vehicles. Therefore, we propose a systemic delivery of chemically modified messenger RNA (mRNA) as an alternative to plasmid DNA (pDNA) in cancer gene therapy. Modified mRNA evaded recognition by the innate immune system and was less immunostimulating than dsDNA or regular mRNA. Moreover, the cytoplasmic delivery of mRNA circumvented the nuclear envelope, which resulted in a higher gene expression level. When formulated in the nanoparticle formulation liposome-protamine-RNA (LPR), modified mRNA showed increased nuclease tolerance and was more effectively taken up by tumor cells after systemic administration. The use of LPR resulted in a substantial increase of the gene expression level compared with the equivalent pDNA in the human lung cancer NCI-H460 carcinoma. In a therapeutic model, when modified mRNA encoding herpes simplex virus 1-thymidine kinase (HSV1-tk) was systemically delivered to H460 xenograft-bearing nude mice, it was significantly more effective in suppressing tumor growth than pDNA.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Alanine Transaminase / analysis
  • Alanine Transaminase / metabolism
  • Animals
  • Apoptosis
  • Aspartate Aminotransferases / analysis
  • Aspartate Aminotransferases / metabolism
  • Blood Urea Nitrogen
  • Cell Line, Tumor
  • Colony-Forming Units Assay
  • Disease Models, Animal
  • Gene Expression
  • Genetic Therapy / methods*
  • Genetic Vectors
  • Herpesvirus 1, Human / genetics*
  • Humans
  • In Situ Nick-End Labeling
  • Liposomes / chemistry
  • Mice
  • Mice, Nude
  • Nanoparticles / chemistry
  • Neoplasms / therapy
  • Nuclear Envelope / genetics
  • Nuclear Envelope / metabolism
  • Plasmids / genetics
  • Protamines / chemistry
  • RNA, Messenger / chemistry
  • RNA, Messenger / genetics*
  • Thymidine Kinase / genetics*
  • Thymidine Kinase / metabolism
  • Transfection
  • Transgenes

Substances

  • Liposomes
  • Protamines
  • RNA, Messenger
  • Aspartate Aminotransferases
  • Alanine Transaminase
  • Thymidine Kinase