title: Systemic delivery of modified mRNA encoding herpes simplex virus 1 thymidine kinase for targeted cancer gene therapy.
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.
authors.affiliation: Division of Molecular Pharmaceutics and Center for Nanotechnology in Drug Delivery, Eshelman School of Pharmacy, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina 27599, USA.
authors.forename: Yuhua
authors.initials: Y
authors.lastname: Wang
authors.forename: Hsing-Hao
authors.initials: HH
authors.lastname: Su
authors.forename: Yang
authors.initials: Y
authors.lastname: Yang
authors.forename: Yunxia
authors.initials: Y
authors.lastname: Hu
authors.forename: Lu
authors.initials: L
authors.lastname: Zhang
authors.forename: Pilar
authors.initials: P
authors.lastname: Blancafort
authors.forename: Leaf
authors.initials: L
authors.lastname: Huang
mesh_terms: D000410:Alanine Transaminase / Q000032:analysis / Q000378:metabolism; D000818:Animals; D017209:Apoptosis; D001219:Aspartate Aminotransferases / Q000032:analysis / Q000378:metabolism; D001806:Blood Urea Nitrogen; D045744:Cell Line, Tumor; D003114:Colony-Forming Units Assay; D004195:Disease Models, Animal; D015870:Gene Expression; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors; D018259:Herpesvirus 1, Human / Q000235:genetics*; D006801:Humans; D020287:In Situ Nick-End Labeling; D008081:Liposomes / Q000737:chemistry; D051379:Mice; D008819:Mice, Nude; D053758:Nanoparticles / Q000737:chemistry; D009369:Neoplasms / Q000628:therapy; D009685:Nuclear Envelope / Q000235:genetics / Q000378:metabolism; D010957:Plasmids / Q000235:genetics; D011479:Protamines / Q000737:chemistry; D012333:RNA, Messenger / Q000737:chemistry / Q000235:genetics*; D013937:Thymidine Kinase / Q000235:genetics* / Q000378:metabolism; D014162:Transfection; D019076:Transgenes
chemical_list: D008081:Liposomes; D011479:Protamines; D012333:RNA, Messenger; D001219:Aspartate Aminotransferases; D000410:Alanine Transaminase; D013937:Thymidine Kinase