title: Comparative analysis of the transduction efficiency of five adeno associated virus serotypes and VSV-G pseudotype lentiviral vector in lung cancer cells.
abstract: BACKGROUND
  Lung cancer is the leading cause of cancer-related deaths in the US. Recombinant vectors based on adeno-associated virus (AAV) and lentivirus are promising delivery tools for gene therapy due to low toxicity and long term expression. The efficiency of the gene delivery system is one of the most important factors directly related to the success of gene therapy.
  
  
  METHODS
  We infected SCLC cell lines, SHP-77, DMS 53, NCI-H82, NCI-H69, NCI-H727, NCI-H1155, and NSCLC cell lines, NCI-H23, NCI-H661, and NCI-H460 with VSV-G pseudo-typed lentivirus or 5 AAV serotypes, AAV2/1, AAV2/2, AAV2/4, AAV2/5, and AAV2/8 expressing the CMV promoter mCherry or green fluorescent protein transgene (EGFP). The transduction efficiency was analyzed by fluorescent microscopy and flow cytometry.
  
  
  RESULTS
  Of all the serotypes of AAV examined, AAV2/1 was the optimal serotype in most of the lung cancer cell lines except for NCI-H69 and NCI-H82. The highest transduction rate achieved with AAV2/1 was between 30-50% at MOI 100. Compared to all AAV serotypes, lentivirus had the highest transduction efficiency of over 50% at MOI 1. Even in NCI-H69 cells resistant to all AAV serotypes, lentivirus had a 10-40% transduction rate. To date, AAV2 is the most widely-used serotype to deliver a transgene. Our results showed the transduction efficiency of AAVs tested was AAV2/1 > AA2/5 = AAV2/2> > AAV2/4 and AAV2/8.
  
  
  CONCLUSIONS
  This study demonstrated that VSV-G pseudotyped lentivirus and AAV2/1 can mediate expression of a transgene for lung cancer gene therapy. Overall, our results showed that lentivirus is the best candidate to deliver a transgene into lung cancer cells for treatment.
authors.affiliation: Research Institute For Children, Children's Hospital New Orleans, New Orleans, LA 70118, USA.
authors.forename: Chiachen
authors.initials: C
authors.lastname: Chen
authors.forename: Victoria
authors.initials: V
authors.lastname: Akerstrom
authors.forename: James
authors.initials: J
authors.lastname: Baus
authors.forename: Michael S
authors.initials: MS
authors.lastname: Lan
authors.forename: Mary B
authors.initials: MB
authors.lastname: Breslin
mesh_terms: D045744:Cell Line, Tumor; D000229:Dependovirus / Q000235:genetics* / Q000502:physiology; D015316:Genetic Therapy / Q000295:instrumentation / Q000379:methods*; D005822:Genetic Vectors / Q000235:genetics / Q000502:physiology; D006801:Humans; D016086:Lentivirus / Q000235:genetics* / Q000378:metabolism; D008175:Lung Neoplasms / Q000235:genetics* / Q000628:therapy* / Q000821:virology; D008562:Membrane Glycoproteins / Q000235:genetics* / Q000378:metabolism; D014161:Transduction, Genetic / Q000295:instrumentation / Q000379:methods*; D014759:Viral Envelope Proteins / Q000235:genetics* / Q000378:metabolism
chemical_list: C028391:G protein, vesicular stomatitis virus; D008562:Membrane Glycoproteins; D014759:Viral Envelope Proteins