Abstract
BackgroundLung 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.MethodsWe 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.ResultsOf 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.ConclusionsThis 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.
Highlights
Lung cancer remains the most common cause of cancerrelated death in USA and worldwide
Transduction efficiency of 5 serotypes recombinant AAV2 (rAAV) vectors in lung cancer cell lines To compare the transduction efficiency of rAAV serotypes in lung cancer cells, six lung cancer cell lines, DMS 53, NCI-H460, SHP-77, NCI-H82, NCI-H1155 and NCI-H69 were infected with Multiplicity of infection (MOI) 100 AAV2/1, AAV2/2, AAV2/4, AAV2/5 and AAV2/8 that carried the CMV promoter–driven mCherry transgene
The results indicate that the lung cancer cells tested were highly resistant to AAV2/4 and AAV2/8 infection
Summary
Lung cancer remains the most common cause of cancerrelated death in USA and worldwide. Adeno-associated virus, AAV, is emerging as one of the most popular gene delivery systems because of a lack of pathogenicity in humans, and its long term and efficient transgene expression in various cells. AAV2 is the most widely used serotype, in cells that lack the necessary cellular receptor components for infection, the transduction efficiency is limited. Many studies suggest that replacement of the AAV2 capsid with the capsid from other AAV serotypes results in increased cell tropism [9]. The transduction efficiency of AAVs in lung cancer cells has not been extensively studied. Several studies indicated AAV1 and AAV5 were the optimal serotypes for infection of lung epithelial cells [11,12,13]. 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
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