A conditionally replicative adenovirus, CRAd-S-pK7, can target endometriosis with a cell-killing effect

Paupoo AA, A A V Paupoo, Arasen A.V. Paupoo, Zhu ZB, Z B Zhu, Zecong Zhu, Wang M, M Wang, Daniel Rein, D T Rein, Rein DT, Starzinski-Powitz A, Anna Starzinski‐Powitz, A Starzinski-Powitz, David T. Curiel, D T Curiel, Curiel DT
Human Reproduction · 2010 · vol. 25(8) , pp. 2068–2083 · doi:10.1093/humrep/deq137 · PMID:20573677 · PMC2907227 · W2144800415
Lab / animal OA: bronze CC0 ⤵ 3 in-corpus citations
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CRAd-S-pK7, a conditionally replicative adenovirus utilizing the survivin promoter and pK7 fiber modification, demonstrated superior infection and cell-killing effects against endometriosis cells in vitro.

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Abstract

BACKGROUND: Novel therapeutic approaches for endometriosis based on molecular strategies may prove to be useful. Conditionally replicative adenoviruses (CRAds) are designed to exploit key differences between target and normal cells. The wild-type adenovirus (Adwt) promoter can be replaced by tissue-specific promoters, allowing viral replication only in target cells. Viral infectivity can be enhanced by altering Ad tropism via fiber modification. We investigated whether CRAds can be used to target endometriosis and determined the most efficient transcriptional- and transductional-targeting strategy. METHODS: An in vitro study was carried out using human endometriotic cell lines, 11Z (epithelial) and 22B (stromal), normal human ovarian surface epithelial cell line (NOSE006) and primary human endometriosis cells. A total of 9 promoters and 12 Ad tropism modifications were screened by means of a luciferase reporter assay. From this screening data, three CRAds (CRAd-S-pK7, CRAd-S-RGD, CRAd-S-F5/3sigma1, all incorporating the survivin promoter but with different fiber modifications) were selected to perform experiments using Adwt and a replication-deficient virus as controls. CRAds were constructed using a plasmid recombination system. Viral-binding capacity, rates of entry and DNA replication were evaluated by quantitative real-time PCR of viral genome copy. Cell-killing effects were determined by crystal violet staining and a cell viability assay for different concentrations of viral particles per cell. RESULTS: Comparison of promoters demonstrated that the survivin promoter exhibited the highest induction in both endometriotic cell lines. Among the fiber-modified viruses, the polylysine modification (pK7) showed the best infection enhancement. CRAd-S-pK7 was validated as the optimal CRAd to target endometriosis in terms of binding ability, entry kinetics, DNA replication and cell-killing effect. CRAd-S-pK7 also exhibited a high level of DNA replication in primary endometriosis cells. CONCLUSIONS: CRAd-S-pK7 has the best infection and cell-killing effect in the context of endometriosis. It could prove to be a useful novel method to target refractory cases of endometriosis.

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Condition tags

endometriosis

MeSH descriptors

Adenoviridae Endometriosis Virus Replication Virus Replication Virus Replication Adenoviridae Adenoviridae Cell Line Cell Line Endometriosis Endometriosis Endometriosis Female Female Humans Humans Promoter Regions, Genetic Promoter Regions, Genetic Virus Attachment Virus Attachment

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