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Modifications to the INSM1 promoter to preserve specificity and activity for use in adenoviral gene therapy of neuroendocrine carcinomas

Cancer Gene Therapy volume 19, pages 828–838 (2012)Cite this article

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Abstract

The INSM1 gene encodes a transcriptional repressor that is exclusively expressed in neuronal and neuroendocrine tissue during embryonic development that is re-activated in neuroendocrine tumors. Using the 1.7 kbp INSM1 promoter, an adenoviral HSV thymidine kinase gene therapy was tested for the treatment of neuroendocrine tumors. An unforeseen interference on the INSM1 promoter specificity from the adenoviral genome was observed. Attempts were made to protect the INSM1 promoter from the influence of essential adenoviral sequences and to further enhance the tissue specificity of the INSM1 promoter region. Using the chicken β-globin HS4 insulator sequence, we eliminated off-target tissue expression from the Ad-INSM1 promoter-luciferase2 constructs in vivo. In addition, inclusion of two copies of the mouse nicotinic acetylcholine receptor (n(AchR)) neuronal-restrictive silencer element (NRSE) reduced nonspecific activation of the INSM1 promoter both in vitro and in vivo. Further, inclusion of both the HS4 insulator with the n(AchR) 2 × NRSE modification showed a two log increase in luciferase activity measured from the NCI-H1155 xenograft tumors compared with the original adenovirus construct. The alterations increase the therapeutic potential of adenoviral INSM1 promoter-driven suicide gene therapy for the treatment of a variety of neuroendocrine tumors.

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Acknowledgements

This work was performed in conjunction with the Diana Helis Henry Medical Research Foundation. This work was supported by the Research Institute for Children, Children’s Hospital, New Orleans, Louisiana and Louisiana State University Health Sciences Center.

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Authors and Affiliations

  1. Research Institute for Children, Children’s Hospital, New Orleans, LA, USA

    V Akerstrom, C Chen, M S Lan & M B Breslin

  2. Diana Helis Henry Medical Research Foundation, New Orleans, LA, USA

    V Akerstrom, C Chen, M S Lan & M B Breslin

  3. Department of Pediatrics and Genetics, LSUHSC, New Orleans, LA, USA

    M S Lan

  4. Departments of Pediatrics and Biochemistry and Molecular Biology, LSUHSC New Orleans, New Orleans, LA, USA

    M B Breslin

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  1. V Akerstrom
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Correspondence to M B Breslin.

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Akerstrom, V., Chen, C., Lan, M. et al. Modifications to the INSM1 promoter to preserve specificity and activity for use in adenoviral gene therapy of neuroendocrine carcinomas. Cancer Gene Ther 19, 828–838 (2012). https://doi.org/10.1038/cgt.2012.66

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