MicroRNA 142-3p attenuates spread of replicating retroviral vector in hematopoietic lineage-derived cells while maintaining an antiviral immune response

Amy H. Lin, Nina Timberlake, Christopher R. Logg, Yanzheng Liu, Shuichi Kamijima, Oscar Diago, Kenneth Wong, Dawn K. Gammon, Derek Ostertag, Katrin Hacke, Emily C. Yang, Harry Gruber, Noriyuki Kasahara, Douglas J. Jolly

Research output: Contribution to journalArticle

4 Scopus citations

Abstract

We are developing a retroviral replicating vector (RRV) encoding cytosine deaminase as an anticancer agent for gliomas. Despite its demonstrated natural selectivity for tumors, and other safety features, such a virus could potentially cause off-target effects by productively infecting healthy tissues. Here, we investigated whether incorporation of a hematopoietic lineage-specific microRNA target sequence in RRV further restricts replication in hematopoietic lineage-derived human cells in vitro and in murine lymphoid tissues in vivo. One or four copies of a sequence perfectly complementary to the guide strand of microRNA 142-3p were inserted into the 3' untranslated region of the RRV genome expressing the transgene encoding green fluorescent protein (GFP). Viral spread and GFP expression of these vectors in hematopoietic lineage cells in vitro and in vivo were measured by qPCR, qRT-PCR, and flow cytometry. In hematopoietic lineage-derived human cell lines and primary human stimulated peripheral blood mononuclear cells, vectors carrying the 142-3pT sequence showed a remarkable decrease in GFP expression relative to the parental vector, and viral spread was not observed over time. In a syngeneic subcutaneous mouse tumor model, RRVs with and without the 142-3pT sequences spread equally well in tumor cells; were strongly repressed in blood, bone marrow, and spleen; and generated antiviral immune responses. In an immune-deficient mouse model, RRVs with 142-3pT sequences were strongly repressed in blood, bone marrow, and spleen compared with unmodified RRV. Tissue-specific microRNA-based selective attenuation of RRV replication can maintain antiviral immunity, and if needed, provide an additional safeguard to this delivery platform for gene therapy applications.

Original languageEnglish (US)
Pages (from-to)759-771
Number of pages13
JournalHuman gene therapy
Volume25
Issue number8
DOIs
StatePublished - Aug 1 2014

ASJC Scopus subject areas

  • Molecular Medicine
  • Molecular Biology
  • Genetics

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    Lin, A. H., Timberlake, N., Logg, C. R., Liu, Y., Kamijima, S., Diago, O., Wong, K., Gammon, D. K., Ostertag, D., Hacke, K., Yang, E. C., Gruber, H., Kasahara, N., & Jolly, D. J. (2014). MicroRNA 142-3p attenuates spread of replicating retroviral vector in hematopoietic lineage-derived cells while maintaining an antiviral immune response. Human gene therapy, 25(8), 759-771. https://doi.org/10.1089/hum.2012.216