830. Determination of the Profile and Site Preference of Lentiviral Vector Integration In Vivo

Caroline V Hacker,Conrad V Vink,Susan M Kingsman,Kyriacos A Mitrophanous,James E Miskin

doi:10.1016/j.ymthe.2006.08.915

Caroline V Hacker, Conrad V Vink + Show 3 more

Open Access

https://doi.org/10.1016/j.ymthe.2006.08.915

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Abstract

Top of pageAbstract Oxford BioMedica (OBM) has developed minimal lentiviral vectors based on Equine Infectious Anaemia Virus (EIAV), and these are currently being developed for use in the clinic. One advantage of lentiviral vectors over other retroviral vector systems is that they can integrate in non-dividing cells such as neurons. OBM have now developed ProSavin|[reg]|, which is an EIAV-based vector for the treatment for Parkinson's disease. ProSavin that has demonstrated efficacy in industry standard models of Parkinson's disease and it is hoped that ProSavin will enter clinical evaluation in the near future. Integration of retroviruses and retroviral vectors within the target cell genome has been shown to be non-random. Human immunodeficiency virus (HIV) and HIV-based vectors have a bias for integrating into genes, and more specifically genes that are active following infection with HIV. Murine leukaemia virus (MLV) integration favours transcriptional start sites whilst Avian sarcoma-leukosis virus (ASLV) shows only a mild preference for transcriptional start sites. OBM has used ligation mediated PCR (LM-PCR) to map 458 integration sites for EIAV based lentiviral vectors and 162 control sites for HIV based lentiviral vectors in vitro. By comparison of these two data sets to a control data set that was generated in silico, we have shown that, like HIV, EIAV based vectors show a preference for active genes. In contrast to published analysis of MLV integration, EIAV and HIV do not show a preference for the transcriptional start site or promoter region of genes. An investigation of the integration profile of EIAV-based vectors in vivo has now been initiated. Template DNA was isolated either from EIAV vector-treated rat striatum tissue (following stereotactic injection into the brain), or from murine ocular tissue following administration of vector into the eye. The results of EIAV integration analysis will be reported.

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