Biosafety in Handling Gene Transfer Vectors

Andrew Braun1

1 Harvard Medical School, Boston, Massachusetts
Publication Name:  Current Protocols in Human Genetics
Unit Number:  Unit 12.1
DOI:  10.1002/0471142905.hg1201s50
Online Posting Date:  August, 2006
GO TO THE FULL TEXT: PDF or HTML at Wiley Online Library

Abstract

This unit is devoted to safety issues that must be considered when generating and working with vectors for gene therapy. A short description of the relevant regulatory bodies and mechanisms is followed by a general discussion of containment and safety procedures. This is followed by vector‐specific guidelines for nine commonly used vector systems, including adenovirus, vaccinia, HSV‐1, lentivirus, and nonviral vectors. Recent tragic experiences with retroviral integration near oncogene loci demonstrate the surprise effects of successful gene therapy. The unit also outlines the problems encountered when taking a promising vector to clinical trials.

Keywords: biosafety; gene transfer; vectors; hazards; risks

     
 
GO TO THE FULL PROTOCOL:
PDF or HTML at Wiley Online Library

Table of Contents

  • Introduction
  • Background
  • Running A BL‐2 Lab
  • Specific Vectors
  • Getting a Useful Vector into Clinical Trials
  • Further Reading
  • Literature Cited
  • Figures
  • Tables
     
 
GO TO THE FULL PROTOCOL:
PDF or HTML at Wiley Online Library

Materials

GO TO THE FULL PROTOCOL:
PDF or HTML at Wiley Online Library

Figures

Videos

Literature Cited

   Abken, H., Butzler, C., and Willecke, K. 1987. Adenovirus type 5 persisting in human lymphocytes is unlikely to be involved in immortalization of lymphoid cells by fusion with cytoplasts or by transfection with DNA of mouse L cells. Anticancer Res. 7:553‐558.
   Bassin, R.H., Tuttle, N., and Fischinger, P.J. 1971. Rapid cell culture assay technique for murine leukaemia viruses. Nature 229:564‐566.
   Büning, H., Ried, M.U, Perabo, L., Gerner, F.M., Huttner N.A., Enssle, J., and Hallek M. 2003. Receptor targeting of adeno‐associated virus vectors. Gene Ther. 10:1142‐1151.
   Burns, P.A., Jack, A., Neilson, F., Haddow, S., and Balmain, A. 1991. Transformation of mouse skin endothelial cells in vivo by direct application of plasmid DNA encoding the human T24 h‐ras oncogene. Oncogene 6:1973‐1978.
   Crystal, R.G., McElvaney, N.G., Rosenfeld, M.A., Chu, C.S., Mastrangeli, A., Hay, J.G., Brody, S.L., Jaffe, H.A., Eissa, N.T., and Danel, C. 1994. Administration of an adenovirus containing the human CFTR cDNA to the respiratory tract of individuals with cystic fibrosis. Nat. Genet. 8:42‐51.
   De Palma, M., Montini, E., de Sio, F.R.S., Benedicenti, F., Gentile, A., Medico, E., and Naldini, L. 2005. Promoter trapping reveals significant differences in integration site selection between MLV And HIV vectors in primary hematopoietic cells Blood 105:2307‐2315.
   Doerfler, W. 1991. Abortive infection and malignant transformation by adenoviruses: Integration of viral DNA and control of viral gene expression by specific patterns of DNA methylation. Adv. Virus Res. 39:89‐128.
   Fallaux, F.J., Bout, A., van der Velde, I., van den Wollenberg, D.J.M., Hehir, K., Keegan, J., Auger, C., Cramer, S.J., van Ormondt, H., van der Eb, A.J., Valerio, D., and Hoeben, R.C. 1998. New helper cells and matched early region 1‐deleted adenovirus vectors prevent generation of replication‐competent adenoviruses. Hum. Gene Ther. 9:1909‐1917.
   Fleming, D.O. and Hunt, D.L. (eds.) 2000. Biological Safety: Principles and Practices, 3rd Edition. ASM Press, Washington, D.C.
   Fox, J.P., Brandt, C.D., Wassermann, F.E., Hall, C.E., Spigland, I., Kogon, A., and Elveback, L.R. 1969. The virus watch program: A continuing surveillance of viral infections in metropolitan New York families. VI. Observations of adenovirus infections: Virus excretion patterns, antibody response, efficiency of surveillance, patterns of infections, and relation to illness. Am. J. Epidemiol. 89:25‐50.
   Fraefel, C., Song, S., Lim, F., Lang, P., Yu, L., Wang, Y., Wild, P., and Geller, A.I. 1996. Helper virus‐free transfer of herpes simplex virus type 1 plasmid vectors into neural cells. J. Virol. 70:7190‐7197.
   Gallimore, P.H. 1972. Tumor production in immunosupressed rats with cells transformed in vitro by Ad2. J. Gen. Virol. 62:414‐4126.
   Glover, D.J., Lipps, H.J., and Jans, D.A. 2005. Towards safe, non‐viral therapeutic gene expression in humans. Nat. Rev. Genet. 6:299‐310.
   Hanawa, H., Persons, D.A., and Nienhuis, A.W. 2005. Mobilization and mechanism of transcription of integrated self‐inactivating lentiviral vectors J. Virol. 79:8410‐8421.
   Harvey, B.G., Maroni, J., O'Donoghue, K.A., Chu, K.W., Muscat, J.C., Pippo, A.L., Wright, C.E., Hollman, C., Wisnivesky, J.P., Kessler, P.D., Rasmussen, H.S., Rosengart, T.K., and Crystal, R.G. 2002. Safety of local delivery of low‐ and intermediate‐dose adenovirus gene transfer vectors to individuals with a spectrum of morbid conditions. Hum. Gene Ther. 13:15‐63.
   Hacein‐Bey‐Abina, S., Le Deist, F., Carlier, F., Bouneaud, C., Hue, C., De Villartay, J.P., Thrasher, A.J., Wulffraat, N., Sorensen, R., Dupuis‐Girod, S., Fischer, A., Davies, E.G., Kuis, W., Leiva, L., and Cavazzana‐Calvo, M. 2002. Sustained correction of X‐linked severe combined immunodeficiency by ex vivo gene therapy. New Engl. J. Med. 346:1185‐1193.
   Hacein‐Bey‐Abina, S., von Kalle, C., Schmidt, M., McCormack, M.P., Wulffraat, N., Leboulch, P., Lim, A., Osborne, C.S., Pawliuk, R., Morillon, E., Sorensen, R., Forster, A., Fraser, P., Cohen, J.I., de Saint Basile, G., Alexander, I., Wintergerst, U., Frebourg, T., Aurias, A., Stoppa‐Lyonnet, D., Romana, S., Radford‐Weiss, I., Gross, F., Valensi, F., Delabesse, E., Macintyre, E., Sigaux, F., Soulier, J., Leiva, L.E., Wissler, M., Prinz, C., Rabbitts, T.H., Le Deist, F., Fischer, A., and Cavazzana‐Calvo, M. LMO2‐associated clonal T cell proliferation in two patients after gene therapy for SCID‐X1. 2003. Science. 302:415‐419.
   Published erratum appears in Science 302:568.
   Harui, A., Suzuki, S., Kochanek, S., and Mitani, K. 1999. Frequency and stability of chromosomal integration of adenovirus vectors. J. Virol. 73:6141‐6146.
   High, K.A. 2003. AAV‐mediated gene transfer for hemophilia. Trans. Am. Clin. Climatol. Assoc. 114:337‐351.
   Ibelgaufts, H. and Jones, K.W. 1983. Papovavirus‐related RNA sequences in human neurogenic tumours. Acta Neuropathol. 56:118‐122.
   Johnston, K.M., Jacoby, D., Pechan, P.A., Fraefel, C., Borghesani, P., Schuback, D., Dunn, R.J., Smith, F.I., and Breakefield, X.O. 1997. HSV/AAV hybrid amplicon vectors extend transgene expression in human glioma cells. Hum. Gene Ther. 8:359‐370.
   Kochanek, S., Clemens, P.R., Mitani, K., Chen, H.H., Chan, S., and Caskey, C.T. 1996. A new adenoviral vector: Replacement of all viral coding sequences with 28 kb of DNA independently expressing both full‐length dystrophin and β‐galactosidase. Proc. Natl. Acad. Sci. U.S.A. 93:5731‐5736.
   Krisky, D.M., Wolfe, D., Goins, W.F., Marconi, P.C., Ramakrishnan, R., Mata, M., Rouse, R.J., Fink, D.J., and Glorioso, J.C. 1998. Deletion of multiple immediate‐early genes from herpes simplex virus reduces cytotoxicity and permits long‐term gene expression in neurons. Gene Ther. 5:1593‐603.
   Laufs, S., Gentner, B., Nagy, K.Z., Jauch, A., Benner, A., Naundorf, S., Kuehlcke, K., Schiedlmeier, B., Ho, A.D., Zeller, W.J, and Fruehauf, S. 2003. Retroviral vector integration occurs in preferred genomic targets of human bone marrow–repopulating cells. Blood 101:2191‐2198.
   Maxfield, L.F., Fraize, C.D., and Coffin, J.M. 2005. Relationship between retroviral DNA‐integration‐site selection and host cell transcription. Proc. Natl. Acad. Sci. U.S.A. 102:1436‐1441.
   McCarty, D.M., Young Jr. S.M., and Samulski, R.J. 2004. Integration of adeno‐associated virus (AAV) and recombinant AAV vectors. Annu. Rev. Genet. 38:819‐845.
   McCormack, M.P. and Rabbitts, T.H. 2004. Activation of the t‐cell oncogene LMO2 after gene therapy for X‐linked severe combined immunodeficiency New Engl. J. Med. 350:913‐922.
   McElvaney, N.G. and Crystal, R.G. 1995. IL‐6 release and airway administration of human CFR cDNA adenovirus vector. Nat. Med. 1:182‐184.
   Meyer, H., Sutte, G., and Mayer, A. 1991. Mapping of deletions in the genome of the highly attenuated vaccinia virus MVA and their influence on virulence. J. Gen. Virol. 72:1031‐1038.
   Mostoslavsky, G., Kotton, D.N., Fabian, A.J., Gray, J.T, Lee, J‐S., and Mulligan, R.C. 2005. Efficiency of transduction of highly purified murine hematopoietic stem cells by lentiviral and oncoretroviral vectors under conditions of minimal in vitro manipulation. Mol. Ther. 11:932‐940.
   Naldini, L., Blomer, U., Gallay, P., Ory, D., Mulligan, R., Gage, F.H., Verma, I.M., and Trono, D. 1996. In vivo gene delivery and stable transduction of non‐dividing cells by lentiviral vector. Science 272:263‐267.
   National Research Council (NRC). 1989. Biosafety in the Laboratory: Prudent Practices for the Handling and Disposal of Infectious Materials. National Academy Press, Washington, D.C.
   NIH Advisory Committee Report. 2002. Assessment of adenoviral vector safety and toxicity: Report of the National Institutes of Health Recombinant DNA Advisory Committee. Hum. Gene Ther. 13:3‐13.
   Printz, M., Reynolds, J., Mento, S.J., Jolly, D., Kowal, K., and Sajjadi, N. 1995. Recombinant retroviral vector interferes with the detection of amphotropic replication competent retrovirus in standard culture assays. Gene Ther. 2:143‐150.
   Raper, S.E., Yudkoff, M., Chirmule, N., Gao, G.P., Nunes, F., Haskal, Z.J., Furth, E.E., Propert, K.J., Robinson, M.B., Magosin, S., Simoes, H., Speicher, L., Hughes, J., Tazelaar, J., Wivel, N.A., Wilson, J.M., and Batshaw, M.L. 2002. A pilot study of in vivo liver‐directed gene transfer with an adenoviral vector in partial ornithine transcarbamylase deficiency. Hum. Gene Ther. 13:163‐175.
   Vogels, R., Zuijdgeest, D., van Rijnsoever, R., Hartkoorn, E., Damen, I., de Bethune, M.P., Kostense, S., Penders, G., Helmus, N., Koudstaal, W., Cecchini, M., Wetterwald, A., Sprangers, M., Lemckert, A., Ophorst, O., Koel, B., van Meerendonk, M., Quax, P., Panitti, L., Grimbergen, J., Bout, A., Goudsmit, J., and Havenga, M. 2003. Replication‐deficient human adenovirus type 35 vectors for gene transfer and vaccination: Efficient human cell infection and bypass of preexisting adenovirus immunity. J. Virol. 77:8263‐8271.
   Wang, S. and Vos, J. 1996. A hybrid herpesvirus infectious vector based on Epstein‐Barr virus and herpes simplex virus type 1 for gene transfer into human cells in vitro and in vivo. J. Virol. 70:8422‐8430.
   Wang, H., Shayakhmetov, D.M., Leege, T., Harkey, M., Li, Q., Papayannopoulou, T., Stamatoyannopolous, G., and Lieber, A.A. 2005. Capsid‐modified helper‐dependent adenovirus vector containing the β‐globin locus control region displays a nonrandom integration pattern and allows stable, erythroid‐specific gene expression. J. Virol. 79:10999‐11013.
   Xiao, X., Li, J., and Samulski, R.J. 1998. Production of high‐titer recombinant adeno‐associated virus vectors in the absence of helper adenovirus. J. Virol. 72:2224‐2232.
   Yohn, D.S., Funk, C.A., and Grace, J.T. Jr. 1967. Sex‐related resistance in hamsters to adenovirus‐12 oncogenesis. II. Influence of virus dose. J. Virol. 1:1186‐1192.
Internet Resources
   http://www.cdc.gov/od/ohs/biosfty/bmbl4/bmbl4toc.htm
  Biosafety in Microbiological and Biomedical Laboratories (BMBL), from the Centers for Disease Control (CDC) Office of Health Safety (OHS) and the NIH.
   http://www.fda.gov/cber/index.html
  FDA Center for Biologics Evaluation and Research (CBER) home page.
   http://www.fda.gov/cber/gdlns/somgene.htm
  FDA guidelines for human somatic cell therapy and gene therapy.
   http://www.fda.gov/cber/gdlns/retrogt1000.pdf
  Guidance for Testing Replication Competent Retrovirus, FDA, October 2000.
   http://www4.od.nih.gov/oba
  NIH Office of Biotechnology Activities (OBA) home page, providing a wide range of guidelines and other useful documents.
   http://www4.od.nih.gov/oba/rac/guidelines_02/APPENDIX_B.htm
  APPENDIX M of the April 2002 NIH Recombinant DNA guidelines, listing recombinant DNA agents by risk groups.
   http://www4.od.nih.gov/oba/rac/guidelines_02/APPENDIX_M.htm
  APPENDIX M of the April 2002 NIH Recombinant DNA Guidelines, consisting of a series of screening questions concerning potential vectors and their application to human gene therapy.
   http://www4.od.nih.gov/oba/rac/PROTOCOL.pdf
  List of human gene transfer studies registered with the NIH.
   http://orf.od.nih.gov/PoliciesAndGuidelines/DesignPolicy/
  NIH Office of Research Facilities' Research Laboratory Design Policy and Guidelines.
   http://www.ehrs.upenn.edu/programs/bio/bsm/pdfs/BioManual.pdf
  Biosafety Manual from the University of Pennsylvania
   http://www.stanford.edu/dept/EHS/prod/researchlab/bio/Biosafety_Manual/Biosafety_Manual_complete.pdf
  Biosafety Manual from Stanford University
   http://web.princeton.edu/sites/ehs/biosafety/biosafetypage/toc.htm
  Biosafety Manual from Princeton University (contains a particularly useful index)
   http://www.ngvl.org/
  Home page of the National Gene Vector Laboratories (NGVL), a group set up under NIH auspices to help researchers obtain clinical‐grade human gene transfer vectors.
   http://agmed.sante.gouv.fr/htm/10/filcoprs/050103en.htm
  Announcement of 3rd SAE in Necker University Hospital X‐SCID series.
GO TO THE FULL PROTOCOL:
PDF or HTML at Wiley Online Library