Chemistry: molecular biology and microbiology – Treatment of micro-organisms or enzymes with electrical or... – Modification of viruses
Patent
1995-09-29
1997-12-02
Stone, Jacqueline M.
Chemistry: molecular biology and microbiology
Treatment of micro-organisms or enzymes with electrical or...
Modification of viruses
4352401, 4352402, 4353201, 514 44, 530345, 530358, A61K 4800, C07K 308, C12N 500
Patent
active
056935099
DESCRIPTION:
BRIEF SUMMARY
The invention relates to the introduction of nucleic acids into higher eukaryotic cells.
There is a need for an efficient system of introducing nucleic acid into living cells particularly in gene therapy. This involves delivering genes into cells in order to achieve, in vivo, the synthesis of therapeutically effective gene products, e.g. in order to replace a defective gene in the event of a genetic defect.
For transferring genes into the cells, viral vectors are used, for example, which make use of the efficient entry mechanisms of their original viruses. By this is meant viruses in which the gene to be expressed in the cell has been integrated in the genome by recombinant methods. This strategy was used in the construction of recombinant retroviral and adenoviral vectors, in order to achieve a highly effective gene transfer in vitro and in vivo. The most advanced technologies for using nucleic acids in the course of gene therapy use retroviral systems for delivering genes into the cell (Wilson et al., 1990; Kasid et al., 1990). Therefore, methods have already been developed for expanding the applicability of the retroviral systems or for making them specific to a defined cell population, e.g. by altering the tropism of the viruses.
Roux et al., 1989, as well as French Patent Application 2 649 119, described a system which changes the tropism of retroviruses by means of bifunctional conjugates which contain on the one hand an antibody against the virus coat and, on the other hand, a specific cell membrane marker for the target cell and thus establish a connection between the virus and the host cell.
The approach suggested by Goud et al., 1988 is also based on the principle of bifunctional conjugates. These conjugates are a construction of two monoclonal antibodies, one of which is directed against the human transferrin receptor whilst the other is directed against the gp70 coat protein of the Moloney retrovirus. With these conjugates the retrovirus was able to penetrate into the target cells but could not replicate therein.
The method of changing the tropism of a virus described in WO 92/06180 consists in providing the surface of a virus with a molecule which binds to a surface receptor of the target cell, thereby giving the virus a specificity for the target cell which would not naturally be present. WO 92/06180 describes the modification of a retrovirus and hepatitis virus B with carbohydrate molecules which bind to the asialoglycoprotein receptor.
For use in gene therapy, recombinant adenoviruses have recently come to replace recombinant retroviruses to an increasing extent (Berkner, 1988; Stratford-Perricaudet et al., 1990; Rosenfeld et al., 1991; Rosenfeld et al., 1992; Stratford-Perricaudet et al., 1992).
Adenoviral vectors have the advantageous ability to penetrate into non-dividing cells and absorb a foreign DNA sequence to an extent of up to 8.5 kb. Moreover, the adenovirus particles can be thoroughly purified without losing any stability and are produced in titres greater than 10.sup.11 PFUs/ml.
One restriction on the use of the recombinant vectors derived from the adenoviruses Ad2 and Ad5 consists in their limited ability to penetrate into blood cells. However, blood cells are a preferred target for applications in gene therapy since they are readily available and can be re-introduced into the patients, in addition, the methods of obtaining and cultivating blood cells are well established. The reason for the poor activity of the adenoviral vectors in blood cells lies in the obviously small number of receptors for adenoviruses on these cells (Horvath and Weber 1988; Silver and Anderson, 1988). Whilst the binding of the virus to these cells is reduced by a factor of two to five, the internalisation of the bound virus is even less. The low number of receptors at the outset would thus appear to be accompanied by a greatly reduced internalisation of the available receptors.
Recently, in a number of studies, it was proposed to use non-recombinant adenoviruses for gene transfer with DNA complexes by m
REFERENCES:
patent: 5192553 (1993-03-01), Boyse et al.
patent: 5354844 (1994-10-01), Beug et al.
Cotten, M., et al., "High-Efficiency Receptor-Mediated Delivery of Small and Large (48 Kilobase) Gene Constructs Using the Endosome-Disruption Activity of Defective or Chemically Inactivated Adenovirus Particles," Proc. Natl. Acad. Sci. USA 89(13):6094-6098 (1992).
Wagner, E., et al., "Coupling of Adenovirus to Transferrin-Polylysine/DNA Complexes Greatly Enhances Receptor-Mediated Gene Delivery and Expression of Transfected Genes," Proc. Natl. Acd. Sci. USA 89(13):6099-6103 (1992).
Gao et al., Hum. Gene Ther., 4, 1993, 17-24.
Wagner et al., P.N.A.S., 89, 1992, 6099-6103.
Wagner et al., Bioconjugate Chem., 2, 1991, 226-231.
Curiel et al., PNAS, 88, 1991, 8856-8854.
Curiel et al., Human Gene Therapy, 3, 1992, 147-154.
Marshall, Science, 269, 1995, 1050-1055.
Miller et al., FASEB J., 9, 1995, 190-199.
Culver et al., TIG, 10(5), 1994, 174-178.
Hodgson, Exp. Opin. Ther. Pat., 5(5), 1995, 459-468.
Cotten Matthew
Wagner Ernst
Boehringer Ingelheim International GmbH
Genentech Inc.
Milne Andrew
Stone Jacqueline M.
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