Chemistry: molecular biology and microbiology – Process of mutation – cell fusion – or genetic modification – Introduction of a polynucleotide molecule into or...
Reexamination Certificate
1999-04-29
2001-10-16
Schwartzman, Robert A. (Department: 1636)
Chemistry: molecular biology and microbiology
Process of mutation, cell fusion, or genetic modification
Introduction of a polynucleotide molecule into or...
C435S320100, C435S325000, C435S357000, C435S367000, C435S372300, C435S456000
Reexamination Certificate
active
06303380
ABSTRACT:
BACKGROUND OF THE INVENTION
The present invention is in the field of retroviral vectors for gene therapy.
Recent progress in human genetics, especially in human genome research and gene transfer techniques, has brought gene therapy closer to clinical reality. Experimental human gene therapy has enjoyed some successes in the treatment of genetic defects, including adenosine deaminase deficiency, (Blaese R M, et al.
Sciene
1995; 270: 470-474; Blaese R M, et al.
Hum Gene Ther
1990; 1: 327-329, Bordignon C, et al.
Science
1995; 270: 470-474) familial hypercholesterolemia, (Grossman M, et al.
Nature Genet
1994; 6: 335-341) hemophilia, (Chuah M K L, et al.
Hum Gene Ther
1995, 6: 1363-1377) and cystic fibrosis, (Colledge W H, et al.
Brit Med Bull
1995; 51: 82-90) and acquired diseases such as cancer (Blaese R M, et al.
Cancer Gene Ther
1995; 2: 291-297), heart disease (Smith L C, et al.
Adv Exp Med Biol
1995; 369: 77-88), kidney disease (Smith L C, et al.
Adv Exp Med Biol
1995; 369: 77-88), and acquired immunodeficiency syndrome (Yu M, et al.
Gene Ther
1994; 1: 13-26).
At present, the primary limitation in the use of gene therapy to treat human disease is the ineffectiveness of gene delivery methods. Among several types of gene delivery systems for human gene therapy in clinical trials, Moloney murine leukemia virus (MLV) based vectors are the most widely used (Afione S A, et al.
Clin Pharmacokinet
1995; 28: 181-189; Morgan R A, et al.
Annu Rev Biochem
1993; 62: 191-217; Smith A E.
Annu Rev Microbiol
1995; 49: 807-838, Vile R G, et al.
Brit Med Bull
1995; 51: 12-30). MLV-based vectors offer highly efficient chromosome integration; thus, the therapeutic genes are transmitted to the progeny cells. However, MLV-based gene delivery methods are largely limited to ex vivo protocols, (Blaese R M, et al.
Science
1995; 270: 470-474; Blaese R M, et al.
Hum Gene Ther
1990; 1: 327-329; Bordignon C, et al.
Science
1995; 270: 470-474; Grossman M, et al.
Nature Genet
1994; 6: 335-341; Chuah M K L, et al.
Hum Gene Ther
1995; 6: 1363-1377) in which the target cells are removed from the patient to receive therapeutic genes from the MLV vectors in vitro. Then the transduced cells are selected, expanded and reimplanted in the patient. The ex vivo procedure is cumbersome and costly, and in most cases, it can transduce only a small fraction of the target cell population (Rettinger S D, et al.
Proc Natl Acad Sci USA
1994; 91: 1460-1464; Salmons B, et al.
Leukemia
1995; 1: S53-S60). More efficient gene delivery protocols need to be developed, for advancing gene therapy to routine clinical practice.
In vivo gene transfer is conceptually attractive and potentially can be more efficient than the ex vivo procedure. However, due to limited titer (Vile R G, et al.
Brit Med Bull
1995; 51: 12-30) and short half life at body temperature, direct administration of retroviral vectors into patients is limited in its applicability. Another alternative is to introduce producer cells directly into patients. In this scenario, gene transfer may continue in vivo for the duration of the life span of the implanted producer cells. Gene therapy using MLV-based producer cells to treat brain tumors (Culver K W, et al.
Science
1992; 256: 1550-1552) has been carried out in clinical trials, but no clear clinical benefit has been reported Murine producer cells are rapidly inactivated in human serum, (Rother R P, et al
J Exp Med
1995; 182: 1345-1355; Welsh R M, et al.
Nature
1975; 257:612-614) thus, the lack of success in this case is not surprising. Implanting established human producer cell lines, however, risks the introduction of malignancy to the recipient. For these reasons, the conversion of human primary cells into producer cells has been proposed (Welsh R M, et al. Human serum lyses RNA tumor viruses.
Nature
1975; 257:612-614). The primary cell-converted “producer cells” do not have the drawbacks discussed above yet they have the advantages of in vivo gene delivery.
Accordingly it is an object of the present invention to provide a method for gene delivery
SUMMARY OF THE INVENTION
A combination of adenoviral and retroviral vectors used to construct second generation packaging cells that deliver marker genes to target cells is described. A vector based upon Moloney murine leukemia virus (MLV) was used to deliver marker genes, and an adenovirus-based delivery system was used to deliver MLV structural genes (gagpol and env) to cultured cells. The procedure transformed the cells into new retroviral producer cells, which generate replication-incompetent retrovirat particles in the culture supernatant for transferring marker genes to target cells. The titer of the retroviral-containing supernatant generated from the second generation producer cells reached above 10
5
cfu/ml, which is comparable to the MLV-based producer cell lines currently used in human gene therapy trials. Additional adenoviral/MLV-based vectors were constructed with increased safety, by replacing the 5′-LTR down to the primer binding site in pPAM3 and replacing it with a CMV promoter
Another viral vector system was developed based on adenovirus and HIV. Adenoviral vectors for delivering HIV-1 gagpol genes and marker genes in a lentivurs construct were made.
The examples demonstrate the construction of these vectors as well as delivery and expression of the thymidine kinase gene and killing of tumors in mice following gancyclovir administration. The vectors and procedures are adaptable for human gene therapy in which the new producer cells are transplanted into patients for continuous gene transfer.
REFERENCES:
patent: 4861719 (1989-08-01), Miller
patent: 5278056 (1994-01-01), Bank et al.
patent: 5449614 (1995-09-01), Danos et al.
patent: 5591624 (1997-01-01), Barber et al.
patent: 5691177 (1997-11-01), Guber et al.
patent: 5716975 (1998-02-01), Bue-Valleskey et al.
patent: 5719137 (1998-02-01), Washburn et al.
patent: 5731284 (1998-03-01), Williams
patent: 5731354 (1998-03-01), Pruss
patent: 5736565 (1998-04-01), Ferrari
patent: 5885806 (1999-03-01), Dropulic et al.
patent: WO 96/33280 A1 (1996-10-01), None
patent: WO 97/30169 A1 (1997-08-01), None
patent: WO 98/13499 A2 (1998-04-01), None
patent: WO 98/13510 A1 (1998-04-01), None
Adam, et al., “Internal initiation of translation in retroviral vectors carrying picornavirus 5′ nontranslated regions,”J Virol.65(9):4985-90 (1991).
Afione, et al., “Gene therapy vectors as drug delivery systems,”Clin Pharmacokinet.28(3):181-9 (1995).
Bett, et al., “An efficient and flexible system for construction of adenovirus vectors with insertions or deletions in early regions 1 and 3,”Proc Natl Acad Sci U S A. 91(19):8802-6 (1994).
Bett, et al., “Packaging capacity and stability of human adenovirus type 5 vectors.”J Virol.Oct. 1993;67(10):5911-21.
Bilbao, et al., “Adenoviral/retroviral vector chimeras: a novel strategy to achieve high-efficiency stable transduction in vivo,”FASEB J.11(8):624-34 (1997).
Blaese, et al., “Vectors in cancer therapy: how will they deliver?”Cancer Gene Ther.2(4):291-7 (1995).
Blaese, et al., “The ADA Human Gene Therapy Clinical Protocol,”Hum Gene Ther1:327-329 (1990).
Bordignon, et al., “Gene therapy in peripheral blood lymphocytes and bone marrow for ADA—immunodeficient patients,”Science.270(5235):470-5 (1995).
Bramson, et al., “The use of adenoviral vectors for gene therapy and gene transfer in vivo,”Curr Opin Biotechnol.6(5):590-5 (1995).
Chuah, et al., “Development and analysis of retroviral vectors expressing human factor VIII as a potential gene therapy for hemophilia A,”Hum Gene Ther.6(11):1363-77 (1995).
Colledge, et al., “Cystic fibrosis gene therapy,”Br Med Bull.51(1):82-90 (1995).
Cosset, et al. “Targeting retrovirus entry,”Gene Therapy3:946-956 (1996).
Cournoyer & Caskey, “Gene therapy of the immune system,”Annu. Rev. Immunol.11:297-329 (1993).
Culver, et al., “In Vivo Gene Transfer with Retroviral Vector-Producer Cells for Treatment of Experimental Brain Tumors,”Science256:1550-1552 (1992).
Felsenstein, et al., “Expres
Lin Xinli
Tang Jordan J. N.
Holland & Knight LLP
Oklahoma Medical Research Foundation
Schwartzman Robert A.
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