Drug – bio-affecting and body treating compositions – Whole live micro-organism – cell – or virus containing – Genetically modified micro-organism – cell – or virus
Patent
1997-08-25
2000-05-30
Crouch, Deborah
Drug, bio-affecting and body treating compositions
Whole live micro-organism, cell, or virus containing
Genetically modified micro-organism, cell, or virus
424 931, 424 932, 424 937, 435325, 435372, 435383, 435455, A01N 6300, A01N 4304, C12N 500, C12N 508
Patent
active
06068836&
DESCRIPTION:
BRIEF SUMMARY
BACKGROUND OF THE INVENTION
The invention relates to the field of stem cell engraftment and human ex vivo gene therapy.
Bone marrow transplantation is often accompanied by marrow cytotoxic therapy to create marrow space or niches. The usual method of creating such niches is by irradiation and/or chemotherapy treatment. This practice has been challenged by Brecher, Saxe and colleagues, who demonstrated that bone marrow cells (e.g., hematopoietic stem cells) engraft into normal non-myeloablated hosts (Brecher, G. et al. (1982) PNAS USA 79:5085; and Saxe, D. F. et al. (1984) Exp. Hematol. 12:277). These investigators showed varying levels of engraftment up to 25% after infusion of normal bone marrow into non-myeloablated hosts.
One theoretical approach to transplantation involves procurement of stem cells for transplantation followed by expansion of the stem cells to increase their number prior to infusion into the host. Cellular expansion and/or induction into active cell cycling is accomplished by contacting cytokines or other agents, such as 5-fluorouracil, with the stem cells in vivo, if expansion occurs prior to collection of stem cells; or in vitro, if expansion occurs following collection of stem cells. Long-term repopulation of bone marrow with cytokine-treated cells has been assessed in vivo (Bodine D. M., Blood, 79:913, 1992; Szilvassy S. J. and Cory, S., Blood 84:74, 1994; Li, C. L. and Johnson, G. R. Blood 84:408, 1994; and Ramshaw, H. S. et al., Experimental Hematology, 22:823, No. 548, 1994).
Additionally, cytokines have been used in various combinations to enhance retroviral integration into stem cells by increasing the percent of stem cells in the process of active cell cycle. In vitro cytokine incubations have been used to induce active cell cycling in hematopoietic cells to facilitate retroviral integration in various gene therapy approaches (Bodine D. M. et al., Exp Hematol 19:206, 1991; Szilvassy S. J. and Cory, S., Blood 84:74, 1994; and Kittler, E. L. W. et al., Blood 84:344, 1994 (abstr. suppl I)). Subsequent to retroviral integration, these actively cycling transfected stem cells are introduced into a mammal so that they engraft in the bone marrow of the mammal. Thus the success of ex vivo gene therapy is dependent not only on the efficiency of retroviral integration, but also on the efficiency of transfected stem cell engraftment.
There is a great need for methodologies to enhance engraftment of stem cells in a host mammal for the purpose of improved human bone marrow transplantation therapy as well as for improved human ex vivo gene therapy.
SUMMARY OF THE INVENTION
Applicant discloses that rendering cells quiescent prior to transplantation results in improved long term engraftment, i.e., for time periods greater than approximately 6 weeks post-transplantation. Accordingly, the invention features stem cell compositions and methods of preparing stem cell compositions for transplantation or ex vivo gene therapy that significantly enhance engraftment of the transplanted stem cells, wherein the stem cells are expanded to produce a population of expanded stem cells. The expanded stem cells are next treated so that a majority of them become quiescent stem cells to form the stem cell composition that is introduced into a mammal by standard stem cell transplantation techniques. In preferred embodiments the mammal is a non-myeloablated or a myeloablated host mammal, preferably a human patient.
Stem cells are expanded and/or treated to enter active cell cycling in vivo or in vitro by methods well known to those skilled in the art and include, but are not limited to, the administration of a cytokine, e.g., stem cell factor (Steel factor), IL-3-CHO, IL-6, IL-11, and the like, to the stem cell in vivo or in vitro; and administration of 5-fluorouracil (5-FU) in vivo.
Expanded and/or actively cycling stem cells are treated to become quiescent according to the invention by methods well known to those of ordinary skill in the relevant arts and include, but are not limited to, serum starvation and
REFERENCES:
patent: 4396601 (1983-08-01), Salser et al.
patent: 5032407 (1991-07-01), Wagner et al.
patent: 5061620 (1991-10-01), Tsukamoto et al.
patent: 5437994 (1995-08-01), Emerson et al.
patent: 5665350 (1997-09-01), Quesenberry
Barlogie, B. "Hemopoietic stem cell transplant for multiple myeloma (MM)", Leukemia, 7:1095, (1993).
Biron, P. et al., "Autologous Bone Marrow Transplantation:Proceedings of the First International Symposium".
Breacher, G. et al. "Special proliferative sites are not needed for seeding and proliferation of . . . syngeneic mice", PNAS USA 79:5085-5087, (1982).
Cheson, B.D. et al., "Autologous bone . . . future directions", Ann. Intern. Med. 110(1):51-56, (1989).
Crittenden R. et al., "Repetitive Marrow . . . Gene Therapy", Experimental Hematology, 21:1016, Abstract 30.
Culver K. et al., "Gene therapy for cancer", TIG, 10:174-178, (1994).
Harrison, D., "Competitive Repopulation in Unirradated Normal Recipients", Blood, 81:2473-2474, (1993).
Hodgson, C., "Advanced in vector systems for gene therapy", Exp. Opin. Ther. Patents, 5(5):459-468, (1995).
Kohn, D., "The current status of gene therapy using hematopoietic stem cells", Current Opinion in Pediatrics 7:56-63, (1995).
Marshall, E., "Gene Therapy's Growing Pains", Science, 269:1050-1055 (1995).
Miller, N. et al., "Targeted vectors for gene therapy", The FASEB Journal, 9:190-198, (1995).
Peters, S.Q. et al., "Cytokine Incubation of Murine Marrow Cells Impairs Repopulating Capacity in normal Non-myeloablated) Hosts", Experimental Hematology, 22(8):823, abstract 547 (1994).
Peters, W.P., "The Rationale for High-Dose . . . in Treating Breast Cancer", AMBT supra, 189-195, (1985).
Ramshaw, H.S. et al., "Effect of cell cycle on the engraftment . . . unmyeloablated hosts", Experimental Hematology, 22:823, abstract 548.
Saxe, D.F. et al., "Transplantation of Chromosomally . . . Hematopietic Stem Cell Depletion", Exp. Hematol., 12:277-283, (1984).
Stewart et al., "Long-Term Engraftment of . . . Nonmyeloablated Mice", Blood, 81(10):2566-2571, (1993).
Sullivan, K.M., "Marrow transplantation . . . hematopoises", Leukemia, 7:1098-1099 (1993).
Takvorian, T. et al., "Prolonged disease-free survival . . . with a poor prognosis", N. Engl. J. Med., 316:1499-1505, (1987).
Wheeler C., et al., "Cyclophosphamide, carmustine, . . . and non-Hodgkin's Lymphoma:a dose-finding study", J. clin. Oncol., 8(4):548-656, (1990).
Yeager, A.M. et al., "Autologous bone marrow . . . with 4-hydroperoxycyclophosphamide", N. Eng. J. Med., 315(3):141-147, (1986).
Orkin and Motulsky, Report and recommendations of the panel to assess the NIH investment in research on gene therapy, Dec. 1995.
Friedmann, T. Overcoming the obstacles to gene therapy. Sci. Am. pp. 96-101, Jun. 1997.
Verma et al. Gene therpy--promises, problems, and prospects. Nature 389:239-242, Sep. 1997.
Marshall, E. Gene therapy's growing pains. Science 269. 1050-1055, Aug. 1995.
Sandstrom et al. Review: Serum-free media for cultures of primitive and mature hematopoietic cells. Biotechnology and Bioengineering 43: 706-733, Apr. 1994.
Stewart et al. Long-term engraftment of normal and post-5-fluorouracil murine marrow into normal nonmyeloablated mice. Blood 81(10) 2566-2571, May 1993.
Brecher et al. Special proliferative sites are not needed for seeding and proliferation of transfused bone marrow cell in normal syngeneic mice. Proc. Natl. Acad. Sci. USA 79: 5085-5087, Aug. 1982.
Baker Anne-Marie
Crouch Deborah
University of Massachusetts
LandOfFree
Cell compositions for use in transplantation does not yet have a rating. At this time, there are no reviews or comments for this patent.
If you have personal experience with Cell compositions for use in transplantation, we encourage you to share that experience with our LandOfFree.com community. Your opinion is very important and Cell compositions for use in transplantation will most certainly appreciate the feedback.
Profile ID: LFUS-PAI-O-1908108