Controlling proliferation of cells before and after encapsulation in a bioartificial organ by gene transformation

• US 5,843,431 A
• Filed: 05/09/1995
• Issued: 12/01/1998
• Est. Priority Date: 07/20/1994
• Status: Expired due to Term

• Current Assignee: Neurotech USA, Inc. (Woodbrook Group Holdings, Inc.)
• Original Assignee: Cytotherapeutics, Inc.
• Inventors: Aebischer, Patrick, Shoichet, Molly S., Hammang, Joseph P., Doherty, Edward J., Gentile, Frank T., Holland, Laura M., Winn, Shelley R., Cain, Brian M., Schinstine, Malcolm
• Primary Examiner(s): Naff, David M.
• Application Number:
  US08/432,698
• Time in Patent Office: 1,302 Days
• Field of Search: 435/174, 435/178, 435/172.3, 435/240.7, 435/240.22, 435/240.23, 435/240.24, 435/240.241, 435/240.242, 435/240.243, 435/377, 435/382, 435/395, 424/93.21, 424/93.7, 424/422
• US Class Current: 424/93.21
• CPC Class Codes: A61K 48/00 Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy
• CPC Class Codes: A61L 27/18 obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds
• CPC Class Codes: A61L 27/227 Other specific proteins or polypeptides not covered by A61L27/222, A61L27/225 or A61L27/24
• CPC Class Codes: A61L 27/38 containing added animal cells organs or tissue containing native cells A61L27/36
• CPC Class Codes: A61L 27/3839 characterised by the site of application in the body materials for artificial blood vessels A61L27/507; materials for use in artificial skin A61L27/60
• CPC Class Codes: C07K 14/82 Translation products from oncogenes
• CPC Class Codes: C08L 83/12 containing polyether sequences
• CPC Class Codes: C12N 2510/00 Genetically modified cells
• CPC Class Codes: C12N 2510/04 Immortalised cells
• CPC Class Codes: C12N 2533/30 Synthetic polymers thermoreactive polymers, e.g. PNIPAm, C12N2539/10
• CPC Class Codes: C12N 2533/54 Collagen; Gelatin
• CPC Class Codes: C12N 2533/74 Alginate
• CPC Class Codes: C12N 5/0012 Cell encapsulation
• CPC Class Codes: C12N 5/0068 General culture methods using substrates for specific animal cell type C12N5/06

Abstract

Methods and compositions are provided for controlling cell distribution within an implantable bioartificial organ by exposing the cells to a treatment that inhibits cell proliferation, promotes cell differentiation, or affects cell attachment to a growth surface within the bioartificial organ. Such treatments include (1) genetically manipulating cells, (2) exposing the cells to a proliferation-inhibiting compound or a differentiation-inducing compound or removing the cells from exposure to a proliferation-stimulating compound or a differentiation-inhibiting compound; exposing the cells to irradiation, and (3) modifying a growth surface of the bioartificial organ with extracellular matrix molecules, molecules affecting cell proliferation or adhesion, or an inert scaffold, or a combination thereof. These treatments may be used in combination. Cells can be transformed with a proliferation-promoting gene such as the oncogene, SV40, linked to a regulatable promoter such as the Mx1 promoter, the promotor is activated in vitro to express the gene to result in cell proliferation, and the promotor is inactivated before or after insertion of the cells in the bioartificial organ to inhibit expression of the gene to reduce or stop cell proliferation in vivo. The promoter can be reactivated in vivo to again express the gene to result in further cell proliferation. The gene may be a proliferation-suppressing gene such as p53 gene or RB gene, or a differentiation-inducing gene such as high mobility group chromosomal protein 14. Inhibiting gene expression in vitro causes cell proliferation, and inducing gene expression reduces or stops cell proliferation in vivo.

Claims

1. A method for controlling distribution of cells within an implantable bioartificial organ comprising the steps of:

(a) transforming the cells with a proliferation-promoting gene operatively linked to a regulatable promoter wherein the regulatable promoter is activated in vitro to achieve expression of the proliferation-promoting gene, resulting in proliferation of the transformed cells in vitro; and

(b) inserting the cells from (a) in a bioartificial organ and inactivating the regulatable promoter either before or after inserting the cells in the bioartificial organ such that expression of the proliferation-promoting gene is inhibited to reduce or stop proliferation of the cells in the bioartificial organ in vivo in a host.

2. The method according to claim 1, wherein the proliferation promoting gene is an oncogene.

3. The method according to claim 2, wherein the oncogene is selected from the group consisting of c-myc, v-mos, v-Ha-ras, SV40 early region and E1-A.

4. The method according to claim 1, wherein the regulatable promoter is selected from the group consisting of tetracycline-responsive promoters, interferon-responsive promoters, glucocorticoid-responsive promoters and retroviral long terminal repeat promoters.

5. The method according to claim 4, wherein the interferon-responsive promoter is the Mx1 promoter and the proliferation-promoting gene is SV40 early region.

6. The method of claim 1 further comprising the step of:

(c) reactivating the regulatable promoter in vivo in the bioartificial organ such that expression of the proliferation-promoting gene is achieved, resulting in further proliferation of the cells in the bioartificial organ.

7. A method for controlling distribution of cells within an implantable bioartificial organ comprising the steps of:

(a) transforming the cells with a proliferation-suppressing gene operatively linked to a regulatable promoter wherein the regulatable promoter is inactivated in vitro to inhibit expression of the proliferation-suppressing gene, resulting in proliferation of the transformed cells in vitro; and

(b) inserting the cells from (a) in a bioartificial organ and activating the regulatable promoter either before or after inserting the cells in the bioartificial organ such that expression of the proliferation-suppressing gene occurs to reduce or stop proliferation of the cells in the bioartificial organ in vivo in a host.

8. The method according to claim 7, wherein the proliferation-suppressing gene is a tumor suppressor gene selected from the group consisting of the p53 gene and the RB gene.

9. A method for controlling distribution of cells within an implantable bioartificial organ comprising the steps of:

(a) transforming the cells with a differentiation-inducing gene operatively linked to a regulatable promoter wherein the regulatable promoter is inactivated in vitro to inhibit expression of the differentiation-inducing gene, resulting in proliferation of the transformed cells invtro; and

(b) inserting the cells from (a) in a bioartificial organ and activating the regulatable promoter either before or after inserting the cells in the bioartificial organ such that expression of the differentiation-inducing gene occurs to reduce or stop proliferation of the cells in the bioartificial organ in vivo in a host.

10. The method according to claim 9, wherein the differentiation-inducing gene is the high mobility group chromosomal protein 14.