Chemistry: molecular biology and microbiology – Treatment of micro-organisms or enzymes with electrical or... – Modification of viruses
Patent
1992-10-22
1996-05-28
Chambers, Jasemine C.
Chemistry: molecular biology and microbiology
Treatment of micro-organisms or enzymes with electrical or...
Modification of viruses
4352402, 435 691, 4353201, 435948, 424 9321, 424 932, 935 70, 935 71, 935 62, 935 57, 935 11, 935 13, 935 32, 935107, C12N 1500, C12N 500
Patent
active
055210767
DESCRIPTION:
BRIEF SUMMARY
BACKGROUND
The liver is of endodermal origin and the largest gland in the human body. It has numerous crucial roles, including bile secretion, participation in carbohydrate; lipid and protein metabolism, fibrinogen production and detoxification of drugs. The liver also serves as the main site at which nutrients absorbed from the gastro-intestinal tract and transported via the blood are processed for use by other body cells.
Hepatocytes, which are the main type of parenchymal or distinguishing cell in the liver, carry out the liver functions and, thus, are responsible for synthesizing, degrading and storing a wide variety of substances. In addition, a system of small channels (canaliculi) and larger ducts connects hepatocytes with the gut lumen. Through this route, hepatocytes secrete bile, an emulsifying agent which helps in absorption of ingested fats. Hepatocytes are also the main location at which lipoprotein particles for export are made; enzymes responsible for synthesis of the lipid constituents of lipoproteins occur in hepatocyte membranes.
Because of the many important functions the liver has, its inability to function normally (e.g., as a result of a genetic defect or damage caused by alcohol or other toxic substances) will often have significant adverse effects on an individual's health. A means by which normal function can be conferred upon or restored to a liver whose function is compromised would be very useful in treating, correcting or preventing such an abnormality.
SUMMARY OF THE INVENTION
The invention described herein relates to genetically engineered or transduced hepatocytes which express genetic material (DNA or RNA) of interest introduced or incorporated into them, as well as to methods of producing, transplanting and using the genetically engineered hepatocytes. The genetic material of interest can be incorporated through the use of a vector, such as a recombinant retrovirus, which contains the genetic material of interest, or by other means.
Hepatocytes of the present invention express the genetic material of interest. Such genetic material of interest can be: 1) genetic material present in and expressed at biologically effective levels by normal hepatocytes, but present in or expressed in less than normal quantities in the hepatocytes prior to transfer of genetic material of interest into them by the method of the present invention; 2) genetic material not present in normal hepatocytes; or 3) genetic material present in normal hepatocytes but not expressed at biologically effective levels in such cells, alone or in any combination thereof.
In hepatocytes of the present invention, the genetic material of interest can be incorporated into the celluar genetic material (e.g., into genomic DNA) Or can be present extrachromosomally (i.e., expressed episomally). The genetic material of interest can be DNA or RNA; the DNA can constitute all or a portion of a gene of interest (i.e., one whose expression in hepatocytes is desired).
The genetic material incorporated into and expressed by hepatocytes of the present invention can additionally include genetic material (e.g., DNA) encoding a selectable marker, which provides a means by which cells expressing the genetic material of interest are identified and selected for. Hepatocytes containing incorporated genetic material (i.e., genetic material of interest and, optionally, genetic material encoding a selectable marker) are referred to as transduced hepatocytes.
Genetic material can be introduced into hepatocytes ex vivo or in vivo. That is, it can be introduced, by means of an appropriate vector, into isolated (cultured) hepatocytes, which are subsequently transplanted into the recipient. Alternatively, it can be introduced directly into the recipient in such a manner that it is directed to and taken up by target cells (hepatocytes), where it is incorporated and expressed. Particularly useful for this purpose are retroviral vectors which have an amphotropic host range and include the genetic material of interest which is to be incorporat
REFERENCES:
patent: 4497796 (1985-02-01), Salser et al.
patent: 4686098 (1987-08-01), Kopchick et al.
patent: 4868116 (1989-09-01), Morgan et al.
patent: 4980286 (1990-12-01), Morgan et al.
patent: 5399346 (1995-03-01), Anderson et al.
Leffert et al., In "Methods for serum-free culture of epithelial and fibroblastic cells," eds Barnes et al., Alan Liss Inc., pp. 43-55 (1984).
Ledley et al., Proc. Natl. Acad. Sci. 83: 409-413 (1986).
Miller et al., In "Cold Spring Harbor Symp. Quant. Biol.", vol. LI, pp. 1013-1019 (1986), Cold Spring Harbor Lab.
Verma et al., Current Opinion in Genetics & Devel. 1: 54-59 (1991).
Ledley, et al., "Retroviral gene transfer into primary hepatocytes: Implications for genetic therapy of liver-specific functions", Proc. Natl. Acad. Sci. USA 84: 5335-5339 (1987).
Wolff, et al., "Expression of retrovirally transduced genes in primary cultures of adult rat hepatocytes", Proc. Natl. Acad. Sci. USA 84: 3344-3348 (1987).
Wilson, et al., "Correction of the genetic defect in hepatocytes from the Watanabe heritable hyperlipidemic rabbit", Proc. Natl. Acad. Sci. USA 85: 4421-4425 (1988).
Wilson, et al., "Retrovirus-mediated transduction of adult hepatocytes", Proc. Natl. Acad. Sci. USA 85: 3014-3018 (1988).
Peng, et al., "Retroviral-mediated gene transfer and expression of human phenylalanine hydroxylase in primary mouse hepatocytes", Proc. Natl. Acad. Sci. USA 85: 8146-8150 (1988).
Wilson, et al., "Temporary amelioration of hyperlipidemia in low density lipoprotein receptor-deficient rabbits transplanted with genetically modified hepatocytes", Proc. Natl. Acad. Sci. USA 87: 8437-8441 (1990).
Friedmann, et al., "Retrovirus Vector-mediated Gene Transfer into Hepatocytes", Mol. Biol. Med. 6: 117-125 (1989).
Morgan, et al., "Expression of an Exogenous Growth Hormone Gene by Transplantable Human Epidermal Cells" Science 237: 1476-1479 (1987).
Yamamoto, et al., "The Human LDL Receptor: A Cysteine-Rich Protein with Multiple Alu Sequences in Its mRNA", Cell 39: 27-38 (1984).
Demetriou, Achilles A. "Hepatocyte Transplantation". Scientific American (Nov./Dec. 1994) pp. 58-67.
Mulligan Richard C.
Wilson James M.
Chambers Jasemine C.
Halluin Albert P.
Howard Hughes Medical Center
Whitehead Institute for Biomedical Research
LandOfFree
Modified hepatocytes and uses therefor does not yet have a rating. At this time, there are no reviews or comments for this patent.
If you have personal experience with Modified hepatocytes and uses therefor, we encourage you to share that experience with our LandOfFree.com community. Your opinion is very important and Modified hepatocytes and uses therefor will most certainly appreciate the feedback.
Profile ID: LFUS-PAI-O-785949