Chemistry: molecular biology and microbiology – Micro-organism – per se ; compositions thereof; proces of... – Bacteria or actinomycetales; media therefor
Patent
1996-05-22
1998-07-14
Kemmerer, Elizabeth C.
Chemistry: molecular biology and microbiology
Micro-organism, per se ; compositions thereof; proces of...
Bacteria or actinomycetales; media therefor
4353201, 435325, 4351723, 530350, 530399, 5303879, 536 235, C07K 14475, C12N 121, C12N 510, C12N 1512
Patent
active
057802912
DESCRIPTION:
BRIEF SUMMARY
BACKGROUND OF THE INVENTION
The Wnt growth factor family is composed of several structurally related proteins. The Wnt-1 proto-oncogene (int-1) was originally identified from mammary tumors induced by mouse mammary tumor virus (MMTV) due to an insertion of viral DNA sequence (Nusse and Varmus, 1982, Cell, 31, pp. 99-109). At least ten Wnt genes were identified in the mouse (Wnt-1, 2, 3, 2319-2332! and seven Wnt genes have been identified in the human (Wnt-1, 2, 3, 4, 5a, 7a, and 7b) by cDNA cloning (Vant Veer et al., 1984, Mol. Cell. Biol., 4, pp. 2532-2534; Wainright et al., 1988, EMBO J., 7, pp. 1743-1748). In situ hybridization studies have shown that the expression of many Wnt growth factor genes were specifically related to patterning and morphogenesis during early neural development in the central nervous system, suggesting that the expression of these genes are required for patterning and development in the Drosophila and invertebrates. In adult mice, the expression level of Wnt-1 mRNA is detected only in the testis during later stages of sperm development. Wnt-1 protein is about 42 KDa and contains an amino terminal hydrophobic region, which may function as a signal sequence for secretion (Nusse and Varmus, supra). The expression of Wnt-2/irp is detected in mouse fetal and adult tissues and its distribution does not overlap with the expression pattern for Wnt-1. Wnt-3 is associated with mouse mammary tumorigenesis. The expression of Wnt-3 in mouse embryos detected in the neural tubes and in the limb buds. Wnt-5a transcripts are detected in the developing fore- and hind limbs at 9.5 through 14.5 days and highest levels are concentrated in apical ectoderm at the distal tip of limbs (Nusse and Varmus, 1992, Cell, 69, pp. 1073-1087).
SUMMARY OF THE INVENTION
A new member of the Wnt growth factor family from humans is disclosed. DNA encoding the new Wnt growth factor, termed Wnt-x, is also disclosed as is the detection of Wnt-x expression in bone tissues and in bone-derived cells. Also disclosed is the role of Wnt-x in the maintenance of mature osteoblasts and the use of the Wnt-x growth factor as a therapeutic agent or in the development of other therapeutic agents to treat bone-related diseases.
BRIEF DESCRIPTION OF THE DRAWINGS
FIGS. 1A-1D--Nucleotide sequence of Wnt-x cDNA (SEQ ID NO:7) with the amino acid sequence of Wnt-x (SEQ ID NO:8) deduced from the cDNA sequence is shown.
DETAILED DESCRIPTION OF THE INVENTION
The present invention relates to cDNA encoding a novel growth factor termed Wnt-x. The present invention is also related to recombinant host cells which express the cloned Wnt-x-encoding DNA contained in a recombinant expression plasmid. The present invention is also related to a method for the screening of substances which modulate Wnt-x protein activity. The DNA of the present invention is isolated from Wnt-x producing cells. Wnt-x, as used herein, refers to a growth factor which is specifically expressed in bone cells. The present invention also relates to a unique growth factor protein, also described as Wnt-x, which is isolated from Wnt-x producing cells. Wnt-x protein, as used herein, refers to a growth factor protein which is specifically produced by bone cells.
Mammalian cells capable of producing Wnt-x include, but are not limited to, cells derived from tissues including heart, brain and bone. Transformed mammalian cell lines which produce Wnt-x include, but are not limited to, giant cell tumor cells. The preferred cells for the present invention include human giant cell tumor cells.
Other cells and cell lines may also be suitable for use to isolate Wnt-x cDNA. Selection of suitable cells may be done by screening for Wnt-x produced by the cells. Methods for detecting Wnt growth factor RNA and 69, pp. 1073-1978! and measure the level of Wnt-x RNA produced by the cells. Cells which possess Wnt-x activity in this assay may be suitable for the isolation of Wnt-x cDNA.
Cells which are responsive to Wnt gene activity are known in the art, and include but are not limited to, t
REFERENCES:
Bowie et al. Science 247:1306-1310, 1990.
Nusse and Varmus, "Wnt Genes", Cell, vol. 69, pp. 1073-1087 (1992).
Wainwright, et al., "Isolation of a human gene with protein sequence similarity to human and murine int-1 . . . ", The EMBO Journal, vol. 7, No. 6, pp. 1743-1748 (1988).
van 't Veer, et al., "Molecular Cloning and Chromosomal Assignment of the Human Homolog of int-1 . . . ", Mol andCell Biology, vol. 4, No. 11, pp. 2532-2534 (1984).
Gavin, et al., "Expression of multiple novel Wnt-1/int-1 related genes during fetal and adult mouse development", Genes & Development, vol. 4, pp. 2532-2534 (1984).
Nusse and Varmus, "Manu Tumors Induced by the Mouse Mammary Tumor Virus Contain a Provirus Integrated . . . ", Cell, vol. 31, pp. 99-109 (1982).
Rijsewink, et al., "Transfection of the int-1 mannary in cuboidal RAC mammary cell line results in morphological . . . ", The EMBO Journal, vol. 6, No. 1, pp. 127-131 (1987).
Brown, et al., "A Retrovirus Vector Expressing the Putative Mammary Oncogene int-1 Causes Partial . . . ", Cell., vol. 46, pp. 1001-1009 (1986).
Savard, "Body Axis Determination During Early Development in Amphibians", Biochem. Cell. Biol., vol. 70, pp. 875-891, (1992).
McMahon, et al., Nucleotide sequence, chromosomal localization and decelopmental expression of the mouse int-1-related gene, Development, vol. 107, pp. 643-650 (1989).
Lin, et al., "Role of Endocrine, Autocrine, and Paracrine Interactions in the Development of Mammary . . . ", Cancer Research, vol. 52, pp. 4413-4419, (1992).
Barton, et al., "Bacillus thuringiensis -Endotoxin Expressed in Transgenic Vicotiana tabacum Provides Resistance . . . ", Plant Physiol., vol. 85, pp. 355-359, (1987).
Gelvin, et al., "Biotechnology News and Views", Plant Mol. Biol., vol. 8, pp. 355-359 (1987).
Rodan Gideon A.
Rutledge Su Jane
Schmidt Azriel
Kemmerer Elizabeth C.
Merck & Co. , Inc.
Tribble Jack L.
Yablonsky Michael D.
LandOfFree
Wnt-x growth factor polypeptide, DNA encoding same, and Wnt-x an does not yet have a rating. At this time, there are no reviews or comments for this patent.
If you have personal experience with Wnt-x growth factor polypeptide, DNA encoding same, and Wnt-x an, we encourage you to share that experience with our LandOfFree.com community. Your opinion is very important and Wnt-x growth factor polypeptide, DNA encoding same, and Wnt-x an will most certainly appreciate the feedback.
Profile ID: LFUS-PAI-O-1880892