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Human prostate specific G-protein receptor HPRAJ70

a prostate specific, gprotein receptor technology, applied in the field of psgr, can solve the problems of increased patient mortality, ineffective methods in a significant percentage of cases, and difficult treatment of prostate cancer, so as to reduce psgr mediated signalling, inhibit psgr mediated signalling, and enhance psgr mediated signalling

Inactive Publication Date: 2005-03-24
HUMAN GENOME SCI INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

[0016] Thus, the invention further provides cells which express the PSGR polypeptide with a candidate compound and its ligand, assaying for inhibiting PSGR mediated signalling induced by said ligand which involves administering to a cell which expresses the PSGR polypeptide an effective amount of a PSGR agonist capable of decreasing PSGR mediated signalling.
[0017] In a further aspect, the present invention is directed to a method for enhancing PSGR mediated signalling induced by its ligand which involves administering to a cell which expresses the PSGR polypeptide an effective amount of a PSGR agonist or antagonist capable of increasing PSGR mediated signalling.
[0018] Whether any candidate “agonist” or “antagonist” of the present invention can enhance or inhibit PSGR mediated signalling can be determined using art-known G-protein coupled ligand / receptor cellular response assays, including those well known in the art. Thus, in a firther aspect, a screening method is provided for determining whether a candidate agonist or antagonist is capable of enhancing or inhibiting a PSGR mediated cellular response to a ligand. The method involves contacting cellular response, and comparing the cellular response to a standard cellular response, the standard being assayed when contact is made with the ligand in absence of the candidate compound, whereby an increased cellular response over the standard indicates that the candidate compound is an agonist of the ligand / receptor signaling pathway and a decreased cellular response compared to the standard indicates that the candidate compound is an antagonist of the ligand / receptor signaling pathway. By the invention, a cell expressing the PSGR polypeptide can be contacted with either an endogenous or exogenously administered ligand.

Problems solved by technology

Overwhelming clinical evidence shows that human prostate cancer has the propensity to metastasize to bone, and the disease appears to progress inevitably from androgen dependent to androgen refractory status, leading to increased patient mortality.
In spite of considerable research into therapies for the disease, prostate cancer remains difficult to treat.
Commonly, treatment is based on surgery and / or radiation therapy, but these methods are ineffective in a significant percentage of cases.
In spite of considerable research into therapies for these and other cancers, prostate cancer remains difficult to diagnose and treat effectively.

Method used

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  • Human prostate specific G-protein receptor HPRAJ70
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  • Human prostate specific G-protein receptor HPRAJ70

Examples

Experimental program
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example 1

[0635] Expression and Purification of a Soluble Portion of PSGR in E. coli

[0636] The bacterial expression vector pQE60 is used for bacterial expression in this example. (QIAGEN, Inc., 9259 Eton Avenue, Chatsworth, Calif., 91311). pQE60 encodes ampicillin antibiotic resistance (“Ampr”) and contains a bacterial origin of replication (“ori”), an IPTG inducible promoter, a ribosome binding site (“RBS”), six codons encoding histidine residues that allow affinity purification using nickel-nitrilo-tri-acetic acid (“Ni-NTA”) affinity resin sold by QIAGEN, Inc., supra, and suitable single restriction enzyme cleavage sites. These elements are arranged such that a DNA fragment encoding a polypeptide may be inserted in such as way as to produce that polypeptide with the six His residues (i.e., a “6×His tag”) covalently linked to the carboxyl terminus of that polypeptide.

[0637] The DNA sequence encoding the desired portion of the PSGR protein is amplified from the deposited cDNA clone using PC...

example 2

[0647] Cloning and Expression of PSGR in a Baculovirus Expression System

[0648] In this illustrative example, the plasmid shuttle vector pA2 is used to insert the cloned DNA encoding the complete protein, including its naturally associated secretary signal (leader) sequence, into a baculovirus to express the mature PSGR protein, using standard methods as described in Summers et al., A Manual of Methods for Baculovirus Vectors and Insect Cell Culture Procedures, Texas Agricultural Experimental Station Bulletin No. 1555 (1987). This expression vector contains the strong polyhedrin promoter of the Autographa californica nuclear polyhedrosis virus (AcMNPV) followed by convenient restriction sites such as Bam-HI and Asp718. The polyadenylation site of the simian virus 40 (“SV40”) is used for efficient polyadenylation. For easy selection of recombinant virus, the plasmid contains the beta-galactosidase gene from E. coli under control of a weak Drosophila promoter in the same orientation, ...

example 3

[0659] Cloning and Expression of the PSGR Receptor in Mammalian Cells

[0660] A typical mammalian expression vector contains the promoter element, which mediates the initiation of transcription of mRNA, the protein coding sequence, and signals required for the termination of transcription and polyadenylation of the transcript. Additional elements include enhancers, Kozak sequences and intervening sequences flanked by donor and acceptor sites for RNA splicing. Highly efficient transcription can be achieved with the early and late promoters from SV40, the long terminal repeats (LTRs) from Retroviruses, e.g. RSV, HTLVI, HIVI and the early promoter of the cytomegalovirus (CMV). However, cellular signals can also be used (e.g., the human actin promoter). Suitable expression vectors for use in practicing the present invention include, for example, vectors such as pSVL and pMSG (Pharmacia, Uppsala, Sweden), pRSVcat (ATCC 37152), pSV2dhfr (ATCC 37146) and pBC12MI (ATCC 67109). Mammalian host...

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Abstract

The present invention relates to PSGR, a novel prostate specific gene with homology to a G-protein coupled receptor overexpressed in prostate cancer. More specifically, the invention relates to PSGR polynucleotides and the polypeptides encoded by these polynucleotides, and the use of PSGR polynucleotides and polypeptides for detecting disorders of the reproductive system, including disorders of the prostate, particularly the presence of cancer. This invention relates to PSGR polynucleotides and polypeptides as well as vectors, host cells, antibodies directed to PSGR polynucleotides and polypeptides and recombinant and synthetic methods for producing the same. Also provided are methods for diagnosing, treating, preventing, and / or prognosing disorders related to the prostate, including cancer. The invention further relates to screening methods for identifying agonists and antagonists of PSGR polynucleotides and polypeptides of the invention and methods and / or compositions for inhibiting or enhancing the production and / or function of the PSGR polypeptides of the present invention.

Description

[0001] This application is a divisional of U.S. application Ser. No. 09 / 968,033, filed Oct. 2, 2001, which claims benefit of U.S. Provisional Application No. 60 / 237,275, filed Oct. 3, 2000, and U.S. application Ser. No. 09 / 968,033 is a Continuation-in-Part of U.S. application No. 09 / 339,115, filed Jun. 24, 1999, which is a divisional of U.S. application No. 09 / 053,303, filed Apr. 1, 1998, now U.S. Pat. No. 5,948,890, issued Sep. 7, 1999, which is a divisional of U.S. application Ser. No. 08 / 465,980, filed Jun. 6, 1995, now U.S. Pat. No. 5,756,309, issued May 26, 1998, each of which is hereby incorporated by reference in its entirety.BACKGROUND OF THE INVENTION [0002] 1. Field of the Invention [0003] The present invention relates to PSGR, a novel prostate specific gene with homology to a G-protein coupled receptor overexpressed in prostate cancer. More specifically, the invention relates to PSGR polynucleotides and the polypeptides encoded by these polynucleotides, and the use of PSG...

Claims

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Application Information

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Patent Type & Authority Applications(United States)
IPC IPC(8): A61K38/00A61K39/00C07K14/705C07K16/28
CPCA61K38/00A61K39/00C07K2317/622C07K14/705C07K16/28A61K2039/505
Inventor SOPPET, DANIELLI, YIROSEN, CRAIGRUBEN, STEVEN
Owner HUMAN GENOME SCI INC
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