Chemistry: molecular biology and microbiology – Micro-organism – tissue cell culture or enzyme using process... – Recombinant dna technique included in method of making a...
Reexamination Certificate
1999-12-13
2001-10-02
Wortman, Donna (Department: 1645)
Chemistry: molecular biology and microbiology
Micro-organism, tissue cell culture or enzyme using process...
Recombinant dna technique included in method of making a...
C435S091100, C435S091400, C435S252300, C435S320100, C435S328000, C435S455000, C536S023100, C536S025300
Reexamination Certificate
active
06297030
ABSTRACT:
FIELD OF THE INVENTION
This invention relates to newly identified polypeptides and polynucleotides encoding such polypeptides, to their use in therapy and in identifying compounds which may be agonists, antagonists and/or inhibitors which are potentially useful in therapy, and to production of such polypeptides and polynucleotides.
BACKGROUND OF THE INVENTION
The drug discovery process is currently undergoing a fundamental revolution as it embraces ‘functional genomics’, that is, high throughput genome- or gene-based biology. This approach as a means to identify genes and gene products as therapeutic targets is rapidly superceding earlier approaches based on ‘positional cloning’. A phenotype, that is a biological function or genetic disease, would be identified and this would then be tracked back to the responsible gene, based on its genetic map position.
Functional genomics relies heavily on high-throughput DNA sequencing technologies and the various tools of bioinformatics to identify gene sequences of potential interest from the many molecular biology databases now available. There is a continuing need to identify and characterize further genes and their related polypeptides/proteins, as targets for drug discovery.
SUMMARY OF THE INVENTION
The present invention relates to Wnt-7a, in particular Wnt-7a polypeptides and Wnt-7a polynucleotides, recombinant materials and methods for their production. In another aspect, the invention relates to methods for using such polypeptides and polynucleotides, including the treatment of cancer; cardiovascular disease; neurological disorders, including, Parkinson's disease, bipolar/unipolar disorder, schizophrenia, Alzheimer's disease; developmental disorders including disorder of the reproductive system, hereinafter referred to as “the Diseases”, amongst others. In a further aspect, the invention relates to methods for identifying agonists and antagonists/inhibitors using the materials provided by the invention, and treating conditions associated with Wnt-7a imbalance with the identified compounds. In a still further aspect, the invention relates to diagnostic assays for detecting diseases associated with inappropriate Wnt-7a activity or levels.
DESCRIPTION OF THE INVENTION
In a first aspect, the present invention relates to Wnt-7a polypeptides. Such peptides include isolated polypeptides comprising an amino acid sequence which has at least 99% identity to that of SEQ ID NO:2 over the entire length of SEQ ID NO:2. Such polypeptides include those comprising the amino acid of SEQ ID NO:2.
Further peptides of the present invention include isolated polypeptides in which the amino acid sequence has at least 99% identity to the amino acid sequence of SEQ ID NO:2 over the entire length of SEQ ID NO:2. Such polypeptides include the polypeptide of SEQ ID NO:2.
Further peptides of the present invention include isolated polypeptides encoded by a polynucleotide comprising the sequence contained in SEQ ID NO:1.
Polypeptides of the present invention are believed to be members of the Wnt ligand family of polypeptides. They are therefore of interest because this protein serves as a transducer molecule for developmental processes during Wnt signal transduction. This is essential for normal morphogenesis and/or differentiated function in diverse tissues, including the brain, kidneys, limbs, somites, reproductive and mammary tissues. Wnt-7a maps to human chromosome 3p25. A number of diseases have been linked to this locus, most particularly schizophrenia and cancer. Pulver, et al. 1995 (Am J Med Genet 60(3):252-60), found significant linkage between schizophrenia and 3p24-p26 in 57 families. In the adult Wnt-7a shows fairly restricted expression in the brain, CNS and reproductive tissues. The involvement of the Wnt signaling cascade in neurological disorders has already been established. Knockout in mice of dishevelled-1 (a downstream transducer of Wnt signaling), shows a phenotype with clear psychiatric, and social interaction abnormalities, which correlate closely to human schizophrenia (Lijam, et al, 1997. Cell 90:895-905). More recently Cotter, et al (Neuroreport. 9:1379-1383. 1998), reported abnormalities in Wnt signaling in Schizophrenics, based upon significant differences in beta and gamma catenin distribution in healthy and schizophrenic individuals. Defective Wnt-7a signaling may therefore induce a similar phenotype to the disheveled knockout phenotype. In addition to neuropsychiatric disorders, Wnt-7a may also play an important role in cancer. The 3p25 region is commonly deleted in a variety of tumors, including breast, prostate and nasopharyngeal tumors(Matsumoto, et al., Genes Chromosomes Cancer 1997 Nov;20(3):268-74). Wnt-7a is a known proto-oncogene, hence it is a strong candidate for some of the 3p deletion tumors. Small molecule agonists/antagonists or antibodies and antisense sequences corresponding to Wnt-7a may be used as pharmaceuticals for the treatment of these and various other disorders.
These properties are hereinafter referred to as Wnt-7a activity” or Wnt-7a polypeptide activity” or “biological activity of Wnt-7a”. Also included amongst these activities are antigenic and immunogenic activities of said Wnt-7a polypeptides, in particular the antigenic and immunogenic activities of the polypeptide of SEQ ID NO:2. Preferably, a polypeptide of the present invention exhibits at least one biological activity of Wnt-7a.
The polypeptides of the present invention may be in the form of the “mature” protein or may be a part of a larger protein such as a precursor or a fusion protein. It is often advantageous to include an additional amino acid sequence which contains secretory or leader sequences, prosequences, sequences which aid in purification such as multiple histidine residues, or an additional sequence for stability during recombinant production.
The present invention also includes variants of the aforementioned polypeptides, that is polypeptides that vary from the referents by conservative amino acid substitutions, whereby a residue is substituted by another with like characteristics. Typical such substitutions are among Ala, Val, Leu and Ile; among Ser and Thr; among the acidic residues Asp and Glu; among Asn and Gln; and among the basic residues Lys and Arg; or aromatic residues Phe and Tyr. Particularly preferred are variants in which several, 5-10, 1-5, 1-3, 1-2 or 1 amino acids are substituted, deleted, or added in any combination.
Polypeptides of the present invention can be prepared in any suitable manner. Such polypeptides include isolated naturally occurring polypeptides, recombinantly produced polypeptides, synthetically produced polypeptides, or polypeptides produced by a combination of these methods. Means for preparing such polypeptides are well understood in the art.
In a further aspect, the present invention relates to Wnt-7a polynucleotides. Such polynucleotides include isolated polynucleotides comprising a nucleotide sequence encoding a polypeptide which has at least 99% identity to SEQ ID NO:2. Such polynucleotides include a polynucleotide comprising the nucleotide sequence contained in SEQ ID NO:1 encoding the polypeptide of SEQ ID NO:2.
Further polynucleotides of the present invention include isolated polynucleotides comprising a nucleotide sequence that has at least 99.5% identity, to a nucleotide sequence encoding a polypeptide of SEQ ID NO:2, over the entire coding region.
Further polynucleotides of the present invention include isolated polynucleotides comprising a nucleotide sequence which has at least 99.5% identity, to SEQ ID NO:1 over the entire length of SEQ ID NO:1. Such polynucleotides include a polynucleotide comprising the polynucleotide of SEQ ID NO:1 as well as the polynucleotide of SEQ ID NO:1.
The invention also provides polynucleotides which are complementary to all the above described polynucleotides.
The nucleotide sequence of SEQ ID NO:1 shows homology with Wnt-7a -Human (U53476-Bui, et al., 1996. Gene 189(1):25-29). The nucleotide sequence of SEQ ID NO:1 is a cDNA sequence and compri
Barnes Michael Robert
Testa Tania Tamson
Han William T.
King William T.
Ratner & Prestia
SmithKline Beecham Plc
Wortman Donna
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