identifying the presence of human mammary carcinoma in mammary tissue from a human; identification of a v-erbB related human gene that is a new member of the tyrosine kinase
Inventors
King, C. Richter • Kraus, Matthias H. • Aaronson, Stuart A.
Assignees
US Department of Health and Human Services
Publication Number
US-7838216-B1
Publication Date
2010-11-23
Expiration Date
2027-11-23
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Abstract
The isolation, cloning and characterization of a human gene related to but distinct from EGF receptor gene has been described. Nucleotide sequence of the gene and amino acid sequence of the polypeptide encoded by the gene have been determined. The use of the nucleic acid probes and antibodies having specific binding affinity with said polypeptide for diagnostic and therapeutic purposes have also been described.
Core Innovation
The invention relates to the isolation, cloning, and characterization of a human gene that is related to, but distinct from, the epidermal growth factor (EGF) receptor gene, identifying it as a new member of the tyrosine kinase encoding family of genes. This gene, discovered through amplification in a human mammary carcinoma designated MAC117, encodes a protein product whose nucleotide and amino acid sequences have been determined. The invention also encompasses nucleic acid probes and antibodies specific to this gene product for diagnostic and therapeutic applications.
The problem addressed is the lack of knowledge about genes other than the known EGF receptor that belong to the tyrosine kinase family and that might play a role in human tumor development. Previous studies had indicated gene amplification or rearrangements in oncogenes like EGF receptor in tumors, but a gene closely related to, yet distinct from, EGF receptor and amplified specifically in human mammary carcinoma had not been identified. The invention solves this by identifying and characterizing this novel v-erbB-related gene, enabling detection of its amplification and expression in human mammary carcinoma.
The invention provides cloned DNA sequences comprising the gene, its RNA and protein products, and antibodies directed against the protein. It also describes methods for detection of gene amplification, mRNA overexpression, and protein overexpression in human mammary carcinoma samples. Additionally, it details the expression of the gene in heterologous systems and demonstrates transformation of NIH/3T3 cells, establishing the gene’s oncogenic potential. These elements collectively enable both diagnostic and therapeutic strategies based on this novel gene.
Claims Coverage
The patent contains one independent claim directed to a method for identifying human mammary carcinoma by detecting amplification of a specific gene encoding a protein comprising SEQ ID NO:1. The inventive features center on detecting gene amplification and its correlation to carcinoma presence.
Method of identifying mammary carcinoma via gene amplification
A method comprising determining whether a gene encoding a protein with amino acid sequence SEQ ID NO:1 is amplified in human mammary tissue. Amplification is defined as a higher gene occurrence in tissue compared to normal mammary tissue. Such amplification indicatively identifies the presence of human mammary carcinoma.
Use of specific nucleic acid sequences for amplification detection
Utilization of the gene’s nucleic acid sequence SEQ ID NO:2 or DNA fragments, including the Bam HI fragment in the deposited pUC12 subclone, to detect gene amplification in mammary tissue by hybridization techniques.
Hybridization-based analysis of gene amplification
Analyzing DNA from human mammary tissue by contacting with a nucleic acid probe that hybridizes to the gene encoding the protein of SEQ ID NO:1, measuring hybridization amount, and determining gene amplification when hybridization exceeds normal tissue levels.
Overexpression as a diagnostic indicator
Determining overexpression of the gene in mammary tissue as further indication of carcinoma presence by measuring mRNA or protein levels encoding or comprising SEQ ID NO:1 and comparing to normal tissue levels.
Protein level detection using antibodies
Measuring protein levels by reacting antibodies prepared against the protein (SEQ ID NO:1) with mammary tissue, detecting elevated protein expression indicative of carcinoma.
The claims collectively cover a diagnostic method for detecting human mammary carcinoma through identifying amplification and overexpression of a specific gene encoding a protein characterized by SEQ ID NO:1, employing nucleic acid probes and antibody-based protein detection.
Stated Advantages
Provides a novel diagnostic tool for detecting human mammary carcinoma by identifying gene amplification.
Enables detection of gene expression abnormalities at nucleic acid and protein levels, facilitating early and accurate cancer diagnosis.
Allows production of antibodies for specific detection and potential therapeutic targeting of the amplified gene product.
Permits expression of the gene in heterologous systems to produce large amounts of protein for research, diagnosis, or treatment.
Offers potential treatment methods involving targeting overexpressed gene product on tumor cells with toxic conjugates.
Documented Applications
Molecular diagnosis of human mammary carcinoma by detecting gene amplification via nucleic acid hybridization and overexpression of mRNA.
Detection of the protein product in tumor cells and histological tissue sections using specific antibodies for diagnostic purposes.
Therapeutic targeting of cancers expressing high levels of the protein product using antibodies or ligands conjugated to toxic substances.
Production of recombinant protein in heterologous expression systems for use in diagnostics, research, or as antigen sources.
Development of diagnostic kits comprising specific nucleic acid probes or antibodies for detection of gene amplification or protein overexpression.
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