Wnt and frizzled receptors as targets for immunotherapy in head and neck squamous cell carcinomas

• US 20040203003A1
• Filed: 03/05/2004
• Published: 10/14/2004
• Est. Priority Date: 05/01/2001
• Status: Active Grant

• Current Assignee: Regents of the University of California (University of California)
• Original Assignee: Regents of the University of California (University of California)
• Inventors: Carson, Dennis A, Leoni, Lorenzo M, Wu, Christina, Rhee, Chae-Seo, Corr, Maripat, Sen, Malini
• Granted Patent: US 7,682,607 B2
• Time in Patent Office: Days
• US Class Current: 435/6
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: C07K 16/28 against receptors, cell surface antigens or cell surface determinants
• CPC Class Codes: C12Q 1/6851 Quantitative amplification
• CPC Class Codes: C12Q 1/6886 for cancer immunoassay for cancer G01N33/574
• CPC Class Codes: C12Q 2600/158 Expression markers

Abstract

The diverse receptor-ligand pairs of the Wnt and frizzled (Fzd) families play important roles during embryonic development, and thus may be overexpressed in cancers that arise from immature cells. The mRNA levels and expression levels of 5 Wnt (Wnt-1, 5a, 7a, 10b, 13) and 2 Fzd (Fzd-2, 5) genes in 10 head and neck squamous carcinoma cell lines (HNSCC) were investigated. In addition, anti-Wnt-1 antibodies were used to study the Wnt/Fzd signalling pathway. These results indicate that HNSCC cell lines overexpress one or more Wnt and Fzd genes, and the growth and survival of a subset of HNSCC may depend on the Wnt/Fzd pathway. Therefore, The Wnt and Fzd receptors may be useful targets for immunotherapy of this common cancer.

Claims

1. A method for determining overexpression of a wnt or frizzled gene in a tumor cell comprising:

(a) isolating messenger RNA from a tumor cell and from a corresponding normal cell from the same source;

(b) performing reverse transcription PCR on the tumor cell and the normal cell utilizing primers directed against regions of the wnt or frizzled gene that are non-homologous with a known wnt gene or a known frizzled gene and obtaining amplicons from the reverse transcription PCR;

(c) assessing the concentrations of the resulting amplicons of step (b) in comparison to a control housekeeping gene; and

(d) identifying tumor cells that overexpress a wnt or frizzled gene at about five fold compared to normal cells tissue from the same source

2. An isolated antibody directed against at least one sequence that corresponds to a non-homologous regions of a known wnt gene or a known frizzled gene.

3. A method of detecting overexpression of at least one wnt and/or frizzled protein in a cancer cell compared to a non-cancer cell comprising:

(a) contacting the cancer cell with an antibody directed against at least one sequence that corresponds to a non-homologous region of a known wnt or a known frizzled gene;

(b) contacting the non-cancer cell with the same antibody as in step (a);

(c) comparing the interaction of the antibody with the cancer cell to the interaction of the antibody with the non-cancer cell; and

(d) correlating the interactions in (c) with the expression levels of the wnt and/or frizzled protein in both the cancer cell and the non-cancer cell.

4. A method for altering the growth of a cell overexpressing at least one wnt and/or frizzled protein comprising:

contacting the cell with a non-crossreactive antibody against the wnt and/or frizzled protein.

5. The method of claim 4, wherein the antibody is coupled to a cytotoxic agent and/or radioisotope.

6. A method for altering the growth of a cell overexpressing at least one wnt and/or frizzled protein comprising:

contacting the cell with a synthetic peptide, a recombinant protein, or a DNA vector, or any combination of a synthetic peptide and a recombinant protein and a DNA vector, comprising at least one non-homologous region of known wnt and/or frizzled proteins.