Method of determining a chemotherapeutic regimen based on ERCC1 expression

• US 7,132,238 B2
• Filed: 06/04/2003
• Issued: 11/07/2006
• Est. Priority Date: 12/01/2000
• Status: Expired due to Term

• Current Assignee: Cancer Genetics
• Original Assignee: Response Genetics, Inc
• Inventors: Danenberg, Kathleen D.
• Primary Examiner(s): Horlick, Kenneth R.
• Assistant Examiner(s): Babic, Christopher M.
• Application Number:
  US10/453,784
• Publication Number:
  US 20040005621A1
• Time in Patent Office: 1,252 Days
• Field of Search: None
• US Class Current: 435/6.12
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: C12Q 1/6886 for cancer immunoassay for cancer G01N33/574
• CPC Class Codes: C12Q 2600/158 Expression markers

Abstract

The present invention relates to prognostic methods which are useful in medicine, particularly cancer chemotherapy. The object of the invention to provide a method for assessing ERCC1 expression levels in fixed or fixed and paraffin embedded tissues and determine a platinum-based chemotherapy by examination of the amount of ERCC1 mRNA in a patient'"'"'s tumor cells and comparing it to a predetermined threshold expression level. More specifically, the invention provides to oligonucleotide primer pair ERCC1 and methods comprising their use for detecting levels of ERCC1 mRNA.

Claims

1. A method of treating a tumor with a platinum-based chemotherapeutic regimen comprising:

(a) obtaining a tissue sample of the tumor and fixing the sample, to obtain a fixed tumor sample;

(b) isolating mRNA from the fixed sample in the presence of an effective amount of a chaotrophic agent by first heating the tissue sample in a solution comprising an effective concentration of a chaotrophic compound to a temperature in the range of about 50 to about 100°

C. for a time period of 5 to 120 minutes and recovering said mRNA from said chaotropic solution; and

(c) subjecting the mRNA to amplification using a pair of oligonucleotide primers capable of amplifying a region of the Excision Repair Cross-Complementing 1 (ERCC1) gene, to obtain an amplified sample;

(d) determining the amount of ERCC1 mRNA relative to the quantity of an internal control gene'"'"'s mRNA; and

(f) treating the tumor, wherein the treatment comprises providing a platinum-based chemotherapeutic regimen comprising a genotoxic agent when the determined gene expression level for ERCC1 gene is below a predetermined threshold value.

2. The method of claim 1 wherein the pair of oligonucleotide primers consists of SEQ lID NO:

1 or an oligonucleotide about 80% identical thereto and SEQ ID NO;

2 or an oligonucleotide primer about 80% identical thereto.

3. The method of claim 1 wherein, the tumor is a non-small-cell lung cancer (NSCLC) tumor.

4. The claim 1 wherein, the genotoxic agent is gemcitabine, cisplatin or a combination thereof.