Telomerase diagnostic methods

• US 5,648,215 A
• Filed: 09/28/1994
• Issued: 07/15/1997
• Est. Priority Date: 05/13/1992
• Status: Expired due to Term

• Current Assignee: Board of Regents of the University of Texas System (University of Texas System)
• Original Assignee: Board of Regents of the University of Texas System (University of Texas System)
• Inventors: West, Michael D., Shay, Jerry, Wright, Woodring E.
• Primary Examiner(s): Myers, Carla J.
• Application Number:
  US08/315,216
• Time in Patent Office: 1,021 Days
• Field of Search: 435/6, 435/91.2, 435/91.5, 435/83, 435/184, 435/94, 436/63, 436/64, 436/501, 935/77, 935/78
• US Class Current: 435/6.18
• CPC Class Codes: A61K 31/70 Carbohydrates; Sugars; Derivatives thereof sorbitol A61K31/047
• CPC Class Codes: A61K 38/00 Medicinal preparations containing peptides peptides containing beta-lactam rings A61K31/00; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, A61K31/00; ergot alkaloids of the cyclic peptide type A61K31/48; containing macromolecular compounds having statistically distributed amino acid units A61K31/74; medicinal preparations containing antigens or antibodies A61K39/00; medicinal preparations characterised by the non-active ingredients, e.g. peptides as drug carriers, A61K47/00
• CPC Class Codes: C12N 15/10 Processes for the isolation, preparation or purification of DNA or RNA chemical preparation of DNA or RNA C07H21/00; preparation of non-structural polynucleotides from microorganisms or with enzymes C12P19/34
• CPC Class Codes: C12N 15/113 Non-coding nucleic acids modulating the expression of genes, e.g. antisense oligonucleotides; Antisense DNA or RNA; Triplex- forming oligonucleotides; Catalytic nucleic acids, e.g. ribozymes; Nucleic acids used in co-suppression or gene silencing when used in plants C12N15/8218
• CPC Class Codes: C12N 15/1137 against enzymes viral enzymes C12N15/1131; receptors C12N15/1138
• CPC Class Codes: C12N 2501/70 Enzymes
• CPC Class Codes: C12N 2510/04 Immortalised cells
• CPC Class Codes: C12N 5/0018 Culture media for cell or tissue culture media for specific animal cell type C12N5/06
• CPC Class Codes: C12N 9/1241 Nucleotidyltransferases (2.7.7)
• CPC Class Codes: C12Q 1/6809 Methods for determination or identification of nucleic acids involving differential detection
• CPC Class Codes: C12Q 1/6886 for cancer immunoassay for cancer G01N33/574
• CPC Class Codes: C12Q 2521/113 Telomerase
• CPC Class Codes: C12Q 2600/106 Pharmacogenomics, i.e. genetic variability in individual responses to drugs and drug metabolism
• CPC Class Codes: C12Q 2600/112 Disease subtyping, staging or classification
• CPC Class Codes: C12Y 207/07049 RNA-directed DNA polymerase (2.7.7.49), i.e. telomerase or reverse-transcriptase

Abstract

The presence of telomerase activity in a human somatic tissue or cell sample is positively correlated with the presence of cancer and can be used to diagnose the course of disease progression in a patient.

Claims

1. A method for detecting whether a human breast, prostate, colon, or lung tissue sample contains cancerous cells, said method comprising

(a) preparing a cell extract from said tissue sample;

(b) incubating an aliquot of said cell extract in a reaction mixture comprising a telomerase substrate and a buffer in which telomerase can catalyze the extension of said telomerase substrate;

(c) determining whether said telomerase substrate has been extended in step (b) by addition of telomeric repeat sequences; and

(d) correlating presence of cancerous cells in said sample with the addition of telomeric repeat sequences to said telomerase substrate and absence of cancerous cells in said sample with no addition of telomeric repeat sequences to said telomerase substrate.

2. The method of claim 1, wherein said breast tissue sample is removed from tissue adjacent to a tumor.

3. The method of claim 1, wherein said breast tissue sample is removed from an individual previously diagnosed as having axillary node negative breast cancer.

4. The method of claim 1, wherein said prostate tissue sample is removed from tissue adjacent to a location at which cancer cells are known to have been present.

5. The method of claim 1, wherein said prostate tissue sample is removed from an individual previously diagnosed as having benign prostatic hyperplasia.

6. The method of claim 1, wherein said prostate tissue sample is removed from an individual previously diagnosed as having prostatic intraepithelial neoplasia.

7. The method of claim 1, wherein step (b) further comprises amplifying any extended telomerase substrates in said reaction mixture by an amplification method selected from the group consisting of polymerase chain reaction and ligation chain reaction.

8. A method for determining prognosis of a patient known to have cancer by detecting whether a tissue sample contains cancerous cells, said method comprising

(a) preparing a cell extract from said tissue sample;

(b) incubating an aliquot of said cell extract in a reaction mixture comprising a telomerase substrate and a buffer in which telomerase can catalyze the extension of said telomerase substrate;

(c) determining whether said telomerase substrate has been extended in step (b) by addition of telomeric repeat sequences;

(d) correlating presence of cancerous cells in said sample with the addition of telomeric repeat sequences to said telomerase substrate and absence of cancerous cells in said sample with no addition of telomeric repeat sequences to said telomerase substrate; and

(e) correlating a negative prognosis in said patient with a presence of cancerous cells in said sample, and a positive prognosis in said patient with an absence of cancerous cells in said patient.

9. The method of claim 8 wherein said tissue sample is an axillary-node breast tissue sample.

10. The method of claim 8 wherein said tissue sample is a prostate tissue sample.

11. The method of claim 10 wherein said prostate tissue sample is removed from a patient previously diagnosed as having prostatic intraepithelial neoplasia.

12. The method of claim 10 wherein said prostate tissue sample is removed from a patient previously diagnosed as having benign prostate hyperplasia.