Methods for cancer diagnosis and prognosis

• US 5,639,613 A
• Filed: 04/18/1995
• Issued: 06/17/1997
• 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: Shay, Jerry, Wright, Woodring E., West, Michael David
• Primary Examiner(s): Jones, W. Gary
• Assistant Examiner(s): MYERS, CARLA J
• Application Number:
  US08/423,403
• Time in Patent Office: 791 Days
• Field of Search: 435/6, 435/194, 435/91.2, 435/183, 435/184, 436/63, 436/64, 935/77, 935/78
• US Class Current: 435/6.11
• CPC Class Codes: C12N 2501/70 Enzymes
• CPC Class Codes: C12N 2510/04 Immortalised cells
• CPC Class Codes: C12Q 1/6886 for cancer immunoassay for cancer G01N33/574
• CPC Class Codes: C12Q 2600/118 Prognosis of disease development
• CPC Class Codes: C12Y 207/07049 RNA-directed DNA polymerase (2.7.7.49), i.e. telomerase or reverse-transcriptase

Abstract

A method for predicting tumor progression and prognosing cancer comprises: (a) collecting a sample suspected of containing cancer cells; (b) analyzing the sample for telomerase activity; (c) correlating the activity with a standard level of telomerase activity; and (d) correlating a high telomerase activity with an indication of unfavorable prognosis and a low telomerase activity with a favorable prognosis.

Claims

1. A method for prognosing cancer in humans, said method comprising:

(a) collecting a sample suspected of containing tumor cells;

(b) analyzing said sample for telomerase activity;

(c) correlating said activity with a standard level of telomerase activity; and

(d) correlating a high telomerase activity with an indication of unfavorable prognosis and a low telomerase activity with a favorable prognosis.

2. The method of claim 1, wherein said analyzing in step (b) comprises preparing a cell extract.

3. The method of claim 2, wherein said analyzing in step (b) is under conditions selected to detect any telomerase activity, and said method further comprises analyzing a second aliquot of said sample for telomerase activity under conditions selected to detect telomerase activity only above said standard level of activity;

and associating high telomerase activity to the presence of telomerase in both said first and second aliquots and low telomerase activity to the presence of telomerase activity only in said first aliquot.

4. The method of claim 3, wherein said second aliquot is diluted at least 10-fold relative to the first aliquot.

5. The method of claim 3, wherein said second aliquot is at least 100-fold diluted relative to the first aliquot.

6. The method of claim 1, wherein said analyzing step comprises incubating said aliquot in a reaction mixture comprising a telomerase substrate and a buffer in which telomerase can catalyze the extension of said telomerase substrate, and determining whether said telomerase substrate has been extended by addition of telomeric repeat sequences.

7. The method of claim 6, wherein said analyzing step further comprises amplifying any extended telomerase substrates in said reaction mixture by a polymerase chain reaction using at least one primer complementary to a telomeric repeat sequence.

8. The method of claim 6, wherein said analyzing step further comprises amplifying a control oligonucleotide in said reaction mixture by a polymerase chain reaction.

9. The method of claim 8, wherein said method further comprises normalizing a first signal obtained with said extended telomerase substrates with a second signal obtained with said control oligonucleotide.

10. The method of claim 1, wherein said tumor cells are neuroblastoma cells.

11. The method of claim 10, wherein said standard level is the level of telomerase activity in a fetal adrenal gland cell extract.

12. The method of claim 1, wherein said sample is an adrenal gland sample.

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

14. The method of claim 1, wherein said sample is of sympathetic chain origin.

15. The method of claim 1, wherein said tumor cells are hepatocellular carcinoma cells.

16. The method of claim 1, wherein said sample is a liver sample.

17. The method of claim 1, wherein said tumor cells are leukemic cells.

18. The method of claim 17, wherein said leukemic cells are from a patient with acute myeloid leukemia.

19. The method of claim 1, wherein said favorable prognosis is tumor regression.

20. The method of claim 1, wherein said favorable prognosis is indicative of longer survival rates relative to patients with unfavorable prognosis.