Therapy and diagnosis of conditions related to telomere length and/or telomerase activity

• US 5,707,795 A
• Filed: 06/07/1995
• Issued: 01/13/1998
• 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., Wright, Woodring, Shay, Jerry
• Primary Examiner(s): Myers, Carla J.
• Application Number:
  US08/487,290
• Time in Patent Office: 951 Days
• Field of Search: 435/6, 435/5, 435/91.2, 435/4, 436/64, 436/63, 436/811, 935/78, 935/8, 536/24.33
• US Class Current: 435/5
• CPC Class Codes: A61K 31/522 having oxo groups directly attached to the heterocyclic ring, e.g. hypoxanthine, guanine, acyclovir
• CPC Class Codes: A61K 31/70 Carbohydrates; Sugars; Derivatives thereof sorbitol A61K31/047
• CPC Class Codes: A61K 31/7076 containing purines, e.g. adenosine, adenylic acid
• CPC Class Codes: A61K 31/711 Natural deoxyribonucleic acids, i.e. containing only 2'-deoxyriboses attached to adenine, guanine, cytosine or thymine and having 3'-5' phosphodiester links
• 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: 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: 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 5/163 one of the fusion partners being a B or a T lymphocyte
• CPC Class Codes: C12N 9/1241 Nucleotidyltransferases (2.7.7)
• CPC Class Codes: C12Q 1/48 involving transferase
• CPC Class Codes: C12Q 1/68 involving nucleic acids
• CPC Class Codes: C12Q 1/6827 for detection of mutation or polymorphism
• CPC Class Codes: C12Q 1/686 Polymerase chain reaction [PCR]
• CPC Class Codes: C12Q 1/6876 Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes
• CPC Class Codes: C12Q 1/6886 : for cancer immunoassay for cancer G01N33/574
• CPC Class Codes: C12Q 1/703 : Viruses associated with AIDS
• CPC Class Codes: C12Q 2521/113 : Telomerase
• CPC Class Codes: C12Q 2600/112 : Disease subtyping, staging or classification
• CPC Class Codes: C12Q 2600/136 : Screening for pharmacological compounds
• CPC Class Codes: C12Y 207/07049 : RNA-directed DNA polymerase (2.7.7.49), i.e. telomerase or reverse-transcriptase
• CPC Class Codes: G01N 2333/91245 : Nucleotidyltransferases (2.7.7)
• CPC Class Codes: Y02A 50/30 : Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
• CPC Class Codes: Y10S 977/773 : Nanoparticle, i.e. structure having three dimensions of 100 nm or less
• CPC Class Codes: Y10S 977/801 : Drug
• CPC Class Codes: Y10S 977/918 : Immunological
• CPC Class Codes: Y10S 977/927 : Diagnostic contrast agent
• CPC Class Codes: Y10T 436/10 : Composition for standardization, calibration, simulation, stabilization, preparation or preservation; processes of use in preparation for chemical testing
• CPC Class Codes: Y10T 436/105831 : Protein or peptide standard or control [e.g., hemoglobin, etc.]
• CPC Class Codes: Y10T 436/11 : Automated chemical analysis
• CPC Class Codes: Y10T 436/115831 : Condition or time responsive

Abstract

Method and compositions are provided for the determination of telomere length and telomerase activity, as well as the ability to inhibit telomerase activity in the treatment of proliferative diseases. Particularly, primers are elongated under conditions which minimize interference from other genomic sequences, so as to obtain accurate determinations of telomeric length or telomerase activity. In addition, compositions are provided for intracellular inhibition of telomerase activity.

Claims

1. A method for diagnosis of a stage of disease progression in an individual having a disease associated with an increased rate of proliferation of a cell population, said method comprising steps of:

(a) measuring telomere lengths of telomeres from a cell or tissue sample obtained from said individual to obtain a measured telomere length;

(b) comparing said measured telomere length to a control telomere length obtained by measuring telomere length ill a control sample; and

(c) correlating differences between said measured telomere length and said control telomere length with said stage of disease progression.

2. The method of claim 1, wherein said disease is HIV-infection or AIDS.

3. The method of claim 1, wherein said disease is atherosclerosis.

4. The method of claim 1, wherein said disease is cancer.

5. The method of claim 4, wherein said cancer is ovarian cancer.

6. The method of claim 4, wherein said cancer is breast cancer.

7. The method of claim 1, wherein said disease is Down'"'"'s Syndrome.

8. The method of claim 1, wherein said disease is liver disease.

9. The method of claim 1, wherein said disease is muscular dystrophy.

10. The method of claim 1, wherein said measuring step comprises steps of:

(a) digesting genomic DNA of said sample to obtain terminal restriction fragments;

(b) separating said terminal restriction fragments by size;

(c) hybridizing an oligonucleotide probe complementary to telomeric DNA under conditions such that said probe hybridizes specifically to telomeric DNA in said terminal restriction fragments;

(d) measuring amount of bound probe; and

(e) correlating amount of bound probe with telomere length.

11. The method of claim 10, wherein said disease is HIV-infection or AIDS.

12. The method of claim 10, wherein said disease atherosclerosis.

13. The method of claim 10, wherein said disease is cancer.

14. The method of claim 10, wherein said condition is Down'"'"'s Syndrome.

15. The method of claim 10, wherein said condition is liver disease.

16. The method of claim 10, wherein said condition is muscular dystrophy.

17. The method of claim 1, wherein said measuring step comprises steps of:

(a) adding a primer sufficiently complementary to a 3'"'"'-end of a telomere in double-stranded chromosomal DNA of said sample to hybridize specifically thereto in a mixture containing only those deoxynucleotides complementary to nucleotides in telomeric DNA under conditions such that said primer is extended by an agent for polymerization until reaching a non-telomeric deoxynucleotide to form a primer extension product complementary to telomeric DNA; and

(b) measuring primer extension product size to provide a measure of telomere length.

18. The method of claim 17, wherein said primer is 5'"'"'-CCCTAACCCTAACCCTAACCCTAA-3'"'"' (Seq. ID No. 6).

19. The method of claim 17, wherein one of said deoxynucleotides is radiolabeled.

20. The method of claim 17, wherein said disease is HIV-infection or AIDS.

21. The method of claim 17, wherein said disease atherosclerosis.

22. The method of claim 17, wherein said disease is cancer.

23. The method of claim 17, wherein said condition is Down'"'"'s Syndrome.

24. The method of claim 17, wherein said condition is liver disease.

25. The method of claim 17, wherein said condition is muscular dystrophy.

26. The method of claim 1, wherein said measuring step comprises the steps of:

(a) denaturing cellular DNA of said sample in situ;

(b) adding an oligonucleotide probe labeled with a detectable label and complementary to telomeric DNA to said denatured DNA under conditions such that said probe anneals to said DNA;

(c) measuring signal intensity of said label from said probe annealed to said denatured DNA; and

(d) correlating said signal intensity with telomere length.

27. The method of claim 1, wherein said cell or tissue sample obtained from said individual is enriched for a particular cell type.

28. The method of claim 1, wherein said control sample is obtained from said individual at an earlier stage of disease progression.

29. The method of claim 1, wherein said stage of disease progression is a stage after the administration of a therapeutic for treating said disease.

30. The method of claim 2, wherein said disease is HIV-infection and said stage of disease progression is a stage after seroconversion and prior to AIDS.

31. The method of claim 2, wherein said sample obtained from said individual is a peripheral lymphocyte cell sample.

32. The method of claim 2, wherein said stage of disease progression is a stage after the administration of a therapeutic for treating said disease.

33. The method of claim 29, wherein said peripheral lymphocyte cell sample is a CD4⁺ cell sample.