Methods for monitoring ELOVL2, KLF14 and PENK gene expression following treatment with vitamin C

• US 10,202,650 B2
• Filed: 05/31/2017
• Issued: 02/12/2019
• Est. Priority Date: 05/31/2016
• Status: Active Grant

• Current Assignee: Regents of the University of California (University of California), Youhealth Biotech, Limited
• Original Assignee: Regents of the University of California (University of California), Youhealth Biotech, Limited
• Inventors: Zhang, Kang, Hou, Rui, Zheng, Lianghong
• Primary Examiner(s): Myers, Carla J
• Application Number:
  US15/610,407
• Publication Number:
  US 20170342496A1
• Time in Patent Office: 622 Days
• Field of Search: None
• CPC Class Codes: C12Q 1/6883 for diseases caused by alterations of genetic material
• CPC Class Codes: C12Q 2600/106 Pharmacogenomics, i.e. genetic variability in individual responses to drugs and drug metabolism
• CPC Class Codes: C12Q 2600/154 Methylation markers
• CPC Class Codes: C12Q 2600/158 Expression markers
• CPC Class Codes: G01N 33/57407 Specifically defined cancers
• CPC Class Codes: G01N 33/57434 of prostate

Abstract

Disclosed herein are methods of increasing the expression rate of epigenetic markers such as ELOVL2, KLF14, and PENK with administration of a therapeutic agent (e.g., vitamin C or its derivatives, analogs, metabolites, prodrugs, or pharmaceutically acceptable salts thereof). Also described herein are methods of modulating the methylation pattern of epigenetic markers such as ELOVL2, KLF14, and PENK with administration of a therapeutic agent (e.g., vitamin C or its derivatives, analogs, metabolites, prodrugs, or pharmaceutically acceptable salts thereof).

Claims

1. A method of detecting an expression level of ELOVL2, KLF14, and PENK in a first subject, comprising:

a) administering to the first subject a therapeutically effective dose of vitamin C or its derivatives, metabolites, or pharmaceutically acceptable salts thereof for a first time period;

b) obtaining a sample from the first subject after the first time period; and

c) detecting the expression level of ELOVL2, KLF14, and PENK in the first subject by contacting the sample with one set and no more than one set of nucleic acid probes, wherein the one set of nucleic acid probes hybridizes to three markers and no more than three markers, wherein the three markers are ELOVL2, KLF14, and PENK and detecting binding between ELOVL2, KLF14, or PENK and the nucleic acid probes.

2. The method of claim 1, wherein vitamin C or its derivatives, metabolites, or pharmaceutically acceptable salts thereof is L-ascorbic acid 2-phosphate.

3. The method of claim 1, wherein at least one of the nucleic acid probes hybridizes to a sequence selected from the group consisting of SEQ ID NOs:

1, 5, 10, 14, 18, 21, 31, 33, 45, 47, and 74.

4. The method of claim 1, wherein the sample is a cell sample.

5. The method of claim 1, wherein the first time period comprises at least one day, two days, three days, four days, five days, one week, two weeks, three weeks, four weeks, one month, two months, three months, four months, five months, six months, 1 year, two years, three years, or more.

6. The method of claim 1, wherein the therapeutically effective dose of vitamin C or its derivatives, metabolites, or pharmaceutically acceptable salts thereof comprises from about 0.1 μ

g/mL to about 200 μ

g/mL, from about 1 μ

g/mL to about 150 μ

g/mL, from about 5 μ

g/mL to about 100 μ

g/mL, from about 10 μ

g/mL to about 100 μ

g/mL, from about 20 μ

g/mL to about 100 μ

g/mL, from about 30 μ

g/mL to about 100 μ

g/mL, from about 50 μ

g/mL to about 100 μ

g/mL, from about 1 μ

g/mL to about 50 μ

g/mL, from about 5 μ

g/mL to about 50 μ

g/mL, from about 10 μ

g/mL to about 50 μ

g/mL, or from about 50 μ

g/mL to about 200 μ

g/mL.

7. The method of claim 1, wherein a dose of vitamin C or its derivatives, metabolites, or pharmaceutically acceptable salts thereof greater than 200 μ

g/mL increases reactive oxidative species.

8. The method of claim 1, wherein the sample is obtained from a subject having a metabolic disease or condition.

9. The method of claim 1, wherein the sample is obtained from a subject having diabetes.

10. The method of claim 1, wherein the sample is obtained from a subject having Werner syndrome, progeria, or post-traumatic stress disorder.

11. A method of detecting the expression level of a plurality of markers in a first subject, comprising:

a) administering to the first subject a therapeutically effective dose of vitamin C or its derivatives, metabolites, or pharmaceutically acceptable salts thereof for a first time period;

b) obtaining a sample from the first subject after the first time period; and

c) detecting the expression level of ELOVL2, KLF14, PENK, and one or more markers selected from the group consisting of FHL2, SMC4, SLC12A5, TEZM151A, TTF2, TRIM45, TRIM59, ACSS3, ARID5A, BLMH, BRD4, CD28, EPHX3, SLX1, BDNF, NDF, and GDNF in the first subject by;

i) contacting the sample with two sets and no more than two sets of nucleic acid probes, wherein the first set of nucleic acid probes hybridizes to three markers and no more than three markers, wherein the three markers are ELOVL2, KLF14, and PENK and wherein the second set of nucleic acid probes hybridizes to at least one but no more than 19 markers selected from the group consisting of FHL2, SMC4, SLC12A5, TEZM151A, TTF2, TRIM45, TRIM59, ACSS3, ARID5A, BLMH, BRD4, CD28, EPHX3, RIN1, SLX1, BDNF, NDF, GDNF, and combinations thereof; and

ii) detecting binding between ELOVL2, KLF14, or PENK and nucleic acid probes in the first set of nucleic acid probes and between one or more markers selected from the group consisting of FHL2, SMC4, SLC12A5, TEZM151A, TTF2, TRIM45, TRIM59, ACSS3, ARID5A, BLMH, BRD4, CD28, EPHX3, RIN1, SLX1, BDNF, NDF, and GDNF and nucleic acid probes in the second set of nucleic acid probes.

12. The method of claim 11, wherein vitamin C or its derivatives, metabolites, or pharmaceutically acceptable salts thereof is L-ascorbic acid 2-phosphate.

13. The method of claim 11, wherein at least one of the nucleic acid probes from the first set and second set of nucleic acid probes hybridizes to a sequence selected from the group consisting of SEQ ID NOs:

1, 3, 5, 6, 7, 9, 10, 14, 15, 18, 20, 21, 22, 31, 33, 45, 47, 74, and 140.

14. The method of claim 11, wherein the first time period comprises at least one day, two days, three days, four days, five days, one week, two weeks, three weeks, four weeks, one month, two months, three months, four months, five months, six months, 1 year, two years, three years, or more.

15. The method of claim 11, wherein the therapeutically effective dose of vitamin C or its derivatives, metabolites, or pharmaceutically acceptable salts thereof comprises from about 0.1 μ

g/mL to about 200 μ

g/mL, from about 1 μ

g/mL to about 150 μ

g/mL, from about 5 μ

g/mL to about 100 μ

g/mL, from about 10 μ

g/mL to about 100 μ

g/mL, from about 20 μ

g/mL to about 100 μ

g/mL, from about 30 μ

g/mL to about 100 μ

g/mL, from about 50 μ

g/mL to about 100 μ

g/mL, from about 1 μ

g/mL to about 50 μ

g/mL, from about 5 μ

g/mL to about 50 μ

g/mL, from about 10 μ

g/mL to about 50 μ

g/mL, or from about 50 μ

g/mL to about 200 μ

g/mL.

16. The method of claim 11, wherein a dose of vitamin C or its derivatives, metabolites, or pharmaceutically acceptable salts thereof greater than 200 μ

g/mL increases reactive oxidative species.

17. The method of claim 11, wherein the method further comprises detecting a level of cortisol in the first subject after administration of the therapeutically effective dose of vitamin C or its derivatives, metabolites, or pharmaceutically acceptable salts thereof.

18. The method of claim 11, wherein the sample is obtained from a subject having a metabolic disease or condition.

19. The method of claim 11, wherein the sample is obtained from a subject having diabetes.

20. The method of claim 11, wherein the sample is obtained from a subject having Werner syndrome, progeria, or post-traumatic stress disorder.

21. The method of claim 11, wherein the sample is a cell sample.