Methods of treating diabetes

• US 20070253904A1
• Filed: 01/16/2007
• Published: 11/01/2007
• Est. Priority Date: 07/15/2004
• Status: Active Grant

• Current Assignee: Joslin Diabetes Center Inc.
• Original Assignee: Joslin Diabetes Center Inc.
• Inventors: Gunton, Jenny, Kahn, C.
• Granted Patent: US 7,700,555 B2
• Time in Patent Office: Days
• US Class Current: 424/9.100
• CPC Class Codes: A61K 31/16 Amides, e.g. hydroxamic acids
• CPC Class Codes: A61K 31/7088 Compounds having three or more nucleosides or nucleotides
• CPC Class Codes: A61K 31/713 Double-stranded nucleic acids or oligonucleotides
• CPC Class Codes: A61K 35/39 Pancreas; Islets of Langerhans Langerhans cells of epidermis A61K35/36
• CPC Class Codes: A61K 38/1709 from mammals
• CPC Class Codes: A61P 3/10 for hyperglycaemia, e.g. antidiabetics
• 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 15/1138 against receptors or cell surface proteins
• CPC Class Codes: C12N 2310/14 interfering N.A.
• CPC Class Codes: G01N 2333/4703 Regulators; Modulating activity
• CPC Class Codes: G01N 2800/042 Disorders of carbohydrate metabolism, e.g. diabetes, glucose metabolism
• CPC Class Codes: G01N 33/6893 related to diseases not provided for elsewhere

Abstract

The invention provides methods of screening for compounds that increase levels or activity of Aryl hydrocarbon Nuclear Receptor Translocator (ARNT) and/or Hypoxia Inducible Factor 1α (HIF1α), for the treatment and prevention of diabetes-related disorders, including type 1 and type 2 diabetes mellitus, impaired glucose tolerance, insulin resistance and beta cell dysfunction; compounds identified by said screening methods; and methods of using said compounds. Also included are methods for treating or preventing diabetes-related diseases using ARNT and/or HIF1α polypeptides and polynucleotides, and for using information regarding the expression, level or activity of ARNT and/or HIF1α in predictive medicine, e.g., diagnostic assays, prognostic assays, monitoring clinical trials, and pharmacogenetics.

Claims

1. A method of evaluating a subject for risk, predisposition, or presence of a diabetes-related disorder, the method comprising:

obtaining a sample from the subject;

evaluating expression, level, or activity of one or both of aryl hydrocarbon nuclear receptor translocator (ARNT) and hypoxia-inducible factor 1α

(HIF1α

) in the sample, and

comparing the expression, level, or activity of one or both of ARNT and HIF1α

in the sample to a reference,

wherein the expression, level or activity in the sample as compared to the control indicates whether the subject has an increased risk, predisposition, or presence of the diabetes-related disorder.

2. The method of claim 1, further comprising:

evaluating the expression, level, or activity of all or a subset of the following genes;

GLUT1, GLUT3, aldolase-B, glyceraldehyde-3-phosphate dehydrogenase and L-type pyruvate-kinase, vascular endothelial growth factor (VEGF), plasminogen activator inhibitor 1 (PAI1), or erythropoietin (EPO); and

comparing the expression, level, or activity of the genes in the sample to a reference,

wherein the expression, level or activity in the sample as compared to the control indicates whether the subject has an increased risk, predisposition, or presence of the diabetes-related disorder.

3. The method of claim 1, wherein the subject is human.

4. The method of claim 1, wherein the diabetes-related disorder is selected from the group consisting of type 1 diabetes, type 2 diabetes, impaired glucose tolerance, insulin resistance, and beta-cell dysfunction.

5. The method of claim 1, wherein the biological sample from the subject comprises tissue from a pancreatic biopsy.

6. The method of claim 1, wherein the evaluating step comprises determining a level of ARNT or HIF1α

serine phosphorylation.

7. The method of claim 1, wherein the evaluating step comprises determining one or more of ARNT or HIF1α

protein level, ARNT or HIF1α

RNA level, or ARNT or HIF1α

gene expression.

8. The method of claim 1, wherein the reference represents a level in a normal control individual, in an individual who has an increased risk of a diabetes-related disorder, in an individual who has a predisposition to develop a diabetes-related disorder, or an individual who has a diabetes-related disorder.

9. A method of treating a subject having or at risk for a diabetes-related disorder, the method comprising:

administering to the subject a therapeutically effective amount of a pharmaceutical composition comprising one or both of;

an ARNT polypeptide or active fragment thereof comprising SEQ ID NO;

8 or an active fragment thereof, and

an HIF1α

polypeptide or active fragment thereof comprising SEQ ID NO;

10 or an active fragment thereof,

thereby treating the subject.

10. A method of treating a subject having or at risk for a diabetes-related disorder, the method comprising:

administering to the subject a therapeutically effective amount of a pharmaceutical composition comprising one or both of;

a nucleic acid encoding an ARNT polypeptide or active fragment thereof comprising SEQ ID NO;

7 that increases expression, levels or activity of ARNT in the subject, and

a nucleic acid encoding an HIF1α

polypeptide or active fragment thereof comprising SEQ ID NO;

9 that increases expression, levels or activity of HIF1α

in the subject,

thereby treating the subject.

14. The method of any of claims 9-11, wherein the diabetes-related disorder is selected from the group consisting of type 1 diabetes, type 2 diabetes, impaired glucose tolerance, insulin resistance, and beta-cell dysfunction.

15. The method of any of claims 9-11, wherein the subject is human.

11. A method of treating a subject having or at risk for a diabetes-related disorder, the method comprising:

administering to the subject a therapeutically effective amount of a pharmaceutical composition comprising a cell expressing one or both of;

an exogenous ARNT polypeptide or active fragment thereof that increases expression, levels or activity of ARNT in the subject, and

an exogenous HIF1α

polypeptide or active fragment that increases expression, levels or activity of HIF1α

in the subject,

thereby treating the subject.

12. The method of claim 11, wherein the cell is a beta cell.

13. The method of claim 12, wherein the cell is derived from the subject.

16. A method of evaluating an effect of a treatment for a diabetes-related disorder, the method comprising:

administering a treatment to the subject;

evaluating expression, level or activity of one or both of ARNT and HIF1α

in a sample from the subject after administration of the treatment; and

comparing the expression, level or activity of one or both of ARNT and HIF1α

in the sample to a reference value,

wherein if the expression, level or activity of one or both of ARNT and HIF1α

in the sample has a predetermined relationship to the reference value, the treatment has a positive effect on the diabetes-related disorder in the subject.

17. The method of claim 16, wherein the reference value is a baseline level for the subject.

18. The method of claim 17, wherein if the expression, level or activity of one or both of ARNT and HIF1α

in the sample is greater than the value to the reference value, the treatment has a positive effect on the diabetes-related disorder in the subject.

19. The method of claim 16, further comprising evaluating expression, level or activity of one or both of ARNT and HIF1α

in a sample from a subject having a diabetes-related disorder before administering the treatment to the subject, to provide a reference that is a baseline level for the subject.

20. The method of claim 16, wherein the diabetes-related disorder is selected from the group consisting of type 1 diabetes, type 2 diabetes, impaired glucose tolerance, insulin resistance, and beta-cell dysfunction.

21. A method of identifying a candidate compound for the treatment of a diabetes-related disorder, the method comprising:

providing a sample comprising one or both of ARNT and HIF1α

,

contacting the sample with a test compound; and

determining if the test compound increases one or both of ARNT and HIF1α

expression, level, or activity in the sample,

wherein a test compound that increases one or both of ARNT and HIF1α

expression, level, or activity is a candidate compound.

22. The method of claim 21, wherein the sample comprises a cell that expresses one or both of ARNT and HIF1α

.

23. The method of claim 22, wherein the cell is a beta cell.

24. The method of claim 21, wherein determining if the test compound increases one or both of ARNT and HIF1α

expression, level, or activity comprises one or more of analyzing ARNT or HIF1α

phosphorylation levels, determining levels of ARNT or HIF1α

polypeptide in the sample, or determining levels of a polynucleotide encoding an ARNT or HIF1α

polypeptide in the sample.

25. The method of claim 21, wherein the test compound is selected from the group consisting of small molecules, polypeptides, and nucleic acids.

26. A method of identifying a candidate therapeutic agent for the treatment of a diabetes related disorder, the method comprising:

providing a model of a diabetes-related disorder;

contacting the model with a candidate compound that increases the expression, level, or activity one or both of ARNT and HIF1α

, wherein the compound was identified by the method of claim 21;

and

evaluating an effect of the candidate compound on the model,

wherein a positive effect on the model indicates that the candidate compound is a candidate therapeutic agent for the treatment of a diabetes-related disorder.

27. The method of claim 26, wherein the model of a diabetes-related disorder is a non-human experimental animal model.

28. The method of claim 27, wherein a positive effect on the model is an improvement in a symptom of the disorder in the animal model.

29. A method of identifying a therapeutic agent for the treatment of a diabetes-related disorder, the method comprising administering a candidate therapeutic agent identified by the method of claim 26 to a subject having a diabetes-related disorder, and evaluating the effect of the candidate therapeutic agent on a symptom of the disorder, wherein a candidate therapeutic agent that has a positive effect on a symptom of the disorder is a therapeutic agent for the treatment of the diabetes-related disorder.

30. The method of claim 26, wherein the subject is human.

31. The method of claim 26, wherein the subject is a non-human animal.