METHODS FOR DETECTING PRE-DIABETES AND DIABETES

• US 20110124022A1
• Filed: 07/23/2009
• Published: 05/26/2011
• Est. Priority Date: 07/23/2008
• Status: Active Grant

• Current Assignee: DiabetOmics, Inc.
• Original Assignee: Diabetomics, LLC
• Inventors: Nagalla, Srinivasa R., Paturi, Vishnupriya Rao, Roberts, Charles T.
• Granted Patent: US 8,476,008 B2
• Time in Patent Office: Days
• US Class Current: 435/15
• CPC Class Codes: G01N 2800/042 Disorders of carbohydrate metabolism, e.g. diabetes, glucose metabolism
• CPC Class Codes: G01N 2800/50 Determining the risk of developing a disease
• CPC Class Codes: G01N 2800/52 Predicting or monitoring the response to treatment, e.g. for selection of therapy based on assay results in personalised medicine; Prognosis
• CPC Class Codes: G01N 33/6893 related to diseases not provided for elsewhere

Abstract

Non-invasive methods are provided herein for the diagnosis of pre-diabetes and diabetes using biomarkers identified in a biological fluid, such as saliva. These biomarkers can be identified using proteomic methods, including but not limited to antibody based methods, such as an enzyme-linked immunosorbant assay (ELISA), a radioimmunoassay (RIA), or a lateral flow immunoassay.

Claims

1. A method for determining if a subject of interest has pre-diabetes or diabetes, is at risk for developing pre-diabetes or diabetes, or monitoring the efficacy of a therapy, comprising

(a) comparing a proteomic profile of a test sample of saliva from a subject of interest with a proteomic profile of a reference sample, wherein the proteomic profile comprises at least one unique expression signature characteristic of pre-diabetes or diabetes, respectively, wherein the test sample proteomic profile and the proteomic profile of the reference proteomic sample comprise information on the expression of at least one of alpha-1-antitrypsin, alpha 1 acid glycoprotein, cystatin C, uteroglobin, carbonic anhydrase 6, pyruvate kinase isozymes M1/M2, neutrophil collagenase, alpha 2-macroglobulin, purine nucleoside phosphorylase, aldehyde dehydrogenase, fatty acid biding protein (epidermal), peroxiredoxin-1, -2, +-6, lamin A/C, apolipoprotein B-100, annexin A2, carbonic anhydrase 1, carbonic anhydrase 2, and lipocalin 2, and wherein:

(i) if the reference sample is a normal sample, and the proteomic profile of the test sample is essentially the same as the proteomic profile of the normal sample the subject is determined not to have pre-diabetes or diabetes, respectively, not to be at risk for developing pre-diabetes or diabetes, or to have an effective therapy, while if the proteomic profile of the test sample has a unique expression signature relative to the proteomic profile of the normal sample the subject is determined to have pre-diabetes or diabetes, respectively, to be at risk for developing pre-diabetes or diabetes, or to have an ineffective therapy;

(ii) if the reference sample is a sample from a subject with pre-diabetes or diabetes, and proteomic profile shares at least one unique expression signature characteristic with the reference sample then the subject is determined to have pre-diabetes or diabetes, respectively, to be at risk for developing pre-diabetes or diabetes, or to have an ineffective therapy, while if the proteomic profile of the test sample has a unique expression signature relative to the reference sample the subject is determined not to have pre-diabetes or diabetes, respectively, not to be at risk for developing pre-diabetes or diabetes, or to have an effective therapy.

4. The method of claim 1 or claim 2, wherein the proteomic profile further comprises information on the expression of one or more of proteasome subunit, aldo-keto reductase family 1 member B 10, cathepsin Z, chitotriosidase isoform 2, 3, +4, transmembrane protease, serine 11D, transthyretin, glycogen phosphorylase, heterogeneous nuclear RNPs A2/B1, leukocyte elastase inhibitor, small proline-rich protein 2F, calmodulin-like protein 5, neuroblast differentiation AHNAK, histone cluster 1, H1e, kallikrein-13, chitinase-3-like protein 1, inter-alpha (Globulin) inhibitor H2, 14-3-3 protein eta, cofilin-1, retinol binding protein 4, plasma, basic proline-rich peptide 1B-8a, isoform 2 of P60953 cdc 42 homolog, actin-related protein 2/3 complex subunit 5, ly6/PLAUR domain-containing protein 3, actin-like protein 2, Rearranged VKA17 V gene segment, brain acid soluble protein 1, golgi phosphoprotein 2, protein FAM49B (L1), and acidic leucine-rich nuclear phosphoprotein 32.

10. The method of claim 1 or claim 2, wherein the diabetes is type 2 diabetes.

11. The method of claim 1 or claim 2, wherein the diabetes is type 1 diabetes.

12. The method of claim 1 or claim 2, wherein the method is a method for monitoring the efficacy of therapy.

13. The method of claim 1 or claim 2, further comprising detecting hemoglobin A1C.

14. The method of claim 1 or claim 2, wherein the subject is obese.

15. The method of any of claims 1 to 14, wherein the proteomic profile is determined using a lateral flow device.

2. A method for determining if a subject of interest has pre-diabetes or diabetes, is at risk for developing pre-diabetes or diabetes or monitoring the efficacy of a therapy, comprising

(a) comparing a proteomic profile of a test sample of saliva from a subject of interest with a proteomic profile of a reference sample, wherein the proteomic profile comprises at least one unique expression signature characteristic of pre-diabetes or diabetes, respectively, wherein the test sample proteomic profile and the proteomic profile of the reference proteomic sample comprise information on the expression of at least one of protein plunc, pancreatic ribonuclease, inter-α

-trypsin inhibitor heavy chain H1, inter-α

-trypsin heavy chain H4, nucleobindin-2, moesin, 14-3-3-epsilon, cystatin A, annexin A3, Protein S100-A7, phosphoglycerate kinase 1, annexin A1, isoform2 of P67936 tropomyosin α

-4, kallikrein-10, desmoplakin, flavin reductase, grancalcin, and calnexin, and wherein;

(i) if the reference sample is a normal sample, and the proteomic profile of the test sample is essentially the same as the proteomic profile of the normal sample the subject is determined not to have pre-diabetes or diabetes, respectively, not to be at risk for developing pre-diabetes or diabetes, or to have an effective therapy, while if the proteomic profile of the test sample has a unique expression signature relative to the proteomic profile of the normal sample the subject is determined to have pre-diabetes or diabetes, respectively, to be at risk for developing pre-diabetes or diabetes, or to have an ineffective therapy;

(ii) if the reference sample is a sample from a subject with pre-diabetes or diabetes, and proteomic profile shares at least one unique expression signature characteristic with the reference sample then the subject is determined to have pre-diabetes or diabetes, respectively, to be at risk for developing pre-diabetes or diabetes, or to have an ineffective therapy, while if the proteomic profile of the test sample has a unique expression signature relative to the reference sample the subject is determined not to have pre-diabetes or diabetes, respectively, not to be at risk for developing pre-diabetes or diabetes, or to have an effective therapy.

3. The method of claim 2, wherein the proteomic profile further comprises information on the expression of one or more of alpha-1-antitrypsin, alpha 1 acid glycoprotein, cystatin C, uteroglobin, carbonic anhydrase 6, pyruvate kinase isozymes M1/M2, neutrophil collagenase, alpha 2-macroglobulin, purine nucleoside phosphorylase, aldehyde dehydrogenase, fatty acid biding protein (epidermal), peroxiredoxin-1, -2, +-6, lamin A/C, apolipoprotein B-100, annexin A2, carbonic anhydrase 1, carbonic anhydrase 2, and lipocalin 2.

5. The method of claim 2, wherein the proteomic profile comprises information on the expression of five or more of protein plunc, pancreatic ribonuclease, inter-α

-trypsin inhibitor heavy chain H1, inter-α

-trypsin heavy chain H4, nucleobindin-2, moesin, 14-3-3-epsilon, cystatin A, annexin A3, Protein 5100-A7, phosphoglycerate kinase 1, annexin A1, isotform2 of P67936 tropomyosin α

-4, kallikrein-10, desmoplakin, flavin reductase, grancalcin, and calnexin.

6. The method of claim 2, wherein the proteomic profile comprises information on the expression of ten or more of protein plunc, pancreatic ribonuclease, inter-α

-trypsin inhibitor heavy chain H1, inter-α

-trypsin heavy chain H4, nucleobindin-2, moesin, 14-3-3-epsilon, cystatin A, annexin A3, Protein S100-A7, phosphoglycerate kinase 1, annexin A1, isotform2 of P67936 tropomyosin α

-4, kallikrein-10, desmoplakin, flavin reductase, grancalcin, and calnexin.

7. The method of claim 2, wherein the proteomic profile comprises information on the expression of fifteen or more of protein plunc, pancreatic ribonuclease, inter-α

-trypsin inhibitor heavy chain H1, inter-α

-trypsin heavy chain H4, nucleobindin-2, moesin, 14-3-3-epsilon, cystatin A, annexin A3, Protein S100-A7, phosphoglycerate kinase 1, annexin A1, isotform2 of P67936 tropomyosin α

-4, kallikrein-10, desmoplakin, flavin reductase, grancalcin, and calnexin.

8. The method of claim 2, wherein the proteomic profile comprises all of protein plunc, pancreatic ribonuclease, inter-α

-trypsin inhibitor heavy chain H1, inter-α

-trypsin heavy chain H4, nucleobindin-2, moesin, 14-3-3-epsilon, cystatin A, annexin A3, Protein S100-A7, phosphoglycerate kinase 1, annexin A1, isotform2 of P67936 tropomyosin α

-4, kallikrein-10, desmoplakin, flavin reductase, grancalcin, and calnexin.

9. The method of claim 2, wherein the proteomic profile further comprises information on the expression of apha-1-antitrypsin, alpha 2-macroglobulin, cystatin C, and transthyretin.