Urine and serum biomarkers associated with diabetic nephropathy
Disclosed is use of urine and serum biomarkers in diagnosing diabetic nephropathy, staging diabetic nephropathy, monitoring diabetic nephropathy progress, and assessing efficacy of diabetic nephropathy treatments. These biomarkers include urine precursor alpha-2-HS-glycoprotein, urine alpha-1 antitrypsin, urine alpha-1 acid glycoprotein, urine osteopontin, serum osteopontin, their fragments, and combinations thereof.
1. A method for assessing efficacy of a diabetic nephropathy treatment in a subject in need thereof, comprising:
determining in the subject a pre-treatment level of a urine biomarker, wherein the biomarker is a fragment of alpha-1 antitrypsin selected from the group consisting of (i) KGKWERPFEVKDTEEEDF (SEQ ID NO:3), (ii) MIEQNTKSPLFMGKVVNPTQK (SEQ ID NO:4), (iii) EDPQGDAAQKTDTSHHDQDHPTFNKITPNLAE (SEQ ID NO:5), and (iv) EDPQGDAAQKTDTSHHDQDHPTFNKITPNLAEFA (SEQ ID NO:6);
determining in the subject a post-treatment level of the urine biomarker; and
assessing efficacy of the diabetic nephropathy treatment by comparing the pre-treatment level and the post-treatment level, wherein the post-treatment level of the biomarker being the same or lower than the pre-treatment level of the biomarker indicates effectiveness of the diabetic nephropathy treatment, the level of the biomarker being determined with an antibody that specifically binds to the biomarker.
2. The method of claim 1 , wherein the fragment of alpha-1 antitrypsin is KGKWERPFEVKDTEEEDF (SEQ ID NO:3).
3. The method of claim 1 , wherein the fragment of alpha-1 antitrypsin is MIEQNTKSPLFMGKVVNPTQK (SEQ ID NO:4).
4. The method of claim 1 , wherein the fragment of alpha-1 antitrypsin is EDPQGDAAQKTDTSHHDQDHPTFNKITPNLAE (SEQ ID NO:5).
5. The method of claim 1 , wherein the fragment of alpha-1 antitrypsin is EDPQGDAAQKTDTSHHDQDHPTFNKITPNLAEFA (SEQ ID NO:6).
6. A method for assessing efficacy of a diabetic nephropathy treatment in a subject in need thereof, comprising:
obtaining a first urine sample, and optionally, a first serum sample, from the subject before treatment;
obtaining a second urine sample, and optionally, a second serum sample, from the subject after treatment;
determining the level of each of a plurality of biomarkers in the first and second samples, and optionally, one or more clinical factors, the one or more clinical factors being selected from the group consisting of age, gender, HbA1c, albumin/creatinine ratio, and glomerular filtration rate, wherein the plurality of biomarkers include:
(a) a first urine biomarker that is a fragment of alpha-1 antitrypsin selected from the group consisting of (i) KGKWERPFEVKDTEEEDF (SEQ ID NO:3), (ii) MIEQNTKSPLFMGKVVNPTQK (SEQ ID NO:4), (iii) EDPQGDAAQKTDTSHHDQDHPTFNKITPNLAE (SEQ ID NO:5), and (iv) EDPQGDAAQKTDTSHHDQDHPTFNKITPNLAEFA (SEQ ID NO:6); and
(b) one or more biomarkers selected from the group consisting of (i) a second urine biomarker that is precursor alpha-2-HS-glycoprotein, VVSLGSPSGEVSHPRKT (SEQ ID NO:1), or MGVVSLGSPSGEVSHPRKT (SEQ ID NO:2), (ii) a third urine biomarker that is alpha-1 acid glycoprotein or GQEHFAHLLILRDTKTYMLAFDVNDEKNWGLS (SEQ ID NO:7), (iii) a fourth urine biomarker that is osteopontin or YPDAVATWLNPDPSQKQNLLAPQNAVSSEETNDFKQETLPSK (SEQ ID NO:8), and (iv) if the optional first and second serum samples are obtained, a serum biomarker that is osteopontin or YPDAVATWLNPDPSQKQNLLAPQNAVSSEETNDFKQETLPSK (SEQ ID NO:8);
calculating a first disease score and a second disease score based on the levels of the plurality of biomarkers in the first and second samples, respectively, and optionally, on the one or more clinical factors; and
assessing efficacy of the diabetic nephropathy treatment in the subject, wherein the second disease score being equal to or lower than the first disease score indicates effectiveness of the diabetic nephropathy treatment, the level of the biomarker being determined with an antibody that specifically binds to the biomarker.
7. The method of claim 6 , wherein the disease score is calculated by an analysis selected from the group consisting of ridge regression analysis, factor analysis, discriminant function analysis, and logistic regression analysis.
8. The method of claim 6 , wherein the plurality of biomarkers include the third urine biomarker.
9. The method of claim 6 , wherein the plurality of biomarkers include the second urine biomarker and the third urine biomarker.
10. The method of claim 6 , wherein the plurality of biomarkers include the second urine biomarker, the third urine biomarker, and the serum biomarker.
11. The method of claim 6 , wherein the plurality of biomarkers include the second urine biomarker, the third urine biomarker, the fourth urine biomarker, and the serum biomarker.
12. The method of claim 6 , wherein the disease score is calculated based on one or more clinical factors.
13. The method of claim 6 , wherein the fragment of alpha-1 antitrypsin is KGKWERPFEVKDTEEEDF.
14. The method of claim 6 , wherein the fragment of alpha-1 antitrypsin is MIEQNTKSPLFMGKVVNPTQK.
15. The method of claim 6 , wherein the fragment of alpha-1 antitrypsin is EDPQGDAAQKTDTSHHDQDHPTFNKITPNLAE.
16. The method of claim 6 , wherein the fragment of alpha-1 antitrypsin is EDPQGDAAQKTDTSHHDQDHPTFNKITPNLAEFA.