Diagnosis, prognosis, and treatment of kidney disease
The present invention relates to the discovery that an increased fraction of albumin is carbamylated in patients suffering from kidney disease (e.g., end-stage renal disease) and that the fraction of carbamylated albumin is also correlated with increased disease severity, particularly risk of mortality. The present invention also relates to the discovery that free amino acids can reduce carbamylation of albumin. Based on these discoveries the present invention provides diagnostic and prognostic methods for patients suffering from, or suspected of suffering from kidney disease. The invention also provides methods for treating kidney disease by administration of a compound or composition that reduced protein carbamylation, such as free amino acids or dipeptides.
1. A method for diagnosing kidney disease, determining the likelihood of developing kidney disease, or determining the severity of kidney disease in a subject, said method comprising:
(a) determining the fraction of lysine-carbamylated albumin in a biological sample of said subject by:
(i) enzymatically digesting serum proteins in the biological sample; and
(ii) detecting carbamylated albumin that has a carbamylated lysine residue, and
(b) comparing the fraction of lysine-carbamylated albumin in the biological sample of the subject to a control sample,
wherein an increased fraction of lysine-carbamylated albumin in the biological sample of the subject relative to the control sample is indicative of the presence of kidney disease, an increased likelihood of developing kidney disease, or an increased kidney disease severity.
2. The method of claim 1 , wherein said kidney disease severity is indicative of an increased risk of near-term mortality.
3. The method of claim 2 , wherein said increased risk of mortality is determined based on said subject having greater than 1.00% lysine-carbamylated albumin in the biological sample of the subject relative to a control sample.
4. The method of claim 2 , wherein said near-term mortality is within one year.
5. The method of claim 1 , wherein the carbamylated lysine residue is lysine 549.
6. The method of claim 1 , further comprising determining the level of free amino acids in said subject, where a decreased level of free amino acids is indicative of increased disease severity.
7. The method of claim 1 , wherein said subject is a human.
8. The method of claim 1 , wherein said biological sample is a blood sample or a blood serum sample.
9. The method of claim 1 , wherein said determining is performed using mass spectroscopy, or wherein said determining is performed using an antibody or an antigen-binding fragment thereof, or wherein said determining is performed using an enzyme-linked immunosorbent assay (ELISA).
10. The method of claim 9 , wherein said antibody or antibody fragment is conjugated to a detectable label.
11. The method of claim 1 , wherein said kidney disease is end-stage renal disease.
12. A method comprising determining the fraction of lysine-carbamylated albumin in a biological sample of a subject by:
(i) enzymatically digesting serum proteins in the biological sample; and
(ii) detecting carbamylated albumin that has a carbamylated lysine residue,
wherein said subject has or is suspected of having kidney disease.
13. The method of claim 12 , wherein the carbamylated lysine residue is lysine 549.
14. The method of claim 12 , wherein said subject is a human.
15. The method of claim 12 , wherein said biological sample is a blood sample or a blood serum sample.
16. The method of claim 12 , wherein said determining is performed using mass spectroscopy, or wherein said determining is performed using an antibody or an antigen-binding fragment thereof, or wherein said determining is performed using an enzyme-linked immunosorbent assay (ELISA).
17. The method of claim 16 , wherein said antibody or antibody fragment is conjugated to a detectable label.
18. The method of claim 12 , wherein said kidney disease is end-stage renal disease.