Chronic kidney disease (CKD) with or without uremia affects approximately 10% of worldwide population and increases the risk of developing cardiovascular diseases to several times that of persons with normal renal function because of accelerated atherosclerosis. Multiple studies have shown that atherosclerosis can be initiated by an endothelial cell injury induced by oxidized LDL (oxLDL). Increased levels oxLDL have been recently shown in uremic patients with end-stage renal disease (ESRD), however a mechanistic link between uremia and oxLDL-induced atherosclerosis has not been established. Current theory, developed by Roxborough and Young, suggests that elevated urea results in the formation of carbamylated LDL (cLDL) which makes LDL more susceptible to oxidation. Our data contradict this theory at different levels. In general, our in vitro experiments showed similarities between oxLDL and cLDL action on endothelial and vascular smooth muscle cells, but significantly different levels of cLDL and oxLDL in humans. We hypothesize that blood plasma cLDL is an oxLDL-independent factor that is associated with existing atherosclerotic disease and with its progression in ESRD patients, and is a cause of endothelial cell injury in vitro. A quantitative cLDL sandwich ELISA method recently developed by us showed that concentration of cLDL in human plasma is 5-10 times higher than oxLDL measured by a similar method. Both cLDL and oxLDL are elevated in uremic patients. However, in uremic patients with atherosclerosis measured by intima-media thickness (IMT), cLDL was elevated approximately 200% while oxLDL was not significantly increased compared to uremic patients without thickened IMT. No significant amount of LDL with both types of modifications was found using the ELISA assay of human plasma. Our preliminary data showed that carbamylation of native LDL partially inhibits its oxidation. In vitro, cLDL produced cell injury relevant to atherosclerosis similar to and almost as intense as oxLDL. Changes included endothelial cell death, induction of cell adhesion molecules, attraction of monocytes, and induction of endothelial and vascular smooth muscle cell proliferation. Produced by in vitro chemical modifications, cLDL aggregated more than oxLDL, and cLDL was much more immunogenic in rabbits than oxLDL. Taken together, our data suggest that, at uremic concentrations, cLDL is likely to produce more atherogenic injury to endothelial cells than oxLDL.
Our Specific Aims are: (1) To determine whether cLDL is associated with atherosclerotic disease measures, independently of oxLDL, in cross-sectional and prospective studies of ESRD hemodialysis patients;and (2) To quantify and to study mechanisms of plasma cLDL- and oxLDL-mediated endothelial cell injury induced in vitro by normal physiological and uremia-associated concentrations of the LDL isoforms. If according to our expectation, cLDL is shown to be relevant to CKD-associated atherosclerosis, it may be used in the future as a reliable and mechanistically linked cardiovascular risk factor in CKD patients.

Agency
National Institute of Health (NIH)
Institute
National Heart, Lung, and Blood Institute (NHLBI)
Type
Exploratory/Developmental Grants (R21)
Project #
5R21HL087405-02
Application #
7615532
Study Section
Pathobiology of Kidney Disease Study Section (PBKD)
Program Officer
Liu, Lijuan
Project Start
2008-05-01
Project End
2011-04-30
Budget Start
2009-05-01
Budget End
2011-04-30
Support Year
2
Fiscal Year
2009
Total Cost
$156,500
Indirect Cost
Name
University of Arkansas for Medical Sciences
Department
Internal Medicine/Medicine
Type
Schools of Medicine
DUNS #
122452563
City
Little Rock
State
AR
Country
United States
Zip Code
72205
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Apostolov, Eugene O; Ray, Debarti; Alobuia, Wilson M et al. (2011) Endonuclease G mediates endothelial cell death induced by carbamylated LDL. Am J Physiol Heart Circ Physiol 300:H1997-2004
Apostolov, Eugene O; Ray, Debarti; Savenka, Alena V et al. (2010) Chronic uremia stimulates LDL carbamylation and atherosclerosis. J Am Soc Nephrol 21:1852-7
Apostolov, Eugene O; Shah, Sudhir V; Ray, Debarti et al. (2009) Scavenger receptors of endothelial cells mediate the uptake and cellular proatherogenic effects of carbamylated LDL. Arterioscler Thromb Vasc Biol 29:1622-30