Increased expression of a disintegrin and metalloproteinase thrombospondin 1 in thrombosed hemodialysis grafts.
OBJECTIVE
To use proteomic analysis to identify upregulated and downregulated proteins in thrombosed hemodialysis graft specimens. One of these significantly upregulated proteins was a disintegrin and metalloproteinase thrombospondin-1 (ADAMTS-1), and its expression and activity were determined in thrombosed hemodialysis grafts.


METHODS
Hemodialysis vascular access samples (thrombosed veins, n = 8; control veins, n = 6) were obtained from patients who required surgical revision. Proteomic analysis was performed with isotope-coded affinity tag labeling with multidimensional liquid chromatography followed by tandem mass spectrometry on four thrombosed hemodialysis graft specimens with control veins. Expression of ADAMTS-1 was confirmed by performing immunoprecipitation followed by Western blot analysis. Finally, immunohistochemistry was used to localize expression in a separate group of patients with thrombosed grafts.


RESULTS
Thirty-nine unique proteins were common to all four patients. ADAMTS-1 was one of the only significantly upregulated protein (>38 fold). ADAMTS-1 expression was confirmed by performing immunoprecipitation and Western blot analysis and was significantly increased. ADAMTS-1 expression was localized to adventitial macrophages and neutrophils of thrombosed grafts.


CONCLUSIONS
ADAMTS-1 was significantly upregulated in thrombosed hemodialysis grafts by mass spectrometric analysis and Western blot analysis. Expression was localized to adventitial macrophages and leukocytes. It is hypothesized that ADAMTS-1 may be related to intimal hyperplasia in hemodialysis vascular access grafts. Future work is planned on inhibiting ADAMTS-1 expression and determining the effect on intimal hyperplasia in hemodialysis grafts.
Misra
Sanjay
S
Department of Radiology, Mayo Clinic, Rochester, MN 55905, USA. misra.sanjay@mayo.edu
Lee
Nathan
N
Fu
Alex A
AA
Raghavakaimal
Sreekumar
S
Mandrekar
Jaywant
J
Bjarnason
Haraldur
H
McKusick
Michael A
MA
Iruela-Arispe
Luisa
L
Mukhopadhyay
Debabrata
D
D051722:ADAM Proteins / Q000032:analysis*; D000071097:ADAMTS1 Protein; D000368:Aged; D001807:Blood Vessel Prosthesis; D019917:Blood Vessel Prosthesis Implantation / Q000009:adverse effects* / Q000295:instrumentation; D015153:Blotting, Western; D016022:Case-Control Studies; D002853:Chromatography, Liquid; D005260:Female; D006083:Graft Occlusion, Vascular / Q000201:enzymology* / Q000209:etiology / Q000473:pathology; D006801:Humans; D007150:Immunohistochemistry; D047468:Immunoprecipitation; D007553:Isotope Labeling; D008264:Macrophages / Q000201:enzymology; D008297:Male; D008875:Middle Aged; D009504:Neutrophils / Q000201:enzymology; D011138:Polytetrafluoroethylene; D011474:Prosthesis Design; D040901:Proteomics / Q000379:methods; D006435:Renal Dialysis; D053719:Tandem Mass Spectrometry; D013927:Thrombosis / Q000201:enzymology* / Q000209:etiology / Q000473:pathology; D016896:Treatment Outcome; D015854:Up-Regulation
D011138:Polytetrafluoroethylene; D051722:ADAM Proteins; D000071097:ADAMTS1 Protein; C103859:ADAMTS1 protein, human