Assessment of gene expression profiles in peripheral occlusive arterial disease.
BACKGROUND
Molecular events responsible for the onset and progression of peripheral occlusive arterial disease (POAD) are incompletely understood. Gene expression profiling may point out relevant features of the disease.


METHODS
Tissue samples were collected as operatory waste from a total of 36 patients with (n = 18) and without (n = 18) POAD. The tissues were histologically evaluated, and the patients with POAD were classified according to Leriche-Fontaine (LF) classification: 11% with stage IIB, 22% with stage III, and 67% with stage IV. Total RNA was isolated from all samples and hybridized onto Agilent 4×44K Oligo microarray slides. The bioinformatic analysis identified genes differentially expressed between control and pathologic tissues. Ten genes with a fold change ≥ 2 (1 with a fold change ≥ 1.8) were selected for quantitative polymerase chain reaction validation (GPC3, CFD, GDF10, ITLN1, TSPAN8, MMP28, NNMT, SERPINA5, LUM, and FDXR). C-reactive protein (CRP) was assessed with a specific assay, while nicotinamide N-methyltransferase (NNMT) was evaluated in the patient serum by enzyme-linked immunosorbent assay.


RESULTS
A multiple regression analysis showed that the level of CRP in the serum is correlated with the POAD LF stages (r(2) = 0.22, P = 0.046) and that serum NNMT is higher in IV LF POAD patients (P = 0.005). The mRNA gene expression of LUM is correlated with the LF stage (r(2) = 0.45, P = 0.009), and the mRNA level of ITLN1 is correlated with the ankle-brachial index (r(2) = 0.42, P = 0.008).


CONCLUSIONS
Our analysis shows that NNMT, ITLN1, LUM, CFD, and TSPAN8 in combination with other known markers, such as CRP, could be evaluated as a panel of biomarkers of POAD.
Prof. C.C. Iliescu Emergency Institute for Cardiovascular Diseases, 258 Fundeni Avenue, Bucharest, Romania. bubenek@alsys.ro
Serban
S
Bubenek
Anca
A
Nastase
Ana Maria
AM
Niculescu
Sorin
S
Baila
Vlad
V
Herlea
Vadimir
V
Lazar
Liliana
L
Paslaru
Anca
A
Botezatu
Dana
D
Tomescu
Irinel
I
Popescu
Simona
S
Dima
D055109:Ankle Brachial Index; D001157:Arterial Occlusive Diseases / Q000175:diagnosis / Q000235:genetics* / Q000378:metabolism; D002097:C-Reactive Protein / Q000378:metabolism; D011508:Chondroitin Sulfate Proteoglycans / Q000096:biosynthesis / Q000235:genetics*; D016207:Cytokines / Q000096:biosynthesis / Q000235:genetics*; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D005263:Femoral Artery / Q000378:metabolism / Q000473:pathology; D005500:Follow-Up Studies; D058851:GPI-Linked Proteins / Q000096:biosynthesis / Q000235:genetics; D005786:Gene Expression Regulation; D006801:Humans; D007632:Keratan Sulfate / Q000096:biosynthesis / Q000235:genetics*; D037102:Lectins / Q000096:biosynthesis / Q000235:genetics*; D000071234:Lumican; D008297:Male; D008875:Middle Aged; D050903:Nicotinamide N-Methyltransferase / Q000097:blood; D012333:RNA, Messenger / Q000235:genetics*; D060888:Real-Time Polymerase Chain Reaction
D011508:Chondroitin Sulfate Proteoglycans; D016207:Cytokines; D058851:GPI-Linked Proteins; C432397:ITLN1 protein, human; C000604306:LUM protein, human; D037102:Lectins; D000071234:Lumican; D012333:RNA, Messenger; D002097:C-Reactive Protein; D007632:Keratan Sulfate; D050903:Nicotinamide N-Methyltransferase