Assessment of gene expression profiles in peripheral occlusive arterial disease

Can J Cardiol. Nov-Dec 2012;28(6):712-20. doi: 10.1016/j.cjca.2012.03.013. Epub 2012 Jun 19.

Abstract

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.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Ankle Brachial Index
  • Arterial Occlusive Diseases / diagnosis
  • Arterial Occlusive Diseases / genetics*
  • Arterial Occlusive Diseases / metabolism
  • C-Reactive Protein / metabolism
  • Chondroitin Sulfate Proteoglycans / biosynthesis
  • Chondroitin Sulfate Proteoglycans / genetics*
  • Cytokines / biosynthesis
  • Cytokines / genetics*
  • Enzyme-Linked Immunosorbent Assay
  • Female
  • Femoral Artery / metabolism
  • Femoral Artery / pathology
  • Follow-Up Studies
  • GPI-Linked Proteins / biosynthesis
  • GPI-Linked Proteins / genetics
  • Gene Expression Regulation*
  • Humans
  • Keratan Sulfate / biosynthesis
  • Keratan Sulfate / genetics*
  • Lectins / biosynthesis
  • Lectins / genetics*
  • Lumican
  • Male
  • Middle Aged
  • Nicotinamide N-Methyltransferase / blood
  • RNA, Messenger / genetics*
  • Real-Time Polymerase Chain Reaction

Substances

  • Chondroitin Sulfate Proteoglycans
  • Cytokines
  • GPI-Linked Proteins
  • ITLN1 protein, human
  • LUM protein, human
  • Lectins
  • Lumican
  • RNA, Messenger
  • C-Reactive Protein
  • Keratan Sulfate
  • Nicotinamide N-Methyltransferase