Periostin, discovered by nano-flow liquid chromatography and mass spectrometry, is a novel marker of diabetic retinopathy

• Published in: Biochemical and biophysical research communications 2010-08, Vol.399 (2), p.221-226
• Main Authors: Takada, Michiya, Ban, Yoshiyuki, Yamamoto, Gou, Ueda, Toshihiko, Saito, Yuta, Nishimura, Eiichi, Fujisawa, Kunimi, Koide, Ryohei, Mizutani, Masakazu, Kozawa, Tadahiko, Shiraishi, Yuji, Bando, Yasuhiko, Tachikawa, Tetsuhiko, Hirano, Tsutomu
• Format: Article
• Language: English
• Subjects: 60 APPLIED LIFE SCIENCES; Adult; Aged; Aged, 80 and over; BIOLOGICAL MARKERS; Biomarkers - analysis; Biomarkers - metabolism; Cell Adhesion Molecules - analysis; Cell Adhesion Molecules - genetics; Cell Adhesion Molecules - metabolism; Chromatography, Liquid - methods; Diabetic retinopathy; Diabetic Retinopathy - metabolism; Epiretinal membrane; Epiretinal Membrane - metabolism; EPITHELIUM; Eye Proteins - analysis; Eye Proteins - metabolism; Female; GENES; Humans; HYPOTHESIS; LICENSES; LIQUID COLUMN CHROMATOGRAPHY; Male; MASS SPECTROSCOPY; MEMBRANES; MESSENGER-RNA; Middle Aged; Nerve Growth Factors - analysis; Nerve Growth Factors - metabolism; PATHOGENESIS; PIGMENTS; POLYMERASE CHAIN REACTION; PROTEINS; Proteomics; Serpins - analysis; Serpins - metabolism; Tandem Mass Spectrometry - methods
• ISSN: 0006-291X; 1090-2104
• DOI: 10.1016/j.bbrc.2010.07.058

► In proliferative membrane and epiretinal membrane specimens, the numbers of proteins are 225 and 154, respectively, and 123 proteins are common to both. ► Periostin and thrombospondin-1 proteins are unique to the proliferative membrane specimens. ► The expression of periostin is significantly up-regulated in proliferative membrane specimens. Diabetes can lead to serious microvascular complications including proliferative diabetic retinopathy (PDR), the leading cause of blindness in adults. Recent studies using gene array technology have attempted to apply a hypothesis-generating approach to elucidate the pathogenesis of PDR, but these studies rely on mRNA differences, which may or may not be related to significant biological processes. To better understand the basic mechanisms of PDR and to identify potential new biomarkers, we performed shotgun liquid chromatography (LC)/tandem mass spectrometry (MS/MS) analysis on pooled protein extracts from neovascular membranes obtained from PDR specimens and compared the results with those from non-vascular epiretinal membrane (ERM) specimens. We detected 226 distinct proteins in neovascular membranes and 154 in ERM. Among these proteins, 102 were specific to neovascular membranes and 30 were specific to ERM. We identified a candidate marker, periostin, as well as several known PDR markers such as pigment epithelium-derived factor (PEDF). We then performed RT-PCR using these markers. The expression of periostin was significantly up-regulated in proliferative membrane specimens. Periostin induces cell attachment and spreading and plays a role in cell adhesion. Proteomic analysis by LC/MS/MS, which permits accurate quantitative comparison, was useful in identifying new candidates such as periostin potentially involved in the pathogenesis of PDR.