Diagnostic area

URIFIB

Liver fibrosis and cirrhosis result from the majority of chronic liver pathologies and represent a common and difficult clinical challenge of worldwide importance. Liver fibrosis is the common response to chronic liver injury, ultimately leading to cirrhosis and its complications: portal hypertension, liver failure, hepatic encephalopathy, hepatocellular carcinoma and others. Thus, there is an imperative need in clinical practice to develop diagnostic strategies capable of detecting and classifying the fibrosis progression. Although several methods have been developed to analyze the extracellular matrix accumulation, they are invasive and not very precise determining the fibrosis degree in early stages.

It is known that pathological processes are associated to quantitative and functional changes in molecules present in body fluids. One of biomedicine main interest is focused in the discovery of biomarkers that could define the liver fibrosis degree. In addition, the identification of proteins associated to fibrosis could lead to development of therapeutical molecules. Our aim is the development of a diagnostic method to determine the presence and progression degree of liver fibrosis, based in the quantification of several peptide markers present in urine samples.

Previous studies allowed detection of 4 potential markers of fibrosis in urine using two dimensional electrophoresis analyses. We are currently extending our proteomic study using the most common and successful techniques in biomarkers research:

1.- DIGE

2.- SELDI-TOF

3.- MicroHPLC (n-dimensional) combined with MALDI TOF MS and ESI-MS/MS to analyze the low molecular weight proteome.

The SELDI TOF technique (Surface Enhanced Laser Desorption/ Ionization-Time of Flight) has emerged as a successful tool that combines chromatography and mass spectrometry, and compares protein expression profiles from a variety of biological and clinical samples. Twenty potential markers of liver fibrosis are under evaluation and probably will lead to the development of the first prototype of URIFIB test in 2008. On the other side, since urine is the result of the blood filtering process, low molecular weight proteome is being analyzed combining MicroHPLC with MALDI TOF MS and ESI-MS/MS