Comparative proteomic analysis of skin fibroblasts of patients with familial Alzheimer's and healthy controls: Searching for new therapeutic targets

Abstract

AbstractBackgroundAlzheimer's disease (AD) is a neurodegenerative disorder characterized by memory loss, neuronal degeneration and synaptic dysfunction, resulting in atrophy of the affected regions. The identification of early biomarkers for AD diagnosis has been a challenge due to the high individual variability in markers expression, thus the use of peripheral cells other than neuronal cells such as bone marrow or dental pulp mesenchymal cells, blood cells and fibroblasts represents an opportunity for the detection of early pathological changes of Alzheimer's disease patients. Moreover, neuronal cells differentiated from reprogram somatic cells, recapitulate brain cell pathological associated changes.MethodWe study the proteome using 2D‐PAGE and protein identification by mass spectrometry. Skin fibroblasts were cultured from patients with a mutation in Presenilin‐1 (A246E or M146L) and from control individuals of the Coriell Institute cell repository in Earl MEM salts medium with 15% non‐inactivated Fetal Bovine Serum. Fibroblast phenotype was confirmed by immunodetection of vimentin and S100A4 markers. We use Western Blot and qPCR techniques to evaluate changes in the expression of genes and proteins of particular pathways involved in the neurodegeneration of AD.ResultDifferences in protein expression in fibroblasts were identified between affected individuals and controls, related to cell adhesion, cytoskeleton, energy and glucose metabolism, ubiquitin‐proteasome pathway, autophagy and signal transduction, which have been previously found affected in neurons from AD patients.ConclusionOur results indicate that samples derived from fibroblasts from AD patients have a unique protein profile with respect to controls, both at expression level and proteins patterns, showing that peripheral cells can be useful in modeling degenerative pathways and biomarker identification for Alzheimer's disease.