Eva Hunyadi-Gulyas (Presenter)
Laboratory of Proteomics Research, BRC, HAS
Bio: Eva Hunyadi-Gulyas works as a scientist at the Proteomics Research Group at the Biological Research Center in Szeged (Hungary), led by Katalin F. Medzihradszky. After she defended her PhD on peptide chemistry, she joined to the proteomics core facility in 2002, to do MS based proteomics.
Authorship: Eva Hunyadi-Gulyas (1), Lajos Kemeny (2), Gergely Groma (2,3)
(1) Laboratory of Proteomics Research, Biological Research Center of the Hungarian Academy of Sciences; (2) University of Szeged, Department of Dermatology and Allergology, Szeged, Hungary; (3) MTA-SZTE Dermatological Research Group, Szeged, Hungary
| Short Abstract Psoriasis is a multifactorial skin disease, 2-4% of the population is affected. The subject of this investigation is its most frequent type, the psoriasis vulgaris. Our aim was to identify consistent alterations present in the non-lesional psoriatic versus healthy skin at the proteomic level. We believe, the results will provide some predetermining factor, possibly some drug target for this disease. Multi-step protein extraction followed by tryptic digestion and extensive fractionation on peptide level were applied. Spectral count based label-free semi quantitative analysis were performed. This non-hypothesis driven analysis identified several known as well as potentially novel differentially expressed psoriasis-associated proteins. |
Long Abstract
Introduction
Psoriasis is a multifactorial, systemic, chronic inflammatory skin disease affecting 1-4% of the population worldwide. The main characteristics of the disease include abnormal keratinocyte proliferation, altered immune response, abnormal angiogenesis as well as changes of the extracellular matrix. One of the most effective ways to study such complex diseases is the comparative proteomic analysis of skin biopsy-derived protein extracts. Several proteomic studies have compared the non-lesional and the lesional skin with high success. At the same time, it is known that, in case of psoriasis, certain potentially susceptible alterations are already present in the asymptomatic non-lesional skin areas of patients and these molecular mechanisms are only partially known.
Therefore, the aim of this work was to compare the protein extracts from healthy and the psoriatic non-lesional skin to identify novel alterations specific for the non-lesional skin.
Methods
Methods
Skin biopsies of age and gender matched patients and healthy donors were pooled and the analysis was repeated on three different sample sets. Because the whole skin proteome was analyzed, intensive fractionation was necessary. On protein level solubility-based four-step sequential protein extraction was used. After tryptic digestion of each extracts, the samples were fractionated on a high pH reversed phase column and fractions collected were subjected to nanoLC-MSMS analysis. The merged peak lists (from 48 LC-MS-MS runs/each sample), were searched against the human Uniprot database using ProteinProspector search engine. In addition to protein identification, spectral count-based label-free semi-quantitative analysis was performed.
Results
Conclusions & Discussion
As the outcome of our comparative proteomic approach, 48 proteins we identified that were significantly (p>0.1) and consistently differentially expressed in all the three biological replicates. Out of these 48 proteins 19 are known to be associated psoriasis based on pubmed search using gene ID and psoriasis as a keyword and are in accordance with the current concepts of the pathomechanism of the disease (APOA2, APOB, ATIC, CAT, CD34, COL1A1, COL3A1, CST6, GPNMB, HLA-C, HSPE1, HNRNPU, IFI16, NCAM1, RAN, RTN4, SERPINA1, TGM3, TYMP). The remaining 29 potentially novel, psoriasis-associated proteins are currently under validation by two independent techniques including immune blotting and immunohistochemistry. Their potential roles in the diseas will be also investigated.
References & Acknowledgements:
Acknowledgments: This work was supported by the Hungarian Scientific Research Fund #116992 for GG; GINOP-2.3.2-15-2016-00001 and GINOP-2.2.1-15-2016-00007 grants.
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