Advances in Clinical and Experimental Medicine

Title abbreviation: Adv Clin Exp Med
JCR Impact Factor (IF) – 2.1 (5-Year IF – 2.0)
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ISSN 1899–5276 (print)
ISSN 2451-2680 (online)
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Advances in Clinical and Experimental Medicine

2017, vol. 26, nr 7, October, p. 1053–1061

doi: 10.17219/acem/65784

Publication type: original article

Language: English

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Application of Fourier transform infrared spectroscopy to biomolecular profiling of cultured fibroblast cells from Gaucher disease patients: A preliminary investigation

Nasit Igci1,B,C,D,E,F, Parisa Sharafi2,B,C, Duygu Ozel Demiralp3,A,D,E,F, Cemil Ozerk Demiralp4,B, Aysel Yuce5,B,E, Serap Dokmeci (emre)2,A,D,E,F

1 Department of Molecular Biology and Genetics, Faculty of Sciences and Arts, Nevsehir Haci Bektas Veli University, Turkey

2 Medical Biology Department, Faculty of Medicine, Hacettepe University, Ankara, Turkey

3 Biomedical Engineering Department, Faculty of Engineering, Ankara University, Turkey

4 Plastic and Reconstructive Surgery Department, Ataturk Training and Research Hospital, Ankara, Turkey

5 Pediatric Gastroenterology, Hepatology and Nutrition Unit, Ihsan Dogramaci Children Hospital, Faculty of Medicine, Hacettepe University, Ankara, Turkey

Abstract

Background. Gaucher disease (GD) is defined as an autosomal recessive disorder resulting from the deficiency of glucocerebrosidase (E.C. 3.2.1.45). Glucocerebrosidase is responsible for the degradation of glucosylceramide into ceramide and glucose. The deficiency of this enzyme results in the accumulation of undegraded glucosylceramide, almost exclusively in macrophages. With Fourier transform infrared (FTIR) spectroscopy, the complete molecular diversity of the samples can be studied comparatively and the amount of the particular materials can be determined. Also, the secondary structure ratios of proteins can be determined by analysing the amide peaks.
Objectives. The primary aim of this study is to introduce FTIR-ATR spectroscopy technique to GD research for the first time in the literature and to assess its potential as a new molecular method.
Material and Methods. Primary fibroblast cell cultures obtained from biopsy samples were used, since this material is widely used for the diagnosis of GD. Intact cells were placed onto a FTIR-ATR crystal and dried by purging nitrogen gas. Spectra were recorded in the mid-infrared region between 4500-850 cm-1 wavenumbers. Each peak in the spectra was assigned to as organic biomolecules according to their chemical bond information. A quantitative analysis was performed using peak areas and we also used a hierarchical cluster analysis as a multivariate spectral analysis.
Results. We obtained FTIR spectra of fibroblast samples and assigned the biomolecule origins of the peaks. We observed individual heterogeneity in FTIR spectra of GD fibroblast samples, confirming the well-known phenotypic heterogeneity in GD at the molecular level. Significant alterations in protein, lipid and carbohydrate levels related to the enzyme replacement therapy were also observed, which is also supported by cluster analysis.
Conclusion. Our results showed that the application of FTIR spectroscopy to GD research deserves more attention and detailed studies with an increased sample size in order to evaluate its potential in the diagnosis and follow-up of GD patients.

Key words

FTIR, Gaucher disease, fibroblast

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