title: Raman microspectroscopy as a diagnostic tool for the non-invasive analysis of fibrillin-1 deficiency in the skin and in the in vitro skin models.
abstract: Fibrillin microfibrils and elastic fibers are critical determinants of elastic tissues where they define as tissue-specific architectures vital mechanical properties such as pliability and elastic recoil. Fibrillin microfibrils also facilitate elastic fiber formation and support the association of epithelial cells with the interstitial matrix. Mutations in fibrillin-1 (FBN1) are causative for the Marfan syndrome, a congenital multisystem disorder characterized by progressive deterioration of the fibrillin microfibril/ elastic fiber architecture in the cardiovascular, musculoskeletal, ocular, and dermal system. In this study, we utilized Raman microspectroscopy in combination with principal component analysis (PCA) to analyze the molecular consequences of fibrillin-1 deficiency in skin of a mouse model (GT8) of Marfan syndrome. In addition, full-thickness skin models incorporating murine wild-type and Fbn1GT8/GT8fibroblasts as well as human HaCaT keratinocytes were generated and analyzed. Skin models containing GT8 fibroblasts showed an altered epidermal morphology when compared to wild-type models indicating a new role for fibrillin-1 in dermal-epidermal crosstalk. Obtained Raman spectra together with PCA allowed to discriminate between healthy and deficient microfibrillar networks in murine dermis and skin models. Interestingly, results obtained from GT8 dermis and skin models showed similar alterations in molecular signatures triggered by fibrillin-1 deficiency such as amide III vibrations and decreased levels of glycan vibrations. Overall, this study indicates that Raman microspectroscopy has the potential to analyze subtle changes in fibrillin-1 microfibrils and elastic fiber networks. Therefore Raman microspectroscopy may be utilized as a non-invasive and sensitive diagnostic tool to identify connective tissue disorders and monitor their disease progression.
  
  
  
  Mutations in building blocks of the fibrillin microfibril/ elastic fiber network manifest in disease conditions such as aneurysms, emphysema or lax skin. Understanding how structural changes induced by fibrillin-1 mutation impact the architecture of fibrillin microfibrils, which then translates into an altered activation state of targeted growth factors, represents a huge challenge in elucidating the genotype-phenotype correlations in connective tissue disorders such as Marfan syndrome. This study shows that Raman microspectroscopy is able to reveal structural changes in fibrillin-1 microfibrils and elastic fiber networks and to discriminate between normal and diseased networks in vivo and in vitro. Therefore Raman microspectroscopy may be utilized as a non-invasive and sensitive diagnostic tool to identify connective tissue disorders and monitor their disease progression.
authors.affiliation: Department of Women's Health, Research Institute for Women's Health, Eberhard Karls University, Tuebingen, Germany; Department of Cell and Tissue Engineering, Fraunhofer Institute for Interfacial Engineering and Biotechnology (IGB), Stuttgart, Germany.
authors.forename: Eva
authors.initials: E
authors.lastname: Brauchle
authors.affiliation: Department of Women's Health, Research Institute for Women's Health, Eberhard Karls University, Tuebingen, Germany.
authors.forename: Hannah
authors.initials: H
authors.lastname: Bauer
authors.affiliation: Department of Women's Health, Research Institute for Women's Health, Eberhard Karls University, Tuebingen, Germany.
authors.forename: Patrick
authors.initials: P
authors.lastname: Fernes
authors.affiliation: Center for Biochemistry, Medical Faculty, University of Cologne, Cologne, Germany.
authors.forename: Alexandra
authors.initials: A
authors.lastname: Zuk
authors.affiliation: Department of Women's Health, Research Institute for Women's Health, Eberhard Karls University, Tuebingen, Germany; Department of Cell and Tissue Engineering, Fraunhofer Institute for Interfacial Engineering and Biotechnology (IGB), Stuttgart, Germany; Department of Medicine/Cardiology, Cardiovascular Research Laboratories, University of California, Los Angeles, CA, USA.
authors.forename: Katja
authors.initials: K
authors.lastname: Schenke-Layland
authors.affiliation: Center for Biochemistry, Medical Faculty, University of Cologne, Cologne, Germany; Center for Molecular Medicine, University of Cologne, Germany. Electronic address: gsengle@uni-koeln.de.
authors.forename: Gerhard
authors.initials: G
authors.lastname: Sengle
mesh_terms: D000465:Algorithms; D000818:Animals; D015415:Biomarkers / Q000032:analysis; D003936:Diagnosis, Computer-Assisted / Q000379:methods*; D000071838:Fibrillin-1 / Q000032:analysis*; D008382:Marfan Syndrome / Q000175:diagnosis* / Q000378:metabolism; D051379:Mice; D025341:Principal Component Analysis; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D012867:Skin / Q000737:chemistry* / Q000473:pathology; D012871:Skin Diseases / Q000175:diagnosis* / Q000378:metabolism; D013059:Spectrum Analysis, Raman / Q000379:methods*
chemical_list: D015415:Biomarkers; D000071838:Fibrillin-1
keywords: Elastic fibers; Fibrillin-1; Raman microspectroscopy; Skin equivalents