title: Proteomic Investigation of Dermal Fibroblasts Isolated from Affected and Unaffected Skin Samples from Patients with Limited Cutaneous Systemic Sclerosis: 2 Distinct Entities?
abstract: To identify using proteomic analysis the proteins of altered abundance in the affected and unaffected limited cutaneous systemic sclerosis (lcSSc) skin fibroblasts.
  
  
  
  Excision biopsies (3 mm) were obtained from the affected and unaffected skin of 5 patients with lcSSc. Dermal fibroblasts were isolated enzymatically. Two-dimensional gel electrophoresis was used to separate and define proteins in affected and unaffected fibroblast lysates. Proteins of altered abundance were identified by mass spectrometry. Differences among skin samples were confirmed also by immunohistochemistry (IHC) and by quantitative real-time PCR (qRT-PCR) for type I collagen (Col-1) and vimentin (VIM).
  
  
  
  Proteomic analysis revealed different expressions of proteins involved in cytoskeleton organization (27%), extracellular matrix remodeling (11%), response to oxidative stress (22%), energy metabolism (19%), protein metabolism (5%), cellular homeostasis (5%), signal transduction (3%), and protein transcription, synthesis, and turnover (8%). IHC analysis showed that SSc-affected epidermis is thickened and the dermis is strongly reactive to Col-1 and VIM (typical markers of activated myofibroblasts) compared to SSc-unaffected skin, whose stainings are comparable to those of control healthy skin. Overexpression of Col-1 and VIM mRNA levels in affected lcSSc fibroblasts compared to unaffected lcSSc ones was confirmed by qRT-PCR.
  
  
  
  Consistent with previous studies, these findings are important for 2 reasons: first, because they reveal the opposite behavior of dermal fibroblasts in the unaffected and affected skin areas of the same patient with lcSSc; second, because they demonstrate the histological/histochemical similarities between unaffected skin from patients with lcSSc and healthy control skin.
authors.affiliation: From the Department of Medicine, Surgery, and Neurosciences; the Department of Biotechnology, Chemistry and Pharmacy; and the Department of Medical Biotechnology, University of Siena, Siena, Italy. corallo.claudio@gmail.com.|C. Corallo, PhD, Department of Medicine, Surgery and Neurosciences, University of Siena; A. Santucci, PhD, Department of Biotechnology, Chemistry and Pharmacy, University of Siena; G. Bernardini, PhD, Department of Biotechnology, Chemistry and Pharmacy, University of Siena; N. Figura, MD, Department of Medicine, Surgery, and Neurosciences, University of Siena; R. Leoncini, PhD, Department of Medical Biotechnology, University of Siena; G. Riolo, PhD, Department of Medical Biotechnology, University of Siena; A. Montella, MD, Department of Medicine, Surgery, and Neurosciences, University of Siena; C. Chirico, MD, Department of Medicine, Surgery, and Neurosciences, University of Siena; R. Nuti, MD, Department of Medicine, Surgery, and Neurosciences, University of Siena; N. Giordano, MD, Department of Medicine, Surgery, and Neurosciences, University of Siena. corallo.claudio@gmail.com.
authors.forename: Claudio
authors.initials: C
authors.lastname: Corallo
authors.affiliation: From the Department of Medicine, Surgery, and Neurosciences; the Department of Biotechnology, Chemistry and Pharmacy; and the Department of Medical Biotechnology, University of Siena, Siena, Italy.|C. Corallo, PhD, Department of Medicine, Surgery and Neurosciences, University of Siena; A. Santucci, PhD, Department of Biotechnology, Chemistry and Pharmacy, University of Siena; G. Bernardini, PhD, Department of Biotechnology, Chemistry and Pharmacy, University of Siena; N. Figura, MD, Department of Medicine, Surgery, and Neurosciences, University of Siena; R. Leoncini, PhD, Department of Medical Biotechnology, University of Siena; G. Riolo, PhD, Department of Medical Biotechnology, University of Siena; A. Montella, MD, Department of Medicine, Surgery, and Neurosciences, University of Siena; C. Chirico, MD, Department of Medicine, Surgery, and Neurosciences, University of Siena; R. Nuti, MD, Department of Medicine, Surgery, and Neurosciences, University of Siena; N. Giordano, MD, Department of Medicine, Surgery, and Neurosciences, University of Siena.
authors.forename: Annalisa
authors.initials: A
authors.lastname: Santucci
authors.affiliation: From the Department of Medicine, Surgery, and Neurosciences; the Department of Biotechnology, Chemistry and Pharmacy; and the Department of Medical Biotechnology, University of Siena, Siena, Italy.|C. Corallo, PhD, Department of Medicine, Surgery and Neurosciences, University of Siena; A. Santucci, PhD, Department of Biotechnology, Chemistry and Pharmacy, University of Siena; G. Bernardini, PhD, Department of Biotechnology, Chemistry and Pharmacy, University of Siena; N. Figura, MD, Department of Medicine, Surgery, and Neurosciences, University of Siena; R. Leoncini, PhD, Department of Medical Biotechnology, University of Siena; G. Riolo, PhD, Department of Medical Biotechnology, University of Siena; A. Montella, MD, Department of Medicine, Surgery, and Neurosciences, University of Siena; C. Chirico, MD, Department of Medicine, Surgery, and Neurosciences, University of Siena; R. Nuti, MD, Department of Medicine, Surgery, and Neurosciences, University of Siena; N. Giordano, MD, Department of Medicine, Surgery, and Neurosciences, University of Siena.
authors.forename: Giulia
authors.initials: G
authors.lastname: Bernardini
authors.affiliation: From the Department of Medicine, Surgery, and Neurosciences; the Department of Biotechnology, Chemistry and Pharmacy; and the Department of Medical Biotechnology, University of Siena, Siena, Italy.|C. Corallo, PhD, Department of Medicine, Surgery and Neurosciences, University of Siena; A. Santucci, PhD, Department of Biotechnology, Chemistry and Pharmacy, University of Siena; G. Bernardini, PhD, Department of Biotechnology, Chemistry and Pharmacy, University of Siena; N. Figura, MD, Department of Medicine, Surgery, and Neurosciences, University of Siena; R. Leoncini, PhD, Department of Medical Biotechnology, University of Siena; G. Riolo, PhD, Department of Medical Biotechnology, University of Siena; A. Montella, MD, Department of Medicine, Surgery, and Neurosciences, University of Siena; C. Chirico, MD, Department of Medicine, Surgery, and Neurosciences, University of Siena; R. Nuti, MD, Department of Medicine, Surgery, and Neurosciences, University of Siena; N. Giordano, MD, Department of Medicine, Surgery, and Neurosciences, University of Siena.
authors.forename: Natale
authors.initials: N
authors.lastname: Figura
authors.affiliation: From the Department of Medicine, Surgery, and Neurosciences; the Department of Biotechnology, Chemistry and Pharmacy; and the Department of Medical Biotechnology, University of Siena, Siena, Italy.|C. Corallo, PhD, Department of Medicine, Surgery and Neurosciences, University of Siena; A. Santucci, PhD, Department of Biotechnology, Chemistry and Pharmacy, University of Siena; G. Bernardini, PhD, Department of Biotechnology, Chemistry and Pharmacy, University of Siena; N. Figura, MD, Department of Medicine, Surgery, and Neurosciences, University of Siena; R. Leoncini, PhD, Department of Medical Biotechnology, University of Siena; G. Riolo, PhD, Department of Medical Biotechnology, University of Siena; A. Montella, MD, Department of Medicine, Surgery, and Neurosciences, University of Siena; C. Chirico, MD, Department of Medicine, Surgery, and Neurosciences, University of Siena; R. Nuti, MD, Department of Medicine, Surgery, and Neurosciences, University of Siena; N. Giordano, MD, Department of Medicine, Surgery, and Neurosciences, University of Siena.
authors.forename: Roberto
authors.initials: R
authors.lastname: Leoncini
authors.affiliation: From the Department of Medicine, Surgery, and Neurosciences; the Department of Biotechnology, Chemistry and Pharmacy; and the Department of Medical Biotechnology, University of Siena, Siena, Italy.|C. Corallo, PhD, Department of Medicine, Surgery and Neurosciences, University of Siena; A. Santucci, PhD, Department of Biotechnology, Chemistry and Pharmacy, University of Siena; G. Bernardini, PhD, Department of Biotechnology, Chemistry and Pharmacy, University of Siena; N. Figura, MD, Department of Medicine, Surgery, and Neurosciences, University of Siena; R. Leoncini, PhD, Department of Medical Biotechnology, University of Siena; G. Riolo, PhD, Department of Medical Biotechnology, University of Siena; A. Montella, MD, Department of Medicine, Surgery, and Neurosciences, University of Siena; C. Chirico, MD, Department of Medicine, Surgery, and Neurosciences, University of Siena; R. Nuti, MD, Department of Medicine, Surgery, and Neurosciences, University of Siena; N. Giordano, MD, Department of Medicine, Surgery, and Neurosciences, University of Siena.
authors.forename: Giulia
authors.initials: G
authors.lastname: Riolo
authors.affiliation: From the Department of Medicine, Surgery, and Neurosciences; the Department of Biotechnology, Chemistry and Pharmacy; and the Department of Medical Biotechnology, University of Siena, Siena, Italy.|C. Corallo, PhD, Department of Medicine, Surgery and Neurosciences, University of Siena; A. Santucci, PhD, Department of Biotechnology, Chemistry and Pharmacy, University of Siena; G. Bernardini, PhD, Department of Biotechnology, Chemistry and Pharmacy, University of Siena; N. Figura, MD, Department of Medicine, Surgery, and Neurosciences, University of Siena; R. Leoncini, PhD, Department of Medical Biotechnology, University of Siena; G. Riolo, PhD, Department of Medical Biotechnology, University of Siena; A. Montella, MD, Department of Medicine, Surgery, and Neurosciences, University of Siena; C. Chirico, MD, Department of Medicine, Surgery, and Neurosciences, University of Siena; R. Nuti, MD, Department of Medicine, Surgery, and Neurosciences, University of Siena; N. Giordano, MD, Department of Medicine, Surgery, and Neurosciences, University of Siena.
authors.forename: Antonio
authors.initials: A
authors.lastname: Montella
authors.affiliation: From the Department of Medicine, Surgery, and Neurosciences; the Department of Biotechnology, Chemistry and Pharmacy; and the Department of Medical Biotechnology, University of Siena, Siena, Italy.|C. Corallo, PhD, Department of Medicine, Surgery and Neurosciences, University of Siena; A. Santucci, PhD, Department of Biotechnology, Chemistry and Pharmacy, University of Siena; G. Bernardini, PhD, Department of Biotechnology, Chemistry and Pharmacy, University of Siena; N. Figura, MD, Department of Medicine, Surgery, and Neurosciences, University of Siena; R. Leoncini, PhD, Department of Medical Biotechnology, University of Siena; G. Riolo, PhD, Department of Medical Biotechnology, University of Siena; A. Montella, MD, Department of Medicine, Surgery, and Neurosciences, University of Siena; C. Chirico, MD, Department of Medicine, Surgery, and Neurosciences, University of Siena; R. Nuti, MD, Department of Medicine, Surgery, and Neurosciences, University of Siena; N. Giordano, MD, Department of Medicine, Surgery, and Neurosciences, University of Siena.
authors.forename: Chiara
authors.initials: C
authors.lastname: Chirico
authors.affiliation: From the Department of Medicine, Surgery, and Neurosciences; the Department of Biotechnology, Chemistry and Pharmacy; and the Department of Medical Biotechnology, University of Siena, Siena, Italy.|C. Corallo, PhD, Department of Medicine, Surgery and Neurosciences, University of Siena; A. Santucci, PhD, Department of Biotechnology, Chemistry and Pharmacy, University of Siena; G. Bernardini, PhD, Department of Biotechnology, Chemistry and Pharmacy, University of Siena; N. Figura, MD, Department of Medicine, Surgery, and Neurosciences, University of Siena; R. Leoncini, PhD, Department of Medical Biotechnology, University of Siena; G. Riolo, PhD, Department of Medical Biotechnology, University of Siena; A. Montella, MD, Department of Medicine, Surgery, and Neurosciences, University of Siena; C. Chirico, MD, Department of Medicine, Surgery, and Neurosciences, University of Siena; R. Nuti, MD, Department of Medicine, Surgery, and Neurosciences, University of Siena; N. Giordano, MD, Department of Medicine, Surgery, and Neurosciences, University of Siena.
authors.forename: Ranuccio
authors.initials: R
authors.lastname: Nuti
authors.affiliation: From the Department of Medicine, Surgery, and Neurosciences; the Department of Biotechnology, Chemistry and Pharmacy; and the Department of Medical Biotechnology, University of Siena, Siena, Italy.|C. Corallo, PhD, Department of Medicine, Surgery and Neurosciences, University of Siena; A. Santucci, PhD, Department of Biotechnology, Chemistry and Pharmacy, University of Siena; G. Bernardini, PhD, Department of Biotechnology, Chemistry and Pharmacy, University of Siena; N. Figura, MD, Department of Medicine, Surgery, and Neurosciences, University of Siena; R. Leoncini, PhD, Department of Medical Biotechnology, University of Siena; G. Riolo, PhD, Department of Medical Biotechnology, University of Siena; A. Montella, MD, Department of Medicine, Surgery, and Neurosciences, University of Siena; C. Chirico, MD, Department of Medicine, Surgery, and Neurosciences, University of Siena; R. Nuti, MD, Department of Medicine, Surgery, and Neurosciences, University of Siena; N. Giordano, MD, Department of Medicine, Surgery, and Neurosciences, University of Siena.
authors.forename: Nicola
authors.initials: N
authors.lastname: Giordano
mesh_terms: D003599:Cytoskeleton / Q000378:metabolism / Q000473:pathology; D004734:Energy Metabolism / Q000502:physiology; D005260:Female; D005347:Fibroblasts / Q000378:metabolism* / Q000473:pathology; D006801:Humans; D008875:Middle Aged; D040901:Proteomics; D045745:Scleroderma, Limited; D015398:Signal Transduction / Q000502:physiology; D012867:Skin / Q000378:metabolism* / Q000473:pathology
keywords: 2-D ELECTROPHORESIS; DERMAL FIBROBLASTS; FIBROSIS; PROTEOMICS; PROTEOMICS OF SCLERODERMIC FIBROBLASTS; SYSTEMIC SCLEROSIS