title: Scleroderma Mesenchymal Stem Cells display a different phenotype from healthy controls; implications for regenerative medicine.
abstract: BACKGROUND
  Vascular involvement is a key feature of Systemic sclerosis (SSc). Although the pericytes/endothelial cells (ECs) cross-talk regulates vessels formation, no evidences about the pericytes contribution to ineffective angiogenesis in SSc are available. Recent findings showed similarities between pericytes and Bone Marrow Mesenchymal Stem Cells (BM-MSCs). Due to difficulties in pericytes isolation, this work explores the possibility to use BM-MSCs as pericytes surrogate, clarifying their role in supporting neo-angiogenesis during SSc.
  
  
  METHODS
  To demonstrate their potential to normally differentiate into pericytes, both SSc and healthy controls (HC) BM-MSCs were treated with TGF-β and PDGF-BB. The expression of pericytes specific markers (α-SMA, NG2, RGS5 and desmin) was assessed by qPCR, western blot, and immunofluorescence; chemioinvasion and capillary morphogenesis were also performed. Cell-sorting of BM-MSCs co-cultured with HC-ECs was used to identify a possible change in contractile proteins genes expression.
  
  
  RESULTS
  We showed that BM-MSCs isolated from SSc patients displayed an up-regulation of α-SMA and SM22α genes and a reduced proliferative activity. Moreover during SSc, both TGF-β and PDGF-BB can specifically modulate BM-MSCs toward pericytes. TGF-β was found interfering with the PDGF-BB effects. Using BM-MSCs/MVECs co-culture system we observed that SSc BM-MSCs improve ECs tube formation in stressed condition, and BM-MSCs, sorted after co-culture, showed a reduced α-SMA and SM22α gene expression.
  
  
  CONCLUSIONS
  BM-MSCs from SSc patients behave as pericytes. They display a more mature and myofibroblast-like phenotype, probably related to microenvironmental cues operating during the disease. After their co-culture with HC-MVECs, SSc BM-MSCs underwent to a phenotypic modulation which re-programs these cells toward a pro-angiogenic behaviour.
authors: Department of Biotechnological and Applied Clinical Science, Rheumatology Unit, School of Medicine, University of L'Aquila, Delta 6 Building, Via dell'Ospedale, 67100 L'Aquila, Italy. paola.cipriani@cc.univaq.it
authors: Paola
authors: P
authors: Cipriani
authors: Alessandra
authors: A
authors: Marrelli
authors: Paola Di
authors: PD
authors: Benedetto
authors: Vasiliki
authors: V
authors: Liakouli
authors: Francesco
authors: F
authors: Carubbi
authors: Piero
authors: P
authors: Ruscitti
authors: Saverio
authors: S
authors: Alvaro
authors: Ilenia
authors: I
authors: Pantano
authors: Antonio Francesco
authors: AF
authors: Campese
authors: Paola
authors: P
authors: Grazioli
authors: Isabella
authors: I
authors: Screpanti
authors: Roberto
authors: R
authors: Giacomelli
mesh_terms: D000199:Actins / Q000378:metabolism; D000077214:Becaplermin; D015153:Blotting, Western; D002454:Cell Differentiation / Q000187:drug effects / Q000502:physiology; D002478:Cells, Cultured; D017931:DNA Primers / Q000235:genetics; D004305:Dose-Response Relationship, Drug; D004730:Endothelium, Vascular / Q000166:cytology*; D005434:Flow Cytometry; D005455:Fluorescent Antibody Technique; D006801:Humans; D059630:Mesenchymal Stem Cells / Q000502:physiology*; D008840:Microfilament Proteins / Q000378:metabolism; D009124:Muscle Proteins / Q000378:metabolism; D018919:Neovascularization, Physiologic / Q000502:physiology*; D020286:Pericytes / Q000502:physiology; D010641:Phenotype; D020574:Proto-Oncogene Proteins c-sis / Q000494:pharmacology; D060888:Real-Time Polymerase Chain Reaction; D044968:Regenerative Medicine / Q000379:methods*; D012595:Scleroderma, Systemic / Q000503:physiopathology* / Q000628:therapy; D016212:Transforming Growth Factor beta / Q000494:pharmacology
chemical_list: C541116:ACTA2 protein, human; D000199:Actins; D017931:DNA Primers; D008840:Microfilament Proteins; D009124:Muscle Proteins; D020574:Proto-Oncogene Proteins c-sis; D016212:Transforming Growth Factor beta; C051615:transgelin; D000077214:Becaplermin