title: Hematopoietic stem/progenitor cells express functional mitochondrial energy-dependent cystic fibrosis transmembrane conductance regulator.
abstract: Bone marrow-derived hematopoietic stem/progenitor cells (HSPCs) encompass a wide array of cell subsets with different capacities of engraftment and injured tissue-regenerating potential. The characterization/isolation of the stem cell subpopulations represents a major challenge to improve the efficacy of transplantation protocols used in regenerative medicine. Cystic fibrosis (CF) is one of the diseases whose hope of cure relies on the successful application of cell-based gene therapy. This study was aimed at characterizing murine HSPCs on the basis of their bioenergetic competence and CF transmembrane conductance regulator (CFTR) expression. Positively immunoselected Sca-1(+) HSPCs encompassed 2 populations distinguished by their different size, Sca-1 expression and mitochondrial content. The smaller were the cells, the higher was Sca-1 expression and the lower was the intracellular density of functional mitochondria. Reverse transcription-polymerase chain reaction and western blotting revealed that HSPCs expressed CFTR mRNA and protein, which was also functional, as assessed by spectrofluorimetric and patch-clamp techniques. Inhibition of mitochondrial oxidative phosphorylation by oligomycin resulted in a 70% decrease of both the intracelluar adenosine triphosphate content and CFTR-mediated channel activity. Finally, HSPCs with lower Sca-1 expression and higher mitochondrial content displayed higher CFTR levels. Our findings identify 2 subpopulations in HSPCs and unveil a so-far unappreciated relationship between bioenergetic metabolism and CFTR in HSPC biology.
authors.affiliation: Department of Biomedical Sciences, University of Foggia, Foggia, Italy.
authors.forename: Donatella
authors.initials: D
authors.lastname: Piro
authors.forename: Claudia
authors.initials: C
authors.lastname: Piccoli
authors.forename: Lorenzo
authors.initials: L
authors.lastname: Guerra
authors.forename: Francesca
authors.initials: F
authors.lastname: Sassone
authors.forename: Annamaria
authors.initials: A
authors.lastname: D'Aprile
authors.forename: Maria
authors.initials: M
authors.lastname: Favia
authors.forename: Stefano
authors.initials: S
authors.lastname: Castellani
authors.forename: Sante
authors.initials: S
authors.lastname: Di Gioia
authors.forename: Silvia
authors.initials: S
authors.lastname: Lepore
authors.forename: Maria Luisa
authors.initials: ML
authors.lastname: Garavaglia
authors.forename: Teresa
authors.initials: T
authors.lastname: Trotta
authors.forename: Angela Bruna
authors.initials: AB
authors.lastname: Maffione
authors.forename: Valeria
authors.initials: V
authors.lastname: Casavola
authors.forename: Giuliano
authors.initials: G
authors.lastname: Meyer
authors.forename: Nazzareno
authors.initials: N
authors.lastname: Capitanio
authors.forename: Massimo
authors.initials: M
authors.lastname: Conese
mesh_terms: D000818:Animals; D000950:Antigens, Ly / Q000096:biosynthesis*; D003550:Cystic Fibrosis / Q000378:metabolism / Q000473:pathology; D019005:Cystic Fibrosis Transmembrane Conductance Regulator / Q000096:biosynthesis; D004734:Energy Metabolism; D005786:Gene Expression Regulation; D006412:Hematopoietic Stem Cells / Q000378:metabolism / Q000473:pathology; D008297:Male; D008565:Membrane Proteins / Q000096:biosynthesis*; D051379:Mice; D008928:Mitochondria / Q000378:metabolism / Q000473:pathology
chemical_list: D000950:Antigens, Ly; C074650:Ly6a protein, mouse; D008565:Membrane Proteins; D019005:Cystic Fibrosis Transmembrane Conductance Regulator