title: Gene and microRNA analysis of neutrophils from patients with polycythemia vera and essential thrombocytosis: down-regulation of micro RNA-1 and -133a.
abstract: BACKGROUND
  Since the V617F mutation in JAK2 may not be the initiating event in myeloprofilerative disorders (MPDs) we compared molecular changes in neutrophils from patients with polycythemia vera (PV) and essential thrombocythosis (ET), to neutrophils stimulated by G-CSF administration and to normal unstimulated neutrophils
  
  
  METHODS
  A gene expression oligonucleotide microarray with more than 35,000 probes and a microRNA (miR) expression array with 827 probes were used to assess neutrophils from 6 MPD patients; 4 with PV and 2 with ET, 5 healthy subjects and 6 healthy subjects given G-CSF. In addition, neutrophil antigen expression was analyzed by flow cytometry and 64 serum protein levels were analyzed by ELISA.
  
  
  RESULTS
  Gene expression profiles of neutrophils from the MPD patients were similar but distinct from those of healthy subjects, either unstimulated or G-CSF-mobilized. The differentially expressed genes in MPD neutrophils were more likely to be in pathways involved with inflammation while those of G-CSF-mobilized neutrophils were more likely to belong to metabolic pathways. In MPD neutrophils the expression of CCR1 was increased and that of several NF-kappaB pathway genes were decreased. MicroRNA miR-133a and miR-1 in MPD neutrophils were down-regulated the most. Levels of 11 serum proteins were increased in MPD patients including MMP-10, MMP-13, VCAM, P-selectin, PDGF-BB and a CCR1 ligand, MIP-1alpha.
  
  
  CONCLUSIONS
  These studies showed differential expression of genes particularly involved in inflammatory pathways including the NF-kappaB pathway and down-regulation of miR-133a and miR-1. These two microRNAs have been previous associated with certain cancers as well as the regulation of hyperthrophy of cardiac and skeletal muscle cells. These changes may contribute to the clinical manifestations of the MPDs.
authors.affiliation: Department of Transfusion Medicine, Clinical Center, National Institutes of Health, Bethesda, Maryland, USA. stefanie.slezak@gmail.com
authors.forename: Stefanie
authors.initials: S
authors.lastname: Slezak
authors.forename: Ping
authors.initials: P
authors.lastname: Jin
authors.forename: Lorraine
authors.initials: L
authors.lastname: Caruccio
authors.forename: Jiaqiang
authors.initials: J
authors.lastname: Ren
authors.forename: Michael
authors.initials: M
authors.lastname: Bennett
authors.forename: Nausheen
authors.initials: N
authors.lastname: Zia
authors.forename: Sharon
authors.initials: S
authors.lastname: Adams
authors.forename: Ena
authors.initials: E
authors.lastname: Wang
authors.forename: Joao
authors.initials: J
authors.lastname: Ascensao
authors.forename: Geraldine
authors.initials: G
authors.lastname: Schechter
authors.forename: David
authors.initials: D
authors.lastname: Stroncek
mesh_terms: D015703:Antigens, CD / Q000378:metabolism; D018821:CD18 Antigens / Q000378:metabolism; D016022:Case-Control Studies; D015536:Down-Regulation / Q000235:genetics; D005260:Female; D058851:GPI-Linked Proteins; D016179:Granulocyte Colony-Stimulating Factor / Q000235:genetics / Q000378:metabolism; D006801:Humans; D007519:Isoantigens / Q000378:metabolism; D053614:Janus Kinase 2 / Q000235:genetics; D016256:Lewis X Antigen / Q000378:metabolism; D008297:Male; D008562:Membrane Glycoproteins / Q000378:metabolism; D035683:MicroRNAs / Q000032:analysis*; D008875:Middle Aged; D009504:Neutrophils / Q000378:metabolism*; D011087:Polycythemia Vera / Q000097:blood / Q000235:genetics / Q000378:metabolism; D011956:Receptors, Cell Surface / Q000378:metabolism; D013920:Thrombocythemia, Essential / Q000097:blood*
chemical_list: D015703:Antigens, CD; C407242:CD177 protein, human; D018821:CD18 Antigens; D058851:GPI-Linked Proteins; D007519:Isoantigens; D016256:Lewis X Antigen; C544547:MIRN1 microRNA, human; D008562:Membrane Glycoproteins; D035683:MicroRNAs; D011956:Receptors, Cell Surface; D016179:Granulocyte Colony-Stimulating Factor; D053614:Janus Kinase 2