title: Ex vivo pretreatment of bone marrow mononuclear cells with endothelial NO synthase enhancer AVE9488 enhances their functional activity for cell therapy.
abstract: Bone marrow mononuclear cells (BMC) from patients with ischemic cardiomyopathy (ICMP) show a reduced neovascularization capacity in vivo. NO plays an important role in neovascularization, and NO bioavailability is typically reduced in patients with ICMP. We investigated whether the impaired neovascularization capacity of ICMP patient-derived progenitor cells can be restored by pretreatment with the novel endothelial NO synthase (eNOS) transcription enhancer AVE9488 (AVE). Ex vivo pretreatment of BMC from patients with ICMP with AVE significantly increased eNOS mRNA expression by 2.1-fold (P < 0.05) and eNOS activity as assessed by ESR by >3-fold (P < 0.05). The increased eNOS expression was associated with an enhanced migratory capacity in vitro (P < 0.01) and improved neovascularization capacity of the infused BMC in an ischemic hind limb model in vivo (P < 0.001). The improvement in ischemic limb perfusion after infusion of AVE-pretreated BMC resulted in an increase in swimming time (P < 0.05). The enhancement of limb perfusion by AVE-treated BMC was abrogated by ex vivo pretreatment with the eNOS inhibitor N(G)-nitro-l-arginine methyl ester. Consistently, AVE showed no effect on the impaired migratory capacity of BMC derived from eNOS-deficient mice, documenting the specific involvement of NO. The reduced neovascularization capacity of BMC from patients with ICMP may limit their therapeutic potential in cell therapy studies. Here, we show that pharmacological enhancement of eNOS expression with AVE at least partially reverses the impaired functional activity of BMC from ICMP patients, highlighting the critical role of NO for progenitor cell function.
authors.affiliation: Department of Internal Medicine III, J. W. Goethe University, 60590 Frankfurt, Germany.
authors.forename: Ken-ichiro
authors.initials: K
authors.lastname: Sasaki
authors.forename: Christopher
authors.initials: C
authors.lastname: Heeschen
authors.forename: Alexandra
authors.initials: A
authors.lastname: Aicher
authors.forename: Thomas
authors.initials: T
authors.lastname: Ziebart
authors.forename: Joerg
authors.initials: J
authors.lastname: Honold
authors.forename: Carmen
authors.initials: C
authors.lastname: Urbich
authors.forename: Lothar
authors.initials: L
authors.lastname: Rossig
authors.forename: Ulrike
authors.initials: U
authors.lastname: Koehl
authors.forename: Masamichi
authors.initials: M
authors.lastname: Koyanagi
authors.forename: Annisuddin
authors.initials: A
authors.lastname: Mohamed
authors.forename: Ralf P
authors.initials: RP
authors.lastname: Brandes
authors.forename: Hans
authors.initials: H
authors.lastname: Martin
authors.forename: Andreas M
authors.initials: AM
authors.lastname: Zeiher
authors.forename: Stefanie
authors.initials: S
authors.lastname: Dimmeler
mesh_terms: D000818:Animals; D001854:Bone Marrow Cells / Q000166:cytology / Q000187:drug effects*; D002465:Cell Movement / Q000187:drug effects; D064987:Cell- and Tissue-Based Therapy / Q000379:methods*; D004791:Enzyme Inhibitors / Q000494:pharmacology*; D015971:Gene Expression Regulation, Enzymologic / Q000235:genetics; D006614:Hindlimb / Q000473:pathology; D006801:Humans; D007511:Ischemia / Q000139:chemically induced; D007963:Leukocytes, Mononuclear / Q000166:cytology / Q000187:drug effects*; D051379:Mice; D008819:Mice, Nude; D009389:Neovascularization, Pathologic; D052250:Nitric Oxide Synthase Type III / Q000235:genetics / Q000378:metabolism*; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism
chemical_list: D004791:Enzyme Inhibitors; D012333:RNA, Messenger; C496314:NOS3 protein, human; D052250:Nitric Oxide Synthase Type III