Augmentation of therapeutic angiogenesis using genetically modified human endothelial progenitor cells with altered glycogen synthase kinase-3beta activity.
Previously we reported that inhibition of glycogen synthase kinase-3beta (GSK3beta), a key regulator in many intracellular signaling pathways, enhances the survival and migration of vascular endothelial cells. Here we investigated the effect of inhibition of GSK3beta activity on the angiogenic function of endothelial progenitor cell (EPC) and demonstrated a new therapeutic angiogenesis strategy using genetically modified EPC. As we previously reported, two biologically distinct types of EPC, spindle-shaped "early EPC" and cobblestone-shaped "late EPC" could be cultivated from human peripheral blood. Catalytically inactive GSK3beta gene was transduced into both EPC. Inhibition of GSK3beta signaling pathway led to increased nuclear translocation of beta-catenin and increased secretion of angiogenic cytokines (vascular endothelial growth factor and interleukin-8). It enhanced the survival and proliferation of early EPC, whereas it promoted the survival and differentiation of late EPC. Transplantation of either of these genetically modified EPC into the ischemic hind limb model of athymic nude mouse significantly improved blood flow, limb salvage, and tissue capillary density compared with nontransduced EPC. Inhibition of GSK3beta signaling of either of these genetically modified EPC augmented the in vitro and in vivo angiogenic potency of these cell populations. These data provide evidence that GSK3beta has a key role in the angiogenic properties of EPC. Furthermore, the genetic modification of EPC to alter this signaling step can improve the efficacy of cell-based therapeutic vasculogenesis.
Choi
Jin-Ho
JH
Cardiovascular Laboratory, Clinical Research Institute, Seoul National University Hospital, Seoul 110-744, Korea.
Hur
Jin
J
Yoon
Chang-Hwan
CH
Kim
Ji-Hyun
JH
Lee
Choon-Soo
CS
Youn
Seock-Won
SW
Oh
Il-Young
IY
Skurk
Carsten
C
Murohara
Toyoaki
T
Park
Young-Bae
YB
Walsh
Kenneth
K
Kim
Hyo-Soo
HS
D021581:Active Transport, Cell Nucleus; D000256:Adenoviridae / Q000235:genetics; D000818:Animals; D015153:Blotting, Western; D002196:Capillaries / Q000378:metabolism; D002384:Catalysis; D049109:Cell Proliferation; D002470:Cell Survival; D003094:Collagen / Q000494:pharmacology; D016207:Cytokines / Q000096:biosynthesis; D003598:Cytoskeletal Proteins / Q000378:metabolism; D004338:Drug Combinations; D004730:Endothelium, Vascular / Q000166:cytology*; D018014:Gene Transfer Techniques; D015316:Genetic Therapy / Q000379:methods*; D038362:Glycogen Synthase Kinase 3 / Q000235:genetics* / Q000378:metabolism; D000071679:Glycogen Synthase Kinase 3 beta; D006801:Humans; D007511:Ischemia / Q000628:therapy*; D007797:Laminin / Q000494:pharmacology; D051379:Mice; D008819:Mice, Nude; D008856:Microscopy, Fluorescence; D018482:Muscle, Skeletal / Q000378:metabolism; D018919:Neovascularization, Physiologic; D010477:Perfusion; D011509:Proteoglycans / Q000494:pharmacology; D015398:Signal Transduction; D013234:Stem Cells / Q000166:cytology*; D013997:Time Factors; D015534:Trans-Activators / Q000378:metabolism; D051176:beta Catenin
C495270:CTNNB1 protein, human; C495267:CTNNB1 protein, mouse; D016207:Cytokines; D003598:Cytoskeletal Proteins; D004338:Drug Combinations; D007797:Laminin; D011509:Proteoglycans; D015534:Trans-Activators; D051176:beta Catenin; C059485:matrigel; D003094:Collagen; C000605506:GSK3B protein, human; D000071679:Glycogen Synthase Kinase 3 beta; C000605507:Gsk3b protein, mouse; D038362:Glycogen Synthase Kinase 3