title: Silencing of eIF3e promotes blood perfusion recovery after limb ischemia through stabilization of hypoxia-inducible factor 2α activity.
abstract: We previously observed that silencing of eukaryotic translation initiation factor 3 subunit e (eIF3e), a hypoxia-independent downregulator of hypoxia-inducible factor 2α (HIF-2α), led to neoangiogenesis by promoting HIF-2α activity under normoxic conditions. In the current study, we investigated whether temporary silencing of eIF3e in muscles affects blood flow recovery in a mouse ischemic limb model.
  
  
  
  eIF3e silencing was performed using small interfering RNA (siRNA), and changes in gene transcription and protein expression were analyzed in vitro using murine primary skeletal muscle myoblast and human primary skeletal muscle myoblast cell cultures. In unilateral femoral artery ligation experiments, eIF3e siRNA-expressing plasmids were injected into the muscles of BALB/c mice near the ligation sites, and tissue damage and loss of limb function were scored for 28 days while serial measurements of limb perfusions were performed with laser Doppler perfusion imaging.
  
  
  
  Silencing of eIF3e in murine primary skeletal muscle myoblasts led to stabilization of HIF-2α and upregulation of angiogenic transcripts, including basic fibroblast growth factor and platelet-derived growth factor B (P < .05), and the supernatant of eIF3e-silenced human primary skeletal muscle myoblasts triggered the tube formation of human umbilical vein endothelial cells. The in vivo mouse model of hindlimb ischemia revealed that single intramuscular injections of eIF3e siRNA-expressing plasmids significantly enhanced perfusion of ischemia-damaged limbs (P < .05) at days 7 and 14 and functional recovery at days 7, 14, and 21 (P < .05).
  
  
  
  eIF3e is an angiogenesis suppressor and may be a therapeutic target for promoting angiogenesis after ischemic injuries.
authors.affiliation: Division of Vascular Surgery, Department of Surgery, Graduate School of Medicine, University of Tokyo, Tokyo, Japan.
authors.forename: Takuya
authors.initials: T
authors.lastname: Hashimoto
authors.affiliation: Department of Molecular Medical Research, Tokyo Metropolitan Institute of Medical Science, Tokyo, Japan.
authors.forename: Li
authors.initials: L
authors.lastname: Chen
authors.affiliation: Division of Vascular Surgery, Department of Surgery, Graduate School of Medicine, University of Tokyo, Tokyo, Japan.
authors.forename: Hideo
authors.initials: H
authors.lastname: Kimura
authors.affiliation: Department of Molecular Medical Research, Tokyo Metropolitan Institute of Medical Science, Tokyo, Japan.
authors.forename: Alexander
authors.initials: A
authors.lastname: Endler
authors.affiliation: Division of Vascular Surgery, Department of Surgery, Graduate School of Medicine, University of Tokyo, Tokyo, Japan; Division of Tissue Engineering, University of Tokyo Hospital, Tokyo, Japan; Department of Vascular Surgery, Saitama Medical Center, Saitama Medical University, Saitama, Japan. Electronic address: hkoyama-tky@umin.ac.jp.
authors.forename: Hiroyuki
authors.initials: H
authors.lastname: Koyama
authors.affiliation: Vascular Center, Sanno Hospital and Sanno Medical Center, Tokyo, Japan.
authors.forename: Tetsuro
authors.initials: T
authors.lastname: Miyata
authors.affiliation: Department of Molecular Medical Research, Tokyo Metropolitan Institute of Medical Science, Tokyo, Japan.
authors.forename: Futoshi
authors.initials: F
authors.lastname: Shibasaki
authors.affiliation: Division of Vascular Surgery, Department of Surgery, Graduate School of Medicine, University of Tokyo, Tokyo, Japan.
authors.forename: Toshiaki
authors.initials: T
authors.lastname: Watanabe
mesh_terms: D000818:Animals; D051792:Basic Helix-Loop-Helix Proteins / Q000235:genetics / Q000378:metabolism*; D001783:Blood Flow Velocity; D002478:Cells, Cultured; D004195:Disease Models, Animal; D039621:Eukaryotic Initiation Factor-3 / Q000235:genetics / Q000378:metabolism*; D016222:Fibroblast Growth Factor 2 / Q000235:genetics / Q000378:metabolism; D006614:Hindlimb; D061307:Human Umbilical Vein Endothelial Cells / Q000378:metabolism; D006801:Humans; D007511:Ischemia / Q000235:genetics / Q000378:metabolism / Q000503:physiopathology / Q000628:therapy*; D008297:Male; D008807:Mice, Inbred BALB C; D018482:Muscle, Skeletal / Q000098:blood supply* / Q000378:metabolism; D032448:Myoblasts, Skeletal / Q000378:metabolism*; D018919:Neovascularization, Physiologic; D055550:Protein Stability; D020574:Proto-Oncogene Proteins c-sis / Q000235:genetics / Q000378:metabolism; D034622:RNA Interference; D000067492:RNAi Therapeutics; D020127:Recovery of Function; D012039:Regional Blood Flow; D013997:Time Factors; D014162:Transfection; D000099162:Endothelial PAS Domain-Containing Protein 1
chemical_list: D051792:Basic Helix-Loop-Helix Proteins; D039621:Eukaryotic Initiation Factor-3; D016222:Fibroblast Growth Factor 2; D020574:Proto-Oncogene Proteins c-sis; D000099162:Endothelial PAS Domain-Containing Protein 1