title: A key role of PGC-1α transcriptional coactivator in production of VEGF by a novel angiogenic agent COA-Cl in cultured human fibroblasts.
abstract: We previously demonstrated a potent angiogenic effect of a newly developed adenosine-like agent namedCOA-Cl.COA-Cl exerted tube forming activity in human umbilical vein endothelial cells in the presence of normal human dermal fibroblasts (NHDF). We therefore explored whether and howCOA-Cl modulates gene expression and protein secretion ofVEGF, a master regulator of angiogenesis, inNHDFRT-PCRandELISArevealed thatCOA-Cl upregulatedVEGF mRNAexpression and protein secretion inNHDFHIF1α(hypoxia-inducible factor 1α), a transcription factor, andPGC-1α(peroxisome proliferator-activated receptor-γcoactivator-1α), a transcriptional coactivator, are known to positively regulate theVEGFgene. Immunoblot andRT-PCRanalyses revealed thatCOA-Cl markedly upregulated the expression ofPGC-1αprotein andmRNACOA-Cl had no effect on the expression ofHIF1αprotein andmRNAin both hypoxia and normoxia. SilencingPGC-1αgene, but notHIF1αgene, by small interferingRNAattenuated the ability ofCOA-Cl to promoteVEGFsecretion. When an N-terminal fragment ofPGC-1αwas cotransfected with its partner transcription factorERRα(estrogen-related receptor-α) inCOS-7 cells,COA-Cl upregulated the expression of the endogenousVEGF mRNA However,COA-Cl had no effect on the expression ofVEGF, whenHIF1αwas transfected.COA-Cl inducesVEGFgene expression and protein secretion in fibroblasts. The transcriptional coactivatorPGC-1α, in concert withERRα, plays a key role in theCOA-Cl-inducedVEGFproduction.COA-Cl-induced activation ofPGC-1α-ERRα-VEGFpathway has a potential as a novel means for therapeutic angiogenesis.
authors.affiliation: Department of Cardiovascular Physiology, Faculty of Medicine, Kagawa University, Kita-Gun, Japan igarashi@med.kagawa-u.ac.jp.
authors.forename: Junsuke
authors.initials: J
authors.lastname: Igarashi
authors.affiliation: Department of Cardiovascular Physiology, Faculty of Medicine, Kagawa University, Kita-Gun, Japan.
authors.forename: Ryuji
authors.initials: R
authors.lastname: Okamoto
authors.affiliation: Department of Cardiovascular Physiology, Faculty of Medicine, Kagawa University, Kita-Gun, Japan.
authors.forename: Tetsuo
authors.initials: T
authors.lastname: Yamashita
authors.affiliation: Department of Cardiovascular Physiology, Faculty of Medicine, Kagawa University, Kita-Gun, Japan.
authors.forename: Takeshi
authors.initials: T
authors.lastname: Hashimoto
authors.affiliation: Department of Cardiovascular Physiology, Faculty of Medicine, Kagawa University, Kita-Gun, Japan.
authors.forename: Sakiko
authors.initials: S
authors.lastname: Karita
authors.affiliation: Department of Dermatology, Faculty of Medicine, Kagawa University, Kita-Gun, Japan.
authors.forename: Kozo
authors.initials: K
authors.lastname: Nakai
authors.affiliation: Department of Dermatology, Faculty of Medicine, Kagawa University, Kita-Gun, Japan.
authors.forename: Yasuo
authors.initials: Y
authors.lastname: Kubota
authors.affiliation: Department of Pharmaco-Bio-Informatics, Faculty of Medicine, Kagawa University, Kita-Gun, Japan.
authors.forename: Maki
authors.initials: M
authors.lastname: Takata
authors.affiliation: Department of Cell Physiology, Faculty of Medicine, Kagawa University, Kita-Gun, Japan.
authors.forename: Fuminori
authors.initials: F
authors.lastname: Yamaguchi
authors.affiliation: Department of Cell Physiology, Faculty of Medicine, Kagawa University, Kita-Gun, Japan.
authors.forename: Masaaki
authors.initials: M
authors.lastname: Tokuda
authors.affiliation: Kagawa School of Pharmaceutical Sciences, Tokushima Bunri University, Sanuki, Japan.
authors.forename: Norikazu
authors.initials: N
authors.lastname: Sakakibara
authors.affiliation: Department of Pharmaco-Bio-Informatics, Faculty of Medicine, Kagawa University, Kita-Gun, Japan.
authors.forename: Ikuko
authors.initials: I
authors.lastname: Tsukamoto
authors.affiliation: Department of Pharmaco-Bio-Informatics, Faculty of Medicine, Kagawa University, Kita-Gun, Japan.
authors.forename: Ryoji
authors.initials: R
authors.lastname: Konishi
authors.affiliation: Department of Cardiovascular Physiology, Faculty of Medicine, Kagawa University, Kita-Gun, Japan.
authors.forename: Katsuya
authors.initials: K
authors.lastname: Hirano
mesh_terms: D000241:Adenosine / Q000031:analogs & derivatives* / Q000494:pharmacology; D043925:Angiogenesis Inducing Agents / Q000494:pharmacology*; D000818:Animals; D019556:COS Cells; D002522:Chlorocebus aethiops; D004305:Dose-Response Relationship, Drug; D005347:Fibroblasts / Q000187:drug effects* / Q000378:metabolism; D006801:Humans; D051795:Hypoxia-Inducible Factor 1, alpha Subunit / Q000235:genetics / Q000378:metabolism; D000071248:Peroxisome Proliferator-Activated Receptor Gamma Coactivator 1-alpha; D034622:RNA Interference; D012333:RNA, Messenger / Q000378:metabolism; D011960:Receptors, Estrogen / Q000235:genetics / Q000378:metabolism; D015398:Signal Transduction / Q000187:drug effects; D013997:Time Factors; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism*; D014158:Transcription, Genetic; D015533:Transcriptional Activation; D014162:Transfection; D015854:Up-Regulation; D042461:Vascular Endothelial Growth Factor A / Q000235:genetics / Q000378:metabolism*; D000097983:ERRalpha Estrogen-Related Receptor
chemical_list: D000241:Adenosine; D043925:Angiogenesis Inducing Agents; D051795:Hypoxia-Inducible Factor 1, alpha Subunit; D000071248:Peroxisome Proliferator-Activated Receptor Gamma Coactivator 1-alpha; D012333:RNA, Messenger; D011960:Receptors, Estrogen; D014157:Transcription Factors; D042461:Vascular Endothelial Growth Factor A; D000097983:ERRalpha Estrogen-Related Receptor; C552550:2Cl-C.OXT-A; C497442:HIF1A protein, human; C486497:PPARGC1A protein, human; C467484:VEGFA protein, human
keywords: Angiogenesis; PGC‐1α; VEGF; gene regulation; nucleic acid; transcriptional coactivator