title: Adenoviral-mediated placental gene transfer of IGF-1 corrects placental insufficiency via enhanced placental glucose transport mechanisms.
abstract: Previous work in our laboratory demonstrated that over-expression of human insulin-like growth factor -1 (hIGF-1) in the placenta corrects fetal weight deficits in mouse, rat, and rabbit models of intrauterine growth restriction without changes in placental weight. The underlying mechanisms of this effect have not been elucidated. To investigate the effect of intra-placental IGF-1 over-expression on placental function we examined glucose transporter expression and localization in both a mouse model of IUGR and a model of human trophoblast, the BeWo Choriocarcinoma cell line.
  
  
  METHODS
  At gestational day 18, animals were divided into four groups; sham-operated controls, uterine artery branch ligation (UABL), UABL+Ad-hIGF-1 (10(8) PFU), UABL+Ad-LacZ (10(8) PFU). At gestational day 20, pups and placentas were harvested by C-section. For human studies, BeWo choriocarcinoma cells were grown in F12 complete medium +10%FBS. Cells were incubated in serum-free control media ± Ad-IGF-1 or Ad-LacZ for 48 hours. MOIs of 10∶1 and 100∶1 were utilized. The RNA, protein expression and localization of glucose transporters GLUT1, 3, 8, and 9 were analyzed by RT-PCR, Western blot and immunohistochemistry.
  
  
  RESULTS
  In both the mouse placenta and BeWo, GLUT1 regulation was linked to altered protein localization. GLUT3, localized to the mouse fetal endothelial cells, was reduced in placental insufficiency but maintained with Ad-I GF-1 treatment. Interestingly, GLUT8 expression was reduced in the UABL placenta but up-regulated following Ad-IGF-1 in both mouse and human systems. GLUT9 expression in the mouse was increased by Ad-IGF-1 but this was not reflected in the BeWo, where Ad-IGF-1 caused moderate membrane relocalization.
  
  
  CONCLUSIONS
  Enhanced GLUT isoform transporter expression and relocalization to the membrane may be an important mechanism in Ad-hIGF-1mediated correction of placental insufficiency.
authors.affiliation: Center for Fetal Cellular and Molecular Therapy, Cincinnati Children's Hospital Medical Center, Cincinnati, Ohio, United States of America.
authors.forename: Helen N
authors.initials: HN
authors.lastname: Jones
authors.forename: Timothy
authors.initials: T
authors.lastname: Crombleholme
authors.forename: Mounira
authors.initials: M
authors.lastname: Habli
mesh_terms: D000256:Adenoviridae / Q000235:genetics*; D000818:Animals; D001483:Base Sequence; D017931:DNA Primers; D005260:Female; D018014:Gene Transfer Techniques; D015316:Genetic Therapy; D005947:Glucose / Q000378:metabolism*; D007334:Insulin-Like Growth Factor I / Q000235:genetics*; D008431:Maternal-Fetal Exchange; D051379:Mice; D008810:Mice, Inbred C57BL; D010920:Placenta / Q000378:metabolism*; D010927:Placental Insufficiency / Q000628:therapy*; D011247:Pregnancy; D020133:Reverse Transcriptase Polymerase Chain Reaction
chemical_list: D017931:DNA Primers; D007334:Insulin-Like Growth Factor I; D005947:Glucose
references.citation: Reprod Sci. 2011 Jan;18(1):20-7
references.pmid: 20926839
references.citation: J Physiol. 2008 Sep 15;586(18):4567-76
references.pmid: 18653658
references.citation: Am J Obstet Gynecol. 1999 Jan;180(1 Pt 1):163-8
references.pmid: 9914598
references.citation: Mol Cell Endocrinol. 2001 Dec 20;185(1-2):109-17
references.pmid: 11738800
references.citation: Diabetologia. 2000 May;43(5):576-82
references.pmid: 10855532
references.citation: Am J Physiol Cell Physiol. 2007 Jul;293(1):C477-85
references.pmid: 17442736
references.citation: Endocrinology. 2012 Mar;153(3):1464-71
references.pmid: 22234467
references.citation: Endocrinology. 1999 Sep;140(9):4236-43
references.pmid: 10465297
references.citation: Biol Reprod. 2012 Aug 30;87(2):49
references.pmid: 22649075
references.citation: Placenta. 2000 Jan;21(1):14-22
references.pmid: 10692246
references.citation: Nucleic Acids Res. 2001 May 1;29(9):e45
references.pmid: 11328886
references.citation: Am J Obstet Gynecol. 2009 Nov;201(5):499.e1-8
references.pmid: 19716119
references.citation: Placenta. 2004 Jan;25(1):70-7
references.pmid: 15013641
references.citation: Endocrinology. 2007 Jun;148(6):2963-72
references.pmid: 17347307
references.citation: J Clin Invest. 1979 Mar;63(3):388-94
references.pmid: 429559
references.citation: Endocrinology. 2012 Aug;153(8):3995-4007
references.pmid: 22700768
references.citation: Am J Physiol Endocrinol Metab. 2009 Apr;296(4):E614-8
references.pmid: 19176349
references.citation: Am J Physiol Endocrinol Metab. 2002 Apr;282(4):E974-6
references.pmid: 11882521
references.citation: J Clin Endocrinol Metab. 1999 Apr;84(4):1445-52
references.pmid: 10199793
references.citation: Placenta. 2011 Dec;32(12):1041-9
references.pmid: 22000473
references.citation: J Clin Endocrinol Metab. 1997 Aug;82(8):2689-94
references.pmid: 9253355
references.citation: Placenta. 2002 May;23(5):392-9
references.pmid: 12061855