title: Protein therapy using heme-oxygenase-1 fused to a polyarginine transduction domain attenuates cerebral vasospasm after experimental subarachnoid hemorrhage.
abstract: A sequence of 11 consecutive arginine residues (11R) is one of the best protein transduction domains for introducing proteins into cell membranes. Heme-oxygenase-1 (HO-1) is involved in heme catabolism and reduces the contractile effect of hemoglobin after subarachnoid hemorrhage (SAH). Therefore, we constructed 11R-fused HO-1 protein to achieve successful transduction of the protein into the cerebral arteries and examined the therapeutic effect of the 11R-HO-1 protein for cerebral vasospasm (CV) after SAH. We injected the 11R-HO-1 protein into the cisterna magna of male rats and, several hours after the injection, performed immunofluorescence staining and western blotting analysis of the rat basilar arteries (BAs) to determine transduction efficacy. We also assessed intraarterial HO-1 activity as cGMP (cyclic guanosine 3', 5'-cyclic monophosphate) accumulation in SAH and determined whether protein transduction of 11R-HO-1 quantified the therapeutic effect in a rat double-hemorrhage model of SAH. The BAs expressed significantly more HO-1 in the group injected with 11R-HO-1 (3.56±0.54 (11R-HO-1) versus control (saline)), and transduction of 11R-HO-1 resulted in higher activity (>3.25-fold) in rat BAs with SAH. Moreover, the results of the rat double-hemorrhage model showed that the 11R-HO-1 protein significantly attenuated CV after SAH (317.59±23.48 μm (11R-HO-1) versus 270.08±14.66 μm (11R-fused enhanced green fluorescent protein), 252.05±13.95 μm (saline), P<0.01).
authors.affiliation: Department of Neurosurgery, Heinrich-Heine University, Düsseldorf, Germany. Tomoyuki.Ogawa@uni-duesseldorf.de
authors.forename: Tomoyuki
authors.initials: T
authors.lastname: Ogawa
authors.forename: Daniel
authors.initials: D
authors.lastname: Hänggi
authors.forename: Yumei
authors.initials: Y
authors.lastname: Wu
authors.forename: Hiroyuki
authors.initials: H
authors.lastname: Michiue
authors.forename: Kazuhito
authors.initials: K
authors.lastname: Tomizawa
authors.forename: Shigeki
authors.initials: S
authors.lastname: Ono
authors.forename: Hideki
authors.initials: H
authors.lastname: Matsui
authors.forename: Isao
authors.initials: I
authors.lastname: Date
authors.forename: Hans-Jakob
authors.initials: HJ
authors.lastname: Steiger
mesh_terms: D000818:Animals; D001488:Basilar Artery / Q000378:metabolism; D015153:Blotting, Western; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects; D002536:Cerebral Arteries / Q000378:metabolism; D004195:Disease Models, Animal; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000235:genetics; D051547:Heme Oxygenase-1 / Q000235:genetics / Q000378:metabolism / Q000627:therapeutic use*; D007150:Immunohistochemistry; D008297:Male; D010455:Peptides / Q000235:genetics / Q000378:metabolism / Q000627:therapeutic use*; D010957:Plasmids; D011487:Protein Conformation; D051381:Rats; D017207:Rats, Sprague-Dawley; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism / Q000627:therapeutic use*; D013345:Subarachnoid Hemorrhage / Q000150:complications*; D014161:Transduction, Genetic; D020301:Vasospasm, Intracranial / Q000188:drug therapy* / Q000209:etiology
chemical_list: D010455:Peptides; D011993:Recombinant Fusion Proteins; C015462:polyarginine; C496541:HMOX1 protein, human; D051547:Heme Oxygenase-1