title: Recombinant fusion protein of albumin-retinol binding protein inactivates stellate cells.
abstract: Quiescent pancreatic- (PSCs) and hepatic- (HSCs) stellate cells store vitamin A (retinol) in lipid droplets via retinol binding protein (RBP) receptor and, when activated by profibrogenic stimuli, they transform into myofibroblast-like cells which play a key role in the fibrogenesis. Despite extensive investigations, there is, however, currently no appropriate therapy available for tissue fibrosis. We previously showed that the expression of albumin, composed of three homologous domains (I-III), inhibits stellate cell activation, which requires its high-affinity fatty acid-binding sites asymmetrically distributed in domain I and III. To attain stellate cell-specific uptake, albumin (domain I/III) was coupled to RBP; RBP-albumin(domain III) (R-III) and albumin(domain I)-RBP-albumin(III) (I-R-III). To assess the biological activity of fusion proteins, cultured PSCs were used. Like wild type albumin, expression of R-III or I-R-III in PSCs after passage 2 (activated PSCs) induced phenotypic reversal from activated to fat-storing cells. On the other hand, R-III and I-R-III, but not albumin, secreted from transfected 293 cells were successfully internalized into and inactivated PSCs. FPLC-purified R-III was found to be internalized into PSCs via caveolae-mediated endocytosis, and its efficient cellular uptake was also observed in HSCs and podocytes among several cell lines tested. Moreover, tissue distribution of intravenously injected R-III was closely similar to that of RBP. Therefore, our data suggest that albumin-RBP fusion protein comprises of stellate cell inactivation-inducing moiety and targeting moiety, which may lead to the development of effective anti-fibrotic drug.
authors.affiliation: Laboratory of Cellular Oncology, Korea University Graduate School of Medicine, Ansan, Gyeonggi do 425-707, Republic of Korea.
authors.forename: Soyoung
authors.initials: S
authors.lastname: Choi
authors.forename: Sangeun
authors.initials: S
authors.lastname: Park
authors.forename: Suhyun
authors.initials: S
authors.lastname: Kim
authors.forename: Chaeseung
authors.initials: C
authors.lastname: Lim
authors.forename: Jungho
authors.initials: J
authors.lastname: Kim
authors.forename: Dae Ryong
authors.initials: DR
authors.lastname: Cha
authors.forename: Junseo
authors.initials: J
authors.lastname: Oh
mesh_terms: D000418:Albumins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D000818:Animals; D002460:Cell Line; D002478:Cells, Cultured; D017077:Culture Media, Conditioned / Q000378:metabolism / Q000494:pharmacology; D003550:Cystic Fibrosis / Q000378:metabolism / Q000473:pathology; D005223:Fats / Q000378:metabolism; D006801:Humans; D008103:Liver Cirrhosis / Q000378:metabolism / Q000473:pathology; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D058954:Pancreatic Stellate Cells / Q000187:drug effects / Q000378:metabolism* / Q000473:pathology*; D017434:Protein Structure, Tertiary; D051381:Rats; D017207:Rats, Sprague-Dawley; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D012177:Retinol-Binding Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D014162:Transfection
chemical_list: D000418:Albumins; D017077:Culture Media, Conditioned; D005223:Fats; D011993:Recombinant Fusion Proteins; D012177:Retinol-Binding Proteins