Recombinant fusion protein of albumin-retinol binding protein inactivates stellate cells

Soyoung Choi, Sangeun Park, Suhyun Kim, Chae Seung Lim, Jungho Kim, Dae-Ryong Cha, Jun Seo Oh

Research output: Contribution to journalArticle

5 Citations (Scopus)

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-albumindomain III (R-III) and albumindomain I-RBP-albuminIII (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.

Original languageEnglish
Pages (from-to)191-197
Number of pages7
JournalBiochemical and Biophysical Research Communications
Volume418
Issue number1
DOIs
Publication statusPublished - 2012 Feb 3

Fingerprint

Recombinant Fusion Proteins
Retinol-Binding Proteins
Albumins
Vitamin A
Fusion reactions
Tissue
Caveolae
Hepatic Stellate Cells
Podocytes
Myofibroblasts
Tissue Distribution
Cell- and Tissue-Based Therapy
Endocytosis
Bioactivity
Adipocytes
Protein Binding
Proteins
Fibrosis
Fatty Acids
Chemical activation

Keywords

  • Albumins
  • Anti-fibrotic drug
  • Fibrosis
  • Pancreatic stellate cells
  • Retinol-binding protein

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Cell Biology
  • Molecular Biology

Cite this

Recombinant fusion protein of albumin-retinol binding protein inactivates stellate cells. / Choi, Soyoung; Park, Sangeun; Kim, Suhyun; Lim, Chae Seung; Kim, Jungho; Cha, Dae-Ryong; Oh, Jun Seo.

In: Biochemical and Biophysical Research Communications, Vol. 418, No. 1, 03.02.2012, p. 191-197.

Research output: Contribution to journalArticle

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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-albumindomain III (R-III) and albumindomain I-RBP-albuminIII (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.",
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