Publication

Targeted Recombinant Fusion Proteins of IFN gamma and Mimetic IFN gamma with PDGF beta R Bicyclic Peptide Inhibits Liver Fibrogenesis In Vivo

Bansal, R., Prakash, J., De Ruiter, M. & Poelstra, K., 24-Feb-2014, In : PLoS ONE. 9, 2, 10 p., e89878.

Research output: Contribution to journalArticleAcademicpeer-review

APA

Bansal, R., Prakash, J., De Ruiter, M., & Poelstra, K. (2014). Targeted Recombinant Fusion Proteins of IFN gamma and Mimetic IFN gamma with PDGF beta R Bicyclic Peptide Inhibits Liver Fibrogenesis In Vivo. PLoS ONE, 9(2), [e89878]. https://doi.org/10.1371/journal.pone.0089878

Author

Bansal, Ruchi ; Prakash, Jai ; De Ruiter, Marieke ; Poelstra, Klaas. / Targeted Recombinant Fusion Proteins of IFN gamma and Mimetic IFN gamma with PDGF beta R Bicyclic Peptide Inhibits Liver Fibrogenesis In Vivo. In: PLoS ONE. 2014 ; Vol. 9, No. 2.

Harvard

Bansal, R, Prakash, J, De Ruiter, M & Poelstra, K 2014, 'Targeted Recombinant Fusion Proteins of IFN gamma and Mimetic IFN gamma with PDGF beta R Bicyclic Peptide Inhibits Liver Fibrogenesis In Vivo', PLoS ONE, vol. 9, no. 2, e89878. https://doi.org/10.1371/journal.pone.0089878

Standard

Targeted Recombinant Fusion Proteins of IFN gamma and Mimetic IFN gamma with PDGF beta R Bicyclic Peptide Inhibits Liver Fibrogenesis In Vivo. / Bansal, Ruchi; Prakash, Jai; De Ruiter, Marieke; Poelstra, Klaas.

In: PLoS ONE, Vol. 9, No. 2, e89878, 24.02.2014.

Research output: Contribution to journalArticleAcademicpeer-review

Vancouver

Bansal R, Prakash J, De Ruiter M, Poelstra K. Targeted Recombinant Fusion Proteins of IFN gamma and Mimetic IFN gamma with PDGF beta R Bicyclic Peptide Inhibits Liver Fibrogenesis In Vivo. PLoS ONE. 2014 Feb 24;9(2). e89878. https://doi.org/10.1371/journal.pone.0089878


BibTeX

@article{1bcb731dfa8940689a16fd14097d96c5,
title = "Targeted Recombinant Fusion Proteins of IFN gamma and Mimetic IFN gamma with PDGF beta R Bicyclic Peptide Inhibits Liver Fibrogenesis In Vivo",
abstract = "Hepatic stellate cells (HSCs), following transdifferentiation to myofibroblasts plays a key role in liver fibrosis. Therefore, attempts to attenuate this myofibroblastic phenotype would be a promising therapeutic approach. Interferon gamma (IFN gamma) is a potent anti-fibrotic cytokine, but its pleiotropic receptor expression leading to severe adverse effects has limited its clinical application. Since, activated HSC express high-level of platelet derived growth factor beta receptor (PDGF beta R), we investigated the potential of PDGFbR-specific targeting of IFN gamma and its signaling peptide that lacks IFN gamma R binding site (mimetic IFN gamma or mimIFNc) in liver fibrosis. We prepared DNA constructs expressing IFN gamma, mimIFN gamma or BiPPB (PDGF beta R-specific bicyclic peptide)-IFN gamma, BiPPB-mimIFN gamma fusion proteins. Both chimeric proteins alongwith IFN gamma and mimIFN gamma were produced in E. coli. The expressed proteins were purified and analyzed for PDGF beta R-specific binding and in vitro effects. Subsequently, these recombinant proteins were investigated for the liver uptake (pSTAT1 alpha signaling pathway), for antifibrotic effects and adverse effects (platelet counts) in CCl4-induced liver fibrogenesis in mice. The purified HSC-targeted IFN gamma and mimIFN gamma fusion proteins showed PDGF beta R-specific binding and significantly reduced TGF beta-induced collagen-I expression in human HSC (LX2 cells), while mouse IFN gamma and mimIFN gamma did not show any effect. Conversely, mouse IFN gamma and BiPPB-IFN gamma induced activation and dose-dependent nitric oxide release in mouse macrophages (express IFN gamma R while lack PDGF beta R), which was not observed with mimIFNc and BiPPB-mimIFN gamma, due to the lack of IFN gamma R binding sites. In vivo, targeted BiPPB-IFN gamma and BiPPB-mimIFN gamma significantly activated intrahepatic IFNc-signaling pathway compared to IFN gamma and mimIFN gamma suggesting increased liver accumulation. Furthermore, the targeted fusion proteins ameliorated liver fibrogenesis in mice by significantly reducing collagen and alpha-SMA expression and potentiating collagen degradation. IFN gamma also induced reduction in fibrogenesis but showed significant decrease in platelet counts, which was restored with targeted proteins. These results suggest that these rationally designed proteins can be further developed as novel anti-fibrotic therapeutics.",
keywords = "HEPATIC STELLATE CELLS, GROWTH-FACTOR RECEPTOR, INTERFERON-GAMMA, FIBROSIS, ACTIVATION, EXPRESSION, CIRRHOSIS, EFFICACY, INJURY",
author = "Ruchi Bansal and Jai Prakash and {De Ruiter}, Marieke and Klaas Poelstra",
year = "2014",
month = "2",
day = "24",
doi = "10.1371/journal.pone.0089878",
language = "English",
volume = "9",
journal = "PLOS-One",
issn = "1932-6203",
publisher = "PUBLIC LIBRARY SCIENCE",
number = "2",

}

RIS

TY - JOUR

T1 - Targeted Recombinant Fusion Proteins of IFN gamma and Mimetic IFN gamma with PDGF beta R Bicyclic Peptide Inhibits Liver Fibrogenesis In Vivo

AU - Bansal, Ruchi

AU - Prakash, Jai

AU - De Ruiter, Marieke

AU - Poelstra, Klaas

PY - 2014/2/24

Y1 - 2014/2/24

N2 - Hepatic stellate cells (HSCs), following transdifferentiation to myofibroblasts plays a key role in liver fibrosis. Therefore, attempts to attenuate this myofibroblastic phenotype would be a promising therapeutic approach. Interferon gamma (IFN gamma) is a potent anti-fibrotic cytokine, but its pleiotropic receptor expression leading to severe adverse effects has limited its clinical application. Since, activated HSC express high-level of platelet derived growth factor beta receptor (PDGF beta R), we investigated the potential of PDGFbR-specific targeting of IFN gamma and its signaling peptide that lacks IFN gamma R binding site (mimetic IFN gamma or mimIFNc) in liver fibrosis. We prepared DNA constructs expressing IFN gamma, mimIFN gamma or BiPPB (PDGF beta R-specific bicyclic peptide)-IFN gamma, BiPPB-mimIFN gamma fusion proteins. Both chimeric proteins alongwith IFN gamma and mimIFN gamma were produced in E. coli. The expressed proteins were purified and analyzed for PDGF beta R-specific binding and in vitro effects. Subsequently, these recombinant proteins were investigated for the liver uptake (pSTAT1 alpha signaling pathway), for antifibrotic effects and adverse effects (platelet counts) in CCl4-induced liver fibrogenesis in mice. The purified HSC-targeted IFN gamma and mimIFN gamma fusion proteins showed PDGF beta R-specific binding and significantly reduced TGF beta-induced collagen-I expression in human HSC (LX2 cells), while mouse IFN gamma and mimIFN gamma did not show any effect. Conversely, mouse IFN gamma and BiPPB-IFN gamma induced activation and dose-dependent nitric oxide release in mouse macrophages (express IFN gamma R while lack PDGF beta R), which was not observed with mimIFNc and BiPPB-mimIFN gamma, due to the lack of IFN gamma R binding sites. In vivo, targeted BiPPB-IFN gamma and BiPPB-mimIFN gamma significantly activated intrahepatic IFNc-signaling pathway compared to IFN gamma and mimIFN gamma suggesting increased liver accumulation. Furthermore, the targeted fusion proteins ameliorated liver fibrogenesis in mice by significantly reducing collagen and alpha-SMA expression and potentiating collagen degradation. IFN gamma also induced reduction in fibrogenesis but showed significant decrease in platelet counts, which was restored with targeted proteins. These results suggest that these rationally designed proteins can be further developed as novel anti-fibrotic therapeutics.

AB - Hepatic stellate cells (HSCs), following transdifferentiation to myofibroblasts plays a key role in liver fibrosis. Therefore, attempts to attenuate this myofibroblastic phenotype would be a promising therapeutic approach. Interferon gamma (IFN gamma) is a potent anti-fibrotic cytokine, but its pleiotropic receptor expression leading to severe adverse effects has limited its clinical application. Since, activated HSC express high-level of platelet derived growth factor beta receptor (PDGF beta R), we investigated the potential of PDGFbR-specific targeting of IFN gamma and its signaling peptide that lacks IFN gamma R binding site (mimetic IFN gamma or mimIFNc) in liver fibrosis. We prepared DNA constructs expressing IFN gamma, mimIFN gamma or BiPPB (PDGF beta R-specific bicyclic peptide)-IFN gamma, BiPPB-mimIFN gamma fusion proteins. Both chimeric proteins alongwith IFN gamma and mimIFN gamma were produced in E. coli. The expressed proteins were purified and analyzed for PDGF beta R-specific binding and in vitro effects. Subsequently, these recombinant proteins were investigated for the liver uptake (pSTAT1 alpha signaling pathway), for antifibrotic effects and adverse effects (platelet counts) in CCl4-induced liver fibrogenesis in mice. The purified HSC-targeted IFN gamma and mimIFN gamma fusion proteins showed PDGF beta R-specific binding and significantly reduced TGF beta-induced collagen-I expression in human HSC (LX2 cells), while mouse IFN gamma and mimIFN gamma did not show any effect. Conversely, mouse IFN gamma and BiPPB-IFN gamma induced activation and dose-dependent nitric oxide release in mouse macrophages (express IFN gamma R while lack PDGF beta R), which was not observed with mimIFNc and BiPPB-mimIFN gamma, due to the lack of IFN gamma R binding sites. In vivo, targeted BiPPB-IFN gamma and BiPPB-mimIFN gamma significantly activated intrahepatic IFNc-signaling pathway compared to IFN gamma and mimIFN gamma suggesting increased liver accumulation. Furthermore, the targeted fusion proteins ameliorated liver fibrogenesis in mice by significantly reducing collagen and alpha-SMA expression and potentiating collagen degradation. IFN gamma also induced reduction in fibrogenesis but showed significant decrease in platelet counts, which was restored with targeted proteins. These results suggest that these rationally designed proteins can be further developed as novel anti-fibrotic therapeutics.

KW - HEPATIC STELLATE CELLS

KW - GROWTH-FACTOR RECEPTOR

KW - INTERFERON-GAMMA

KW - FIBROSIS

KW - ACTIVATION

KW - EXPRESSION

KW - CIRRHOSIS

KW - EFFICACY

KW - INJURY

U2 - 10.1371/journal.pone.0089878

DO - 10.1371/journal.pone.0089878

M3 - Article

C2 - 24587093

VL - 9

JO - PLOS-One

JF - PLOS-One

SN - 1932-6203

IS - 2

M1 - e89878

ER -

ID: 16221544