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Neutralizing antibody against osteopontin attenuates non-alcoholic steatohepatitis in mice

Abstract

Previously, we reported that an extracellular matrix protein, osteopontin (OPN), is involved in various autoimmune diseases using a neutralizing polyclonal antibody against OPN generated in rabbits. However, the antibody cannot be used for long-term mouse models of chronic inflammatory disease because of the induction of antibodies against anti-OPN rabbit IgG. In this study, we generated a new antibody, anti-mouse OPN mouse IgG (35B6). 35B6 inhibited the cell adhesion of mouse and human OPN to Chinese Hamster Ovary (CHO) cells or CHO cells expressing α4 or α9 integrin. It was reported that OPN is highly expressed and has an important role in a chronic liver disease, non-alcoholic steatohepatitis (NASH). 35B6 injection twice a week for 8 weeks attenuated liver inflammation and fibrosis in a NASH mouse model, suggesting 35B6 is beneficial for the treatment of NASH. 35B6 was preferable to the rabbit anti-OPN antibody for investigating the in vivo function of OPN in mouse models of long-term disease.

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Abbreviations

CDAHFD:

0.1% methionine on a high fat diet background

col1a1:

type I collagen α1 chain

CHO:

Chinese Hamster Ovary

ECM:

extracellular matrix

HE:

hematoxylin-eosin

HSC:

hepatic stellate cells

RGD:

arginine-glycine-aspartate

NAFLD:

non-alcoholic fatty liver disease

NASH:

non-alcoholic steatohepatitis in mice

OPN:

osteopontin

SMA:

smooth muscle actin

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Acknowledgements

We would like to thank to Junko Miyazaki (Fukuyama University), Kazutaka Matsunami (Immuno-Biological Laboratories Co., Ltd.) and Masaya Takeuchi (Sapporo General Pathology Laboratory Co., Ltd.) for excellent support in the in vivo studies. This study was supported by research grants from JSPS KAKENHI (Grant Numbers 16 K08221 and 19 K07492), the Japan Rheumatism Foundation, the SUHARA Memorial Foundation, the FUGAKU TRUST for Medical Research, and academic support from Pfizer, Chugai, Astellas and Eli Lilly to SK. We thank Edanz Group (www.edanzediting.com/ac) for editing a draft of this manuscript.

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TU and SK designed the study. MH, CK, and SK performed the experiments. SK wrote the manuscript. All authors read and approved the final manuscript.

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Correspondence to Shigeyuki Kon.

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Honda, M., Kimura, C., Uede, T. et al. Neutralizing antibody against osteopontin attenuates non-alcoholic steatohepatitis in mice. J. Cell Commun. Signal. 14, 223–232 (2020). https://doi.org/10.1007/s12079-020-00554-7

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Keywords

  • Osteopontin
  • Antibody
  • Non-alcoholic steatohepatitis in mice
  • Inflammation
  • Fibrosis