A new fluorescence-based optical imaging method to non-invasively monitor hepatic myofibroblasts in vivo

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• 作者：Saimir Luli¹ ; Daniela Di Paolo² ; Patrizia Perri² ; Chiara Brignole² ; Stephen J. Hill¹ ; Helen Brown¹ ; Jack Leslie¹ ; H.L. Marshall¹ ; Matthew C. Wright¹ ; Derek A. Mann¹ ; Mirco Ponzoni² ; Fiona Oakley¹ ; ^{fiona.oakley@newcastle.ac.uk" class="auth_mail" title="E-mail the corresponding author}
• 关键词：HM ; hepatic myofibroblast ; αSMA ; alpha-smooth muscle actin ; WT ; wild-type ; ScAb ; single chain antibody ; CCl4 ; chronic carbon tetrachloride ; NIR ; near infra-red ; ROI ; region of interest ; ALT ; alanine transaminase ; AST ; aspartate aminotransferase ; PBS ; phosphate buffered saline ; BDL ; bile duct ligation ; ALP ; alkaline phosphatase ; DOX ; Doxorubin ; SYN ; synaptophysin
• 刊名：Journal of Hepatology
• 出版年：2016
• 出版时间：July 2016
• 年：2016
• 卷：65
• 期：1
• 页码：75-83
• 全文大小：2918 K

文摘

Currently, staging of fibrosis in preclinical rodent liver fibrosis models is achieved histologically. Many animals are used at multiple time-points to assess disease progression or therapeutic responses. Hepatic myofibroblasts promote liver fibrosis therefore quantifying these cells in vivo could assess disease or predict therapeutic responses in mice. We fluorescently labelled a single chain antibody (C1-3) that binds hepatic myofibroblasts to monitor fibrogenesis in vivo.

Methods

CCl₄ was used to induce acute liver injury in WT and cRel^−/− mice. Bile duct ligation was used to model chronic fibrosis. Hepatic myofibroblasts were depleted using a liposome-drug delivery system or chemically with sulfasalazine. An IVIS® spectrum visualised fluorophore-conjugated C1-3 in vivo.

Results

IVIS detection of fluorescently labelled-C1-3 but not a control antibody discriminates between fibrotic and non-fibrotic liver in acute and chronic liver fibrosis models. cRel^−/− mice have a fibro-protective phenotype and IVIS signal is reduced in CCl₄ injured cRel^−/− mice compared to wild-type. In vivo imaging of fluorescently labelled-C1-3 successfully predicts reductions in hepatic myofibroblast numbers in fibrotic liver disease in response to therapy.

Conclusions

We report a novel fluorescence imaging method to assess murine hepatic myofibroblast numbers in vivo during liver fibrosis and after therapy. We also describe a novel liposomal antibody targeting system to selectively deliver drugs to hepatic myofibroblasts in vivo. C1-3 binds human hepatic myofibroblast therefore imaging labelled-C1-3 could be used for clinical studies in man to help stage fibrosis, demonstrate efficacy of drugs that promote hepatic myofibroblast clearance or predict early therapeutic responses.

Lay summary

In response to damage and injury scars develop in the liver and the main cell that makes the scar tissue is the hepatic myofibroblast (HM). C1-3 is an antibody fragment that binds to the scar forming HM. We have fluorescently labelled C1-3 and given it to mice that have either normal or scarred livers (which contain HM) and then used a machine called an in vivo imaging system (IVIS) that takes pictures of different wavelengths of light, to visualise the antibody binding to HM inside the living mouse. Using fluorescently labelled C1-3 we can assess HM numbers in the injured liver and monitor response to therapy. We have also used C1-3 to target drugs encapsulated in lipid carriers (liposomes) to the HM to kill the HM and reduce the liver disease.