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Synthesis and Preclinical Evaluation of a 89Zr-labelled Human Single Chain Antibody for Non-invasive Detection of Hepatic Myofibroblasts in Acute Liver Injury

Lookup NU author(s): Toni Pringle, Erik Ramon Gil, Dr Jack LeslieORCiD, Professor Fiona OakleyORCiD, Professor Matthew Wright, Dr James KnightORCiD, Dr Saimir Luli

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This work is licensed under a Creative Commons Attribution 4.0 International License (CC BY 4.0).


Abstract

Synaptophysin is expressed on fibrogenic hepatic myofibroblasts. C1-3 is a single chain human antibody (scAb) that binds specifically to synaptophysin on hepatic myofibroblasts, providing a targeting vector for novel in vivo imaging agents of chronic liver disease. C1-3 and a negative control scAb, CSBD9, were radiolabelled with zirconium-89 via desferrioxamine chelation to enable non-invasive molecular imaging with Positron Emission Tomography (PET). DFO-scAb conjugates were characterised by gel electrophoresis (SDS-PAGE) and MALDI-TOF spectrometry, and 89Zr-labelled with high radiolabelling efficiency (99%). [89Zr]Zr-DFO-C1-3 exhibited high in vitro stability (>99%) in mouse and human sera over three days at 25 and 37°C. Activated hepatic myofibroblasts incubated with [89Zr]Zr-DFO-C1-3 displayed significantly higher internalised activity (59.46%, P = 0.001) compared to the [89Zr]Zr-DFO-CSBD9 control, indicating synaptophysin-mediated uptake and high binding specificity of [89Zr]Zr-DFO-C1-3. Mice with CCl4-induced acute liver damage exhibited significantly higher liver uptake of [89Zr]Zr-DFO-C1-3, compared to controls, confirmed by both Cerenkov imaging and ex vivo gamma counting (4.41 ± 0.19 %ID/g, P < 0.0001). CCl4-induced liver damage and the number of hepatic myofibroblasts was confirmed by aSMA staining of liver sections. These findings indicate that [89Zr]Zr-DFO-C1-3 has promising utility as a PET imaging agent for non-invasive detection of hepatic myfibroblasts following acute liver injury.


Publication metadata

Author(s): Pringle TA, Ramon-Gil E, Leslie J, Oakley F, Wright MC, Knight JC, Luli S

Publication type: Article

Publication status: Published

Journal: Scientific Reports

Year: 2024

Volume: 14

Online publication date: 05/01/2024

Acceptance date: 25/12/2023

Date deposited: 20/12/2023

ISSN (electronic): 2045-2322

Publisher: Nature Publishing Group

URL: https://doi.org/10.1038/s41598-023-50779-w

DOI: 10.1038/s41598-023-50779-w


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Funding

Funder referenceFunder name
MR/K0019494/1
MRC
MR/R023026/1Medical Research Council (MRC)
Wellcome Trust
WE Harker Foundation

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