Design Synthesis and Validation of Axl-TargetedMonoclonal Antibody Probe for microPET Imaging in Human Lung CancerXenograft

摘要

Accumulating experimental evidence indicates that overexpression of the oncogenic receptor tyrosine kinase, Axl, plays a key role in the tumorigenesis and metastasis of various types of cancer. The objective of this study is to design a novel imaging probe based on the monoclonal antibody, h173, for microPET imaging of Axl expression in human lung cancer. A bifunctional chelator, DOTA, was conjugated to h173, followed by radiolabeling with ⁶⁴Cu. The binding of DOTA-h173 to the Axl receptor was first evaluated by a cell uptake assay and flow cytometry analysis using human lung cancer cell lines. The probe ⁶⁴Cu-DOTA-h173 was further evaluated by microPET imaging, and ex vivo histology studies in the Axl-positive A549 tumors. In vitro cellular study showed that Axl probe, ⁶⁴Cu-DOTA-h173, was highly immuno-reactive with A549 cells. Western blot analysis confirmed that Axl is highly expressed in the A549 cell line. For microPET imaging, the A549 xenografts demonstrated a significantly higher ⁶⁴Cu-DOTA-h173 uptake compared to the NCI-H249 xenograft (a negative control model). Furthermore, ⁶⁴Cu-DOTA-h173 uptake in A549 is significantly higher than that of ⁶⁴Cu-DOTA-hIgG. Immuno-fluorescence staining was consistent with the in vivo micro-PET imaging results. In conclusion, ⁶⁴Cu-DOTA-h173 could be potentially used as a probe for noninvasive imaging of Axlexpression, which could collect important information regarding tumorresponse to Axl-targeted therapeutic interventions.