Journal: bioRxiv
Article Title: NECTIN-4 PET FOR OPTIMIZING ENFORTUMAB VEDOTIN DOSE-RESPONSE IN UROTHELIAL CARCINOMA
doi: 10.1101/2024.12.25.630315
Figure Lengend Snippet: A) Schematic representation of competition assays between [ 68 Ga]AJ647 and EV for binding to Nectin-4. On the left, the schematic illustrates the impact of varying EV concentrations on Nectin-4 levels, with accessible Nectin-4 quantified using [ 68 Ga]AJ647. On the right, the schematic depicts how varying concentrations of AJ647 affect Nectin-4 levels, with accessible Nectin-4 determined by EV-FITC binding. B) In vitro binding of [ 68 Ga]AJ647 to HT1376 cells with varying concentrations of EV. The data show concentration-dependent inhibition of [ 68 Ga]AJ647 binding, with an IC 50 of 2.7 nM, indicating the competitive nature of EV in blocking [ 68 Ga]AJ647 binding to Nectin-4. C) In vitro binding of EV-FITC to HT1376 cells, assessed by flow cytometry, with varying concentrations of AJ647. The results demonstrate concentration-dependent inhibition of EV-FITC binding, with an IC 50 of 35 nM, highlighting that AJ647 competes with EV-FITC for Nectin-4 binding. data represented as mean±SD (n=3–4). D) In vitro binding of [ 68 Ga]AJ647 to HT1376, SCaBER and T24 cells with and without 60 nM EV. E) In vivo PET-CT imaging in mice harboring SCaBER (left) and HT1376 (right) tumors pre- and post- 20 mpk EV treatment (n=2). PET-CT was acquired 60-minutes after 7.4 MBq (~200 μCi) [ 68 Ga]AJ647 injection. E) In vivo PET-CT imaging in mice harboring SCaBER (left) and HT1376 (right) tumors at saturating dose of 20 mpk or saline (n=2). PET-CT was acquired at 60-minutes after 7.4 MBq (~200 μCi) [ 68 Ga]AJ647 injection. F) Ex vivo biodistribution in mice harboring SCaBER (left) and HT1376 (right) tumors treated with 20 mpk or saline control (n=5). Mice were sacrificed at 60-minutes after 1.85 MBq (~50 μCi) [ 68 Ga]AJ647 injection.
Article Snippet: Fluorescein isothiocyanate (FITC) analog of EV was made using manufacturer recommended protocol (Thermo #46409).
Techniques: Binding Assay, In Vitro, Concentration Assay, Inhibition, Blocking Assay, Flow Cytometry, In Vivo, Positron Emission Tomography-Computed Tomography, Imaging, Injection, Saline, Ex Vivo, Control