Figure 6.

BAT1 decoration of dox-liposomes leads to increased uptake in primary CLL cells, causing significantly increased rates of death compared with bare liposomes. (A) Schematic illustrating the postinsertion process for decorating dox-liposomes. (B) Primary CLL cells from 10 subjects were incubated with dox-liposomes, with and without BAT1 decoration. (Bi) Liposomal uptake and doxorubicin delivery were determined by assessing the doxorubicin-associated fluorescence in dead cells using flow cytometry. Each point represents the median fluorescence from an individual case. Median doxorubicin-associated fluorescence in dead CLL cells is significantly higher when CLL cells are incubated with BAT1-decorated dox-liposomes than with bare dox-liposomes. P = .0020, Wilcoxon matched-pairs signed-rank test. (Bii) Dox-liposome–associated death was measured in primary CLL cells across 10 cases using flow cytometry. The number of dead cells was calculated using forward/side scatter. BAT1-decorated liposomes consistently led to increased levels of cell death across all cases tested. P = .0039, Wilcoxon matched-pairs signed-rank test (**P < .01). (C) Cellular localization of the doxorubicin was assessed using laser deconvolution microscopy. Images were prepared by building a z-projection (intensity sum) from the image stack comprising the central region of the cell. Colocalization of the doxorubicin-associated red fluorescence and the DAPI staining was used as a measure of delivery to the cell nucleus. Doxorubicin is delivered to the cell nucleus in large quantities when cells are incubated for 3 hours with BAT1-decorated liposomes. Scale bar, 20 µm.