Journal: EMBO Molecular Medicine

Article Title: A versatile drug delivery system targeting senescent cells

doi: 10.15252/emmm.201809355

Figure Lengend Snippet: A SK‐MEL‐103 melanoma cells were treated with palbociclib (1 μM) for 1 or 14 days, and senescence induction was assessed by SAβgal staining. Next, cultures were exposed to free doxorubicin (50 μM) or GalNP(dox) (1 mg/ml, filtered) for 30 min. Pictures show representative images illustrating doxorubicin fluorescence by confocal microscopy. Scale bar: 50 μm. B SK‐MEL‐103 melanoma cells were treated with palbociclib (5 μM) for 14 days, cultures were exposed to GalNP(dox) (0.06 mg/ml, filtered), and annexin V signal was quantified over time. Representative pictures are shown in Appendix Fig S3F . C Athymic nude female mice carrying subcutaneous SK‐MEL‐103 xenografts were treated daily with palbociclib (oral gavage, 50 mg/kg) and/or GalNP(dox) (tail vein injection, 200 μl of a solution with 4 mg/ml of GalNP containing a total of 1 mg/kg of deliverable doxorubicin), alone or in combination, as indicated. For each tumor, the relative tumor volume change was calculated relative to its baseline prior to treatment. Values are expressed as mean ± SEM. Individual tumor size measurements are shown in Appendix Fig S3G . D Similar to (B) but using GalNP(nav) (tail vein injection, 200 μl of a solution with 4 mg/ml of GalNP containing a total of 1 mg/kg of deliverable navitoclax), as indicated. Individual tumor size measurements are shown in Appendix Fig S3F . E Left, fold change of tumor size of SK‐MEL‐103 xenografts, after the indicated daily treatments. Data for palbociclib, and for palbociclib plus GalNP(dox), correspond to the same data in panel (C), at day 17. Data for free doxorubicin (daily tail vein injection, 1 mg/kg, for 17 days) were obtained in parallel. Right, mRNA levels of cardiotoxicity markers in hearts from the same mice. Actb and Gapdh were used for input normalization. Values are relative to control mice and are expressed as mean ± SD, and statistical significance was assessed by one‐way ANOVA and Dunnett's multiple comparisons test (versus palbociclib‐alone treated group). F Left, fold change of tumor size, as in (C), after the indicated daily treatments. Data for palbociclib, and for palbociclib plus GalNP(nav), correspond to the same data in panel (D), at day 13. Data for free navitoclax (daily oral gavage, 25 mg/kg, for 13 days) were obtained in parallel. Right, platelet levels in the blood of the same mice. Values are expressed as mean ± SEM in the case of tumor size, and as mean ± SD in the case of platelet counting, and statistical significance was assessed by one‐way ANOVA and Dunnett's multiple comparisons test (versus palbociclib‐alone treated group).

Article Snippet: Loading of the NP scaffolds with the different compounds was performed as follows: For NP(rho), 200 mg of NPs were suspended in a solution of 16 ml of EtOH together with 76.65 mg of rhodamine B (Sigma, #R6226); for NP(icg), 200 mg of mesoporous silica nanoparticles (MSNs) were suspended in a solution of 5 ml of water together with 5 mg of indocyanine green (Sigma, #I2633); for NP(dox), 200 mg of MSNs were suspended in a solution of 12.5 ml of water together with 110 mg of doxorubicin (Carbosynth, #AD15377); for NP(nav), 250 mg of NPs were suspended in 27 ml EtOH together with 91.3 mg of navitoclax (Active BioChem, #A‐1001).

Techniques: Staining, Fluorescence, Confocal Microscopy, Injection