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Letters to the Editor
cells with cHL-MSC,5 it could cooperate with trabectedin not only by enhancing DNA damage in CCR5+ cells but also by counteracting the protective effects of the cross- talk between HRS and cHL-MSC. We evaluated the effi- cacy of the maraviroc-trabectedin combination in formed
heterospheroids (direct contact). We cultured HRS cells (L-1236 or HDLM-2 cells) with cHL-MSC under non- adherent conditions. After 24 h (the time necessary to obtain cell aggregation as heterospheroids), we added maraviroc and different concentrations of trabectedin9
A
BC
D
Figure 3. Maraviroc cooperated with trabectedin to decrease cell viability in three-dimensional heterospheroids formed by Hodgkin and Reed-Sternberg cells and classical Hodgkin lymphoma mesenchymal stromal cells. (A) Schedule of treatment of Hodgkin and Reed-Sternberg (HRS) cells and classical Hodgkin lym- phoma mesenchymal stromal cells (cHL-MSC). (B) L-1236, or (C) HDLM-2 cells were cultured in non-adherent conditions with cHL-MSC (1.0 x 104/mL of each cell type) in 24-well plates and treated with trabectedin (360 pM) alone, maraviroc (100 mM) alone or in combination. Drugs were added immediately (add. T= 0) and after heterospheroid formation (add. T=24h). After 6 days, cell viability was evaluated using the PrestoBlue Cell Viability Reagent (Invitrogen). Values are mean and standard deviation of three experiments. °P<0.05 for T=0 vs. T=24h, Student t-test. *P<0.05 treatments vs. medium, One-way analysis of vari- ance followed by the Dunnett test. (D) Representative phase contrast micrographs of heterospheroids cultured with trabectedin, maraviroc and their combina- tion (T=0). MVC, maraviroc; TB, trabectedin. add., added.
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haematologica | 2022; 107(1)