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. 2022 Oct 2;13:5789. doi: 10.1038/s41467-022-33461-z

Fig. 3. Plasma cells from patients with AL amyloidosis become more primed for apoptosis and more dependent on pro-survival BCL-2 family proteins when treated with bortezomib, which induces Noxa expression.

Fig. 3

a, b Plasma cells were isolated from patient bone marrow aspirates, cultured ex vivo, and treated with a bortezomib and b ixazomib as single agents or in combination with BH3 mimetics at 100 nM. Apoptosis was measured via Annexin V positivity after 24 h. c, d Ex vivo sensitivity of the clonal plasma cell population in response to c bortezomib and d ixazomib treatments measured via Annexin V positivity after 24 h correlated to apoptotic priming of clonal plasma cells at the time of isolation, measured by sensitivity to the BIM BH3 peptide via BH3 profiling. Spearman rho was used to test for correlation between responses to BH3 peptides and cellular sensitivity to indicated agents. e, f Plasma cells isolated from unique patients were cultured ex vivo with or without 100 nM bortezomib and BH3 profiled after 16 h. Data are presented as mean values + /- SEM (n = 3). f Treatment-induced changes in apoptotic dependencies. g Lysates were prepared from patient-derived bone marrow mononuclear cells and western blot analysis was performed looking at changes in expression of BCL-2 family proteins in response to 24 h of ex vivo culture with bortezomib treatment as a single agent or in combination with ABT-199 and S632845. h BCL-2 family protein expression was measured in response to 24 h ex vivo treatment with bortezomib, ixazomib, lenalidomide, or pomalidomide. i, j BH3 profiling and k chemosensitivity analysis of clonal plasma isolated from patients on active treatment with bortezomib. l–n Plasma cells were isolated from patient bone marrow aspirates, cultured ex vivo, and treated with l lenalidomide, m pomalidomide, or n dexamethasone as single agents or in combination with BH3 mimetics. P-values were calculated using one-way ANOVA with Holm-Sidak’s adjustment for a, b, l–n, two-way ANOVA with Holm–Sidak’s adjustment for e, j, k, and two-way ANOVA for i.