PMC8357369_01
(A) Morphology of lymphoblasts at diagnosis (original magnification, x 1000). (B) Flow cytometry result. (C) FISH results using a break-apart probe for TCF3. Rearrangement of TCF3 was shown as one red and one green signal. (D) FISH results using a break-apart probe for ZNF384. (E) Drug sensitivity screening test in vitro with a panel of 156 kinds of cytotoxic drugs, molecular targeted therapy drugs, and combination chemotherapy regimens. Relative inhibition rates higher than 80% were listed. (F) Relapsed lymphoblasts showed moderate sensitivity to both an ALL scheme [VDLP, CAM, hyper-CVAD (B)] and AML schemes (HAD, DA, MA). (G) Relapsed lymphoblasts were sensitive to multiple targeted therapeutic drugs. (H) The hyper- CVAD (B) scheme showed multiple synergistic effects with multiple targeted compounds.
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FAB classification (Figure 1A). Flow cytometry showed malignant B lymphoblasts (P3 group, 92.1%) mainly positive for CD34, CD19, CD10, cCD79a, TdT, CD22, HLA-DR, CD58, and CD13; partially positive for CD38 and CD123; weakly positive for CD33; and negative for CD117, CD7, CD3, MPO, cCD3, CD56, CD15, CD79a, cIgM, CD25, CD20, surface Kappa and Lambda, in accordance with com-B-ALL (Figure 1B) (Experiment was conducted by CANTO II, 6 color flow cytometry, Becton, Dickinson and Company(BD) and all antibodies were purchased from BD company). Karyotype analysis showed that the patient had a 46,XY karyotype [20]. TCF3 mutation (c.1061delC|p.Ser354fs), a NOTCH2 mutation (c.1274A>G|p.Asn425Ser), a CARD11 mutation (c.2735G>A|p.Arg912Gln), and an SH2B3 mutation (c.1307A>G|p.Asp436Gly). The TCF3-ZNF384 fusion was further confirmed by qPCR and fluorescence in situ hybridization (FISH) using two break-apart probes for TCF3 and ZNF384 (Figures 1C, D) (Wuhan Kanglu Biotechnology Co., Ltd). Relative inhibition rates higher than 80% were listed in Figure 1E. Interestingly, the lymphoblasts showed moderate sensitivity to both ALL schemes (VDLP, CAM, hyper-CVAD (B), etc.) and AML schemes (HAD, DA, MA, etc.) (Figure 1F), as well as to multiple targeted therapeutic drugs, such as the Jak2 inhibitor ruxolitinib, the tyrosine kinase inhibitors sorafenib, midostaurin, and imatinib, the proteasome inhibitors carfilzomib (Figure 1G). Furthermore, the hyper-CVAD (B) scheme showed multiple synergistic effects with multiple targeted compounds (bortezomib, decitabine, sorafenib and chidamide) (Figure 1H) and had the highest inhibition rates (Figure 1E).