Summary of therapy, clinical course, and cytogenetic and molecular analyses of sequential marrows. (A) Timeline with scale in months (mos) from neuroblastoma diagnosis. Percentages indicate maximum abnormal marrow cells by karyotype, FISH with dual color probe for MLL (Vysis, Downers Grove, IL), or chromosome 4 and 11 painting probes (Vysis). Abbreviations: CAV, cyclophosphamide (2100 mg/m2 per day × 2), doxorubicin (25 mg/m2 per day × 3), vincristine (0.66 mg/m2 per day × 3); PVP, cisplatin (50 mg/m2 per day × 4), etoposide (200 mg/m2 per day × 3); PBSCH, autologous peripheral blood stem cell harvest; 3F8, anti-GD2 antibodies; TCT, thiotepa (300 mg/m2 per day × 3), carboplatin (500 mg/m2 per day × 3), topotecan (2 mg/m2 per day × 5); PBSCT, autologous peripheral blood stem cell transplant; GMCSF, granulocyte macrophage colony stimulating factor; oral VP, oral etoposide (50 mg/m2 per day × 21 days). (B) Representative bone marrow aspirate smear from 45.5 months after neuroblastoma diagnosis at low-power magnification showing normal morphology and trilinear hematopoiesis (top left), and higher power magnification of normal erythroid and myeloid cells in same bone marrow aspirate (top right). Representative bone marrow aspirate smear from 57.2 months after neuroblastoma diagnosis at high-power magnification showing blasts, immature monocytes, a hypolobulated megakaryocyte, and a dysplastic myeloid cell (bottom left and right). Bone marrow aspirates were visualized using a Leica DMLB microscope (20×/0.40 aperature; 50×/0.90 aperature; 100×/1.25 aperature) captured with a Leica DFC420 camera (imaging medium of air for 20×, immersion oil for 50× and 100×), acquired using Leica Application Suite and processed using Microsoft (Redmond, WA) PowerPoint. (C) Tracing of der(11) genomic breakpoint junction in sequential marrows. The clonotypic der(11) genomic breakpoint junction was detectable by PCR in sample from 17 months after neuroblastoma diagnosis and all specimens thereafter. Time represents months from neuroblastoma diagnosis. All positive samples were sequenced to verify the breakpoint junction.
