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. 2015 Jun 12;5:11295. doi: 10.1038/srep11295

Figure 3. Conventional Biomarkers Provide Limited Information; Chemotherapy is a Confounder.

Figure 3

(a) Molm-14-engrafted NSG chimerism by flow. Flow cytometry for human CD45 was performed on the peripheral blood and bone marrow of Molm-14-engrafted NSG mice, both without and with cytarabine treatment (300 mg/kg, Q3Dx3 starting 14d post-engraftment). (b) mRNA markers of AML in serum exosomes. Exosomes isolated from the peripheral blood of Molm-14-engrafted NSG mice were tested for human FLT3 and GAPDH and murine GAPDH using 45 cycles of PCR (cropped; full gels in Figure S3). NTC: null-template control. M14: Molm-14 stock culture. PB: Peripheral blood chimerism; BM: Bone marrow chimerism by flow cytometry for human CD45. (c) CBC of Molm-engrafted NSG. Peripheral blood CBC of mice, comparing baseline NSG (n = 31), NSG engrafted with Molm-14 at 2 (n = 16) and 3 (n = 5) weeks post-engraftment, NSG engrafted with Molm-14 and treated with cytarabine (n = 8) at 3 weeks, control NSG given cytarabine (n = 4), and NSG given nonmalignant CD34+ human cells (n = 4). WBC, white blood cells; LY, lymphocytes; NE, neutrophils; Hb, hemoglobin; PLT, platelets (×1000/uL). *, p < 0.05 by Student’s t. Error bars represent SEM. (d,e) In vitro chemotherapy and exosomal miRNA. Molm-14, HL-60, and U937 cells were exposed to cytarabine (25 ng/mL, d) overnight, and Molm-14 were exposed to quizartinib (0.1 nM, e) overnight. Cellular (on the x-axis) and exosomal (on the y-axis) levels of miRNA were measured by qRT-PCR, and are presented as ddCT (a -1 ddCT represents a 2-fold increase in miRNA).