BALB/c (IMSR Cat# JAX:000651, RRID:IMSR_JAX:000651) mice were sub-lethally X-ray irradiated with 300 cGy (CIXD irradiator, Xstrahl) and injected with 5 × 105 Ba/F3-FLT3/ITD-Luc cells via tail vein. Intraperitoneal injections were performed using PBS vehicle with human AGP (Sigma) at 20 mg/mL. Midostaurin was suspended in 30% (w/v) Cremophor EL, 30% (w/v) PEG 400, 10% ethanol, and 10% glucose (all Sigma), and instilled once daily by oral gavage (10 (link), 11 (link)). These drug doses have previously been demonstrated to be effective in mice. Mifepristone (Sigma) was dissolved in 50% ethanol and 50% (2-hydroxypropyl)-β-cyclodextrin (Sigma) and gavaged every 48 hours. Mice were imaged by intraperitoneal injection of luciferin (3 mg) and visualizing on an IVIS Spectrum imager (Caliper LifeSciences) using Living Image software for analysis on day 5 (to monitor engraftment) and on day 9 (to assess drug effect). Animal husbandry and procedures were conducted in accordance with the policy of the Johns Hopkins University School of Medicine Animal Care and Use Committee.
Protocol detail
Evaluating FLT3/ITD Leukemia Treatment
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Animal
Cells
Cremophor el
Cyclodextrin
Ethanol
Flt3
Glucose
Human
Hydroxypropyl
Intraperitoneal injection
Luciferin
Medicine
Mice
Midostaurin
Mifepristone
Peg 400
Tail
Vein
X ray
Corresponding Organization : Johns Hopkins Medicine
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Variable analysis
independent variables
- X-ray irradiation (300 cGy)
- Injection of Ba/F3-FLT3/ITD-Luc cells (5 × 10^5 cells)
- Intraperitoneal injection of human AGP (20 mg/mL)
- Oral gavage of Midostaurin (10 and 11 mg/kg)
- Oral gavage of Mifepristone
- Tumor engraftment (monitored by bioluminescence imaging on day 5)
- Tumor response to drug treatment (assessed by bioluminescence imaging on day 9)
- Mouse strain (BALB/c)
- PBS vehicle for human AGP injections
- Vehicle for Midostaurin (30% Cremophor EL, 30% PEG 400, 10% ethanol, 10% glucose)
- Vehicle for Mifepristone (50% ethanol, 50% (2-hydroxypropyl)-β-cyclodextrin)
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