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Blood, Vol. 113, Issue 24, 6172-6181, June 11, 2009
Targeting the Notch1 and mTOR pathways in a mouse T-ALL model
Blood Cullion et al.
113: 6172
Supplemental materials for: Cullion et al
Files in this Data Supplement:
- Table S1. Body weight is maintained in mice treated with successive cycles of intermittent GSI dosing (PDF, 54.4 KB)
- Figure S1. MRK-003 is a potent gamma secretase inhibitor that represses Notch1 target gene expression and induces apoptosis in vitro (JPG, 273 KB)
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(A) MRK-003 treatment represses Hes1 and Deltex1 expression. Mouse leukemic cell lines were treated with 1µM MRK-003 for the times periods indicated and Hes1 and Deltex1 expression examined using RT-PCR. Glyceraldehyde-3-phosphate dehydrogenase (Gapdh) was used as an internal control. (B) MRK-003 treatment of mouse leukemic cell lines results in growth arrest and apoptosis. Three murine leukemic lines (720,135,5151) were treated for 72 hours with 1µM DAPT, 1µM MRK-003, or DMSO at a final concentration of 0.01%. Cells were harvested and stained with PI and DNA content measured by flow cytometry. (C) MRK-003 treatment is effective against primary mouse tumors that harbor Notch1 mutations. Thymic masses isolated directly from Tal1/Ink4a/Arf+∕− transgenic animals were left untreated or were treated ex vivo with 1µM DAPT, 1µM MRK-003, or DMSO for 3 days. Cells were then stained with FITC-Annexin V/PI and analyzed by flow cytometry. Three independent tumors were analyzed, one representative experiment is shown. Statistics were analyzed using a Kruskal-Wallis test.
- Figure S2. GSI-induced gut metaplasia is reduced during the four-day rest period (JPG, 322 KB)
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FVB/N mice were treated with either vehicle (0.5% methylcellulose) or MRK-003 at 150mg/kg, once daily for three consecutive days. Mouse duodenum was harvested at 6, 24, 48, and 96 hours following the last treatment and stained with PAS. The presence of goblet cells (visualized as red clusters) was then compared. Scale bar, 100µM.
- Figure S3. GSI plasma concentrations do not accumulate upon successive treatment cycles (JPG, 31.5 KB)
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TALL-1 tumor bearing xenograft mice were treated with either vehicle or MRK-003 at 75, 150, 300 or 450mg/kg once a week for 1, 2, or 3 successive weeks. Plasma was isolated four hours following MRK-003 administration during each cycle (n=4 mice/group).
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