Supplementary Materials

Supplementary Material for:

Exploiting an Asp-Glu "switch" in glycogen synthase kinase 3 to design paralog-selective inhibitors for use in acute myeloid leukemia

Florence F. Wagner,* Lina Benajiba, Arthur J. Campbell, Michel Weïwer, Joshua R. Sacher, Jennifer P. Gale, Linda Ross, Alexandre Puissant, Gabriela Alexe, Amy Conway, Morgan Back, Yana Pikman, Ilene Galinsky, Daniel J. DeAngelo, Richard M. Stone, Taner Kaya, Xi Shi, Matthew B. Robers, Thomas Machleidt, Jennifer Wilkinson, Olivier Hermine, Andrew Kung, Adam J. Stein, Damodharan Lakshminarasimhan, Michael T. Hemann, Edward Scolnick, Yan-Ling Zhang, Jen Q. Pan, Kimberly Stegmaier,* Edward B. Holson

*Corresponding author. Email: fwagner{at}broadinstitute.org (F.F.W.); kimberly_stegmaier{at}dfci.harvard.edu (K.S.)

Published 7 March 2018, Sci. Transl. Med. 10, eaam8460 (2018)
DOI: 10.1126/scitranslmed.aam8460

This PDF file includes:

  • Materials and Methods
  • Fig. S1. GSK3 conservation across species.
  • Fig. S2. Compounds’ electron density maps.
  • Fig. S3. X-ray crystal structure of GSK3 from U. maydis.
  • Fig. S4. Schematic of backend interactions.
  • Fig. S5. Hinge backend interaction measurements and energy plots.
  • Fig. S6. Synthesis of pyrazolodihydroquinolinones.
  • Legend for fig. S7
  • Fig. S8. X-ray crystal structures of hGSK3B bound to BRD3731 and BRD0705 and hGSK3B (D133E) bound to BRD0705.
  • Fig. S9. Live cell target engagement analysis for GSK3α and GSK3β using NanoBRET.
  • Fig. S10. Heterogeneous effects of BRD3731 on differentiation in AML cell lines.
  • Fig. S11. Differentiation and impaired stemness and mitochondria transcriptional programs triggered by BRD0705.
  • Fig. S12. Heterogeneous effects of BRD3731 on colony formation in AML cell lines.
  • Fig. S13. Pharmacokinetic, efficacy, and tolerability properties of BRD0705.
  • Fig. S14. Glycogen accumulation induced by BRD0705 in AML.
  • Table S1. X-ray data reduction statistics for x-ray co-crystal structures of hGSK3β with BRD0209, BRD3731, and BRD0705.
  • Table S2. Crystallographic refinement statistics for x-ray co-crystal structures of hGSK3β with BRD0209, BRD3731, and BRD0705.
  • Table S7. Final diffraction statistics for the mutant hGSK3β (D133E)/BRD0705 complex crystal used in structure determination.
  • Table S8. Final refinement statistics for the mutant hGSK3β (D133E)/BRD0705 complex structure.
  • Legends for tables S3 to S6 and S9 to 13

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Other Supplementary Material for this manuscript includes the following:

  • Fig. S7 (.pdf format). Spectra for compounds in Fig. 2B.
  • Table S3 (Microsoft Excel format). Kinome selectivity for BRD0320 (Carna Biosciences).
  • Table S4 (Microsoft Excel format). Kinome selectivity for BRD5648 (Carna Biosciences).
  • Table S5 (Microsoft Excel format). Kinome selectivity for BRD0705 (Carna Biosciences).
  • Table S6 (Microsoft Excel format). Kinome selectivity for BRD3731 (Carna Biosciences).
  • Table S9 (Microsoft Excel format). Lists of genes differentially expressed between control and BRD0705-treated U937 cells.
  • Table S10 (Microsoft Excel format). Lists of genes differentially expressed between control and BRD3731-treated U937 cells.
  • Table S11 (Microsoft Excel format). Lists of genes differentially expressed between control and BRD0320-treated U937 cells.
  • Table S12 (Microsoft Excel format). Top enriched gene sets from the functional groups predicted in BRD0705 versus DMSO enrichment map.
  • Table S13 (Microsoft Excel format). Cytogenetics and molecular and clinical characteristics of patient samples used in this study.

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