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Glycogen synthase kinase-3 is an in vivo regulator of hematopoietic stem cell repopulation

Abstract

The in vivo regulation of hematopoietic stem cell (HSC) function is poorly understood. Here, we show that hematopoietic repopulation can be augmented by administration of a glycogen synthase kinase-3 (GSK-3) inhibitor to recipient mice transplanted with mouse or human HSCs. GSK-3 inhibitor treatment improved neutrophil and megakaryocyte recovery, recipient survival and resulted in enhanced sustained long-term repopulation. The output of primitive Linc-Kit+Sca-1+ cells and progenitors from HSCs increased upon GSK-3 inhibitor treatment without altering secondary repopulating ability, suggesting that the HSC pool is maintained while overall hematopoietic reconstitution is increased. GSK-3 inhibitors were found to modulate gene targets of Wnt, Hedgehog and Notch pathways in cells comprising the primitive hematopoietic compartment without affecting mature cells. Our study establishes GSK-3 as a specific in vivo modulator of HSC activity, and suggests that administration of GSK-3 inhibitors may provide a clinical means to directly enhance the repopulating capacity of transplanted HSCs.

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Figure 1: In vivo administration of GSK-3 inhibitor augments long-term repopulating capacity of wild-type HSCs and short-term recovery of peripheral blood counts.
Figure 2: In vivo administration of GSK-3 inhibitor augments human neonatal and adult HSC capacity.
Figure 3: In vivo administration of GSK-3 inhibitor expands a subset of Linc-Kit+Sca-1+ cells with progenitor capacity but not secondary reconstitution potential.
Figure 4: Short-term in vivo administration of GSK-3 inhibitor expands and increases cycling of primitive Linc-Kit+Sca-1+ cells.
Figure 5: In vivo administration of GSK-3 inhibitor regulates targets of the Wnt, Notch and Hedgehog pathways in Linc-Kit+Sca-1+ cells.
Figure 6: In vitro effects of GSK-3 inhibitor on purified Linc-Kit+Sca-1+ cells and summary model.

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Acknowledgements

We would like to thank Chiron Corporation for providing the GSK-3 inhibitors for these studies, and M. Scott for providing the Ptc1+/−lacZ transgenic mice. We acknowledge L. Gallacher for technical assistance, K. Levac and P. Menendez for critical review of the manuscript, D. Sheerar for cell isolation, the nurses of the labor and delivery division of St. Joseph's Health Care and London Health Sciences Centre for collection of cord blood samples. Funding for this research was provided by a research grant from the Canadian Institutes of Health Research, US National Institutes of Health grant P01 GM069983 (to R.T.M.), the Krembil Foundation, Ontario Research Fund, Canada Research Chair in Stem Cell Biology and Regenerative Medicine (to M.B.), and postgraduate scholarship awards from the Ontario Graduate Society and National Cancer Institute of Canada (to J.J.T.). R.T.M. is an investigator of the Howard Hughes Medical Institute.

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Correspondence to Mickie Bhatia.

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Supplementary information

Supplementary Fig. 1

Progenitor (CFU) capacity and NOD/SCID repopulation activity of primitive mouse bone marrow subsets. (PDF 67 kb)

Supplementary Fig. 2

Administration of GSK-3 inhibitor and survival following sublethal irradiation in the absence of transplanted mouse HSCs. (PDF 30 kb)

Supplementary Table 1

Primer sequences used for real-time PCR amplification. (PDF 58 kb)

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Trowbridge, J., Xenocostas, A., Moon, R. et al. Glycogen synthase kinase-3 is an in vivo regulator of hematopoietic stem cell repopulation. Nat Med 12, 89–98 (2006). https://doi.org/10.1038/nm1339

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