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Dipeptidylpeptidase 4 negatively regulates colony-stimulating factor activity and stress hematopoiesis

Nature Medicine volume 18, pages 1786–1796 (2012)Cite this article

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Abstract

Enhancement of hematopoietic recovery after radiation, chemotherapy, or hematopoietic stem cell (HSC) transplantation is clinically relevant. Dipeptidylpeptidase (DPP4) cleaves a wide variety of substrates, including the chemokine stromal cell-derived factor-1 (SDF-1). In the course of experiments showing that inhibition of DPP4 enhances SDF-1–mediated progenitor cell survival, ex vivo cytokine expansion and replating frequency, we unexpectedly found that DPP4 has a more general role in regulating colony-stimulating factor (CSF) activity. DPP4 cleaved within the N-termini of the CSFs granulocyte-macrophage (GM)-CSF, G-CSF, interleukin-3 (IL-3) and erythropoietin and decreased their activity. Dpp4 knockout or DPP4 inhibition enhanced CSF activities both in vitro and in vivo. The reduced activity of DPP4-truncated versus full-length human GM-CSF was mechanistically linked to effects on receptor-binding affinity, induction of GM-CSF receptor oligomerization and signaling capacity. Hematopoiesis in mice after radiation or chemotherapy was enhanced in Dpp4−/− mice or mice receiving an orally active DPP4 inhibitor. DPP4 inhibition enhanced engraftment in mice without compromising HSC function, suggesting the potential clinical utility of this approach.

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Figure 1: Deletion or inhibition of Dpp4 enhances SDF-1's effects on survival and ex vivo expansion of HPCs.
Figure 2: Inhibition of DPP4 enhances the in vitro activity of selected CSFs with DPP4 truncation sites.
Figure 3: Influence of soluble DPP4 on activities of recombinant mouse CSFs in vitro, and effects of full-length and truncated CSFs alone and in combination on hematopoiesis in vivo in mice.
Figure 4: Influence of DPP4 inhibition on colony formation, receptor binding and signaling in the TF-1 factor-dependent human cell line and in CD34+ cord blood cells.
Figure 5: Modeling of the GM-CSF GM-CSFR interaction, and effects of DPP4 deficiency or inhibition on hematopoietic recovery.
Figure 6: Influence of DPP4 inhibition in vivo on engraftment of HSCs and of CXCR4 deficiency on sitagliptin-enhanced hematopoietic recovery.

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Acknowledgements

These studies were supported by US Health Service National Institutes of Health (NIH) grants R01 HL056416, R01 HL067284 and R01 HL112669 and a Center of Excellence in Hematology grant P01 DK090948 to H.E.B., and HL69669 and HL96305 to L.M.P. J.H. was supported as a doctoral candidate on NIH T32 DK07519 (to H.E.B.) and as a postdoc on NIH T32 HL07910 (to H.E.B.); H.A.O. was supported as a postdoc on NIH T32 DK07519; S.M.-G. was supported as a doctoral candidate on NIH R25 GM079657 (to H.E.B.); and T.B.C. was supported as a doctoral candidate on NIH T32 DK07519. S.L.R. was supported by NIH T32 CA111198 for parts of this study. We thank Y.-R. Zhou, The Feinstein Insitute for Medical Research, for the Cre+Cxcr4Fl/Fl mice.

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Author notes

  1. Jonathan Hoggatt & Timothy B Campbell

    Present address: Current addresses: Massachusetts General Hospital, Boston, Massachusetts, USA (J.H.) and University of California–San Francisco, San Francisco, California, USA (T.B.C.).,

  2. Hal E Broxmeyer, Jonathan Hoggatt and Heather A O'Leary: These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Microbiology and Immunology, Indiana University School of Medicine, Indianapolis, Indiana, USA

    Hal E Broxmeyer, Jonathan Hoggatt, Heather A O'Leary, Charlie Mantel, Scott Cooper, Steven Messina-Graham, Giao Hangoc, Sara L Rohrabaugh, Xuan Ou, Jennifer Speth, Louis M Pelus, Edward F Srour & Timothy B Campbell

  2. Department of Medicine, Indiana University School of Medicine, Indianapolis, Indiana, USA

    Brahmananda R Chitteti, Sherif Farag & Edward F Srour

  3. Department of Pediatrics, Indiana University School of Medicine, Indianapolis, Indiana, USA

    Edward F Srour

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Contributions

H.E.B. initiated the study, supervised experiments, contributed to concepts and experimental design, scored experiments, interpreted experimental data, did data analysis and wrote the manuscript. J.H. supervised experiments, contributed to concepts and experimental design, conducted and scored experiments, interpreted experimental data, did data analysis and helped in the writing of the manuscript. H.A.O. supervised experiments, contributed to concepts and experimental design, conducted and scored experiments, interpreted experimental data, did data analysis and helped in the writing of the manuscript. C.M. contributed to concepts and experimental design, conducted and scored experiments, interpreted experimental data, did data analysis and helped in the writing of the manuscript. B.R.C. conducted and scored experiments. S.C. conducted experiments and did data analysis. S.M.-G. conducted and scored experiments and did data analysis. G.H. contributed to concepts and experimental design, conducted and scored experiments and did data analysis. S.F. contributed to concepts and experimental design. S.L.R. and X.O. conducted and scored experiments. J.S. contributed to concepts and experimental design. L.M.P. contributed to concepts and experimental design, interpreted experimental data and helped in the writing of the manuscript. E.F.S. contributed to concepts and experimental design and interpreted experimental data. T.B.C. contributed to concepts and experimental design.

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Correspondence to Hal E Broxmeyer.

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Competing interests

H.E.B. is a founder of the CordUse family cord blood bank and a member of the Medical Scientific Advisory Board of CordUse.

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Broxmeyer, H., Hoggatt, J., O'Leary, H. et al. Dipeptidylpeptidase 4 negatively regulates colony-stimulating factor activity and stress hematopoiesis. Nat Med 18, 1786–1796 (2012). https://doi.org/10.1038/nm.2991

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