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Notch-mediated expansion of human cord blood progenitor cells capable of rapid myeloid reconstitution

Abstract

Delayed myeloid engraftment after cord blood transplantation (CBT) is thought to result from inadequate numbers of progenitor cells in the graft and is associated with increased early transplant–related morbidity and mortality. New culture strategies that increase the number of cord blood progenitors capable of rapid myeloid engraftment after CBT would allow more widespread use of this stem cell source for transplantation. Here we report the development of a clinically relevant Notch-mediated ex vivo expansion system for human CD34+ cord blood progenitors that results in a marked increase in the absolute number of stem/progenitor cells, including those capable of enhanced repopulation in the marrow of immunodeficient nonobese diabetic–severe combined immunodeficient (NOD-SCID) mice. Furthermore, when cord blood progenitors expanded ex vivo in the presence of Notch ligand were infused in a clinical setting after a myeloablative preparative regimen for stem cell transplantation, the time to neutrophil recovery was substantially shortened. To our knowledge, this is the first instance of rapid engraftment derived from ex vivo expanded stem/progenitor cells in humans.

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Figure 1: Culture of CD34+ cord blood progenitors with Notch ligand results in enhanced and more rapid human engraftment as well as increased SRC frequency.
Figure 2: Clinical-grade culture of cord blood progenitors with Delta1ext-IgG results in marked in vitro expansion of CD34+ cells and more rapid neutrophil recovery in a myeloablative double-CBT setting.

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Acknowledgements

This work was supported by grants R24 HL74445, K23 HL077446, K12 CA076930, 5R01HL080245-02 (from the National Institutes of Health), CA15704 and DK56465. C.D. is a Damon Runyon Clinical Investigator supported in part by the Damon Runyon Cancer Research Foundation (CI# 35-07). I.D.B. is an American Cancer Society–F.M. Kirby Clinical Research Professor. We would like to thank F. Appelbaum for his contribution to clinical trial design and clinical consultation, J. Gutman for help in manuscript editing and clinical data coordination and D. Woodle for expert technical assistance. We also acknowledge the work of the clinical faculty and staff at the Seattle Cancer Care Alliance for the excellent clinical care provided to the patients.

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C.D. conceived of the project, designed the experiments, analyzed the data, wrote the manuscript, performed in vitro and mouse studies, was principal investigator of clinical studies and prepared the figures; S.H. contributed to experimental design, data analysis and manuscript editing; C.B.-S. and H.V. performed in vitro studies, data analysis and mouse studies; R.L.M. contributed methods for large-scale production of cGMP-engineered Delta1; I.D.B. conceived of the project and contributed to experimental design and manuscript editing.

Corresponding author

Correspondence to Irwin D Bernstein.

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The authors declare no competing financial interests.

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Supplementary Figures 1 and 2, Supplementary Tables 1–3, Supplementary Methods (PDF 173 kb)

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Delaney, C., Heimfeld, S., Brashem-Stein, C. et al. Notch-mediated expansion of human cord blood progenitor cells capable of rapid myeloid reconstitution. Nat Med 16, 232–236 (2010). https://doi.org/10.1038/nm.2080

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