Article | Published:

Reprogramming human endothelial cells to haematopoietic cells requires vascular induction

Nature volume 511, pages 312318 (17 July 2014) | Download Citation

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Abstract

Generating engraftable human haematopoietic cells from autologous tissues is a potential route to new therapies for blood diseases. However, directed differentiation of pluripotent stem cells yields haematopoietic cells that engraft poorly. Here, we have devised a method to phenocopy the vascular-niche microenvironment of haemogenic cells, thereby enabling reprogramming of human endothelial cells into engraftable haematopoietic cells without transition through a pluripotent intermediate. Highly purified non-haemogenic human umbilical vein endothelial cells or adult dermal microvascular endothelial cells were transduced with the transcription factors FOSB, GFI1, RUNX1 and SPI1 (hereafter referred to as FGRS), and then propagated on serum-free instructive vascular niche monolayers to induce outgrowth of haematopoietic colonies containing cells with functional and immunophenotypic features of multipotent progenitor cells (MPPs). These endothelial cells that have been reprogrammed into human MPPs (rEC-hMPPs) acquire colony-forming-cell potential and durably engraft into immune-deficient mice after primary and secondary transplantation, producing long-term rEC-hMPP-derived myeloid (granulocytic/monocytic, erythroid, megakaryocytic) and lymphoid (natural killer and B cell) progenies. Conditional expression of FGRS transgenes, combined with vascular induction, activates endogenous FGRS genes, endowing rEC-hMPPs with a transcriptional and functional profile similar to that of self-renewing MPPs. Our approach underscores the role of inductive cues from the vascular niche in coordinating and sustaining haematopoietic specification and may prove useful for engineering autologous haematopoietic grafts to treat inherited and acquired blood disorders.

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Primary accessions

Gene Expression Omnibus

Data deposits

Data have been deposited in the GEO database under accession number GSE57662.

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Acknowledgements

V.M.S. is supported by Empire State Stem Cell Board (ESSCB) and New York State Department of Health (NYSDH C026878). S.R. is supported by Ansary Stem Cell Institute (ASCI), HHMI, ESSCB/NYSDH (C024180, C026438, C026878, C028117), NHLBI (R01HL097797, R01HL119872, U01 HL099997), NIDDK (R01DK095039) , NCI (U54CA163167), Qatar National Priorities Research Foundation grant NPRP08-663-3-140 and the Qatar Foundation BioMedical Research Program. J.M.S. is supported by grants from NCI (CA159175 and CA163167), NHLBI (HL119872 and HL055748), Starr Foundation and a Leukemia & Lymphoma Society Scholar award. J.M.B. is supported by an American Society of Hematology Scholar Award, NHLBI U01-HL099997 and Angiocrine Bioscience and ASCI. We acknowledge the contribution of J. Z. Xiang and Agnes J. Viale for enabling and executing molecular profiling, and E. Gars for technical support. We appreciate W. Schachterle for recommendations and edits of the manuscript.

Author information

Affiliations

  1. Ansary Stem Cell Institute, Department of Genetic Medicine, and Howard Hughes Medical Institute, Weill Cornell Medical College, New York, New York 10065, USA

    • Vladislav M. Sandler
    • , Raphael Lis
    • , Ying Liu
    • , Alon Kedem
    • , Daylon James
    • , Jason M. Butler
    •  & Shahin Rafii
  2. Ronald O. Perelman and Claudia Cohen Center for Reproductive Medicine, Weill Cornell Medical College, New York, New York 10065, USA

    • Raphael Lis
    • , Alon Kedem
    •  & Daylon James
  3. HRH Prince Alwaleed Bin Talal Bin Abdulaziz Alsaud Institute for Computational Biomedicine, Weill Cornell Medical College, New York, New York 10065, USA

    • Olivier Elemento
  4. Department of Medicine, Hematology-Oncology, Weill Cornell Medical College and the New York Presbyterian Hospital, New York, New York 10065, USA

    • Joseph M. Scandura

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Contributions

V.M.S. and S.R. conceived and designed the project, performed experiments, analysed the data and wrote the manuscript. R.L., Y.L. and J.M.B. performed experiments, interpreted and analysed data. J.M.S. interpreted, analysed data and wrote the manuscript. D.J., O.E. and A.K. performed the experiments and analysed the data. All authors commented on the paper.

Competing interests

S.R. is the founder of and consultant to Angiocrine Bioscience New York, NY. J.M.B. received sponsored research funding from Angiocrine Bioscience New York, NY.

Corresponding author

Correspondence to Shahin Rafii.

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DOI

https://doi.org/10.1038/nature13547

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