Letter | Published:

PPAR-α and glucocorticoid receptor synergize to promote erythroid progenitor self-renewal

Nature volume 522, pages 474477 (25 June 2015) | Download Citation

Abstract

Many acute and chronic anaemias, including haemolysis, sepsis and genetic bone marrow failure diseases such as Diamond–Blackfan anaemia, are not treatable with erythropoietin (Epo), because the colony-forming unit erythroid progenitors (CFU-Es) that respond to Epo are either too few in number or are not sensitive enough to Epo to maintain sufficient red blood cell production1,2,3,4,5,6,7,8,9. Treatment of these anaemias requires a drug that acts at an earlier stage of red cell formation and enhances the formation of Epo-sensitive CFU-E progenitors. Recently, we showed that glucocorticoids specifically stimulate self-renewal of an early erythroid progenitor, burst-forming unit erythroid (BFU-E), and increase the production of terminally differentiated erythroid cells10,11. Here we show that activation of the peroxisome proliferator-activated receptor α (PPAR-α) by the PPAR-α agonists GW7647 and fenofibrate synergizes with the glucocorticoid receptor (GR) to promote BFU-E self-renewal. Over time these agonists greatly increase production of mature red blood cells in cultures of both mouse fetal liver BFU-Es and mobilized human adult CD34+ peripheral blood progenitors, with a new and effective culture system being used for the human cells that generates normal enucleated reticulocytes. Although Ppara−/− mice show no haematological difference from wild-type mice in both normal and phenylhydrazine (PHZ)-induced stress erythropoiesis, PPAR-α agonists facilitate recovery of wild-type but not Ppara−/− mice from PHZ-induced acute haemolytic anaemia. We also show that PPAR-α alleviates anaemia in a mouse model of chronic anaemia. Finally, both in control and corticosteroid-treated BFU-E cells, PPAR-α co-occupies many chromatin sites with GR; when activated by PPAR-α agonists, additional PPAR-α is recruited to GR-adjacent sites and presumably facilitates GR-dependent BFU-E self-renewal. Our discovery of the role of PPAR-α agonists in stimulating self-renewal of early erythroid progenitor cells suggests that the clinically tested PPAR-α agonists we used may improve the efficacy of corticosteroids in treating Epo-resistant anaemias.

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Accessions

Primary accessions

Gene Expression Omnibus

Data deposits

RNA-seq and ChIP-seq data have been deposited in the Gene Expression Omnibus under accession numbers GSE63836 and GSE63837, respectively.

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Acknowledgements

We thank the Whitehead Institute Flow Cytometry Facility, Genome Technology Core and Bioinformatics & Research Computing Facility, as well as the Massachusetts Institute of Technology Koch Institute Flow Cytometry Core. We thank V. Sankaran for haemoglobin HPLC and J. Flygare for the plasmid encoding the RPS19 shRNA, and are grateful to animal technicians F. Reinhardt and T. E. Chavarria for their assistance. We thank T. DiCesare for assistance with graphics. This study was supported by grants to H.F.L. (Defense Advanced Research Projects Agency HR0011-14-2-0005; Department of Defense/US Army Medical Research and Materiel Command W81WH-12-1-0449, National institutes of Health (NIH)/National Heart, Lung, and Blood Institute 2 P01 HL032262-25; as well as research support from the Diamond-Blackfan Anemia Foundation and Diamond Blackfan Anemia Canada. L.L.P. was supported by NIH grant DK100692. X.G. was supported by a postdoctoral fellowship from the Leukemia and Lymphoma Society.

Author information

Author notes

    • Hsiang-Ying Lee
    •  & Xiaofei Gao

    These authors contributed equally to this work.

Affiliations

  1. Whitehead Institute for Biomedical Research, Nine Cambridge Center, Cambridge, Massachusetts 02142, USA

    • Hsiang-Ying Lee
    • , Xiaofei Gao
    • , M. Inmaculada Barrasa
    • , Russell R. Elmes
    •  & Harvey F. Lodish
  2. Center for Individualized Medicine, Department of Molecular Pharmacology and Experimental Therapeutics, Mayo Clinic, Rochester, Minnesota 55905, USA

    • Hu Li
  3. The Jackson Laboratory, 600 Main Street, Bar Harbor, Maine 04609, USA

    • Luanne L. Peters
  4. Departments of Biology and Biological Engineering, Massachusetts Institute of Technology, 77 Massachusetts Avenue, Cambridge, Massachusetts 02139, USA

    • Harvey F. Lodish

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Contributions

H.-Y.L., X.G., L.L.P. and H.F.L. designed the experiments. H.-Y.L., X.G. and R.R.E. performed the experiments. M.I.B. and H.L. conducted bioinformatic analyses of ChIP-seq and RNA-seq. H.-Y.L., X.G. and H.F.L. wrote the manuscript with input from M.I.B. All authors discussed the results and commented on the manuscript.

Competing interests

The authors declare no competing financial interests.

Corresponding author

Correspondence to Harvey F. Lodish.

Extended data

Supplementary information

PDF files

  1. 1.

    Supplementary Information

    This file contains a Supplementary Discussion and Supplementary References.

Excel files

  1. 1.

    Supplementary Table 1

    This table contains RNA-Seq analysis from previous studies (ref. 10). Genes with gene expression changes above or below 2 fold are included here. Gene ID, symbols and description are also included.

  2. 2.

    Supplementary Table 2

    This table shows gene expression changes in mouse BFU-Es treated with GW7647 and DEX versus DEX alone. Mouse BFU-Es were isolated from E14.5 embryos. BFU-Es were treated with or without DEX plus or minus GW7647 for 12 hrs. Cells were harvested and RNA were extracted for RNA-Seq analysis.

  3. 3.

    Supplementary Table 3

    This table contains complete blood count analysis of wild-type and PPARα-/- mice.

  4. 4.

    Supplementary Table 4

    This table shows gene expression changes in mouse BFU-Es treated with GW7647 and DEX versus DEX alone. Mouse BFU-Es were isolated from E14.5 embryos. BFU-Es were treated with or without DEX plus or minus GW7647 for 12 hrs. Cells were harvested and RNA were extracted for RNA-Seq analysis.

  5. 5.

    Supplementary table 5

    This table contains lists of up- or down-regulated genes (DEX and GW7647 versus DEX) with 10 kb of GR and PPARa binding peaks.

  6. 6.

    Supplementary Table 6

    This table contains raw data for Figure 3a and Extended Data Figure 4b.

  7. 7.

    Supplementary Table 7

    This table contains raw data for Figure 3b and Extended Data Figure 5c.

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DOI

https://doi.org/10.1038/nature14326

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