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CXCL12 in early mesenchymal progenitors is required for haematopoietic stem-cell maintenance

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Abstract

Haematopoietic stem cells (HSCs) primarily reside in the bone marrow where signals generated by stromal cells regulate their self-renewal, proliferation and trafficking. Endosteal osteoblasts1,2 and perivascular stromal cells including endothelial cells3, CXCL12-abundant reticular cells4,5, leptin-receptor-positive stromal cells6, and nestin–green fluorescent protein (GFP)-positive mesenchymal progenitors7 have all been implicated in HSC maintenance. However, it is unclear whether specific haematopoietic progenitor cell (HPC) subsets reside in distinct niches defined by the surrounding stromal cells and the regulatory molecules they produce. CXCL12 (chemokine (C–X–C motif) ligand 12) regulates both HSCs and lymphoid progenitors and is expressed by all of these stromal cell populations7,8,9,10,11. Here we selectively deleted Cxcl12 from candidate niche stromal cell populations and characterized the effect on HPCs. Deletion of Cxcl12 from mineralizing osteoblasts has no effect on HSCs or lymphoid progenitors. Deletion of Cxcl12 from osterix-expressing stromal cells, which include CXCL12-abundant reticular cells and osteoblasts, results in constitutive HPC mobilization and a loss of B-lymphoid progenitors, but HSC function is normal. Cxcl12 deletion from endothelial cells results in a modest loss of long-term repopulating activity. Strikingly, deletion of Cxcl12 from nestin-negative mesenchymal progenitors using Prx1–cre (Prx1 also known as Prrx1) is associated with a marked loss of HSCs, long-term repopulating activity, HSC quiescence and common lymphoid progenitors. These data suggest that osterix-expressing stromal cells comprise a distinct niche that supports B-lymphoid progenitors and retains HPCs in the bone marrow, and that expression of CXCL12 from stromal cells in the perivascular region, including endothelial cells and mesenchymal progenitors, supports HSCs.

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Figure 1: Targeting Cxcl12 deletion in bone marrow stromal cell populations.
Figure 2: Deletion of Cxcl12 in defined stromal cell population results in the selective loss of HSCs and lymphoid progenitors.
Figure 3: Deletion of Cxcl12 in defined stromal cell populations results in HPC mobilization and a selective loss of repopulating activity and HSC quiescence.
Figure 4: Prx1–cre differentially targets a PDGFRα + SCA1 + CXCL12-expressing mesenchymal progenitor cell population.

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Primary accessions

Gene Expression Omnibus

Data deposits

The RNA expression profiling data have been deposited in Gene Expression Omnibus under accession number GSE43613.

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Acknowledgements

We thank J. Woloszynek, F. Liu, A. Khalaf and M. Romine for technical assistance; G. Callis, S. Oh and M. Vig for technical advice; J. Tucker-Davis for animal care; and T. Clemens for the Oc–cre mice. This work was supported by NIH grants RO1 HL60772 (D.C.L.) and F30 HL097423 (A.G.).

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Authors

Contributions

A.G. and Y.-M.S.H. designed and performed the research, analysed the data and wrote the manuscript. L.G.S., J.N.B. and R.B.D. performed experiments characterizing haematopoiesis in the conditional Cxcl12-deficient mice. M.J.C. designed and cloned the CXCL12 conditional knockout construct. D.C.L. supervised all of the research and edited the manuscript, which was approved by all co-authors.

Corresponding author

Correspondence to Daniel C. Link.

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The authors declare no competing financial interests.

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Greenbaum, A., Hsu, YM., Day, R. et al. CXCL12 in early mesenchymal progenitors is required for haematopoietic stem-cell maintenance. Nature 495, 227–230 (2013). https://doi.org/10.1038/nature11926

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