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HMGA2 promotes long-term engraftment and myeloerythroid differentiation of human hematopoietic stem and progenitor cells

Kumar, Praveen, Beck, Dominik, Galeev, Roman, Thoms, Julie A. I., Talkhoncheh, Mehrnaz Safaee, de Jong, Ineke, Unnikrishnan, Ashwin, Baudet, Aurélie, Subramaniam, Agatheeswaran, Pimanda, John E., Larsson, Jonas·2019·Blood Advances
DOI 10.1182/bloodadvances.2018023986
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Abstract

Identification of determinants of fate choices in hematopoietic stem cells (HSCs) is essential to improve the clinical use of HSCs and to enhance our understanding of the biology of normal and malignant hematopoiesis. Here, we show that high-mobility group AT hook 2 (HMGA2), a nonhistone chromosomal-binding protein, is highly and preferentially expressed in HSCs and in the most immature progenitor cell subset of fetal, neonatal, and adult human hematopoiesis. Knockdown of HMGA2 by short hairpin RNA impaired the long-term hematopoietic reconstitution of cord blood (CB)–derived CB CD34+ cells. Conversely, overexpression of HMGA2 in CB CD34+ cells led to overall enhanced reconstitution in serial transplantation assays accompanied by a skewing toward the myeloerythroid lineages. RNA-sequencing analysis showed that enforced HMGA2 expression in CD34+ cells induced gene-expression signatures associated with differentiation toward megakaryocyte-erythroid and myeloid lineages, as well as signatures associated with growth and survival, which at the protein level were coupled with strong activation of AKT. Taken together, our findings demonstrate a key role of HMGA2 in regulation of both proliferation and differentiation of human HSPCs.

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Crossref · 10.1182/bloodadvances.2018023986

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Figures

Figure 1 Preferential expression of HMGA2 within the primitive compartment of human hematopoietic cells. (A-C) HMGA2 expression measured by quantitative PCR in FACS-purified populations, HSCs (CD34 1 CD38 2 CD45RA 2 CD90 1 ), MPPs (CD34 1 CD38 2 CD45RA 2 CD90 2 ), MLP (CD34 1 CD38 2 CD45RA 1 CD90 2 ), and committed progenitors (CP; CD34 1 CD38 1 ) obtained from fetal liver (FL) (A), CB (B), and adult BM (C). (D) Relative expression of HMGA2 mRNA in purified HSCs from different developmental sources. (E) HMGA2 protein expression was assessed by western blot in FACS-purified CB populations. * P , .05; ** P , .01; *** P , .001; **** P , .0001. ns, not significant.
Figure 1 Preferential expression of HMGA2 within the primitive compartment of human hematopoietic cells. (A-C) HMGA2 expression measured by quantitative PCR in FACS-purified populations, HSCs (CD34 1 CD38 2 CD45RA 2 CD90 1 ), MPPs (CD34 1 CD38 2 CD45RA 2 CD90 2 ), MLP (CD34 1 CD38 2 CD45RA 1 CD90 2 ), and committed progenitors (CP; CD34 1 CD38 1 ) obtained from fetal liver (FL) (A), CB (B), and adult BM (C). (D) Relative expression of HMGA2 mRNA in purified HSCs from different developmental sources. (E) HMGA2 protein expression was assessed by western blot in FACS-purified CB populations. * P , .05; ** P , .01; *** P , .001; **** P , .0001. ns, not significant.CC-BY-NC-ND · From the PDF
Figure 2 shRNA-mediated knockdown of HMGA2
Figure 2 shRNA-mediated knockdown of HMGA2CC-BY-NC-ND · From the PDF
Figure 3 Knockdown of HMGA2 impairs the in vivo reconstitution potential of CB-derived CD34 1 cells. (A) Overview of the in vivo transplantation assay for HMGA2-deficient cells. (B) Human CD45 1 GFP 1 chimerism in mice transplanted with shHMGA2-1 -transduced cells as analyzed in peripheral blood after 7 and 14 weeks. (C) Representative FACS plots showing BM engraftment of primary recipient NSG mice 14 weeks after transplantation. (D) Accumulated data for huCD45 1 GFP 1 cell engraftment, myeloid (CD15/CD33 1 ), and lymphoid (CD19 1 ) lineage distribution, as well as levels of HSC engraftment (CD34 1 CD38 2 cells) measured in huCD45 1 GFP 1 graft in BM of primary recipient NSG. (E) Human CD45 1 GFP 1 engraftment in secondary recipients analyzed 20 weeks posttransplantation. * P , .05; ** P , .01.
Figure 3 Knockdown of HMGA2 impairs the in vivo reconstitution potential of CB-derived CD34 1 cells. (A) Overview of the in vivo transplantation assay for HMGA2-deficient cells. (B) Human CD45 1 GFP 1 chimerism in mice transplanted with shHMGA2-1 -transduced cells as analyzed in peripheral blood after 7 and 14 weeks. (C) Representative FACS plots showing BM engraftment of primary recipient NSG mice 14 weeks after transplantation. (D) Accumulated data for huCD45 1 GFP 1 cell engraftment, myeloid (CD15/CD33 1 ), and lymphoid (CD19 1 ) lineage distribution, as well as levels of HSC engraftment (CD34 1 CD38 2 cells) measured in huCD45 1 GFP 1 graft in BM of primary recipient NSG. (E) Human CD45 1 GFP 1 engraftment in secondary recipients analyzed 20 weeks posttransplantation. * P , .05; ** P , .01.CC-BY-NC-ND · From the PDF
Image 4
CC-BY-NC-ND · From the PDF
Figure 5 HMGA2 overexpression in CB CD34 1 leads to altered hematopoietic differentiation. (A) Distribution of hematopoietic lineages as measured by expression of CD15/CD33, CD19, and CD3 within the engrafted human CD45 1 ZsGreen 1 in primary and secondary recipient mice. (B) Frequency of GlyA 1 cells in the BM of primary and secondary recipients. Representative FACS plots for GlyA 1 cells in the BM of secondary recipients are shown to the right. * P , .05; ** P , .01.
Figure 5 HMGA2 overexpression in CB CD34 1 leads to altered hematopoietic differentiation. (A) Distribution of hematopoietic lineages as measured by expression of CD15/CD33, CD19, and CD3 within the engrafted human CD45 1 ZsGreen 1 in primary and secondary recipient mice. (B) Frequency of GlyA 1 cells in the BM of primary and secondary recipients. Representative FACS plots for GlyA 1 cells in the BM of secondary recipients are shown to the right. * P , .05; ** P , .01.CC-BY-NC-ND · From the PDF
Figure 6 HMGA2-overexpressing HSPCs show gene-expression signatures associated with myeloerythroid differentiation. (A-B) GSEA to identify gene sets enriched or depleted in HMGA2-overexpressing CD34 1 cells compared with controls. GSEA plots were generated using population-specific gene signatures described in previous studies for human hematopoietic cells. (C) Top 20 upregulated genes in HMGA2-overexpressing cells. CMP, common myeloid progenitor; EryUP, erythroid upregulated; FDR, false discovery rate; NES, normalized enrichment score.
Figure 6 HMGA2-overexpressing HSPCs show gene-expression signatures associated with myeloerythroid differentiation. (A-B) GSEA to identify gene sets enriched or depleted in HMGA2-overexpressing CD34 1 cells compared with controls. GSEA plots were generated using population-specific gene signatures described in previous studies for human hematopoietic cells. (C) Top 20 upregulated genes in HMGA2-overexpressing cells. CMP, common myeloid progenitor; EryUP, erythroid upregulated; FDR, false discovery rate; NES, normalized enrichment score.CC-BY-NC-ND · From the PDF
Figure 7 Increased activation of AKT signaling in HMGA2-overexpressing cells. (A) IPA showing cellular functions associated with upregulated or downregulated genes in HMGA2-overexpressing cells. (B) IPA analysis showing cellular pathways enriched in upregulated genes in HMGA2-overexpressing cells. (C) Western blot for the components of the AKT-mTOR pathways in HMGA2-overexpressing cells. Exp., experiment; NO, nitric oxide; ROS, reactive oxygen species.
Figure 7 Increased activation of AKT signaling in HMGA2-overexpressing cells. (A) IPA showing cellular functions associated with upregulated or downregulated genes in HMGA2-overexpressing cells. (B) IPA analysis showing cellular pathways enriched in upregulated genes in HMGA2-overexpressing cells. (C) Western blot for the components of the AKT-mTOR pathways in HMGA2-overexpressing cells. Exp., experiment; NO, nitric oxide; ROS, reactive oxygen species.CC-BY-NC-ND · From the PDF

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Research works printed but not linked: 20

  1. Pfannkuche, 2009, The high mobility group protein HMGA2: a co-regulator of chromatin structure and pluripotency in stem cells?, Stem Cell Rev, 5, 224, 10.1007/s12015-009-9078-9 · DOI
  2. Li, 2012, An HMGA2-IGF2BP2 axis regulates myoblast proliferation and myogenesis, Dev Cell, 23, 1176, 10.1016/j.devcel.2012.10.019 · DOI
  3. Nishino, 2008, Hmga2 promotes neural stem cell self-renewal in young but not old mice by reducing p16Ink4a and p19Arf Expression, Cell, 135, 227, 10.1016/j.cell.2008.09.017 · DOI
  4. Ikeda, 2011, 3'UTR-truncated Hmga2 cDNA causes MPN-like hematopoiesis by conferring a clonal growth advantage at the level of HSC in mice, Blood, 117, 5860, 10.1182/blood-2011-02-334425 · DOI
  5. Copley, 2013, The Lin28b-let-7-Hmga2 axis determines the higher self-renewal potential of fetal haematopoietic stem cells, Nat Cell Biol, 15, 916, 10.1038/ncb2783 · DOI
  6. Lam, 2014, Hmga2 is a direct target gene of RUNX1 and regulates expansion of myeloid progenitors in mice, Blood, 124, 2203, 10.1182/blood-2014-02-554543 · DOI
  7. Rowe, 2016, Developmental regulation of myeloerythroid progenitor function by the Lin28b-let-7-Hmga2 axis, J Exp Med, 213, 1497, 10.1084/jem.20151912 · DOI
  8. Kim, 2013, TopHat2: accurate alignment of transcriptomes in the presence of insertions, deletions and gene fusions, Genome Biol, 14, R36, 10.1186/gb-2013-14-4-r36 · DOI · Paper page
  9. Anders, 2010, Differential expression analysis for sequence count data, Genome Biol, 11, R106, 10.1186/gb-2010-11-10-r106 · DOI · Paper page
  10. Laurenti, 2013, The transcriptional architecture of early human hematopoiesis identifies multilevel control of lymphoid commitment, Nat Immunol, 14, 756, 10.1038/ni.2615 · DOI
  11. Kumar, 2007, Impaired microRNA processing enhances cellular transformation and tumorigenesis, Nat Genet, 39, 673, 10.1038/ng2003 · DOI
  12. Kumar, 2008, Suppression of non-small cell lung tumor development by the let-7 microRNA family, Proc Natl Acad Sci USA, 105, 3903, 10.1073/pnas.0712321105 · DOI
  13. Viswanathan, 2009, Lin28 promotes transformation and is associated with advanced human malignancies, Nat Genet, 41, 843, 10.1038/ng.392 · DOI
  14. Wang, 2010, Dynamics of gene-modified progenitor cells analyzed by tracking retroviral integration sites in a human SCID-X1 gene therapy trial, Blood, 115, 4356, 10.1182/blood-2009-12-257352 · DOI
  15. Yu, 2013, HMGA2 regulates the in vitro aging and proliferation of human umbilical cord blood-derived stromal cells through the mTOR/p70S6K signaling pathway, Stem Cell Res, 10, 156, 10.1016/j.scr.2012.11.002 · DOI
  16. Tan, 2016, Amplified HMGA2 promotes cell growth by regulating Akt pathway in AML, J Cancer Res Clin Oncol, 142, 389, 10.1007/s00432-015-2036-9 · DOI
  17. Buitenhuis, 2009, The role of the PI3K-PKB signaling module in regulation of hematopoiesis, Cell Cycle, 8, 560, 10.4161/cc.8.4.7654 · DOI
  18. Juntilla, 2010, AKT1 and AKT2 maintain hematopoietic stem cell function by regulating reactive oxygen species, Blood, 115, 4030, 10.1182/blood-2009-09-241000 · DOI · Paper page
  19. Li, 2011, Quiescence regulators for hematopoietic stem cell, Exp Hematol, 39, 511, 10.1016/j.exphem.2011.01.008 · DOI
  20. Lechman, 2012, Attenuation of miR-126 activity expands HSC in vivo without exhaustion, Cell Stem Cell, 11, 799, 10.1016/j.stem.2012.09.001 · DOI · Paper page

Cited by

Metadata

Authors
Kumar, P., Beck, D., Galeev, R., Thoms, J. A. I., Talkhoncheh, M. S., de Jong, I., Unnikrishnan, A., Baudet, A., Subramaniam, A., Pimanda, J. E., & Larsson, J.
Venue
Blood Advances
Publisher
American Society of Hematology
Published
2019-02-26
Volume
3
Issue
4
Pages
681-691
ISSN
  • 2473-9529
  • 2473-9537
DOI
10.1182/bloodadvances.2018023986
Cited by
30
References
3
Language
en

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