Distinct hematopoietic stem cell subtypes are differentially regulated by TGF-beta1 - PubMed (original) (raw)
Distinct hematopoietic stem cell subtypes are differentially regulated by TGF-beta1
Grant A Challen et al. Cell Stem Cell. 2010.
Abstract
The traditional view of hematopoiesis has been that all the cells of the peripheral blood are the progeny of a unitary homogeneous pool of hematopoietic stem cells (HSCs). Recent evidence suggests that the hematopoietic system is actually maintained by a consortium of HSC subtypes with distinct functional characteristics. We show here that myeloid-biased HSCs (My-HSCs) and lymphoid-biased HSCs (Ly-HSCs) can be purified according to their capacity for Hoechst dye efflux in combination with canonical HSC markers. These phenotypes are stable under natural (aging) or artificial (serial transplantation) stress and are exacerbated in the presence of competing HSCs. My- and Ly-HSCs respond differently to TGF-beta1, presenting a possible mechanism for differential regulation of HSC subtype activation. This study demonstrates definitive isolation of lineage-biased HSC subtypes and contributes to the fundamental change in view that the hematopoietic system is maintained by a continuum of HSC subtypes, rather than a functionally uniform pool.
2010 Elsevier Inc. All rights reserved.
Figures
Figure 1
Phenotypes of SP subtypes. Left column shows surface marker HSC-identification strategy from whole bone marrow based on lineage-negative, c-kit, Scal-1 expression, as well as Flk-2 and CD34. Top middle panel demonstrates delineation of SP. Top right shows regions designated as lower-SP and upper-SP and below that, c-kit and Sca-1 cells expression of cells gated on these and through a lineage-negative gate. Surface markers on these cells, designated as SPKLS are shown below. CD150 expression is heterogeneous, with CD150+ cells more prevalent in the lower-SP (see also Figure S1).
Figure 2
Repopulation kinetics of transplanted SP subtypes. (A) Competitive transplant scheme for lower- and upper-SPKLS cells individually (200 per recipient) or in combination (100 of each subtype per recipient), with 200,000 whole bone marrow competitor cells in all transplants. (B) Overall peripheral blood contribution (Engraft) and proportional contribution to myeloid, B- and T-cells at 4- and 16-weeks after transplantation of SP fractions into separate mice. (C) Peripheral blood analysis after transplantation of both upper- and lower-SP into the same recipients. (D) Scheme for re-purification of primary HSCs and transplantation into secondary recipients. (E) Contribution to the blood of SP sub-populations after secondary transplantation at 4- and 16-weeks. (F) Reconstitution of blood in secondary recipients by SP originally co-transplanted in primary transplants (see also Figure S2A).
Figure 3
Donor-derived contribution to stem and progenitor cell compartments of transplant recipients. Analysis of the bone marrow of recipient mice to determine donor cell contribution to stem and progenitor cell compartments 18-weeks post-transplant of SP subtypes transplanted separately (A) and into the same recipients (B). (C) Analysis of the donor cell contribution to SP of recipient mice. In reference to the whole bone marrow competitor cells which serve as an internal control, each SP subfraction tended to regenerate itself. Data presented are averages of three separate pooled experiments (see also Figure S2B).
Figure 4
Clonal analysis of HSC subtypes using single-cell transplantation. (A) Tracking of eight individual lower- and upper-SPKLS clones showing overall hematopoietic chimerism in recipient mice (engraftment; top panels in grey) and lineage distribution of test cells 12-weeks post-transplant. (B) Analysis of overall engraftment and lineage contribution of lower- and upper-SPKLS cells also fractionated with respect to CD150. (C) Distribution of lineage contribution 12-weeks after transplant from mice transplanted with single HSCs with the indicated phenotypes. (D) Average level of hematopoietic chimerism from single-cell transplantation experiments. (E) Lineage contribution measured 12 weeks after secondary transplantation of cells derived from primary mice reconstituted with a single HSC. Bone marrow from each clone was tested in four new recipients. All clones met the assigned threshold of at least 0.1% overall contribution to all 4 secondary recipients at 12-weeks post-transplant, with the exception of USP-3, in which 3/4 recipients fell just below the assigned threshold (see also Figure S3).
Figure 5
Aging SP subtypes retain lineage-differentiation biases. (A) Contour plot comparisons of young (10-week old) and old (90-week old) SPs showing an accumulation of lower-SP cells in aged mice. (B) Overall peripheral blood contribution (Engraft) and proportional contribution to myeloid, B- and T-cells at 4- and 16-weeks after transplantation of old and young SP fractions. (C) Peripheral blood contribution after secondary transplantation from recipients of aged lower- and upper-SPKLS cells.
Figure 6
HSC subtypes show differential responses to TGFβ1. (A) Low (10 pg/μL) concentrations of TGFβ1 accelerated colony formation of My-HSCs (more colonies at day 7), but did not change the total number of colonies formed (at day 15), while TGFβ1 proved inhibitory to Ly-HSC colony formation. The lines indicate the total number of colonies per plate, while the bar graphs represent the number of CFU-GM colonies at each timepoint. Data presented are cumulative of four individual experiments each comprising two replicate plates for each condition. (B) The CFU-GEMM colonies from My-HSCs in the presence of TGFβ1 were markedly larger in size than control colonies. (C) Analysis of Pyronin Y staining of My- and Ly-HSCs following 5-hour in vitro exposure to TGFβ1. Data are averages of three separate experiments. (D) BrdU uptake after in vivo TGFβ1 exposure by progeny of transplanted My- and Ly-HSCs. In analyzing the progeny of transplanted HSCs, CD150 expression was used as a surrogate marker for myeloid- (CD150+) and lymphoid-biased (CD150-) daughter HSCs. (E) Real-time PCR analysis of purified My- and Ly-HSCs following 5-hour in vitro TGFβ1 exposure. Data presented are averages for three separate experiments. Stars indicate the genes that exhibit statistically significant differences in response between My- and Ly-HSCs (see also Figure S5-7).
Figure 7
Model for HSC clonal diversity and its relation to the SP phenotype. (A) The traditional clonal succession model (left) in which all mature blood cells are the progeny of a single uniform pool of LT-HSCs, and the clonal diversity model (right), supported by our data, in which distinct HSC subtypes are capable of contributing to all lineages, but are stably programmed to do so in a highly biased fashion (B) The SP allows visual representation of the continuum of HSC subtypes encompassing the spectrum from the most myeloid-biased CD150+ lower-SPKLS to the most lymphoid-biased CD150− upper-SPKLS. The HSC subtypes exhibit additional cellular, molecular, and functional distinctions. A parental unbiased HSC likely exists during development, and conceivably in the adult.
Similar articles
- CD41 expression marks myeloid-biased adult hematopoietic stem cells and increases with age.
Gekas C, Graf T. Gekas C, et al. Blood. 2013 May 30;121(22):4463-72. doi: 10.1182/blood-2012-09-457929. Epub 2013 Apr 5. Blood. 2013. PMID: 23564910 - Tif1γ regulates the TGF-β1 receptor and promotes physiological aging of hematopoietic stem cells.
Quéré R, Saint-Paul L, Carmignac V, Martin RZ, Chrétien ML, Largeot A, Hammann A, Pais de Barros JP, Bastie JN, Delva L. Quéré R, et al. Proc Natl Acad Sci U S A. 2014 Jul 22;111(29):10592-7. doi: 10.1073/pnas.1405546111. Epub 2014 Jul 7. Proc Natl Acad Sci U S A. 2014. PMID: 25002492 Free PMC article. - Clonal-level responses of functionally distinct hematopoietic stem cells to trophic factors.
Mallaney C, Kothari A, Martens A, Challen GA. Mallaney C, et al. Exp Hematol. 2014 Apr;42(4):317-327.e2. doi: 10.1016/j.exphem.2013.11.015. Epub 2013 Dec 25. Exp Hematol. 2014. PMID: 24373928 Free PMC article. - Molecular mechanisms underlying lineage bias in aging hematopoiesis.
Elias HK, Bryder D, Park CY. Elias HK, et al. Semin Hematol. 2017 Jan;54(1):4-11. doi: 10.1053/j.seminhematol.2016.11.002. Epub 2016 Nov 20. Semin Hematol. 2017. PMID: 28088987 Review. - Hematopoietic Stem Cells, Their Niche, and the Concept of Co-Culture Systems: A Critical Review.
Vaidya A, Kale V. Vaidya A, et al. J Stem Cells. 2015;10(1):13-31. J Stem Cells. 2015. PMID: 26665935 Review.
Cited by
- Clonal dynamics and somatic evolution of haematopoiesis in mouse.
Kapadia CD, Williams N, Dawson KJ, Watson C, Yousefzadeh MJ, Le D, Nyamondo K, Cagan A, Waldvogel S, De La Fuente J, Leongamornlert D, Mitchell E, Florez MA, Aguilar R, Martell A, Guzman A, Harrison D, Niedernhofer LJ, King KY, Campbell PJ, Blundell J, Goodell MA, Nangalia J. Kapadia CD, et al. bioRxiv [Preprint]. 2024 Sep 21:2024.09.17.613129. doi: 10.1101/2024.09.17.613129. bioRxiv. 2024. PMID: 39345649 Free PMC article. Preprint. - Aging is associated with functional and molecular changes in distinct hematopoietic stem cell subsets.
Su TY, Hauenstein J, Somuncular E, Dumral Ö, Leonard E, Gustafsson C, Tzortzis E, Forlani A, Johansson AS, Qian H, Månsson R, Luc S. Su TY, et al. Nat Commun. 2024 Sep 11;15(1):7966. doi: 10.1038/s41467-024-52318-1. Nat Commun. 2024. PMID: 39261515 Free PMC article. - Reprogramming hematopoietic stem cell metabolism in lung cancer: glycolysis, oxidative phosphorylation, and the role of 2-DG.
Guo Z, Liu Y, Li X, Huang Y, Zhou Z, Yang C. Guo Z, et al. Biol Direct. 2024 Aug 24;19(1):73. doi: 10.1186/s13062-024-00514-w. Biol Direct. 2024. PMID: 39182128 Free PMC article. - Decoding Clonal Hematopoiesis: Emerging Themes and Novel Mechanistic Insights.
Pendse S, Loeffler D. Pendse S, et al. Cancers (Basel). 2024 Jul 24;16(15):2634. doi: 10.3390/cancers16152634. Cancers (Basel). 2024. PMID: 39123361 Free PMC article. Review. - Repression of SMAD3 by STAT3 and c-Ski induces conventional dendritic cell differentiation.
Yoon JH, Bae E, Nagafuchi Y, Sudo K, Han JS, Park SH, Nakae S, Yamashita T, Ju JH, Matsumoto I, Sumida T, Miyazawa K, Kato M, Kuroda M, Lee IK, Fujio K, Mamura M. Yoon JH, et al. Life Sci Alliance. 2024 Jul 3;7(9):e201900581. doi: 10.26508/lsa.201900581. Print 2024 Sep. Life Sci Alliance. 2024. PMID: 38960622 Free PMC article.
References
- Adolfsson J, Borge OJ, Bryder D, Theilgaard-Monch K, Astrand-Grundstrom I, Sitnicka E, Sasaki Y, Jacobsen SE. Upregulation of Flt3 expression within the bone marrow Lin(-)Sca1(+)c-kit(+) stem cell compartment is accompanied by loss of self-renewal capacity. Immunity. 2001;15:659–669. - PubMed
- Akashi K, Traver D, Miyamoto T, Weissman IL. A clonogenic common myeloid progenitor that gives rise to all myeloid lineages. Nature. 2000;404:193–197. - PubMed
- Benveniste P, Frelin C, Janmohamed S, Barbara M, Herrington R, Hyam D, Iscove NN. Intermediate-term hematopoietic stem cells with extended but time-limited reconstitution potential. Cell Stem Cell. 2010;6:48–58. - PubMed
- Buonamici S, Chakraborty S, Senyuk V, Nucifora G. The role of EVI1 in normal and leukemic cells. Blood Cells Mol Dis. 2003;31:206–212. - PubMed
Publication types
MeSH terms
Substances
Grants and funding
- R01 HL096360/HL/NHLBI NIH HHS/United States
- R21 AG034451-01/AG/NIA NIH HHS/United States
- DK58192/DK/NIDDK NIH HHS/United States
- R01 EB005173/EB/NIBIB NIH HHS/United States
- R01 EB005173-05/EB/NIBIB NIH HHS/United States
- EB005173/EB/NIBIB NIH HHS/United States
- R21 AG034451/AG/NIA NIH HHS/United States
- U54 HL081007/HL/NHLBI NIH HHS/United States
- R01 DK058192/DK/NIDDK NIH HHS/United States
- R01 DK058192-10/DK/NIDDK NIH HHS/United States
- HL081007/HL/NHLBI NIH HHS/United States
- U54 HL081007-05/HL/NHLBI NIH HHS/United States
LinkOut - more resources
Full Text Sources
Other Literature Sources
Medical