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Interactions of Hematopoietic Stem Cells with Bone Marrow Niche

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Stem Cell Renewal and Cell-Cell Communication

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2346))

Abstract

Hematopoietic stem cells (HSCs) are ultimately responsible for the lifelong renewal of all blood cell lineages. In the bone marrow (BM), HSCs reside in specialized microenvironments referred to as the “niche.” HSC niche consists of complex components including heterogeneous cell populations, growth factors, and extracellular matrix molecules. The crosstalk between HSCs and their niche is essential to regulate the survival, self-renewal, migration, quiescence, and differentiation of HSCs. The application of mice models with endogenous ablation of specific cell types, advanced imaging technologies, high-throughput single-cell RNA sequencing, and single-cell mass cytometry methods have provided deep insights into communications between HSCs and niche cells. In this chapter, we have focused on three important cell types in the BM niche: mesenchymal stem cells (MSCs), osteoblasts (OBs), and endothelial cells (ECs). In order to address the interaction between HSCs and these three cell populations in BM niche, we have described methodology for (1) collecting total BM from femur and tibia of C57BL/6 mice; (2) analyzing or sorting of MSCs, OBs, and ECs based on the selection of surface markers CD45, Ter119, CD31, Sca1, and CD51 with flow cytometry; and (3) co-culturing the sorted cells with purified HSCs for further functional assays of HSCs.

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References

  1. Laurenti E, Gottgens B (2018) From haematopoietic stem cells to complex differentiation landscapes. Nature 553(7689):418–426. https://doi.org/10.1038/nature25022

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  2. Passegue E, Jamieson CH, Ailles LE, Weissman IL (2003) Normal and leukemic hematopoiesis: are leukemias a stem cell disorder or a reacquisition of stem cell characteristics? Proc Natl Acad Sci U S A 100(Suppl 1):11842–11849. https://doi.org/10.1073/pnas.2034201100

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  3. Morita Y, Ema H, Nakauchi H (2010) Heterogeneity and hierarchy within the most primitive hematopoietic stem cell compartment. J Exp Med 207(6):1173–1182. https://doi.org/10.1084/jem.20091318

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  4. Pang WW, Price EA, Sahoo D, Beerman I, Maloney WJ, Rossi DJ, Schrier SL, Weissman IL (2011) Human BM hematopoietic stem cells are increased in frequency and myeloid-biased with age. Proc Natl Acad Sci U S A 108(50):20012–20017. https://doi.org/10.1073/pnas.1116110108

    Article  PubMed  PubMed Central  Google Scholar 

  5. Muller-Sieburg CE, Cho RH, Karlsson L, Huang JF, Sieburg HB (2004) Myeloid-biased hematopoietic stem cells have extensive self-renewal capacity but generate diminished lymphoid progeny with impaired IL-7 responsiveness. Blood 103(11):4111–4118. https://doi.org/10.1182/blood-2003-10-3448

    Article  CAS  PubMed  Google Scholar 

  6. Kiel MJ, Morrison SJ (2008) Uncertainty in the niches that maintain haematopoietic stem cells. Nat Rev Immunol 8(4):290–301. https://doi.org/10.1038/nri2279

    Article  CAS  PubMed  Google Scholar 

  7. Pinho S, Frenette PS (2019) Haematopoietic stem cell activity and interactions with the niche. Nat Rev Mol Cell Biol 20(5):303–320. https://doi.org/10.1038/s41580-019-0103-9

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  8. He N, Zhang L, Cui J, Li Z (2014) BM vascular niche: home for hematopoietic stem cells. BM Res 2014:128436. https://doi.org/10.1155/2014/128436

    Article  Google Scholar 

  9. Rashidi NM, Lo Celso C (2014) Flying back to the nest: Intravital microscopy reveals how the niche can induce stemness. Intravital 3(2):e29653. https://doi.org/10.4161/intv.29653

    Article  PubMed  PubMed Central  Google Scholar 

  10. Schofield R (1978) The relationship between the spleen colony-forming cell and the haemopoietic stem cell. Blood Cells 4(1–2):7–25

    CAS  PubMed  Google Scholar 

  11. Lo Celso C, Scadden DT (2011) The haematopoietic stem cell niche at a glance. J Cell Sci 124(Pt 21):3529–3535. https://doi.org/10.1242/jcs.074112

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  12. Morrison SJ, Scadden DT (2014) The BM niche for haematopoietic stem cells. Nature 505(7483):327–334. https://doi.org/10.1038/nature12984

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  13. Mendez-Ferrer S, Bonnet D, Steensma DP, Hasserjian RP, Ghobrial IM, Gribben JG, Andreeff M, Krause DS (2020) BM niches in haematological malignancies. Nat Rev Cancer. https://doi.org/10.1038/s41568-020-0245-2

  14. Zhang P, Zhang C, Li J, Han J, Liu X, Yang H (2019) The physical microenvironment of hematopoietic stem cells and its emerging roles in engineering applications. Stem Cell Res Ther 10(1):327. https://doi.org/10.1186/s13287-019-1422-7

    Article  PubMed  PubMed Central  Google Scholar 

  15. Ho YH, Mendez-Ferrer S (2020) Microenvironmental contributions to hematopoietic stem cell aging. Haematologica 105(1):38–46. https://doi.org/10.3324/haematol.2018.211334

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  16. Tamma R, Ribatti D (2017) Bone niches, hematopoietic stem cells, and vessel formation. Int J Mol Sci 18(1). https://doi.org/10.3390/ijms18010151

  17. Cordeiro-Spinetti E, Taichman RS, Balduino A (2015) The BM endosteal niche: how far from the surface? J Cell Biochem 116(1):6–11. https://doi.org/10.1002/jcb.24952

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  18. Nakamura Y, Arai F, Iwasaki H, Hosokawa K, Kobayashi I, Gomei Y, Matsumoto Y, Yoshihara H, Suda T (2010) Isolation and characterization of endosteal niche cell populations that regulate hematopoietic stem cells. Blood 116(9):1422–1432. https://doi.org/10.1182/blood-2009-08-239194

    Article  CAS  PubMed  Google Scholar 

  19. Crane GM, Jeffery E, Morrison SJ (2017) Adult haematopoietic stem cell niches. Nat Rev Immunol 17(9):573–590. https://doi.org/10.1038/nri.2017.53

    Article  CAS  PubMed  Google Scholar 

  20. Yu VW, Scadden DT (2016) Heterogeneity of the BM niche. Curr Opin Hematol 23(4):331–338. https://doi.org/10.1097/moh.0000000000000265

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  21. Joseph C, Quach JM, Walkley CR, Lane SW, Lo Celso C, Purton LE (2013) Deciphering hematopoietic stem cells in their niches: a critical appraisal of genetic models, lineage tracing, and imaging strategies. Cell Stem Cell 13(5):520–533. https://doi.org/10.1016/j.stem.2013.10.010

    Article  CAS  PubMed  Google Scholar 

  22. Morikawa T, Takubo K (2017) Use of imaging techniques to illuminate dynamics of hematopoietic stem cells and their niches. Front Cell Dev Biol 5:62. https://doi.org/10.3389/fcell.2017.00062

    Article  PubMed  PubMed Central  Google Scholar 

  23. Kunisaki Y, Bruns I, Scheiermann C, Ahmed J, Pinho S, Zhang D, Mizoguchi T, Wei Q, Lucas D, Ito K, Mar JC, Bergman A, Frenette PS (2013) Arteriolar niches maintain haematopoietic stem cell quiescence. Nature 502(7473):637–643. https://doi.org/10.1038/nature12612

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  24. Pinho S, Marchand T, Yang E, Wei Q, Nerlov C, Frenette PS (2018) Lineage-biased hematopoietic stem cells are regulated by distinct niches. Dev Cell 44(5):634–641.e4. https://doi.org/10.1016/j.devcel.2018.01.016

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  25. Christodoulou C, Spencer JA, Yeh SA, Turcotte R, Kokkaliaris KD, Panero R, Ramos A, Guo G, Seyedhassantehrani N, Esipova TV, Vinogradov SA, Rudzinskas S, Zhang Y, Perkins AS, Orkin SH, Calogero RA, Schroeder T, Lin CP, Camargo FD (2020) Live-animal imaging of native haematopoietic stem and progenitor cells. Nature 578(7794):278–283. https://doi.org/10.1038/s41586-020-1971-z

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  26. Kokkaliaris KD (2020) Dissecting the spatial BM microenvironment of hematopoietic stem cells. Curr Opin Oncol 32(2):154–161. https://doi.org/10.1097/cco.0000000000000605

    Article  PubMed  Google Scholar 

  27. Tikhonova AN, Dolgalev I, Hu H, Sivaraj KK, Hoxha E, Cuesta-Dominguez A, Pinho S, Akhmetzyanova I, Gao J, Witkowski M, Guillamot M, Gutkin MC, Zhang Y, Marier C, Diefenbach C, Kousteni S, Heguy A, Zhong H, Fooksman DR, Butler JM, Economides A, Frenette PS, Adams RH, Satija R, Tsirigos A, Aifantis I (2019) The BM microenvironment at single-cell resolution. Nature 569(7755):222–228. https://doi.org/10.1038/s41586-019-1104-8

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  28. Baryawno N, Przybylski D, Kowalczyk MS, Kfoury Y, Severe N, Gustafsson K, Kokkaliaris KD, Mercier F, Tabaka M, Hofree M, Dionne D, Papazian A, Lee D, Ashenberg O, Subramanian A, Vaishnav ED, Rozenblatt-Rosen O, Regev A, Scadden DT (2019) A cellular taxonomy of the BM stroma in homeostasis and leukemia. Cell 177(7):1915–1932.e16. https://doi.org/10.1016/j.cell.2019.04.040

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  29. Vinchi F, Mendelson A, Yazdanbakhsh K, An X (2019) Uncovering the BM microenvironment cell by cell. Hemasphere 3(6):e299. https://doi.org/10.1097/hs9.0000000000000299

    Article  PubMed  PubMed Central  Google Scholar 

  30. Wolock SL, Krishnan I, Tenen DE, Matkins V, Camacho V, Patel S, Agarwal P, Bhatia R, Tenen DG, Klein AM, Welner RS (2019) Mapping distinct BM niche populations and their differentiation paths. Cell Rep 28(2):302–311.e5. https://doi.org/10.1016/j.celrep.2019.06.031

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  31. Severe N, Karabacak NM, Gustafsson K, Baryawno N, Courties G, Kfoury Y, Kokkaliaris KD, Rhee C, Lee D, Scadden EW, Garcia-Robledo JE, Brouse T, Nahrendorf M, Toner M, Scadden DT (2019) Stress-induced changes in BM stromal cell populations revealed through single-cell protein expression mapping. Cell Stem Cell 25(4):570–583.e77. https://doi.org/10.1016/j.stem.2019.06.003

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  32. Caplan AI (2017) Mesenchymal stem cells: time to change the name! Stem Cells Transl Med 6(6):1445–1451. https://doi.org/10.1002/sctm.17-0051

    Article  PubMed  PubMed Central  Google Scholar 

  33. Pittenger MF, Mackay AM, Beck SC, Jaiswal RK, Douglas R, Mosca JD, Moorman MA, Simonetti DW, Craig S, Marshak DR (1999) Multilineage potential of adult human mesenchymal stem cells. Science 284(5411):143–147. https://doi.org/10.1126/science.284.5411.143

    Article  CAS  PubMed  Google Scholar 

  34. Andrzejewska A, Lukomska B, Janowski M (2019) Concise review: mesenchymal stem cells: from roots to boost. Stem Cells 37(7):855–864. https://doi.org/10.1002/stem.3016

    Article  PubMed  PubMed Central  Google Scholar 

  35. Wei Q, Frenette PS (2018) Niches for hematopoietic stem cells and their progeny. Immunity 48(4):632–648. https://doi.org/10.1016/j.immuni.2018.03.024

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  36. Frenette PS, Pinho S, Lucas D, Scheiermann C (2013) Mesenchymal stem cell: keystone of the hematopoietic stem cell niche and a stepping-stone for regenerative medicine. Annu Rev Immunol 31:285–316. https://doi.org/10.1146/annurev-immunol-032712-095919

    Article  PubMed  Google Scholar 

  37. Crippa S, Bernardo ME (2018) Mesenchymal stromal cells: role in the BM niche and in the support of hematopoietic stem cell transplantation. Hema 2(6):e151. https://doi.org/10.1097/hs9.0000000000000151

    Article  Google Scholar 

  38. Sacchetti B, Funari A, Michienzi S, Di Cesare S, Piersanti S, Saggio I, Tagliafico E, Ferrari S, Robey PG, Riminucci M, Bianco P (2007) Self-renewing osteoprogenitors in BM sinusoids can organize a hematopoietic microenvironment. Cell 131(2):324–336. https://doi.org/10.1016/j.cell.2007.08.025

    Article  CAS  PubMed  Google Scholar 

  39. Chan CK, Chen CC, Luppen CA, Kim JB, DeBoer AT, Wei K, Helms JA, Kuo CJ, Kraft DL, Weissman IL (2009) Endochondral ossification is required for haematopoietic stem-cell niche formation. Nature 457(7228):490–494. https://doi.org/10.1038/nature07547

    Article  CAS  PubMed  Google Scholar 

  40. Mendez-Ferrer S, Michurina TV, Ferraro F, Mazloom AR, Macarthur BD, Lira SA, Scadden DT, Ma'ayan A, Enikolopov GN, Frenette PS (2010) Mesenchymal and haematopoietic stem cells form a unique BM niche. Nature 466(7308):829–834. https://doi.org/10.1038/nature09262

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  41. Crippa S, Santi L, Bosotti R, Porro G, Bernardo ME (2019) BM-derived mesenchymal stromal cells: a novel target to optimize hematopoietic stem cell transplantation protocols in hematological malignancies and rare genetic disorders. J Clin Med 9(1). https://doi.org/10.3390/jcm9010002

  42. Greenbaum A, Hsu YM, Day RB, Schuettpelz LG, Christopher MJ, Borgerding JN, Nagasawa T, Link DC (2013) CXCL12 in early mesenchymal progenitors is required for haematopoietic stem-cell maintenance. Nature 495(7440):227–230. https://doi.org/10.1038/nature11926

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  43. Nilsson SK, Johnston HM, Coverdale JA (2001) Spatial localization of transplanted hemopoietic stem cells: inferences for the localization of stem cell niches. Blood 97(8):2293–2299. https://doi.org/10.1182/blood.v97.8.2293

    Article  CAS  PubMed  Google Scholar 

  44. Lo Celso C, Fleming HE, Wu JW, Zhao CX, Miake-Lye S, Fujisaki J, Cote D, Rowe DW, Lin CP, Scadden DT (2009) Live-animal tracking of individual haematopoietic stem/progenitor cells in their niche. Nature 457(7225):92–96. https://doi.org/10.1038/nature07434

    Article  CAS  PubMed  Google Scholar 

  45. Bowers M, Zhang B, Ho Y, Agarwal P, Chen CC, Bhatia R (2015) Osteoblast ablation reduces normal long-term hematopoietic stem cell self-renewal but accelerates leukemia development. Blood 125(17):2678–2688. https://doi.org/10.1182/blood-2014-06-582924

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  46. Taichman RS, Emerson SG (1994) Human osteoblasts support hematopoiesis through the production of granulocyte colony-stimulating factor. J Exp Med 179(5):1677–1682. https://doi.org/10.1084/jem.179.5.1677

    Article  CAS  PubMed  Google Scholar 

  47. Galan-Diez M, Kousteni S (2017) The osteoblastic niche in hematopoiesis and hematological myeloid malignancies. Curr Mol Biol Rep 3(2):53–62. https://doi.org/10.1007/s40610-017-0055-9

    Article  PubMed  PubMed Central  Google Scholar 

  48. Nilsson SK, Johnston HM, Whitty GA, Williams B, Webb RJ, Denhardt DT, Bertoncello I, Bendall LJ, Simmons PJ, Haylock DN (2005) Osteopontin, a key component of the hematopoietic stem cell niche and regulator of primitive hematopoietic progenitor cells. Blood 106(4):1232–1239. https://doi.org/10.1182/blood-2004-11-4422

    Article  CAS  PubMed  Google Scholar 

  49. Haylock DN, Nilsson SK (2006) Osteopontin: a bridge between bone and blood. Br J Haematol 134(5):467–474. https://doi.org/10.1111/j.1365-2141.2006.06218.x

    Article  CAS  PubMed  Google Scholar 

  50. Stier S, Ko Y, Forkert R, Lutz C, Neuhaus T, Grunewald E, Cheng T, Dombkowski D, Calvi LM, Rittling SR, Scadden DT (2005) Osteopontin is a hematopoietic stem cell niche component that negatively regulates stem cell pool size. J Exp Med 201(11):1781–1791. https://doi.org/10.1084/jem.20041992

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  51. Guidi N, Sacma M, Standker L, Soller K, Marka G, Eiwen K, Weiss JM, Kirchhoff F, Weil T, Cancelas JA, Florian MC, Geiger H (2017) Osteopontin attenuates aging-associated phenotypes of hematopoietic stem cells. EMBO J 36(7):840–853. https://doi.org/10.15252/embj.201694969

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  52. Perlin JR, Sporrij A, Zon LI (2017) Blood on the tracks: hematopoietic stem cell-endothelial cell interactions in homing and engraftment. J Mol Med (Berl) 95(8):809–819. https://doi.org/10.1007/s00109-017-1559-8

    Article  CAS  Google Scholar 

  53. Poulos MG, Ramalingam P, Gutkin MC, Llanos P, Gilleran K, Rabbany SY, Butler JM (2017) Endothelial transplantation rejuvenates aged hematopoietic stem cell function. J Clin Invest 127(11):4163–4178. https://doi.org/10.1172/jci93940

    Article  PubMed  PubMed Central  Google Scholar 

  54. Ramalingam P, Poulos MG, Butler JM (2017) Regulation of the hematopoietic stem cell lifecycle by the endothelial niche. Curr Opin Hematol 24(4):289–299. https://doi.org/10.1097/moh.0000000000000350

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  55. Rafii S, Shapiro F, Pettengell R, Ferris B, Nachman RL, Moore MA, Asch AS (1995) Human BM microvascular endothelial cells support long-term proliferation and differentiation of myeloid and megakaryocytic progenitors. Blood 86(9):3353–3363

    Article  CAS  Google Scholar 

  56. Yao L, Yokota T, Xia L, Kincade PW, McEver RP (2005) BM dysfunction in mice lacking the cytokine receptor gp130 in endothelial cells. Blood 106(13):4093–4101. https://doi.org/10.1182/blood-2005-02-0671

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  57. Ding L, Saunders TL, Enikolopov G, Morrison SJ (2012) Endothelial and perivascular cells maintain haematopoietic stem cells. Nature 481(7382):457–462. https://doi.org/10.1038/nature10783

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  58. Ding L, Morrison SJ (2013) Haematopoietic stem cells and early lymphoid progenitors occupy distinct BM niches. Nature 495(7440):231–235. https://doi.org/10.1038/nature11885

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  59. Poulos MG, Guo P, Kofler NM, Pinho S, Gutkin MC, Tikhonova A, Aifantis I, Frenette PS, Kitajewski J, Rafii S, Butler JM (2013) Endothelial Jagged-1 is necessary for homeostatic and regenerative hematopoiesis. Cell Rep 4(5):1022–1034. https://doi.org/10.1016/j.celrep.2013.07.048

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  60. Winkler IG, Sims NA, Pettit AR, Barbier V, Nowlan B, Helwani F, Poulton IJ, van Rooijen N, Alexander KA, Raggatt LJ, Levesque JP (2010) BM macrophages maintain hematopoietic stem cell (HSC) niches and their depletion mobilizes HSCs. Blood 116(23):4815–4828. https://doi.org/10.1182/blood-2009-11-253534

    Article  CAS  PubMed  Google Scholar 

  61. Morikawa S, Mabuchi Y, Kubota Y, Nagai Y, Niibe K, Hiratsu E, Suzuki S, Miyauchi-Hara C, Nagoshi N, Sunabori T, Shimmura S, Miyawaki A, Nakagawa T, Suda T, Okano H, Matsuzaki Y (2009) Prospective identification, isolation, and systemic transplantation of multipotent mesenchymal stem cells in murine BM. J Exp Med 206(11):2483–2496. https://doi.org/10.1084/jem.20091046

    Article  CAS  PubMed  PubMed Central  Google Scholar 

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Acknowledgments

The authors are supported by the National Heart, Lung, and Blood Institute of the National Institutes of Health under awards R01HL124015 (YL), R21HL140213 (YL), and R43AI45726 (YL) and the Markey Cancer Center’s Flow Cytometry and Immune Monitoring Core Shared Resource Facility (P30CA177558). We thank the Markey Cancer Center’s Research Communications Office for editing and graphics support.

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  1. Xinghui Zhao and Cuiping Zhang contributed equally to this work.

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  1. Department of Toxicology and Cancer Biology, University of Kentucky, Lexington, KY, USA

    Xinghui Zhao, Cuiping Zhang, Xiaojing Cui & Ying Liang

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  1. Xinghui Zhao
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Correspondence to Ying Liang .

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    Kursad Turksen

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Zhao, X., Zhang, C., Cui, X., Liang, Y. (2020). Interactions of Hematopoietic Stem Cells with Bone Marrow Niche. In: Turksen, K. (eds) Stem Cell Renewal and Cell-Cell Communication. Methods in Molecular Biology, vol 2346. Humana, New York, NY. https://doi.org/10.1007/7651_2020_298

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