Hematopoietic stem cell (HSC) transplantation therapy is one of the most effective treatments for life-threatening hematopoietic diseases. Bone marrow (BM) and mobilized peripheral blood are the major sources of HSCs, but these resources are limited by a paucity of human leukocyte antigen (HLA)-matched donors. Umbilical cord blood (UCB) is the most promising alternative to obtain HSCs for transplantation therapy. However, UCB transplantation therapy is limited by low numbers of HSCs per unit of UCB. In vitro HSC expansion is believed to be the most effective and applicable strategy to address this issue. Here we report that a moderate concentration of GSK3 inhibitor promotes HSC expansion by inducing moderate levels of β-catenin activity in HSCs. However, such a concentration of GSK3 inhibitor also stimulates myeloid cells to produce inflammatory cytokines, which attenuate HSC expansion by inducing p38 activation. Thus, when unpurified HSCs were used in culture, inhibition of p38-induced inflammatory cytokine signaling was required to ensure HSC expansion induced by the low concentration of GSK3 inhibitor. Our study suggests that the combination of a moderate concentration of p38 inhibitor plus a GSK3 inhibitor synergistically promotes the expansion of both murine BM HSCs and human UCB HSCs.
Get full access to this article
View all access options for this article.
References
1.
LiHW and SykesM. (2012). Emerging concepts in haematopoietic cell transplantation. Nat Rev Immunol, 12:403–416.
2.
PiemonteseS, CiceriF, LabopinM, ArceseW, Kyrcz-KrzemienS, SantaroneS, HuangH, BeelenD, GorinNC, et al. (2017). A comparison between allogeneic stem cell transplantation from unmanipulated haploidentical and unrelated donors in acute leukemia. J Hematol Oncol, 10:24–31.
3.
GragertL, EapenM, WilliamsE, FreemanJ, SpellmanS, BaittyR, HartzmanR, RizzoJD, HorowitzM, ConferD and MaiersM. (2014). HLA match likelihoods for hematopoietic stem-cell grafts in the U.S. registry. N Engl J Med, 371:339–348.
4.
BallenKK, GluckmanE and BroxmeyerHE. (2013). Umbilical cord blood transplantation: the first 25 years and beyond. Blood, 122:491–498.
5.
StiffP, ChenB, FranklinW, OldenbergD, HsiE, BayerR, ShpallE, DouvilleJ, MandalamR, et al. (2000). Autologous transplantation of ex vivo expanded bone marrow cells grown from small aliquots after high-dose chemotherapy for breast cancer. Blood, 95:2169–2174.
6.
de LimaM, McNieceI, RobinsonSN, MunsellM, EapenM, HorowitzM, AlousiA, SalibaR, McMannisJD, et al. (2012). Cord-blood engraftment with ex vivo mesenchymal-cell coculture. N Engl J Med, 367:2305–2315.
7.
DahlbergA, DelaneyC and BernsteinID. (2011). Ex vivo expansion of human hematopoietic stem and progenitor cells. Blood, 117:6083–6090.
8.
DelaneyC, HeimfeldS, Brashem-SteinC, VoorhiesH, MangerRL and BernsteinID. (2010). Notch-mediated expansion of human cord blood progenitor cells capable of rapid myeloid reconstitution. Nat Med, 16:232–236.
9.
Flores-GuzmanP, Fernandez-SanchezV and MayaniH. (2013). Concise review: ex vivo expansion of cord blood-derived hematopoietic stem and progenitor cells: basic principles, experimental approaches, and impact in regenerative medicine. Stem Cell Transl Med, 2:830–838.
10.
WalasekMA, van OsR and de HaanG. (2012). Hematopoietic stem cell expansion: challenges and opportunities. Ann N Y Acad Sci, 1266:138–150.
11.
HeX and AxelrodJD. (2006). A WNTer wonderland in Snowbird. Development, 133:2597–2603.
12.
FamiliF, BrugmanMH, TaskesenE, NaberBE, FoddeR and StaalFJ. (2016). High levels of canonical Wnt signaling lead to loss of stemness and increased differentiation in hematopoietic stem cells. Stem Cell Rep, 6:652–659.
13.
LuisTC, NaberBA, RoozenPP, BrugmanMH, de HaasEF, GhazviniM, FibbeWE, van DongenJJ, FoddeR and StaalFJ. (2011). Canonical wnt signaling regulates hematopoiesis in a dosage-dependent fashion. Cell Stem Cell, 9:345–356.
14.
XiaoY, LiH, ZhangJ, VolkA, ZhangS, WeiW, ZhangS, BreslinP and ZhangJ. (2011). TNF-alpha/Fas-RIP-1-induced cell death signaling separates murine hematopoietic stem cells/progenitors into 2 distinct populations. Blood, 118:6057–6067.
15.
TangM, WeiX, GuoY, BreslinP, ZhangS, ZhangS, WeiW, XiaZ, DiazM, AkiraS and ZhangJ. (2008). TAK1 is required for the survival of hematopoietic cells and hepatocytes in mice. J Exp Med, 205:1611–1619.
XinJ, YouD, BreslinP, LiJ, ZhangJ, WeiW, CannovaJ, VolkA, GutierrezR, et al. (2017). Sensitizing acute myeloid leukemia cells to induced differentiation by inhibiting the RIP1/RIP3 pathway. Leukemia, 31:1154–1165.
20.
LeeJY, NakadaD, YilmazOH, TothovaZ, JosephNM, LimMS, GillilandDG and MorrisonSJ. (2010). mTOR activation induces tumor suppressors that inhibit leukemogenesis and deplete hematopoietic stem cells after Pten deletion. Cell Stem Cell, 7:593–605.
21.
HuangJ, ZhangY, BersenevA, O'BrienWT, TongW, EmersonSG and KleinPS. (2009). Pivotal role for glycogen synthase kinase-3 in hematopoietic stem cell homeostasis in mice. J Clin Invest, 119:3519–3529.
22.
KoKH, HolmesT, PalladinettiP, SongE, NordonR, O'BrienTA and DolnikovA. (2011). GSK-3beta inhibition promotes engraftment of ex vivo-expanded hematopoietic stem cells and modulates gene expression. Stem Cells, 29:108–118.
23.
JiangJ, ZhaoM, ZhangA, YuM, LinX, WuM, WangX, LuH, ZhuS, et al. (2010). Characterization of a GSK-3 inhibitor in culture of human cord blood primitive hematopoietic cells. Biomed Pharmacother, 64:482–486.
FlemingHE, JanzenV, Lo CelsoC, GuoJ, LeahyKM, KronenbergHM and ScaddenDT. (2008). Wnt signaling in the niche enforces hematopoietic stem cell quiescence and is necessary to preserve self-renewal in vivo. Cell Stem Cell, 2:274–283.
26.
LuisTC, WeerkampF, NaberBA, BaertMR, de HaasEF, NikolicT, HeuvelmansS, De KrijgerRR, van DongenJJ and StaalFJ. (2009). Wnt3a deficiency irreversibly impairs hematopoietic stem cell self-renewal and leads to defects in progenitor cell differentiation. Blood, 113:546–554.
27.
LentoW, ItoT, ZhaoC, HarrisJR, HuangW, JiangC, OwzarK, PiryaniS, RacioppiL, ChaoN and ReyaT. (2014). Loss of beta-catenin triggers oxidative stress and impairs hematopoietic regeneration. Genes Dev, 28:995–1004.
28.
ZhaoC, BlumJ, ChenA, KwonHY, JungSH, CookJM, LagooA and ReyaT. (2007). Loss of beta-catenin impairs the renewal of normal and CML stem cells in vivo. Cancer Cell, 12:528–541.
29.
DuncanAW, RattisFM, DiMascioLN, CongdonKL, PazianosG, ZhaoC, YoonK, CookJM, WillertK, GaianoN and ReyaT. (2005). Integration of Notch and Wnt signaling in hematopoietic stem cell maintenance. Nat Immunol, 6:314–322.
HolmesT, YanF, KoKH, NordonR, SongE, O'BrienTA and DolnikovA. (2012). Ex vivo expansion of cord blood progenitors impairs their short-term and long-term repopulating activity associated with transcriptional dysregulation of signalling networks. Cell Prolif, 45:266–278.
32.
TrowbridgeJJ, XenocostasA, MoonRT and BhatiaM. (2006). Glycogen synthase kinase-3 is an in vivo regulator of hematopoietic stem cell repopulation. Nat Med, 12:89–98.
33.
HuangJ, Nguyen-McCartyM, HexnerEO, Danet-DesnoyersG and KleinPS. (2012). Maintenance of hematopoietic stem cells through regulation of Wnt and mTOR pathways. Nat Med, 18:1778–1785.
34.
GoesslingW, NorthTE, LoewerS, LordAM, LeeS, Stoick-CooperCL, WeidingerG, PuderM, DaleyGQ, MoonRT and ZonLI. (2009). Genetic interaction of PGE2 and Wnt signaling regulates developmental specification of stem cells and regeneration. Cell, 136:1136–1147.
35.
ItoK, HiraoA, AraiF, TakuboK, MatsuokaS, MiyamotoK, OhmuraM, NakaK, HosokawaK, IkedaY and SudaT. (2006). Reactive oxygen species act through p38 MAPK to limit the lifespan of hematopoietic stem cells. Nat Med, 12:446–451.
36.
WangY, KellnerJ, LiuL and ZhouD. (2011). Inhibition of p38 mitogen-activated protein kinase promotes ex vivo hematopoietic stem cell expansion. Stem Cells Dev, 20:1143–1152.
37.
ZouJ, ZouP, WangJ, LiL, WangY, ZhouD and LiuL. (2012). Inhibition of p38 MAPK activity promotes ex vivo expansion of human cord blood hematopoietic stem cells. Ann Hematol, 91:813–823.
38.
KariganeD, KobayashiH, MorikawaT, OotomoY, SakaiM, NagamatsuG, KubotaY, GodaN, MatsumotoM, et al. (2016). p38alpha Activates purine metabolism to initiate hematopoietic stem/progenitor cell cycling in response to stress. Cell Stem Cell, 19:192–204.
39.
DobleBW, PatelS, WoodGA, KockeritzLK and WoodgettJR. (2007). Functional redundancy of GSK-3alpha and GSK-3beta in Wnt/beta-catenin signaling shown by using an allelic series of embryonic stem cell lines. Dev Cell, 12:957–971.
40.
VermaA, DebDK, SassanoA, KambhampatiS, WickremaA, UddinS, MohindruM, Van BesienK and PlataniasLC. (2002). Cutting edge: activation of the p38 mitogen-activated protein kinase signaling pathway mediates cytokine-induced hemopoietic suppression in aplastic anemia. J Immunol, 168:5984–5988.
41.
NavasT, MohindruM, EstesM, MaJY, SokolL, PahanishP, ParmarS, HaghnazariE, ZhouL, et al. (2006). Inhibition of overactivated p38 MAPK can restore hematopoiesis in myelodysplastic syndrome progenitors. Blood, 108:4170–4177.
42.
NavasT, ZhouL, EstesM, HaghnazariE, NguyenAN, MoY, PahanishP, MohindruM, CaoT, et al. (2008). Inhibition of p38alpha MAPK disrupts the pathological loop of proinflammatory factor production in the myelodysplastic syndrome bone marrow microenvironment. Leuk Lymphoma, 49:1963–1975.
43.
KatsoulidisE, LiY, YoonP, SassanoA, AltmanJ, Kannan-ThulasiramanP, BalasubramanianL, ParmarS, VargaJ, et al. (2005). Role of the p38 mitogen-activated protein kinase pathway in cytokine-mediated hematopoietic suppression in myelodysplastic syndromes. Cancer Res, 65:9029–9037.
Supplementary Material
Please find the following supplemental material available below.
For Open Access articles published under a Creative Commons License, all supplemental material carries the same license as the article it is associated with.
For non-Open Access articles published, all supplemental material carries a non-exclusive license, and permission requests for re-use of supplemental material or any part of supplemental material shall be sent directly to the copyright owner as specified in the copyright notice associated with the article.
