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Transient inhibition of NF-κB signaling enhances ex vivo propagation of human hematopoietic stem cells

Talkhoncheh, Mehrnaz Safaee, Subramaniam, Agatheeswaran, Magnusson, Mattias, Kumar, Praveen, Larsson, Jonas, Baudet, Aurélie·2018·Haematologica
DOI 10.3324/haematol.2018.188466PMC PMC6119158
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Figure 1. The sh758-dependent ex vivo expansion effect is associated with a downmodulation of the NF-κB pathway. (A) Growth of GFP+ cells was assessed by FACS and normalized to day 2-transduction efficiency. (B) The percentage of CD34+ 90+ cells in the GFP+ fraction was quantified by FACS seven days after transduction of CD34+CD38− CD90+CD45RA− cells (n=2). (C) Gene expression data mining showed modulation of members of NF-κB signaling in sh758 expressing CD34+ CD90+ cells. (D) The expression of total and phosphorylated ΙκΒα was quantified by western blot in CD34+ cells at indicated culture time points. Actin expression is shown as loading control.
Figure 1. The sh758-dependent ex vivo expansion effect is associated with a downmodulation of the NF-κB pathway. (A) Growth of GFP+ cells was assessed by FACS and normalized to day 2-transduction efficiency. (B) The percentage of CD34+ 90+ cells in the GFP+ fraction was quantified by FACS seven days after transduction of CD34+CD38− CD90+CD45RA− cells (n=2). (C) Gene expression data mining showed modulation of members of NF-κB signaling in sh758 expressing CD34+ CD90+ cells. (D) The expression of total and phosphorylated ΙκΒα was quantified by western blot in CD34+ cells at indicated culture time points. Actin expression is shown as loading control.CC BY-NC · PubMed Central
Figure 2. Targeting of the NF-κB regulator IKKβ increases the number of human hematopoietic stem and progenitor cells (HSPCs) in vitro without affecting cell proliferation and survival. (A) The protein level of total and phosphorylated IκBα were quantified in CD34+ cells after 4-hour treatment with STF/PF/TP in culture. Actin expression is shown as loading control (n=2). (B) FACS plots and graph show cell surface expression of CD34 and CD90 and number of CD34+CD90+ cells respectively at day 7 (n=4). (C) Graph displays total live (7AAD−) cell number at day 7 (n=4). (D) Cell cycle distribution, and (E) apoptosis (7AAD−/AnnexinV+) of CD34+CD90+ cells were assessed by FACS at days 4 and 7, respectively (n=3). p-IκBa: phosphorylated IκBa; t-IκBa: total IκBa; PF: PF184; TP: TPCA1.
Figure 2. Targeting of the NF-κB regulator IKKβ increases the number of human hematopoietic stem and progenitor cells (HSPCs) in vitro without affecting cell proliferation and survival. (A) The protein level of total and phosphorylated IκBα were quantified in CD34+ cells after 4-hour treatment with STF/PF/TP in culture. Actin expression is shown as loading control (n=2). (B) FACS plots and graph show cell surface expression of CD34 and CD90 and number of CD34+CD90+ cells respectively at day 7 (n=4). (C) Graph displays total live (7AAD−) cell number at day 7 (n=4). (D) Cell cycle distribution, and (E) apoptosis (7AAD−/AnnexinV+) of CD34+CD90+ cells were assessed by FACS at days 4 and 7, respectively (n=3). p-IκBa: phosphorylated IκBa; t-IκBa: total IκBa; PF: PF184; TP: TPCA1.CC BY-NC · PubMed Central
Figure 3. Targeting of the NF-κB regulator IKKβ improves the in vitro and in vivo function of cultured cord blood (CB)-derived hematopoietic stem and progenitor cells (HSPCs). (A) Colonies per 300 CD34+ input cells were scored after 14 days (n=3). (B) Schematic representation of in vivo transplantation experiment. (C) Human engraftment was assessed by quantifying the percentage of human CD45+ cells in the bone marrow (BM) of NSG recipients four months post transplantation (data from 2 independent experiments). (D) Lineage distribution was quantified by FACS analysis of human B cells (CD19), T cells (CD3), and myeloid (CD33/CD15) in CD45+ cells of the BM of recipients. (E) Representative FACS plots for gating strategy of lineage reconstitution. PF: PF184; TP: TPCA1.
Figure 3. Targeting of the NF-κB regulator IKKβ improves the in vitro and in vivo function of cultured cord blood (CB)-derived hematopoietic stem and progenitor cells (HSPCs). (A) Colonies per 300 CD34+ input cells were scored after 14 days (n=3). (B) Schematic representation of in vivo transplantation experiment. (C) Human engraftment was assessed by quantifying the percentage of human CD45+ cells in the bone marrow (BM) of NSG recipients four months post transplantation (data from 2 independent experiments). (D) Lineage distribution was quantified by FACS analysis of human B cells (CD19), T cells (CD3), and myeloid (CD33/CD15) in CD45+ cells of the BM of recipients. (E) Representative FACS plots for gating strategy of lineage reconstitution. PF: PF184; TP: TPCA1.CC BY-NC · PubMed Central
Figure 4. The benefit of NF-κB pathway inhibition is limited to a short time window, and results in downregulation of inflammatory signaling. (A) Number of CD34+CD90+ cells was assessed after five days by FACS, when TP/STF was added directly or 24 hours after culture start or removed after 24 hours (n=3). (B) Graph shows number of CD34+CD90+ cells after treatment with STF, TP, SR1, UM171 and combination of TP with either SR1 or UM171 at day 6 (representative data from 2 experiments). (C) Graph shows number of CD34+CD90+EPCR+ cells after treatment with STF, TP, SR1, UM171 and combination of TP with either SR1 or UM171 at day 6 (representative data from 2 experiments). (D and E) CD34+ cells were treated with or without TP for six hours and subjected to transcriptome analysis by microarrays after sorting for CD34+90+ population. Volcano plot and GO classification of 2-fold modulated (down-regulated) genes are shown for each condition. See also Online Supplementary Figure S3 and Online Supplementary Table S1. (F) The log ratio (TP/STF) of factors present in supernatant after six hours of culture of CB-derived CD34+ cells is shown (n=3). (G) Each graph displays individual data point for significantly modulated cytokines (IL6, TNFa, IL8, and IP10), extracted from the experiments presented in (F). The stars in (F) indicate that those factors were not detectable. PF: PF184; TP: TPCA1.
Figure 4. The benefit of NF-κB pathway inhibition is limited to a short time window, and results in downregulation of inflammatory signaling. (A) Number of CD34+CD90+ cells was assessed after five days by FACS, when TP/STF was added directly or 24 hours after culture start or removed after 24 hours (n=3). (B) Graph shows number of CD34+CD90+ cells after treatment with STF, TP, SR1, UM171 and combination of TP with either SR1 or UM171 at day 6 (representative data from 2 experiments). (C) Graph shows number of CD34+CD90+EPCR+ cells after treatment with STF, TP, SR1, UM171 and combination of TP with either SR1 or UM171 at day 6 (representative data from 2 experiments). (D and E) CD34+ cells were treated with or without TP for six hours and subjected to transcriptome analysis by microarrays after sorting for CD34+90+ population. Volcano plot and GO classification of 2-fold modulated (down-regulated) genes are shown for each condition. See also Online Supplementary Figure S3 and Online Supplementary Table S1. (F) The log ratio (TP/STF) of factors present in supernatant after six hours of culture of CB-derived CD34+ cells is shown (n=3). (G) Each graph displays individual data point for significantly modulated cytokines (IL6, TNFa, IL8, and IP10), extracted from the experiments presented in (F). The stars in (F) indicate that those factors were not detectable. PF: PF184; TP: TPCA1.CC BY-NC · PubMed Central

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Cites

Research works printed but not linked: 32

  1. [1]Brunstein CG Gutman JA Weisdorf DJ Allogeneic hematopoietic cell transplantation for hematologic malignancy: relative risks and benefits of double umbilical cord blood. Blood. 2010; 116(22): 4693– 4699. 20686119 PMC2996124
  2. [2]Wagner JE Gluckman E Umbilical cord blood transplantation: the first 20 years. Semin Hematol. 2010; 47(1): 3– 12. 20109607
  3. [3]Ballen KK Gluckman E Broxmeyer HE Umbilical cord blood transplantation: the first 25 years and beyond. Blood. 2013; 122(4): 491– 498. 23673863 PMC3952633
  4. [4]Broxmeyer HE Enhancing the efficacy of engraftment of cord blood for hematopoietic cell transplantation. Transfus Apher Sci. 2016; 54(3): 364– 372. 27211041 PMC4899263
  5. [5]Farahbakhshian E Verstegen MM Visser TP Angiopoietin-like protein 3 promotes preservation of stemness during ex vivo expansion of murine hematopoietic stem cells. PLoS One. 2014; 9(8): e105642. 25170927 PMC4149469
  6. [6]Kirouac DC Ito C Csaszar E Dynamic interaction networks in a hierarchically organized tissue. Mol Syst Biol. 2010; 6: 417. 20924352 PMC2990637
  7. [7]Magnusson M Sierra MI Sasidharan R Expansion on stromal cells preserves the undifferentiated state of human hematopoietic stem cells despite compromised reconstitution ability. PLoS One. 2013; 8(1): e53912. 23342037 PMC3547050
  8. [8]Csaszar E Kirouac DC Yu M Rapid expansion of human hematopoietic stem cells by automated control of inhibitory feedback signaling. Cell Stem Cell. 2012; 10(2): 218– 229. 22305571
  9. [9]Galeev R Baudet A Kumar P Genome-wide RNAi Screen Identifies Cohesin Genes as Modifiers of Renewal and Differentiation in Human HSCs. Cell Rep. 2016; 14(12): 2988– 3000. 26997282 PMC7616965
  10. [10]Baudet A Karlsson C Safaee Talkhoncheh M RNAi screen identifies MAPK14 as a druggable suppressor of human hematopoietic stem cell expansion. Blood. 2012; 119(26): 6255– 6258. 22555972
  11. [11]Ali N Karlsson C Aspling M Forward RNAi screens in primary human hematopoietic stem/progenitor cells. Blood. 2009; 113(16): 3690– 3695. 19188664 PMC2670788
  12. [12]Hayden MS Ghosh S Shared principles in NF-kappaB signaling. Cell. 2008; 132(3): 344– 362. 18267068
  13. [13]Oeckinghaus A Hayden MS Ghosh S Crosstalk in NF-kappaB signaling pathways. Nat Immunol. 2011; 12(8): 695– 708. 21772278
  14. [14]Mora E Guglielmotti A Biondi G Sassone-Corsi P Bindarit: an anti-inflammatory small molecule that modulates the NFkappaB pathway. Cell Cycle. 2012; 11(1): 159– 169. 22189654 PMC3356824
  15. [15]Podolin PL Callahan JF Bolognese BJ Attenuation of murine collagen-induced arthritis by a novel, potent, selective small molecule inhibitor of IkappaB Kinase 2, TPCA-1 (2-[(aminocarbonyl)amino]-5-(4-fluorophenyl)-3-thiophenecarboxamide), occurs via reduction of proinflammatory cytokines and antigen-induced T cell Proliferation. J Pharmacol Exp Ther. 2005; 312(1): 373– 381. 15316093
  16. [16]Barkett M Gilmore TD Control of apoptosis by Rel/NF-kappaB transcription factors. Oncogene. 1999; 18(49): 6910– 6924. 10602466
  17. [17]Naugler WE Karin M NF-kappaB and cancer-identifying targets and mechanisms. Curr Opin Genet Dev. 2008; 18(1): 19– 26. 18440219 PMC2587362
  18. [18]Boitano AE Wang J Romeo R Aryl hydrocarbon receptor antagonists promote the expansion of human hematopoietic stem cells. Science. 2010; 329(5997): 1345– 1348. 20688981 PMC3033342
  19. [19]Fares I Chagraoui J Gareau Y Cord blood expansion. Pyrimidoindole derivatives are agonists of human hematopoietic stem cell self-renewal. Science. 2014; 345(6203): 1509– 1512. 25237102 PMC4372335
  20. [20]Fares I Chagraoui J Lehnertz B EPCR expression marks UM171-expanded CD34(+) cord blood stem cells. Blood. 2017; 129(25): 3344– 3351. 28408459
  21. [21]Morgan MJ Liu ZG Crosstalk of reactive oxygen species and NF-kappaB signaling. Cell Res. 2011; 21(1): 103– 115. 21187859 PMC3193400
  22. [22]Stylianou E O’Neill LA Rawlinson L Interleukin 1 induces NF-kappa B through its type I but not its type II receptor in lymphocytes. J Biol Chem. 1992; 267(22): 15836– 15841. 1386364
  23. [23]Baud V Karin M Signal transduction by tumor necrosis factor and its relatives. Trends Cell Biol. 2001; 11(9): 372– 377. 11514191
  24. [24]Espin-Palazon R Stachura DL Campbell CA Proinflammatory signaling regulates hematopoietic stem cell emergence. Cell. 2014; 159(5): 1070– 1085. 25416946 PMC4243083
  25. [25]Oeckinghaus A Ghosh S The NF-kappaB family of transcription factors and its regulation. Cold Spring Harb Perspect Biol. 2009; 1(4): a000034. 20066092 PMC2773619
  26. [26]Stein SJ Baldwin AS Deletion of the NF-kappaB subunit p65/RelA in the hematopoietic compartment leads to defects in hematopoietic stem cell function. Blood. 2013; 121(25): 5015– 5024. 23670180 PMC3689248
  27. [27]Nakata S Matsumura I Tanaka H NF-kappaB family proteins participate in multiple steps of hematopoiesis through elimination of reactive oxygen species. J Biol Chem. 2004; 279(53): 55578– 55586. 15485843
  28. [28]Schepers H Eggen BJ Schuringa JJ Vellenga E Constitutive activation of NF-kappa B is not sufficient to disturb normal steady-state hematopoiesis. Haematologica. 2006; 91(12): 1710– 1711. 17145612
  29. [29]Radhakrishnan SK Kamalakaran S Proapoptotic role of NF-kappaB: implications for cancer therapy. Biochim Biophys Acta. 2006; 1766(1): 53– 62. 16563635
  30. [31]Baldridge MT King KY Goodell MA Inflammatory signals regulate hematopoietic stem cells. Trends Immunol. 2011; 32(2): 57– 65. 21233016 PMC3042730
  31. [32]Dybedal I Bryder D Fossum A Rusten LS Jacobsen SE Tumor necrosis factor (TNF)-mediated activation of the p55 TNF receptor negatively regulates maintenance of cycling reconstituting human hematopoietic stem cells. Blood. 2001; 98(6): 1782– 1791. 11535512
  32. [33]Pietras EM Inflammation: a key regulator of hematopoietic stem cell fate in health and disease. Blood. 2017; 130(15): 1693– 1698. 28874349 PMC5639485

Cited by

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Metadata

Authors
Talkhoncheh, M. S., Subramaniam, A., Magnusson, M., Kumar, P., Larsson, J., & Baudet, A.
Venue
Haematologica
Publisher
Ferrata Storti Foundation (Haematologica)
Published
2018-06-07
Volume
103
Issue
9
Pages
1444-1450
ISSN
  • 0390-6078
  • 1592-8721
DOI
10.3324/haematol.2018.188466
PMC
PMC6119158
Cited by
9
References
1
Language
en

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