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NF-κB inhibition sensitizes to starvation-induced cell death in high-risk myelodysplastic syndrome and acute myeloid leukemia

Oncogene volume 26, pages 4071–4083 (2007)Cite this article

Abstract

CD34+ bone marrow blasts from high-risk myelodysplastic syndrome (MDS) patients as well as MDS patient-derived cell lines (P39 and MOLM13) constitutively activate the nuclear factor-κB (NF-κB) pathway and undergo apoptosis when NF-κB is inhibited. Here, we show that the combination of conventional chemotherapeutic agents (daunorubicin, mitoxantrone, 5-azacytidine or camptothecin) with the NF-κB inhibitor BAY11-7082 did not yield a synergistic cytotoxicity. In contrast, BAY11-7082 (which targets the NF-κB-activating I-κB kinase (IKK) complex) or knockdown of essential components of the NF-κB system (such as the IKK1 and IKK2 subunits of the IKK complex and the p65 subunit of NF-κB), by small interfering RNAs sensitized MDS cell lines to starvation-induced apoptosis. The combination of BAY11-7082 and nutrient depletion synergistically killed the acute myeloid leukemia (AML) cell line U937 as well as primary CD34+ bone marrow blasts from AML and high-risk MDS patients. The synergistic killing by BAY11-7082, combined with nutrient depletion, led to cell death accompanied by all hallmarks of apoptosis, including an early loss of the mitochondrial transmembrane potential, the release of cytochrome c and apoptosis-inducing factor (AIF) from mitochondria, activation of caspase-3, phosphatidylserine exposure on the plasma membrane surface and nuclear chromatin condensation. Transmission electron microscopy revealed the presence of numerous autophagic vacuoles in the cytoplasm before cells underwent nuclear apoptosis. Nonetheless, cell death was neither inhibited by the pan-caspase inhibitor z-VAD-fmk nor by knockdown of AIF or of essential components of the autophagy pathway (ATG5, ATG6/Beclin-1, ATG10, ATG12). In contrast, external supply of glucose, insulin or insulin-like growth factor-I could retard the cell death induced by BAY11-7082 combined with starvation. These results suggest that in MDS cells, NF-κB inhibition can precipitate a bioenergetic crisis that leads to an autophagic stress response followed by apoptotic cell death.

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Accession codes

Accessions

GenBank/EMBL/DDBJ

Abbreviations

AIF:

apoptosis-inducing factor

AML:

acute myeloid leukemia

AnnV:

annexin V

Atg:

autophagy relevant genes

AV:

autophagic vacuole

BAY:

BAY11-7082

Casp-3a:

activated caspase-3

Cyt. c:

cytochrome c

ΔΨm:

mitochondrial transmembrane potential

DAPI:

4′,6-diaminidino-2-phenylindole

DiOC6(3):

3,3′ dihexyloxacarbocyanine iodide

EBSS:

Earle's balanced salt solution

Endo G:

endonuclease G

FACS:

fluorescence-activated cell sorter

FCS:

fetal calf serum

GAPDH:

glyceraldehyde-3-phosphate dehydrogenase

Hsp60:

heat shock protein 60

IGF-I:

insulin-like growth factor-I

IκB:

inhibitor of NF-κB

IKK:

IκB kinase

MAIF:

mouse AIF

MDS:

myelodysplastic syndrome

NF-κB:

nuclear factor-κB

PI:

propidium iodide

siRNA:

small interfering RNA

z-VAD-fmk:

N-benzyloxycarbonyl-Val-Ala-Asp-fluoromethylketone.

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Acknowledgements

CF, SB, ET, JG received fellowships from Fondation pour la Recherche Médicale, Deutsche Forschungsgemeinschaft and Institut National du Cancer, respectively. This work has been supported by grants from Association pour la Recherche contre le Cancer (to GK) and the European Union (TransDeath, RIGHT), Fondation de France (Comité Leucémies), Cancéropôle Ile-de-France, Ligue contre le Cancer (Département de Seine Saint Denis), Association Laurette Fugain (to GK) and the MDS Foundation (to TB).

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Authors and Affiliations

  1. INSERM, Unit ‘Apoptosis, Cancer and Immunity’, Villejuif, France

    C Fabre, G Carvalho, E Tasdemir, T Braun, L Adès, J Grosjean, S Boehrer, D Métivier, P Fenaux & G Kroemer

  2. Institut Gustave Roussy, Villejuif, France

    C Fabre, G Carvalho, E Tasdemir, T Braun, L Adès, J Grosjean, S Boehrer, D Métivier & G Kroemer

  3. Faculté Paris Sud-Université Paris XI, Villejuif, France

    C Fabre, G Carvalho, E Tasdemir, T Braun, L Adès, J Grosjean, S Boehrer, D Métivier & G Kroemer

  4. INSERM U542, Hôpital Paul Brousse, Université Paris XI, Villejuif, France

    G Carvalho

  5. Service d'Hématologie Clinique, Hôpital Avicenne, AP-HP, Université Paris XIII, Bobigny, France

    L Adès & P Fenaux

  6. CNRS, Institut André Lwoff, Villejuif, France

    S Souquère & G Pierron

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  1. C Fabre
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  2. G Carvalho
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Correspondence to G Kroemer.

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Fabre, C., Carvalho, G., Tasdemir, E. et al. NF-κB inhibition sensitizes to starvation-induced cell death in high-risk myelodysplastic syndrome and acute myeloid leukemia. Oncogene 26, 4071–4083 (2007). https://doi.org/10.1038/sj.onc.1210187

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