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Rac1 signaling protects monocytic AML cells expressing the MLL-AF9 oncogene from caspase-mediated apoptotic death

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Abstract

We investigated the relevance of signaling mechanisms regulated by the Ras-homologous GTPase Rac1 for survival of acute myeloid leukemia (AML) cells harbouring the MLL-AF9 oncogene due to t(9;11)(p21;q23) translocation. Monocytic MLL-AF9 expressing cells (MM6, THP-1) were hypersensitive to both small-molecule inhibitors targeting Rac1 (EHT 1864, NSC 23766) (IC50EHT ~12.5 μM) and lipid lowering drugs (lovastatin, atorvastatin) (IC50Lova ~7.5 μM) as compared to acute myelocytic leukemia (NOMO-1, HL60) and T cell leukemia (Jurkat) cells (IC50EHT >30 μM; IC50Lova >25 μM). Hypersensitivity of monocytic cells following Rac1 inhibition resulted from caspase-driven apoptosis as shown by profound activation of caspase-8,-9,-7,-3 and substantial (~90 %) decrease in protein expression of pro-survival factors (survivin, XIAP, p-Akt). Apoptotic death was preceded by S139-posphorylation of histone H2AX (γH2AX), a prototypical surrogate marker of DNA double-strand breaks (DSBs). Taken together, abrogation of Rac1 signaling causes DSBs in acute monocytic leukemia cells harbouring the MLL-AF9 oncogene, which, together with downregulation of survivin, XIAP and p-Akt, results in massive induction of caspase-driven apoptotic death. Apparently, Rac1 signaling is required for maintaining genetic stability and maintaining survival in specific subtypes of AML. Hence, targeting of Rac1 is considered a promising novel strategy to induce lethality in MLL-AF9 expressing AML.

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Abbreviations

Akt:

Protein kinase B

ALL:

Acute lymphoblastic leukemia

AML:

Acute myeloid leukemia

ATM:

Ataxia telangiectasia mutated

ATR:

ATM and Rad3-related

BER:

Base excision repair

BRCA:

Breast cancer susceptibility protein

Chk:

Checkpoint kinase

CRIB:

Cdc42/Rac interactive binding region

DDR:

DNA damage response

DSBs:

DNA double-strand breaks

GEF:

Guanine exchange factor

IC50 :

Inhibitory concentration 50 %

EMT:

Epithelial-to-mesenchymal transition

HMG-CoA:

3-Hydroxy-3-methyl-glutaryl coenzyme A

H2AX:

Histone H2AX

γH2AX:

S139 phosphorylated H2AX

IC50 :

Inhibitory concentration 50 %

IC50EHT :

IC50 after EHT 1864 treatment

IC50Lova :

IC50 after lovastatin treatment

MLL:

Mixed lineage leukemia

MLLT3 (=AF9):

Mixed lineage leukemia, tanslocated to, 3

PARP:

Poly (ADP-ribose) polymerase

Rac1:

Ras-related C3 botulinum substrate 1

Rho:

Ras-homologous

XIAP:

X-linked inhibitor of apoptosis

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Acknowledgments

We would like to thank C. Brachetti for excellent technical assistance. This work was supported by the José Carreras Leukämie Stiftung (SP 10/07) and the Deutsche Krebshilfe (109188).

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The authors declare no conflicts of interest.

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Authors and Affiliations

  1. Institute of Toxicology, Heinrich-Heine University Düsseldorf, Moorenstrasse 5, 40225, Düsseldorf, Germany

    C. Henninger, F. Wartlick, A. Schorr & G. Fritz

  2. Institute of Toxicology, University Medical Center of the Johannes Gutenberg University Mainz, Obere Zahlbacher Str 67, 55131, Mainz, Germany

    C. Hinterleitner, J. Huelsenbeck & B. Kaina

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  1. C. Hinterleitner
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  2. J. Huelsenbeck
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  3. C. Henninger
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  7. G. Fritz
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Correspondence to G. Fritz.

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Hinterleitner, C., Huelsenbeck, J., Henninger, C. et al. Rac1 signaling protects monocytic AML cells expressing the MLL-AF9 oncogene from caspase-mediated apoptotic death. Apoptosis 18, 963–979 (2013). https://doi.org/10.1007/s10495-013-0842-6

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