Apoptosis by leukemia cell-targeted diphtheria toxin occurs via receptor-independent activation of Fas-associated death domain protein

Jacqueline Thorburn, Arthur E. Frankel, Andrew Thorburn

Research output: Contribution to journalArticle

42 Citations (Scopus)

Abstract

Purpose: We examined the mechanism of action of a targeted fusion toxin consisting of diphtheria toxin fused to granulocyte macrophage colony stimulating factor (GMCSF) (DT388-GMCSF), which was designed to selectively kill acute myeloid leukemia cells. Experimental design and Results: U937 cells treated with DT388-GMCSF underwent apoptosis as shown by chromatin degradation and cellular and nuclear fragmentation. This apoptosis was prevented by a general caspase inhibitor. DT388-GMCSF treatment resulted in activation of the initiator caspases 8 and 9 and effector caspases. A selective caspase 8 inhibitor prevented activation of caspase 9, whereas a selective caspase 9 inhibitor did not prevent activation of caspase 8, indicating that caspase 8 activation is the proximal event in DT388-GMCSF-induced apoptosis. Caspase 8 was activated through a Fas-associated death domain protein (FADD)-dependent mechanism as demonstrated by inhibition of DT388-GMCSF-induced apoptosis on expression of a dominant negative FADD molecule. However, unlike most FADD-dependent apoptosis, this pathway may not involve death receptors, including Fas, tumor necrosis factor receptor 1, or tumor necrosis factor-related apoptosis-inducing ligand receptors, because inhibitors of the receptors did not prevent DT388-GMCSF-induced apoptosis. Conclusions: These data indicate that targeted toxins induce apoptosis by activating components of the death receptor pathway in a receptor-independent manner.

Original languageEnglish (US)
Pages (from-to)861-865
Number of pages5
JournalClinical Cancer Research
Volume9
Issue number2
StatePublished - Feb 1 2003

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Fas-Associated Death Domain Protein
Diphtheria Toxin
Immunotoxins
Granulocyte-Macrophage Colony-Stimulating Factor
Leukemia
Caspase 8
Apoptosis
Caspase Inhibitors
Caspase 9
Death Domain Receptors
Initiator Caspases
Effector Caspases
U937 Cells
Tumor Necrosis Factor Receptors
Myeloid Cells
Acute Myeloid Leukemia
Chromatin
Research Design
Tumor Necrosis Factor-alpha
Ligands

ASJC Scopus subject areas

  • Cancer Research
  • Oncology

Cite this

Apoptosis by leukemia cell-targeted diphtheria toxin occurs via receptor-independent activation of Fas-associated death domain protein. / Thorburn, Jacqueline; Frankel, Arthur E.; Thorburn, Andrew.

In: Clinical Cancer Research, Vol. 9, No. 2, 01.02.2003, p. 861-865.

Research output: Contribution to journalArticle

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