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Dz13, a DNAzyme targeting c-jun, induces off-target cytotoxicity in endothelial cells with features of nonapoptotic

Mary Margaret Gozar1, Amber Goodchild, Toby Passioura

  • 1Johnson and Johnson Research Pty Limited, Eveleigh, New South Wales, 1430, Australia. mgozar@medau.jnj.com

Oligonucleotides
|August 14, 2008
PubMed

Insights

Dz13, a DNAzyme targeting c-jun, induces programmed cell death (PCD) in endothelial cells. This novel cytotoxicity involves mitochondrial changes and autophagy, independent of classical apoptosis or ER stress, with potential PARP-1 involvement.

Area of Science:

  • Molecular Biology
  • Cell Biology
  • Biochemistry

Background:

  • Dz13 is a DNAzyme designed to target c-jun.
  • Previous studies indicated Dz13 cytotoxicity is independent of c-jun mRNA cleavage.
  • Programmed cell death (PCD) pathways require further elucidation for Dz13's mechanism of action.

Purpose of the Study:

  • To investigate the programmed cell death (PCD) pathways involved in Dz13-mediated cytotoxicity.
  • To understand the interplay of apoptosis, autophagy, and necrosis in response to Dz13.
  • To explore the potential role of PARP-1 in Dz13's mechanism of action.

Main Methods:

  • Utilized human dermal microvascular endothelial cells (HMEC-1).
  • Assessed cell death markers including mitochondrial depolarization, caspase activation, lysosomal changes, and autophagosome formation.
  • Employed inhibitors, protein extraction, and pull-down assays to investigate molecular players like PARP-1.

Main Results:

  • Dz13 induced cell death characterized by mitochondrial depolarization, caspase-8 activation, lysosomal increase, and autophagosome formation.
  • Classical apoptosis markers (DNA laddering, mitochondrial protein translocation) were absent.
  • Inhibitor studies showed caspases and ER stress were not primary drivers; PARP-1 showed potential involvement.

Conclusions:

  • Dz13 triggers a novel form of oligonucleotide-mediated cytotoxicity with features of PCD.
  • The mechanism involves mitochondrial and autophagic pathways, distinct from classical apoptosis.
  • PARP-1 may play a role in Dz13-induced cell death, warranting further investigation.

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