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Mechanisms of apoptosis-induction by rottlerin: therapeutic implications for B-CLL
I Ringshausen1, M Oelsner, K Weick
13rd Department of Medicine, Technical University of Munich, Munich, Germany. iringshausen@cc.ucsf.edu
Abstract:
Constitutively activated signaling pathways contribute to the apoptosis-defect of B-CLL cells. Protein kinase C-delta is a permanently activated kinase and a putative downstream target of phosphatidylinositol-3 kinase in B-CLL. Blockade of protein kinase C-delta (PKC-delta) by the highly specific inhibitor rottlerin induces apoptosis in chronic lymphocytic leukaemia (CLL) cells. By co-culturing bone marrow stromal and CLL cells, we determined that the proapoptotic effect of rottlerin is not abolished in the presence of survival factors, indicating that a targeted therapy against PKC-delta might be a powerful approach for the treatment of CLL patients. The downstream events following rottlerin treatment engage mitochondrial and non-mitochondrial pathways and ultimately activate caspases that execute the apoptotic cell death. Herein we report that the inhibition of PKC-delta decreases the expression of the important antiapoptotic proteins Mcl-1 and XIAP accompanied by a loss of the mitochondrial membrane potential Deltapsi. In addition, we discovered that ZAP-70-expressing cells are significantly more susceptible to rottlerin-induced cell death than ZAP-70 negative cells. We finally observed that rottlerin can augment cell toxicity induced by standard chemotherapeutic drugs. Conclusively, PKC-delta is a promising new target in the combat against CLL.
Insights
Targeting protein kinase C-delta (PKC-delta) with rottlerin induces apoptosis in chronic lymphocytic leukemia (CLL) cells, even with survival factors present. This suggests PKC-delta is a promising therapeutic target for CLL treatment.
Area of Science:
- Oncology
- Molecular Biology
- Cell Death Research
Background:
- Chronic lymphocytic leukemia (CLL) cells exhibit resistance to apoptosis due to constitutively activated signaling pathways.
- Protein kinase C-delta (PKC-delta), a permanently activated kinase, is implicated as a downstream target of phosphatidylinositol-3 kinase in B-CLL.
Purpose of the Study:
- To investigate the therapeutic potential of inhibiting PKC-delta in CLL.
- To elucidate the downstream apoptotic mechanisms induced by PKC-delta inhibition.
Main Methods:
- Co-culture of bone marrow stromal and CLL cells to assess rottlerin's efficacy in a survival-factor-rich environment.
- Analysis of downstream events including mitochondrial membrane potential (Deltapsi) and expression of antiapoptotic proteins (Mcl-1, XIAP).
- Evaluation of rottlerin's effect on ZAP-70-expressing versus ZAP-70-negative CLL cells and its synergy with chemotherapeutics.
Main Results:
- The specific PKC-delta inhibitor rottlerin effectively induces apoptosis in CLL cells, irrespective of supportive survival factors.
- Rottlerin treatment leads to decreased expression of antiapoptotic proteins Mcl-1 and XIAP and a loss of mitochondrial membrane potential.
- ZAP-70-expressing CLL cells demonstrated significantly higher susceptibility to rottlerin-induced apoptosis.
- Rottlerin enhanced the cytotoxic effects of conventional chemotherapeutic drugs.
Conclusions:
- PKC-delta is a viable and promising therapeutic target for chronic lymphocytic leukemia.
- Targeting PKC-delta triggers both mitochondrial and non-mitochondrial apoptotic pathways, culminating in caspase activation.
- The combination of rottlerin with standard chemotherapy may offer a more effective treatment strategy for CLL patients.
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