The multifunctional oncoprotein MDM2 plays critical roles in cancer initiation, progression and the development of resistance to therapy. We studied pediatric cancer patients, including those with acute lymphoblastic leukemia (ALL) and neuroblastoma (NB), finding that patients with a poor prognosis commonly had cancer cells that expressed constitutively high levels of MDM2. An important function of MDM2 is the ubiquitination (as an E3 ligase) and degradation of the tumor suppressor p53, due to interactions with DAXX and HAUSP. Interestingly, disruption of these interactions can result in self-ubiquitination of MDM2 and p53 activation. Thus, we believe targeting MDM2 by disrupting MDM2-DAXX-HAUSP interactions is an innovative approach to develop new therapeutics for MDM2-overexpressing cancer patients. Berberine (BBR), an isoquinoline alkaloid derived from a traditional Chinese medicinal herb, has been shown to have anti-proliferative and pro-apoptotic effects on human cancer. Our preliminary data supports a previously unrecognized mechanism of action for BBR: It downregulates MDM2 by inhibiting DAXX, disrupting MDM2-DAXX-HAUSP interactions. In contrast to the conventional chemotherapeutic drug doxorubicin (Dox) that induces prior p53 activation and a subsequent upregulation of MDM2, we discovered that BBR strongly induced persistent downregulation of MDM2, followed by a steady-state activation of p53, resulting in potent apoptosis of MDM2-overexpressing cancer cells, including those that are Dox-resistant. This proposal is designed to test our central hypothesis that regulation of MDM2 by DAXX inhibition is the major mechanism by which BBR differs from Dox in exerting its anti-cancer effect. The project's long term objectives are: characterization of the critical role BBR plays in the downregulation of MDM2 to overcome chemoresistance and determination of BBR's potential as a therapeutic reagent for refractory cancer patients.
The specific aims of this study are: 1) To fully elucidate the cellular/molecular mechanisms by which BBR downregulates MDM2;2) To establish that downregulation of MDM2 is the primary mechanism by which BBR exerts its anticancer effects;and 3) To assess the ability of BBR to reverse chemoresistance and its potential as a treatment for refractory cancer patients. Successful completion of these specific aims will generate knowledge with respect to the mechanisms of action for BBR prior to future clinical trials of this natural product in pediatric cancer patients, should introduce BBR as a highly useful new drug candidate for overcoming chemoresistance, and also will provide a basic framework for the rational design of other therapeutic approaches targeting the DAXX-MDM2 pathway.

Public Health Relevance

We have found that BBR specifically and strongly inhibits MDM2 expression and induces activation of p53, which results in potent apoptosis of wt-p53/MDM2-overexpressing cancer cells, including those that were resistant to a conventional chemotherapeutic drug. In this study, we will elucidate the molecular mechanism by which BBR downregulates MDM2, which we have reason to believe involves inhibition of DAXX and eventual initiation of apoptosis through a previously unutilized drug development corridor. Importantly, we will investigate whether the natural product BBR could be used as an effective therapeutic reagent in the treatment of pediatric and refractory cancer patient populations.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Project (R01)
Project #
5R01CA143107-03
Application #
8204584
Study Section
Drug Discovery and Molecular Pharmacology Study Section (DMP)
Program Officer
Fu, Yali
Project Start
2010-01-01
Project End
2014-12-31
Budget Start
2012-01-01
Budget End
2012-12-31
Support Year
3
Fiscal Year
2012
Total Cost
$311,976
Indirect Cost
$110,701
Name
Emory University
Department
Pediatrics
Type
Schools of Medicine
DUNS #
066469933
City
Atlanta
State
GA
Country
United States
Zip Code
30322
Zhang, Hailong; Gu, Lubing; Liu, Tao et al. (2014) Inhibition of MDM2 by nilotinib contributes to cytotoxicity in both Philadelphia-positive and negative acute lymphoblastic leukemia. PLoS One 9:e100960
Li, Jiansha; Gu, Lubing; Zhang, Hailong et al. (2013) Berberine represses DAXX gene transcription and induces cancer cell apoptosis. Lab Invest 93:354-64
Huang, Mei; Zhang, Hailong; Liu, Tao et al. (2013) Triptolide inhibits MDM2 and induces apoptosis in acute lymphoblastic leukemia cells through a p53-independent pathway. Mol Cancer Ther 12:184-94
Zhang, Hailong; He, Jing; Li, Jiansha et al. (2013) Methylation of RASSF1A gene promoter is regulated by p53 and DAXX. FASEB J 27:232-42
Gu, L; Zhang, H; He, J et al. (2012) MDM2 regulates MYCN mRNA stabilization and translation in human neuroblastoma cells. Oncogene 31:1342-53
He, Jing; Gu, Lubing; Zhang, Hailong et al. (2011) Crosstalk between MYCN and MDM2-p53 signal pathways regulates tumor cell growth and apoptosis in neuroblastoma. Cell Cycle 10:2994-3002
Yang, Lin; Gu, Lubing; Li, Zhuoya et al. (2010) Translation of TRAF1 is regulated by IRES-dependent mechanism and stimulated by vincristine. Nucleic Acids Res 38:4503-13
Zhang, Xiaoling; Gu, Lubing; Li, Jiansha et al. (2010) Degradation of MDM2 by the interaction between berberine and DAXX leads to potent apoptosis in MDM2-overexpressing cancer cells. Cancer Res 70:9895-904