Novel Cancer Therapeutics with Allosteric Modulation of the Mitochondrial C-Raf–DAPK Complex by Raf Inhibitor Combination Therapy
| Autor: | Ming-Rong Chen, Hwei-Jen Lee, Dah-Shyong Yu, Jar-Yi Ho, Tai-Lung Cha, Shih-Ming Huang, Yi-Ta Tsai, Shou-Hung Tang, Yu-Chi Chen, Sun-Yran Chang, Hui Kuan Lin, Sheng-Tang Wu, Victor C. Lin, Pei-Wen Hsiao, Cheng-Ping Yu, Chung-Chih Lin, Mei-Jen Chuang, Guang-Huan Sun |
|---|---|
| Rok vydání: | 2015 |
| Předmět: |
Male
Niacinamide Sorafenib Cancer Research Indoles Apoptosis Mitochondrion Biology Disease-Free Survival Gene Knockout Techniques Mice Phenols Cell Line Tumor medicine Animals Humans c-Raf Phosphorylation Aged Phenylurea Compounds Cancer Drug Synergism Protein phosphatase 2 Middle Aged medicine.disease Xenograft Model Antitumor Assays Kidney Neoplasms Mitochondria Cell biology Proto-Oncogene Proteins c-raf Death-Associated Protein Kinases Oncology Cancer cell Female Reactive Oxygen Species Signal Transduction medicine.drug |
| Zdroj: | Cancer Research. 75:3568-3582 |
| ISSN: | 1538-7445 0008-5472 |
| DOI: | 10.1158/0008-5472.can-14-3264 |
| Popis: | Mitochondria are the powerhouses of cells. Mitochondrial C-Raf is a potential cancer therapeutic target, as it regulates mitochondrial function and is localized to the mitochondria by its N-terminal domain. However, Raf inhibitor monotherapy can induce S338 phosphorylation of C-Raf (pC-RafS338) and impede therapy. This study identified the interaction of C-Raf with S308 phosphorylated DAPK (pDAPKS308), which together became colocalized in the mitochondria to facilitate mitochondrial remodeling. Combined use of the Raf inhibitors sorafenib and GW5074 had synergistic anticancer effects in vitro and in vivo, but targeted mitochondrial function, rather than the canonical Raf signaling pathway. C-Raf depletion in knockout MEFC-Raf−/− or siRNA knockdown ACHN renal cancer cells abrogated the cytotoxicity of combination therapy. Crystal structure simulation showed that GW5074 bound to C-Raf and induced a C-Raf conformational change that enhanced sorafenib-binding affinity. In the presence of pDAPKS308, this drug–target interaction compromised the mitochondrial targeting effect of the N-terminal domain of C-Raf, which induced two-hit damages to cancer cells. First, combination therapy facilitated pC-RafS338 and pDAPKS308 translocation from mitochondria to cytoplasm, leading to mitochondrial dysfunction and reactive oxygen species (ROS) generation. Second, ROS facilitated PP2A-mediated dephosphorylation of pDAPKS308 to DAPK. PP2A then dissociated from the C-Raf–DAPK complex and induced profound cancer cell death. Increased pDAPKS308 modification was also observed in renal cancer tissues, which correlated with poor disease-free survival and poor overall survival in renal cancer patients. Besides mediating the anticancer effect, pDAPKS308 may serve as a predictive biomarker for Raf inhibitors combination therapy, suggesting an ideal preclinical model that is worthy of clinical translation. Cancer Res; 75(17); 3568–82. ©2015 AACR. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|