NNMT Silencing Activates Tumor Suppressor PP2A, Inactivates Oncogenic STKs, and Inhibits Tumor Forming Ability.
Purpose:To identify potential molecular hubs that regulate oncogenic kinases and target them to improve treatment outcomes for glioblastoma patients.Experimental Design:Data mining of The Cancer Genome Atlas datasets identified nicotinamide-N-methyl transferase (NNMT) as a prognostic marker for glioblastoma, an enzyme linked to the reorganization of the methylome. We tested our hypothesis that NNMT plays a crucial role by modulating protein methylation, leading to inactivation of tumor suppressors and activation of oncogenes. Further experiments were performed to understand the underlying biochemical mechanisms using glioblastoma patient samples, established, primary, and isogenic cells.Results:We demonstrate that NNMT outcompetes leucine carboxyl methyl transferase 1 (LCMT1) for methyl transfer from principal methyl donor SAM in biological systems. Inhibiting NNMT increased the availability of methyl groups for LCMT1 to methylate PP2A, resulting in the inhibition of oncogenic serine/threonine kinases (STK). Further, NNMT inhibition retained the radiosensitizer nicotinamide and enhanced radiation sensitivity. We have provided the biochemical rationale of how NNMT plays a vital role in inhibiting tumor suppressor PP2A while concomitantly activating STKs.Conclusions:We report the intricate novel mechanism in which NNMT inhibits tumor suppressor PP2A by reorganizing the methylome both at epigenome and proteome levels and concomitantly activating prosurvival STKs. In glioblastoma tumors with NNMT expression, activation of PP2A can be accomplished by FDA approved perphenazine (PPZ), which is currently used to treat mood disorders such as schizophrenia, bipolar disorder, etc. This study forms a foundation for further glioblastoma clinical trials using PPZ with standard of care treatment.Clin Cancer Res; 23(9); 2325-34. ©2016 AACR.
Department of Radiation Oncology, James Cancer Hospital and Solove Research Institute, The Ohio State University College of Medicine and Comprehensive Cancer Center, Columbus, Ohio. Kamalakannan.palanichamy@osumc.edu.
Kamalakannan
K
Palanichamy
Department of Radiation Oncology, James Cancer Hospital and Solove Research Institute, The Ohio State University College of Medicine and Comprehensive Cancer Center, Columbus, Ohio.
Suman
S
Kanji
Department of Radiation Oncology, James Cancer Hospital and Solove Research Institute, The Ohio State University College of Medicine and Comprehensive Cancer Center, Columbus, Ohio.
Nicolaus
N
Gordon
Department of Radiation Oncology, James Cancer Hospital and Solove Research Institute, The Ohio State University College of Medicine and Comprehensive Cancer Center, Columbus, Ohio.
Krishnan
K
Thirumoorthy
Department of Radiation Oncology, James Cancer Hospital and Solove Research Institute, The Ohio State University College of Medicine and Comprehensive Cancer Center, Columbus, Ohio.
John R
JR
Jacob
Department of Radiation Oncology, James Cancer Hospital and Solove Research Institute, The Ohio State University College of Medicine and Comprehensive Cancer Center, Columbus, Ohio.
Kevin T
KT
Litzenberg
Department of Radiation Oncology, James Cancer Hospital and Solove Research Institute, The Ohio State University College of Medicine and Comprehensive Cancer Center, Columbus, Ohio.
Disha
D
Patel
Department of Radiation Oncology, James Cancer Hospital and Solove Research Institute, The Ohio State University College of Medicine and Comprehensive Cancer Center, Columbus, Ohio.
Arnab
A
Chakravarti
D000818:Animals; D002265:Carboxylic Ester Hydrolases / Q000235:genetics*; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects; D020868:Gene Silencing; D016147:Genes, Tumor Suppressor; D005909:Glioblastoma / Q000188:drug therapy* / Q000235:genetics / Q000473:pathology; D006801:Humans; D008745:Methylation / Q000187:drug effects; D051379:Mice; D050903:Nicotinamide N-Methyltransferase / Q000037:antagonists & inhibitors / Q000235:genetics*; D010546:Perphenazine / Q000008:administration & dosage*; D011497:Protein O-Methyltransferase / Q000235:genetics; D017346:Protein Serine-Threonine Kinases / Q000235:genetics
C524761:LCMT1 protein, human; D011497:Protein O-Methyltransferase; C494406:NNMT protein, human; D050903:Nicotinamide N-Methyltransferase; D017346:Protein Serine-Threonine Kinases; D002265:Carboxylic Ester Hydrolases; C119357:protein phosphatase methylesterase-1; D010546:Perphenazine