Overactivated neddylation pathway as a therapeutic target in lung cancer

Lihui Li; Mingsong Wang; Guangyang Yu; Ping Chen; Hui Li; Dongping Wei; Ji Zhu; Li Xie; Huixun Jia; Jieyi Shi; Chunjie Li; Wantong Yao; Yanchun Wang; Qiang Gao; Lak Shin Jeong; Hyuk Woo Lee; Jinha Yu; Fengqing Hu; Ju Mei; Ping Wang; Yiwei Chu; Hui Qi; Meng Yang; Ziming Dong; Yi Sun; Robert M Hoffman; Lijun Jia

doi:10.1093/jnci/dju083

Overactivated neddylation pathway as a therapeutic target in lung cancer

J Natl Cancer Inst. 2014 May 22;106(6):dju083. doi: 10.1093/jnci/dju083. Print 2014 Jun.

Authors

Lihui Li¹, Mingsong Wang¹, Guangyang Yu¹, Ping Chen¹, Hui Li¹, Dongping Wei¹, Ji Zhu¹, Li Xie¹, Huixun Jia¹, Jieyi Shi¹, Chunjie Li¹, Wantong Yao¹, Yanchun Wang¹, Qiang Gao¹, Lak Shin Jeong¹, Hyuk Woo Lee¹, Jinha Yu¹, Fengqing Hu¹, Ju Mei¹, Ping Wang¹, Yiwei Chu¹, Hui Qi¹, Meng Yang¹, Ziming Dong¹, Yi Sun¹, Robert M Hoffman¹, Lijun Jia²

Affiliations

¹ Affiliations of authors: Cancer Institute, Fudan University Shanghai Cancer Center (LL, GY, PC, DW, CL, WY, YW, LJ), Department of Oncology, Shanghai Medical College (LL, GY, PC, DW, CL, WY, YW, LJ), Department of Immunology, School of Basic Medical Sciences (LL, GY, CL, YW, YC, LJ), Clinical Statistics Center, Department of Radiation Oncology, Fudan University Shanghai Cancer Center (JZ, LX, HJ), and Liver Cancer Institute, Zhongshan Hospital (JS, QG), Fudan University, Shanghai, 200032, China; Department of Thoracic Cardiovascular Surgery, Xinhua Hospital of Shanghai Jiaotong University School of Medicine, Shanghai, China (MW, FH, JM); College of Basic Medical Sciences, Zhengzhou University, Zhengzhou, China (PC, ZD); Shanghai Key Laboratory of Regulatory Biology, Institute of Biomedical Sciences and School of Life Sciences, East China Normal University, Shanghai, China (HL, PW); College of Pharmacy, Seoul National University, Seoul, Korea (LSJ); College of Pharmacy, Ewha Womans University, Seoul, Korea (LSJ, HWL, JY); AntiCancer Biotech Beijing Co. Ltd., Beijing, China (HQ, MY); Division of Radiation and Cancer Biology, Department of Radiation Oncology, University of Michigan, Ann Arbor, MI (YS); Department of Surgery, University of California, San Diego, CA (RMH); AntiCancer, Inc., San Diego, CA (RMH, MY).
² Affiliations of authors: Cancer Institute, Fudan University Shanghai Cancer Center (LL, GY, PC, DW, CL, WY, YW, LJ), Department of Oncology, Shanghai Medical College (LL, GY, PC, DW, CL, WY, YW, LJ), Department of Immunology, School of Basic Medical Sciences (LL, GY, CL, YW, YC, LJ), Clinical Statistics Center, Department of Radiation Oncology, Fudan University Shanghai Cancer Center (JZ, LX, HJ), and Liver Cancer Institute, Zhongshan Hospital (JS, QG), Fudan University, Shanghai, 200032, China; Department of Thoracic Cardiovascular Surgery, Xinhua Hospital of Shanghai Jiaotong University School of Medicine, Shanghai, China (MW, FH, JM); College of Basic Medical Sciences, Zhengzhou University, Zhengzhou, China (PC, ZD); Shanghai Key Laboratory of Regulatory Biology, Institute of Biomedical Sciences and School of Life Sciences, East China Normal University, Shanghai, China (HL, PW); College of Pharmacy, Seoul National University, Seoul, Korea (LSJ); College of Pharmacy, Ewha Womans University, Seoul, Korea (LSJ, HWL, JY); AntiCancer Biotech Beijing Co. Ltd., Beijing, China (HQ, MY); Division of Radiation and Cancer Biology, Department of Radiation Oncology, University of Michigan, Ann Arbor, MI (YS); Department of Surgery, University of California, San Diego, CA (RMH); AntiCancer, Inc., San Diego, CA (RMH, MY). ljjia@fudan.edu.cn.

PMID: 24853380
DOI: 10.1093/jnci/dju083

Abstract

Background: A number of oncoproteins and tumor suppressors are known to be neddylated, but whether the neddylation pathway is entirely activated in human cancer remains unexplored.

Methods: NEDD8-activating enzyme (NAE) (E1) and NEDD8-conjugating enzyme (E2) expression and global-protein neddylation were examined by immunohistochemistry, immunoblotting, and real-time polymerase chain reaction analysis. Cell proliferation, clonogenic survival, migration, and motility in vitro, as well as tumor formation and metastasis in vivo, were determined upon neddylation inhibition by MLN4924, an investigational NEDD8-activating enzyme inhibitor. Survival was analyzed with Kaplan-Meier methods and compared by the log-rank test. All statistical tests were two-sided.

Results: The entire neddylation pathway, including NEDD8-activating enzyme E1, NEDD8-conjugating enzyme E2, and global-protein neddylation, is overactivated in both lung adenocarcinoma and squamous-cell carcinoma. Compared with lung adenocarcinoma patients with low expression, those with high expression had worse overall survival (NEDD8-activating enzyme E1 subunit 1 [NAE1]: hazard ratio [HR] = 2.07, 95% confidence interval [CI] = 0.95 to 4.52, P = .07; ubiquitin-conjugating enzyme E2M (UBC12): HR = 13.26, 95% CI = 1.77 to 99.35, P = .01; global protein neddylation: HR = 3.74, 95% CI = 1.65 to 8.47, P = .002). Moreover, inhibition of neddylation by the NAE inhibitor MLN4924 statistically significantly suppressed proliferation, survival, migration, and motility of lung cancer cells in vitro and tumor formation and metastasis in vivo. At the molecular level, MLN4924 inactivated Cullin-RING E3 ligases, led to accumulation of tumor-suppressive Cullin-RING E3 ligase substrates and induced phorbol-12-myristate-13-acetate-induced protein 1 (NOXA)-dependent apoptosis or cellular senescence.

Conclusions: Our study highlights the overactivated neddylation pathway in lung cancer development and as a promising therapeutic target.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Adenocarcinoma / drug therapy
Adenocarcinoma / enzymology
Adenocarcinoma of Lung
Antineoplastic Agents / pharmacology*
Carcinoma, Squamous Cell / drug therapy
Carcinoma, Squamous Cell / enzymology
Cell Line, Tumor
Cell Movement / drug effects
Cell Proliferation / drug effects
Cell Survival / drug effects
Cyclopentanes / pharmacology*
Flow Cytometry
Gene Expression Regulation, Enzymologic / drug effects
Gene Expression Regulation, Neoplastic / drug effects
Humans
Immunoblotting
Immunohistochemistry
Lung Neoplasms / drug therapy*
Lung Neoplasms / enzymology*
Lung Neoplasms / pathology
Molecular Targeted Therapy / methods*
NEDD8 Protein
Pyrimidines / pharmacology*
Real-Time Polymerase Chain Reaction
Signal Transduction / drug effects*
Tumor Stem Cell Assay
Ubiquitins / antagonists & inhibitors*
Ubiquitins / metabolism*

Substances

Antineoplastic Agents
Cyclopentanes
NEDD8 Protein
NEDD8 protein, human
Pyrimidines
Ubiquitins
pevonedistat