DOCSLIB.ORG

Proteomics Identifies Neddylation As a Potential Therapy Target in Small

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Proteomics Identifies Neddylation As a Potential Therapy Target in Small Oncogene (2019) 38:6881–6897 https://doi.org/10.1038/s41388-019-0938-8 ARTICLE Proteomics identifies neddylation as a potential therapy target in small intestinal neuroendocrine tumors 1 2 1,3 1,4 1,4 1 Omid Fotouhi ● Hanna Kjellin ● C. Christofer Juhlin ● Yanbo Pan ● Mattias Vesterlund ● Mehran Ghaderi ● 5 4 6 7 2,8 Abdelhamid Yousef ● Hillevi Andersson-Sand ● Pedram Kharaziha ● Stefano Caramuta ● Magnus Kjellman ● 2,8 1 1,4 Jan Zedenius ● Catharina Larsson ● Lukas M. Orre Received: 26 October 2018 / Revised: 30 April 2019 / Accepted: 15 May 2019 / Published online: 12 August 2019 © The Author(s) 2019. This article is published with open access Abstract Patients with small intestinal neuroendocrine tumors (SI-NETs) frequently develop spread disease; however, the underlying molecular mechanisms of disease progression are not known and effective preventive treatment strategies are lacking. Here, protein expression profiling was performed by HiRIEF-LC-MS in 14 primary SI-NETs from patients with and without liver metastases detected at the time of surgery and initial treatment. Among differentially expressed proteins, overexpression of the ubiquitin-like protein NEDD8 was identified in samples from patients with liver metastasis. Further, NEDD8 correlation 1234567890();,: 1234567890();,: analysis indicated co-expression with RBX1, a key component in cullin-RING ubiquitin ligases (CRLs). In vitro inhibition of neddylation with the therapeutic agent pevonedistat (MLN4924) resulted in a dramatic decrease of proliferation in SI- NET cell lines. Subsequent mass spectrometry-based proteomics analysis of pevonedistat effects and effects of the proteasome inhibitor bortezomib revealed stabilization of multiple targets of CRLs including p27, an established tumor suppressor in SI-NET. Silencing of NEDD8 and RBX1 using siRNA resulted in a stabilization of p27, suggesting that the cellular levels of NEDD8 and RBX1 affect CRL activity. Inhibition of CRL activity, by either NEDD8/RBX1 silencing or pevonedistat treatment of cells resulted in induction of apoptosis that could be partially rescued by siRNA-based silencing of p27. Differential expression of both p27 and NEDD8 was confirmed in a second cohort of SI-NET using immunohistochemistry. Collectively, these findings suggest a role for CRLs and the ubiquitin proteasome system in suppression of p27 in SI-NET, and inhibition of neddylation as a putative therapeutic strategy in SI-NET. Introduction Small intestinal neuroendocrine tumor (SI-NET) is a rare disease with an incidence of around 1 per 100,000, but still the most common small intestinal malignancy [1]. SI-NETs fi Supplementary information The online version of this article (https:// develop from enterochromaf n cells, are positive for neu- doi.org/10.1038/s41388-019-0938-8) contains supplementary roendocrine markers, and the diagnosis is based on material, which is available to authorized users. * Catharina Larsson 4 SciLifeLab, Stockholm, Sweden [email protected] 5 * Lukas M. Orre Department of Pharmacology, Cambridge University, [email protected] Cambridge, UK 6 Department of Clinical Genetics, Linköping University Hospital, 1 Department of Oncology-Pathology, Karolinska Institutet, Linköping, Sweden Stockholm, Sweden 7 Uppsala Clinical Research Center, Uppsala University, 2 Department of Molecular Medicine and Surgery, Karolinska Uppsala, Sweden Institutet, Stockholm, Sweden 8 Department of Breast, Endocrine Tumours and Sarcoma, 3 Department of Pathology and Cytology, Karolinska University Karolinska University Hospital Solna, Stockholm, Sweden Hospital Solna, Stockholm, Sweden 6882 O. Fotouhi et al. histopathological examination [2]. The patients typically Importantly, it has been shown that substrate recognition present with one or more small primary tumors, often and activity of CRLs (such as SCFSkp2) is strongly depen- together with mesenteric lymph node metastases, and later dent on neddylation of the Cullin subunit [13]. The ned- development of distant metastases frequently to the liver. dylation process is similar to ubiquitination and includes Although the tumors are commonly slow growing based on E1, E2, and E3 enzymes that catalyze conjugation of target a low Ki-67 proliferation index, the clinical picture suggests proteins with the ubiquitin-like protein NEDD8 (neural that tumor cell disconnection and spreading occur early in precursor cell expressed, developmentally downregulated 8) the tumor development [3]. [14]. Cullin neddylation results in a conformational change The primary treatment of SI-NET is surgical. In addition, of the CRL complex, eliminating the binding site for the the frequent expression of somatostatin receptors in SI- CRL inhibitor Cand1 and, at the same time, providing a NETs has led to the development and successful application more open structure favoring polyubiquitination [15]. of somatostatin analog treatment. Although this treatment Here we performed in-depth proteomics profiling of 14 may improve symptoms and also in some patients delay primary SI-NET tumors with focus on proteome-level tumor progression, additional therapeutic strategies need to differences between tumors from patients with and without be identified and developed for patients with metastatic liver metastasis. This clinical proteomics analysis impli- disease. cated differences in the ubiquitination machinery, with SI-NETs are characterized by a few recurrent genetic higher expression of NEDD8 and RBX1 in tumors from aberrations such as copy number loss of chromosome 18, patients with liver metastasis. To further investigate the which is seen in >70% of the cases [4]. The high frequency importance of these findings, inhibition of neddylation was of this event spawned a search for putative tumor supressors performed in SI-NET cell lines resulting in reduced pro- located on chromosome 18 in SI-NET, resulting in the liferation and induction of apoptosis. Proteomics profiling identification of DCC (deleted in colorectal cancer) as the so of the effects of neddylation inhibition in two SI-NET cell far best candidate [5]. Even though SI-NETs are in general lines resulted in identification of multiple upregulated mutationally quiet, the application of genome-wide proteins known to be substrates of CRLs including p27. sequencing identified CDKN1B as a recurrently mutated Our findings thus suggest deregulation of CRLs as well as gene in SI-NET with a frequency of 8% [6]. In addition, a an alternative route for CDKN1B/p27 inactivation in SI- broad molecular subtyping of SI-NET was recently reported NET with possible connections to metastatic spread and where genomic, epigenomic, and transcriptomic profiles therapy. were used to suggest three molecular subtypes of SI-NET with differences in progression-free survival [7]. However, the molecular background of SI-NET development, specific Results oncogenic drivers, and mechanisms of metastatic spread still remain largely unknown. In addition, no study so far Proteomics profiling of primary SI-NETs indicate has integrated proteome-level information in the analysis of NEDD8 and RBX1 overexpression in patients with SI-NET from primary tumors. liver metastases CDKN1B encodes p27, a bona-fide tumor suppressor that controls cell cycle entry through inhibition of cyclin/cyclin In order to investigate the proteomic landscape of SI-NET, dependent kinase (CDK) activity. It has been thoroughly and especially to identify proteins connected to disease demonstrated that the CDK inhibitor p27 is regulated pri- progression and metastatic spread, we performed in-depth marily by ubiquitination and subsequent proteasomal proteomics profiling of 14 primary SI-NETs of which 7 degradation [8], and that the ubiquitin E3-ligase responsible presented with liver metastases, while 7 were without any for the ubiquitination is the SCFSkp2 Cullin Ring Ligase detectable liver metastasis at the time of surgery (Supple- (CRL) complex [9]. The SCFSkp2 complex was later shown mentary Table S1). High-resolution isolectric focusing to include Skp1, Cullin-1, Rbx1, and Skp2 as the core liquid chromatography–tandem mass spectrometry (HiR- complex components [10]. Increased degradation of p27 in IEF-LC-MS)-based proteomics combined with relative cancer has also been firmly established beginning with a quantification of proteins between samples (iTRAQ-label- seminal study where increased p27-specific proteolytic ing) resulted in the identification and quantification of 6775 activity was coupled to low abundance of p27, which in turn proteins across all 14 SI-NET samples (Supplementary was demonstrated as a powerful negative prognostic factor Table S2). Unsupervised analysis using principal compo- in colorectal cancer [11]. An overwhelming abundance of nent analysis of the 14 samples revealed no major batch reports since then connects overexpression of Skp2 (the F- effects between the 2 separate quantitative proteomics box protein that recognize p27) with loss of p27 protein and experiments (2 iTRAQ 8-plex kits), but indicated 2 poten- poor prognosis in a long list of different cancer types [12]. tial outliers in the analysis (sample Screen-8 and -12; Proteomics identifies neddylation as a potential therapy target in small intestinal neuroendocrine tumors 6883 Supplementary Fig. S1a, b). Unsupervised Spearman’s correlation across the different cancer types was RBX1 (Fig. hierarchical clustering, however, did not indicate the two 1e, f). These results indicate a co-regulation of NEDD8 and samples as outliers. Instead, this clustering resulted
Load more

Recommended publications
  • Blockage of Neddylation Modification Stimulates Tumor Sphere Formation
    Blockage of neddylation modification stimulates tumor PNAS PLUS sphere formation in vitro and stem cell differentiation and wound healing in vivo Xiaochen Zhoua,b,1, Mingjia Tanb,1, Mukesh K. Nyatib, Yongchao Zhaoc,d, Gongxian Wanga,2, and Yi Sunb,c,e,2 aDepartment of Urology, The First Affiliated Hospital of Nanchang University, Nanchang, Jiangxi 330006, China; bDivision of Radiation and Cancer Biology, Department of Radiation Oncology, University of Michigan, Ann Arbor, MI 48109; cInstitute of Translational Medicine, Zhejiang University School of Medicine, Hangzhou, Zhejiang 310029, China; dKey Laboratory of Combined Multi-Organ Transplantation, Ministry of Public Health, First Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou 310058, China; and eCollaborative Innovation Center for Diagnosis and Treatment of Infectious Diseases, Zhejiang University, Hangzhou 310058, China Edited by Vishva M. Dixit, Genentech, San Francisco, CA, and approved March 10, 2016 (received for review November 13, 2015) MLN4924, also known as pevonedistat, is the first-in-class inhibitor acting alone or in combination with current chemotherapy of NEDD8-activating enzyme, which blocks the entire neddylation and/or radiation (6, 11). One of the seven clinical trials of MLN4924 modification of proteins. Previous preclinical studies and current (NCT00911066) was published recently, concluding a modest effect clinical trials have been exclusively focused on its anticancer property. of MLN4924 against acute myeloid leukemia (AML) (12). Unexpectedly, we show here, to our knowledge for the first time, To further elucidate the role of blocking neddylation in cancer that MLN4924, when applied at nanomolar concentrations, signif- treatment, we thought to study the effect of MLN4924 on cancer icantly stimulates in vitro tumor sphere formation and in vivo stem cells (CSCs) or tumor-initiating cells (TICs), a small group tumorigenesis and differentiation of human cancer cells and mouse of tumor cells with stem cell properties that have been claimed to embryonic stem cells.
    [Show full text]
  • The Anti-Tumor Activity of the NEDD8 Inhibitor Pevonedistat in Neuroblastoma
    International Journal of Molecular Sciences Article The Anti-Tumor Activity of the NEDD8 Inhibitor Pevonedistat in Neuroblastoma Jennifer H. Foster 1,* , Eveline Barbieri 1, Linna Zhang 1, Kathleen A. Scorsone 1, Myrthala Moreno-Smith 1, Peter Zage 2,3 and Terzah M. Horton 1,* 1 Texas Children’s Cancer and Hematology Centers, Department of Pediatrics, Section of Hematology and Oncology, Baylor College of Medicine, Houston, TX 77030, USA; [email protected] (E.B.); [email protected] (L.Z.); [email protected] (K.A.S.); [email protected] (M.M.-S.) 2 Department of Pediatrics, Division of Hematology-Oncology, University of California San Diego, La Jolla, CA 92024, USA; [email protected] 3 Peckham Center for Cancer and Blood Disorders, Rady Children’s Hospital, San Diego, CA 92123, USA * Correspondence: [email protected] (J.H.F.); [email protected] (T.M.H.); Tel.: +1-832-824-4646 (J.H.F.); +1-832-824-4269 (T.M.H.) Abstract: Pevonedistat is a neddylation inhibitor that blocks proteasomal degradation of cullin–RING ligase (CRL) proteins involved in the degradation of short-lived regulatory proteins, including those involved with cell-cycle regulation. We determined the sensitivity and mechanism of action of pevonedistat cytotoxicity in neuroblastoma. Pevonedistat cytotoxicity was assessed using cell viability assays and apoptosis. We examined mechanisms of action using flow cytometry, bromod- eoxyuridine (BrDU) and immunoblots. Orthotopic mouse xenografts of human neuroblastoma were generated to assess in vivo anti-tumor activity. Neuroblastoma cell lines were very sensitive to pevonedistat (IC50 136–400 nM). The mechanism of pevonedistat cytotoxicity depended on p53 Citation: Foster, J.H.; Barbieri, E.; status.
    [Show full text]
  • Neddylation: a Novel Modulator of the Tumor Microenvironment Lisha Zhou1,2*†, Yanyu Jiang3†, Qin Luo1, Lihui Li1 and Lijun Jia1*
    Zhou et al. Molecular Cancer (2019) 18:77 https://doi.org/10.1186/s12943-019-0979-1 REVIEW Open Access Neddylation: a novel modulator of the tumor microenvironment Lisha Zhou1,2*†, Yanyu Jiang3†, Qin Luo1, Lihui Li1 and Lijun Jia1* Abstract Neddylation, a post-translational modification that adds an ubiquitin-like protein NEDD8 to substrate proteins, modulates many important biological processes, including tumorigenesis. The process of protein neddylation is overactivated in multiple human cancers, providing a sound rationale for its targeting as an attractive anticancer therapeutic strategy, as evidence by the development of NEDD8-activating enzyme (NAE) inhibitor MLN4924 (also known as pevonedistat). Neddylation inhibition by MLN4924 exerts significantly anticancer effects mainly by triggering cell apoptosis, senescence and autophagy. Recently, intensive evidences reveal that inhibition of neddylation pathway, in addition to acting on tumor cells, also influences the functions of multiple important components of the tumor microenvironment (TME), including immune cells, cancer-associated fibroblasts (CAFs), cancer-associated endothelial cells (CAEs) and some factors, all of which are crucial for tumorigenesis. Here, we briefly summarize the latest progresses in this field to clarify the roles of neddylation in the TME, thus highlighting the overall anticancer efficacy of neddylaton inhibition. Keywords: Neddylation, Tumor microenvironment, Tumor-derived factors, Cancer-associated fibroblasts, Cancer- associated endothelial cells, Immune cells Introduction Overall, binding of NEDD8 molecules to target proteins Neddylation is a reversible covalent conjugation of an can affect their stability, subcellular localization, conform- ubiquitin-like molecule NEDD8 (neuronal precursor ation and function [4]. The best-characterized substrates cell-expressed developmentally down-regulated protein of neddylation are the cullin subunits of Cullin-RING li- 8) to a lysine residue of the substrate protein [1, 2].
    [Show full text]
  • Phase Ib Study of Pevonedistat, a NEDD8-Activating Enzyme Inhibitor
    Investigational New Drugs (2019) 37:87–97 https://doi.org/10.1007/s10637-018-0610-0 PHASE I STUDIES Phase Ib study of pevonedistat, a NEDD8-activating enzyme inhibitor, in combination with docetaxel, carboplatin and paclitaxel, or gemcitabine, in patients with advanced solid tumors A Craig Lockhart1 & Todd M. Bauer2 & Charu Aggarwal3 & Carrie B. Lee4 & RDonaldHarvey5 & Roger B. Cohen6 & Farhad Sedarati7 & Tsz Keung Nip8 & Hélène Faessel9 & Ajeeta B. Dash10 & Bruce J. Dezube7 & Douglas V. Faller7 & Afshin Dowlati11 Received: 1 March 2018 /Accepted: 11 May 2018 /Published online: 21 May 2018 # The Author(s) 2018 Summary Purpose This phase Ib study (NCT01862328) evaluated the maximum tolerated dose (MTD), safety, and efficacy of pevonedistat in combination with standard-of-care chemotherapies in patients with solid tumors. Methods Patients received pevonedistat with docetaxel (arm 1, n = 22), carboplatin plus paclitaxel (arm 2, n = 26), or gemcitabine (arm 3, n = 10) in 21-days (arms 1 and 2) or 28-days (arm 3) cycles. A lead-in cohort (arm 2a, n = 6) determined the arm 2 carboplatin dose. Dose escalation proceeded via continual modified reassessment. Results Pevonedistat MTD was 25 mg/m2 (arm 1) or 20 mg/m2 (arm 2); arm 3 was discontinued due to poor tolerability. Fifteen (23%) patients experienced dose-limiting toxicities during cycle 1 (grade ≥3 liver enzyme elevations, febrile neutropenia, and thrombocytopenia), managed with dose holds or reductions. Drug-related adverse events (AEs) occurred in 95% of patients. Most common AEs included fatigue (56%) and nausea (50%). One drug-related death occurred in arm 3 (febrile neutropenia). Pevonedistat exposure increased when co-administered with carboplatin plus paclitaxel; no obvious changes were observed when co-administered with docetaxel or gemcitabine.
    [Show full text]
  • Pevonedistat (MLN4924), a First-In-Class NEDD8-Activating Enzyme Inhibitor, in Patients with Acute Myeloid Leukaemia and Myelodysplastic Syndromes: a Phase 1 Study
    research paper Pevonedistat (MLN4924), a First-in-Class NEDD8-activating enzyme inhibitor, in patients with acute myeloid leukaemia and myelodysplastic syndromes: a phase 1 study Ronan T. Swords,1 Harry P. Erba,2 Summary Daniel J. DeAngelo,3 Dale L. Bixby,2 This trial was conducted to determine the dose-limiting toxicities (DLTs) Jessica K. Altman,4 Michael Maris,5 Zhaowei Hua,6 Stephen J. Blakemore,6 and maximum tolerated dose (MTD) of the first in class NEDD8-activating Helene Faessel,6 Farhad Sedarati,6 enzyme (NAE) inhibitor, pevonedistat, and to investigate pevonedistat Bruce J. Dezube,6 Francis J. Giles4 and pharmacokinetics and pharmacodynamics in patients with acute myeloid Bruno C. Medeiros7 leukaemia (AML) and myelodysplastic syndromes (MDS). Pevonedistat was 1Leukemia Program, Sylvester Comprehensive administered via a 60-min intravenous infusion on days 1, 3 and 5 (sche- Cancer Center, Miami, FL, 2Division of Hema- dule A, n = 27), or days 1, 4, 8 and 11 (schedule B, n = 26) every 21-days. tology/Oncology, University of Michigan, Ann Dose escalation proceeded using a standard ‘3 + 3’ design. Responses were 3 Arbor, MI, Department of Medical Oncology, assessed according to published guidelines. The MTD for schedules A and Dana-Farber Cancer Institute, Boston, MA, B were 59 and 83 mg/m2, respectively. On schedule A, hepatotoxicity was 4 Northwestern Medicine Developmental Thera- dose limiting. Multi-organ failure (MOF) was dose limiting on schedule B. peutics Institute, Northwestern University, The overall complete (CR) and partial (PR) response rate in patients trea- Chicago, IL, 5Colorado Blood Cancer Institute, ted at or below the MTD was 17% (4/23, 2 CRs, 2 PRs) for schedule A Denver, CO, 6Takeda Pharmaceuticals Interna- tional Co., Cambridge, MA, and 7Division of and 10% (2/19, 2 PRs) for schedule B.
    [Show full text]
  • Study Protocol and Statistical Analysis Plan
    STUDY00008815 Approval: 4/1/2019 Protocol Title: PSCI 17-056 A Phase II Study of Pevonedistat (MLN4924, TAK924) and Azacitidine as Maintenance Therapy after Allogeneic Stem Cell Transplantation for Non-Remission Acute Myelogenous Leukemia Principal Investigator: Shin Mineishi, M.D. Professor and Director, Blood and Marrow Transplant Program 500 University Drive, CH46 Hershey, PA 17111 717-531-8678 (Office) 717-531-4969 (Fax) Email:[email protected] Site PIs Additional sites may be added in the future. Co-Investigators: David Claxton, M.D. Hong Zheng, M.D. Ph.D. Seema Naik, M.D. Muhammad Khawaja, M.D. Jason Liao, Ph.D. Co-Investigator/MD Coordinator Kevin Rakszawski, MD Fellow, Hematology & Oncology Email:[email protected] Version Date: Version 1.8.5, 07-26-2018 Clinicaltrials.gov Registration #: 03709576 Page 1 of 75 (V.1.8.5, 07/26/2018) STUDY00008815 Approval: 4/1/2019 Contents LIST OF ABBREVIATIONS AND GLOSSARY OF TERMS......................................................................................................6 1.0 Objectives .......................................................................................................................................................10 1.1. Primary Objective.................................................................................................................................................10 1.2. Secondary Objectives ...........................................................................................................................................10
    [Show full text]
  • Clinical, Laboratory Advances Highlighted at 2018 Mantle Cell
    Summer 2018 Clinical, Laboratory Advances Highlighted at 2018 Mantle Cell Lymphoma Workshop convening leaders in the field across a wide range of research environments, funding innovative studies and creating needed resources. Programmatic efforts of the MCLC include MCL investigator research grants, establishment of an MCL cell bank that contains research tools available for conducting laboratory research, patient education programs and the Focus on Lymphoma app, and the biennial MCL Scientific Workshop. This report provides an overview of each presentation given at the LRF Scholars attend the 2018 MCL Workshop (L to R): 2015 Scholar Victor Yazbeck, MD Virginia Common- 13th MCL Scientific Workshop and identifies wealth University, Meghan Gutierrez, LRF CEO, 2016 Scholar Lapo Alinari, MD, PhD, The Ohio State University, key emergent themes. 2018 Scholar Shalin Kothari, MD Roswell Park Comprehensive Cancer Center, 2016 Scholar Connie Batlevi, MD, PhD, Memorial Sloan Kettering Cancer Center, and 2014 Scholars Jonathon Cohen, MD, Emory Universi- “The Lymphoma Research Foundation’s MCL ty and Anita Kumar, MD, Memorial Sloan Kettering Cancer Center. The 2018 Scholar class is profiled on pg. 3. Consortium and its Scientific Workshop have n April 26 and 27, 2018, nearly 100 deliberate communication between scien- been key contributors to the advancement of O lymphoma researchers gathered tists and clinicians across this wide scope our understanding of MCL both clinically and in Atlanta, Georgia, for the Lymphoma of research is unique to this forum and the biologically,” says Thomas M. Habermann, Research Foundation’s (LRF) 13th Mantle cross-pollination between research fields MD, of Mayo Clinic, Chair of the Foundation’s Cell Lymphoma (MCL) Scientific Workshop.
    [Show full text]
  • Neddylation, a Novel Paradigm in Liver Cancer
    Review Article Neddylation, a novel paradigm in liver cancer Teresa Cardoso Delgado1, Lúcia Barbier-Torres1,2, Imanol Zubiete-Franco1,3, Fernando Lopitz-Otsoa1, Marta Varela-Rey1, David Fernández-Ramos1, María-Luz Martínez-Chantar1 1CIC bioGUNE, Centro de Investigación Biomédica en Red de Enfermedades Hepáticas y Digestivas (CIBERehd), Derio, Bizkaia, Spain; 2Division of Gastroenterology, Cedars-Sinai Medical Center, Los Angeles, CA, USA; 3Cold Spring Harbor Laboratory, Cold Spring Harbor, New York, NY, USA Contributions: (I) Conception and design: TC Delgado, ML Martínez-Chantar; (II) Administrative support: None; (III) Provision of study materials or patients: None; (IV) Collection and assembly of data: None; (V) Data analysis and interpretation: None; (VI) Manuscript writing: All authors; (VII) Final approval of manuscript: All authors. Correspondence to: Teresa Cardoso Delgado. CIC bioGUNE, Ed. 801A Parque Tecnológico de Bizkaia, 48160 Derio, Bizkaia, Spain. Email: [email protected]. Abstract: Liver cancer is the sixth most prevailing cancer worldwide. Hepatocellular carcinoma (HCC), the most common form of primary liver cancer, has a rather heterogeneous pathogenesis making it highly refractive to current therapeutic approaches. Hence, HCC patients have a poor and gloomy prognosis making liver cancer the second leading cause of global cancer-related deaths. On this basis, a more global mechanism, such as post-translational modifications (PTMs) of proteins, may provide a valuable therapeutic approach for HCC clinical management by simultaneously regulating multiple disrupted signaling pathways. In the last years, the ubiquitin-like molecule NEDD8 (Neural precursor cell-expressed developmentally downregulated-8) conjugation pathway, neddylation, was shown to be aberrant in HCC patients with a significant positive correlation found among global levels of neddylation and poorer prognosis.
    [Show full text]
  • Neddylation Regulates Class Iia and III Histone Deacetylases to Mediate Myoblast Differentiation
    International Journal of Molecular Sciences Article Neddylation Regulates Class IIa and III Histone Deacetylases to Mediate Myoblast Differentiation Hongyi Zhou 1,* , Huabo Su 2 and Weiqin Chen 1 1 Department of Physiology, Medical College of Georgia at Augusta University, Augusta, GA 30912, USA; [email protected] 2 Vascular Biology Center, Medical College of Georgia at Augusta University, Augusta, GA 30912, USA; [email protected] * Correspondence: [email protected]; Tel.: +1-706-721-8779 Abstract: As the largest tissue in the body, skeletal muscle has multiple functions in movement and energy metabolism. Skeletal myogenesis is controlled by a transcriptional cascade including a set of muscle regulatory factors (MRFs) that includes Myogenic Differentiation 1 (MYOD1), Myocyte En- hancer Factor 2 (MEF2), and Myogenin (MYOG), which direct the fusion of myogenic myoblasts into multinucleated myotubes. Neddylation is a posttranslational modification that covalently conjugates ubiquitin-like NEDD8 (neural precursor cell expressed, developmentally downregulated 8) to protein targets. Inhibition of neddylation impairs muscle differentiation; however, the underlying molec- ular mechanisms remain less explored. Here, we report that neddylation is temporally regulated during myoblast differentiation. Inhibition of neddylation through pharmacological blockade using MLN4924 (Pevonedistat) or genetic deletion of NEDD8 Activating Enzyme E1 Subunit 1 (NAE1), a subunit of the E1 neddylation-activating enzyme, blocks terminal myoblast differentiation par- tially through repressing MYOG expression. Mechanistically, we found that neddylation deficiency enhances the mRNA and protein expressions of class IIa histone deacetylases 4 and 5 (HDAC4 Citation: Zhou, H.; Su, H.; Chen, W. and 5) and prevents the downregulation and nuclear export of class III HDAC (NAD-Dependent Neddylation Regulates Class IIa and Protein Deacetylase Sirtuin-1, SIRT1), all of which have been shown to repress MYOD1-mediated III Histone Deacetylases to Mediate MYOG transcriptional activation.
    [Show full text]
  • Neddylation and Anti-Tumor Immunity
    www.oncotarget.com Oncotarget, Advance Publications 2021 Neddylation and anti-tumor immunity Xiaoguang Wang1, Scott Best2 and Alexey V. Danilov1 1Department of Hematology and Hematopoietic Stem Cell Transplant, City of Hope National Medical Center, Duarte, CA, USA 2Molecular and Cellular Biology, University of Washington, Seattle, WA, USA Correspondence to: Alexey V. Danilov, email: [email protected] Keywords: neddylation; T cell; pevonedistat Received: June 10, 2021 Accepted: June 21, 2021 Published: Copyright: © 2021 Wang et al. This is an open access article distributed under the terms of the Creative Commons Attribution License (CC BY 3.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. ABSTRACT Contrary to chemotherapy, novel targeted therapies are associated with diverse immunomodulatory effects. Nedd8 is a small ubiquitin-like modifier that is involved in regulation of protein degradation. Neddylation is a promising target in cancer. Pevonedistat, a small molecule inhibitor of the Nedd8-activating enzyme, demonstrates pre-clinical activity in multiple tumor types. Recent studies have revealed that neddylation is important in immunity. We and others have shown that interfering with neddylation causes downstream immunomodulatory effects potentially leading to enhanced anti-tumor immunity. Thus, Nedd8 is a promising target in immuno-oncology. The ubiquitin-proteasome system (UPS) is of the CRL [6, 7]. Pevonedistat is a small molecule which a common pathway which controls degradation of covalently adducts with NEDD8, leading to effective unwanted or misfolded proteins in cells. Proteins targeted inhibition of NAE via competitive binding. Consequently, for degradation are labeled with ubiquitin chains by E3 NEDD8 is prevented from conjugation to CRLs, thus ubiquitin ligases [1].
    [Show full text]
  • Pevonedistat and Azacitidine Upregulate NOXA (PMAIP1) to Increase Sensitivity to Venetoclax in Preclinical Models of Acute Myeloid Leukemia by Dan Cojocari, Brianna N
    Pevonedistat and azacitidine upregulate NOXA (PMAIP1) to increase sensitivity to venetoclax in preclinical models of acute myeloid leukemia by Dan Cojocari, Brianna N. Smith, Julie J. Purkal, Maria P. Arrate, Jason D. Huska, Yu Xiao, Agnieszka Gorska, Leah J. Hogdal, Haley E. Ramsey, Erwin R. Boghaert, Darren C. Phillips, and Michael R. Savona Haematologica 2021 [Epub ahead of print] Citation: Dan Cojocari, Brianna N. Smith, Julie J. Purkal, Maria P. Arrate, Jason D. Huska, Yu Xiao, Agnieszka Gorska, Leah J. Hogdal, Haley E. Ramsey, Erwin R. Boghaert, Darren C. Phillips and Michael R. Savona. Pevonedistat and azacitidine upregulate NOXA (PMAIP1) to increase sensitivity to venetoclax in preclinical models of acute myeloid leukemia. Haematologica. 2021; 106:xxx doi:10.3324/haematol.2020.272609 Publisher's Disclaimer. E-publishing ahead of print is increasingly important for the rapid dissemination of science. Haematologica is, therefore, E-publishing PDF files of an early version of manuscripts that have completed a regular peer review and have been accepted for publication. E-publishing of this PDF file has been approved by the authors. After having E-published Ahead of Print, manuscripts will then undergo technical and English editing, typesetting, proof correction and be presented for the authors' final approval; the final version of the manuscript will then appear in print on a regular issue of the journal. All legal disclaimers that apply to the journal also pertain to this production process. Pevonedistat and azacitidine upregulate NOXA (PMAIP1) to increase sensitivity to venetoclax in preclinical models of acute myeloid leukemia Authors: Dan Cojocari1*, Brianna N Smith2-4*, Julie J Purkal1, Maria P Arrate3, Jason D Huska1, Yu Xiao1, Agnieszka Gorska3, Leah J Hogdal5, Haley E.
    [Show full text]
  • Harnessing DNA Replication Stress for Novel Cancer Therapy
    G C A T T A C G G C A T genes Review Harnessing DNA Replication Stress for Novel Cancer Therapy 1, 2, 1 1,3, Huanbo Zhu y, Umang Swami y , Ranjan Preet and Jun Zhang * 1 Division of Medical Oncology, Department of Internal Medicine, University of Kansas Medical Center, 3901 Rainbow Blvd, Kansas City, KS 66160, USA; [email protected] (H.Z.); [email protected] (R.P.) 2 Division of Oncology, Department of Internal Medicine, Huntsman Cancer Institute, University of Utah, 2000 Circle of Hope, Salt Lake City, UT 84112, USA; [email protected] 3 Department of Cancer Biology, University of Kansas Cancer Center, 3901 Rainbow Blvd, Kansas City, KS 66160, USA * Correspondence: [email protected]; Tel.: +1-(913)-588-8150; Fax: +1-(913)-588-4085 H.Z. and U.S. contributed equally. y Received: 2 July 2020; Accepted: 20 August 2020; Published: 25 August 2020 Abstract: DNA replication is the fundamental process for accurate duplication and transfer of genetic information. Its fidelity is under constant stress from endogenous and exogenous factors which can cause perturbations that lead to DNA damage and defective replication. This can compromise genomic stability and integrity. Genomic instability is considered as one of the hallmarks of cancer. In normal cells, various checkpoints could either activate DNA repair or induce cell death/senescence. Cancer cells on the other hand potentiate DNA replicative stress, due to defective DNA damage repair mechanism and unchecked growth signaling. Though replicative stress can lead to mutagenesis and tumorigenesis, it can be harnessed paradoxically for cancer treatment.
    [Show full text]