Login Register Cart 0
Generic placeholder image

CNS & Neurological Disorders - Drug Targets

Editor-in-Chief

ISSN (Print): 1871-5273
ISSN (Online): 1996-3181

Back
Journal Home About Journal Editorial Board Journal Insight Current Issue Volumes/Issues
Subscribe
Research Article

Dyngo-4a Induces Neuroblastoma Cell Differentiation Through The AKT and ERK1/2 Pathway

Author(s): Jinxi Huang*, Yi Zhou, Si Zeng, Jihong Xu, Lilian Liu, John Grothusen and Renyu Liu*

Volume 22, Issue 10, 2023

Published on: 04 January, 2023

Page: [1526 - 1534] Pages: 9

DOI: 10.2174/1871527322666221202145437

Price: $65

Abstract

Aim: The aim of the study is to check whether dyngo-4a can inhibit neuroblastoma (NB) proliferation and induce NB cell differentiation

Background: Dynamin plays a role in regulating neurotransmission, signaling pathways, nutrient uptake, and pathogen infection, enhancing cell proliferation, tumor invasion, and metastasis. Studies have reported that dyngo-4a, a dynamin inhibitor, can be used to identify potential biomarkers and promising novel therapeutic targets for cancer treatment.

Objective: To our knowledge, no published reports are showing that dynamin inhibitors can reduce NB cell proliferation and induce differentiation. In this study, we report that dyngo-4a can inhibit NB proliferation and induce NB cell differentiation.

Methods: In this study, mouse neuroblastoma (Neuro-2a) cells were cultured in the presence or absence of dyngo-4a or retinoic acid (RA), or in the presence of both dyngo-4a and RA, or in the presence of sequential administration of dyngo-4a and RA to compare the effects on the inhibition of cell proliferation and effects on neuroblastoma cell differentiation induction. The neural cell markers, Nestin and Tuj 1 (Neuron-specific class III beta-tubulin), were used to demonstrate that the differentiated cells have neuronal cell features. The phosphorylation of Protein Kinase B (AKT), extracellular signalregulated kinases1/2 (ERK1/2), and epidermal growth factor receptor (EGFR) were determined to examine the potential mechanisms of induced differentiation.

Results: Dyngo-4a or RA or dyngo-4a with subsequent RA administration induced Neuro-2a cell differentiation. However, RA with subsequent dyngo-4a administration results in almost total death of the Neuro-2a cells. The differentiation rate induced by dyngo-4a was significantly higher than the rate by RA treatment (72.5 ± 1.4% vs. 52.9 ± 3.1% with neuron features, P<0.05; 39.0 ± 0.8% vs. 29.9 ± 1.8% for axons under light microscopy, p<0.05). The differentiation rate of cells treated with dyngo-4a first, followed by RA, was greater than when they were added together (74.8 ± 3.8% vs. 10.6 ± 3.6%; 45.5 ± 1.6% vs. 12.4 ± 0.6%, p<0.01). Co-administration of dyngo-4a and RA at the same time diminished differentiation efficacy significantly. Dyngo-4a induced Neuro-2a cell differentiation and increased Tuj-1 positive staining by the 6th day post- treatment. Dyngo-4a also inhibited Neuro-2a cell proliferation in a dose-dependent manner. Regarding the mechanism, dyngo-4a treatment showed a significant increase in p-AKT and p-ERK1/2 but not in p-EGFR.

Conclusion: At a level comparable to RA, dynamin inhibition with dyngo-4a lowers proliferation and causes differentiation of Neuro-2a mouse NB cells in vitro. The AKT pathway is activated by dynago- 4a, which results in differentiation. The combination of RA with dynago-4a reduces the efficiency of differentiation. The application of dynago-4a followed by RA, on the other hand, enhances the differentiating effect, implying alternative mechanistic roles in the process.

Keywords: Dyngo-4a, cell proliferation, retinoic acid, pathways, AKT, ERK1/2 pathway.

« Previous Next »
Graphical Abstract

[1]
Evangelopoulos ME, Weis J, Krüttgen A. Signalling pathways leading to neuroblastoma differentiation after serum withdrawal: HDL blocks neuroblastoma differentiation by inhibition of EGFR. Oncogene 2005; 24(20): 3309-18.
[http://dx.doi.org/10.1038/sj.onc.1208494] [PMID: 15735700]
[2]
Dong R, Yang R, Zhan Y, et al. Single-cell characterization of malignant phenotypes and developmental trajectories of adrenal neuroblas-toma. Cancer Cell 2020; 38(5): 716-733.e6.
[http://dx.doi.org/10.1016/j.ccell.2020.08.014] [PMID: 32946775]
[3]
Zafar A, Wang W, Liu G, et al. Molecular targeting therapies for neuroblastoma: Progress and challenges. Med Res Rev 2021; 41(2): 961-1021.
[http://dx.doi.org/10.1002/med.21750] [PMID: 33155698]
[4]
Zafar A, Wang W, Liu G, et al. Targeting the p53-MDM2 pathway for neuroblastoma therapy: Rays of hope. Cancer Lett 2021; 496: 16-29.
[http://dx.doi.org/10.1016/j.canlet.2020.09.023] [PMID: 33007410]
[5]
Qiao J, Paul P, Lee S, et al. PI3K/AKT and ERK regulate retinoic acid-induced neuroblastoma cellular differentiation. Biochem Biophys Res Commun 2012; 424(3): 421-6.
[http://dx.doi.org/10.1016/j.bbrc.2012.06.125] [PMID: 22766505]
[6]
Kyeong Lee M, Nikodem VM. Differential role of ERK in cAMP-induced Nurr1 expression in N2A and C6 cells. Neuroreport 2004; 15(1): 99-102.
[http://dx.doi.org/10.1097/00001756-200401190-00020] [PMID: 15106839]
[7]
Clark O, Daga S, Stoker AW. Tyrosine phosphatase inhibitors combined with retinoic acid can enhance differentiation of neuroblastoma cells and trigger ERK- and AKT-dependent, p53-independent senescence. Cancer Lett 2013; 328(1): 44-54.
[http://dx.doi.org/10.1016/j.canlet.2012.09.014] [PMID: 23022267]
[8]
Reynolds CP, Matthay KK, Villablanca JG, Maurer BJ. Retinoid therapy of high-risk neuroblastoma. Cancer Lett 2003; 197(1-2): 185-92.
[http://dx.doi.org/10.1016/S0304-3835(03)00108-3] [PMID: 12880980]
[9]
Matthay KK, Reynolds CP, Seeger RC, et al. Long-term results for children with high-risk neuroblastoma treated on a randomized trial of myeloablative therapy followed by 13-cis-retinoic acid: A children’s oncology group study. J Clin Oncol 2009; 27(7): 1007-13.
[http://dx.doi.org/10.1200/JCO.2007.13.8925] [PMID: 19171716]
[10]
Preis PN, Saya H, Nádasdi L, Hochhaus G, Levin V, Sadée W. Neuronal cell differentiation of human neuroblastoma cells by retinoic acid plus herbimycin A. Cancer Res 1988; 48(22): 6530-4.
[PMID: 2846152]
[11]
Abazeed ME, Blanchette JM, Fuller RS. Cell-free transport from the trans-golgi network to late endosome requires factors involved in formation and consumption of clathrin-coated vesicles. J Biol Chem 2005; 280(6): 4442-50.
[http://dx.doi.org/10.1074/jbc.M412553200] [PMID: 15572353]
[12]
Cook T, Mesa K, Urrutia R. Three dynamin-encoding genes are differentially expressed in developing rat brain. J Neurochem 1996; 67(3): 927-31.
[http://dx.doi.org/10.1046/j.1471-4159.1996.67030927.x] [PMID: 8752097]
[13]
Meng J. Distinct functions of dynamin isoforms in tumorigenesis and their potential as therapeutic targets in cancer. Oncotarget 2017; 8(25): 41701-16.
[http://dx.doi.org/10.18632/oncotarget.16678] [PMID: 28402939]
[14]
Gundu C, Arruri VK, Yadav P, et al. Dynamin-independent mechanisms of endocytosis and receptor trafficking. Cells 2022; 11(16): 2557.
[http://dx.doi.org/10.3390/cells11162557] [PMID: 36010634]
[15]
Dutta D, Donaldson JG. Search for inhibitors of endocytosis. Cell Logist 2012; 2(4): 203-8.
[http://dx.doi.org/10.4161/cl.23967] [PMID: 23538558]
[16]
Park R, Shen H, Liu L, Liu X, Ferguson SM, De Camilli P. Dynamin triple knockout cells reveal off target effects of commonly used dynamin inhibitors. J Cell Sci 2013; 126(Pt 22): jcs.138578.
[http://dx.doi.org/10.1242/jcs.138578] [PMID: 24046449]
[17]
Yamada H, Abe T, Li SA, et al. N′-[4-(dipropylamino)benz-ylidene]-2-hydroxybenzohydrazide is a dynamin GTPase inhibitor that sup-presses cancer cell migration and invasion by inhibiting actin polymerization. Biochem Biophys Res Commun 2014; 443(2): 511-7.
[http://dx.doi.org/10.1016/j.bbrc.2013.11.118] [PMID: 24316215]
[18]
Yamada H, Abe T, Li SA, et al. Dynasore, a dynamin inhibitor, suppresses lamellipodia formation and cancer cell invasion by destabi-lizing actin filaments. Biochem Biophys Res Commun 2009; 390(4): 1142-8.
[http://dx.doi.org/10.1016/j.bbrc.2009.10.105] [PMID: 19857461]
[19]
Eleniste PP, Huang S, Wayakanon K, Largura HW, Bruzzaniti A. Osteoblast differentiation and migration are regulated by dynamin GTPase activity. Int J Biochem Cell Biol 2014; 46: 9-18.
[http://dx.doi.org/10.1016/j.biocel.2013.10.008] [PMID: 24387844]
[20]
Macia E, Ehrlich M, Massol R, Boucrot E, Brunner C, Kirchhausen T. Dynasore, a cell-permeable inhibitor of dynamin. Dev Cell 2006; 10(6): 839-50.
[http://dx.doi.org/10.1016/j.devcel.2006.04.002] [PMID: 16740485]
[21]
Eschenburg S, Reubold TF. Modulation of dynamin function by small molecules. Biol Chem 2018; 399(12): 1421-32.
[http://dx.doi.org/10.1515/hsz-2018-0257] [PMID: 30067507]
[22]
Lima AR, Santos L, Correia M, et al. Dynamin-Related Protein 1 at the Crossroads of Cancer. Genes 2018; 9(2): 115.
[http://dx.doi.org/10.3390/genes9020115] [PMID: 29466320]
[23]
Preta G, Cronin JG, Sheldon IM. Dynasore - not just a dynamin inhibitor. Cell Commun Signal 2015; 13(1): 24.
[http://dx.doi.org/10.1186/s12964-015-0102-1] [PMID: 25889964]
[24]
McCluskey A, Daniel JA, Hadzic G, et al. Building a better dynasore: The dyngo compounds potently inhibit dynamin and endocytosis. Traffic 2013; 14(12): 1272-89.
[http://dx.doi.org/10.1111/tra.12119] [PMID: 24025110]
[25]
Wu PY, Lin YC, Chang CL, et al. Functional decreases in P2X7 receptors are associated with retinoic acid-induced neuronal differentia-tion of Neuro-2a neuroblastoma cells. Cell Signal 2009; 21(6): 881-91.
[http://dx.doi.org/10.1016/j.cellsig.2009.01.036] [PMID: 19385050]
[26]
Hynds DL, Spencer ML, Andres DA, Snow DM. Rit promotes MEK-independent neurite branching in human neuroblastoma cells. J Cell Sci 2003; 116(10): 1925-35.
[http://dx.doi.org/10.1242/jcs.00401] [PMID: 12668729]
[27]
Eom HS, Park HR, Jo SK, Kim YS, Moon C, Jung U. Ionizing radiation induces neuronal differentiation of Neuro-2a cells via PI3-kinase and p53-dependent pathways. Int J Radiat Biol 2015; 91(7): 585-95.
[http://dx.doi.org/10.3109/09553002.2015.1029595] [PMID: 25912236]
[28]
Min JY, Park MH, Park MK, et al. Staurosporin induces neurite outgrowth through ROS generation in HN33 hippocampal cell lines. J Neural Transm 2006; 113(11): 1821-6.
[http://dx.doi.org/10.1007/s00702-006-0500-z] [PMID: 16715208]
[29]
Singh RK. Key Heterocyclic Cores for Smart Anticancer Drug–Design Part II. Bentham Science Publishers 2022.
[http://dx.doi.org/10.2174/97898150400741220101]
[30]
Singh RK, Prasad DN, Bhardwaj TR. Design, synthesis and evaluation of aminobenzophenone derivatives containing nitrogen mustard moiety as potential central nervous system antitumor agent. Med Chem Res 2013; 22(12): 5901-11.
[http://dx.doi.org/10.1007/s00044-013-0582-8]
[31]
Singh RK, Prasad DN, Bhardwaj TR. Design, synthesis, chemical and biological evaluation of brain targeted alkylating agent using re-versible redox prodrug approach. Arab J Chem 2017; 10(3): 420-9.
[http://dx.doi.org/10.1016/j.arabjc.2013.12.008]
[32]
Singh RK, Kumar S, Prasad DN, Bhardwaj TR. Therapeutic journery of nitrogen mustard as alkylating anticancer agents: Historic to fu-ture perspectives. Eur J Med Chem 2018; 151: 401-33.
[http://dx.doi.org/10.1016/j.ejmech.2018.04.001] [PMID: 29649739]
[33]
Singh RK, Ed. Protein Kinases - Promising Targets for Anticancer Drug Research. IntechOpen 2021.
[http://dx.doi.org/10.5772/intechopen.82939]
[34]
Nakata H. Mitogen-activated protein kinase signaling is involved in suramin-induced neurite outgrowth in a neuronal cell line. Biochem Biophys Res Commun 2007; 355(3): 842-8.
[http://dx.doi.org/10.1016/j.bbrc.2007.02.045] [PMID: 17321499]
[35]
Miloso M, Villa D, Crimi M, et al. Retinoic acid‐induced neuritogenesis of human neuroblastoma SH‐SY5Y cells is ERK independent and PKC dependent. J Neurosci Res 2004; 75(2): 241-52.
[http://dx.doi.org/10.1002/jnr.10848] [PMID: 14705145]
[36]
Rössler J, Geoerger B, Taylor M, Vassal G. Small molecule tyrosine kinase inhibitors: Potential role in pediatric malignant solid tumors. Curr Cancer Drug Targets 2008; 8(1): 76-85.
[http://dx.doi.org/10.2174/156800908783497113] [PMID: 18288945]
[37]
Sigismund S, Argenzio E, Tosoni D, Cavallaro E, Polo S, Di Fiore PP. Clathrin-mediated internalization is essential for sustained EGFR signaling but dispensable for degradation. Dev Cell 2008; 15(2): 209-19.
[http://dx.doi.org/10.1016/j.devcel.2008.06.012] [PMID: 18694561]
[38]
Jo U, Park KH, Whang YM, et al. EGFR endocytosis is a novel therapeutic target in lung cancer with wild-type EGFR. Oncotarget 2014; 5(5): 1265-78.
[http://dx.doi.org/10.18632/oncotarget.1711] [PMID: 24658031]
[39]
Cheung YT, Lau WKW, Yu MS, et al. Effects of all-trans-retinoic acid on human SH-SY5Y neuroblastoma as in vitro model in neuro-toxicity research. Neurotoxicology 2009; 30(1): 127-35.
[http://dx.doi.org/10.1016/j.neuro.2008.11.001] [PMID: 19056420]
[40]
Nuttall JR, Oteiza PI. Zinc and the ERK kinases in the developing brain. Neurotox Res 2012; 21(1): 128-41.
[http://dx.doi.org/10.1007/s12640-011-9291-6] [PMID: 22095091]
[41]
Pucilowska J, Puzerey PA, Karlo JC, Galán RF, Landreth GE. Disrupted ERK signaling during cortical development leads to abnormal progenitor proliferation, neuronal and network excitability and behavior, modeling human neuro-cardio-facial-cutaneous and related syndromes. J Neurosci 2012; 32(25): 8663-77.
[http://dx.doi.org/10.1523/JNEUROSCI.1107-12.2012] [PMID: 22723706]
[42]
Feng X, Yu W, Liang R, Shi C, Zhao Z, Guo J. Receptor-interacting protein 140 overexpression promotes neuro-2a neuronal differentia-tion by ERK1/2 signaling. Chin Med J 2015; 128(1): 119-24.
[http://dx.doi.org/10.4103/0366-6999.147850] [PMID: 25563324]
[43]
Han NLR, Wen J, Lin Q, Tan PL, Liou YC, Sheu FS. Proteomics analysis of the expression of neurogranin in murine neuroblastoma (Neuro-2a) cells reveals its involvement for cell differentiation. Int J Biol Sci 2007; 3(5): 263-73.
[http://dx.doi.org/10.7150/ijbs.3.263] [PMID: 17505539]
[44]
Sousa LP, Lax I, Shen H, Ferguson SM, Camilli PD, Schlessinger J. Suppression of EGFR endocytosis by dynamin depletion reveals that EGFR signaling occurs primarily at the plasma membrane. Proc Natl Acad Sci 2012; 109(12): 4419-24.
[http://dx.doi.org/10.1073/pnas.1200164109] [PMID: 22371560]
[45]
Izycka-Swieszewska E, Brzeskwiniewicz M, Wozniak A, et al. EGFR, PIK3CA and PTEN gene status and their protein product expres-sion in neuroblastic tumours. Folia Neuropathol 2010; 48(4): 238-45.
[PMID: 21225506]

Rights & Permissions Print Cite
Article Metrics
33
3
© 2024 Bentham Science Publishers | Privacy Policy