Co-delivery of GOLPH3 siRNA and gefitinib by cationic lipid-PLGA nanoparticles improves EGFR-targeted therapy for glioma
Glioblastoma is one of the most aggressive types of brain tumor. Epidermal growth factor receptors (EGFRs) are overexpressed in glioma, and EGFR amplifications and mutations lead to rapid proliferation and invasion. EGFR-targeted therapy might be an effective treatment for glioma. Gefitinib (Ge) is an EGFR tyrosine kinase inhibitor (TKI), and Golgi phosphoprotein 3 (GOLPH3) expression is associated with worse glioma prognosis. Downregulation of GOLPH3 could promote EGFR degradation. Here, an angiopep-2 (A2)-modified cationic lipid-poly (lactic-co-glycolic acid) (PLGA) nanoparticle (A2-N) was developed that can release Ge and GOLPH3 siRNA (siGOLPH3) upon entering glioma cells and therefore acts as a combinatorial anti-tumor therapy. The in vitro and in vivo studies proved that A2-N/Ge/siGOLPH3 successfully crossed the blood-brain barrier (BBB) and targeted glioma. Released siGOLPH3 effectively silenced GOLPH3 mRNA expression and further promoted EGFR and p-EGFR degradation. Released Ge also markedly inhibited EGFR signaling. This combined EGFR-targeted action achieved remarkable anti-glioma effects and could be a safe and effective treatment for glioma.
Angiopep-2-modified cationic lipid polymer can penetrate the BBB.
Gefitinib can inhibit EGFR signaling and block the autophosphorylation of critical tyrosine residues on EGFR.
GOLPH3 siRNA can be transfected into glioma and downregulate GLOPH3 expression.
A2-N/Ge/siGOLPH3 can inhibit glioma growth.
KeywordsGlioma Nanoparticles EGFR GOLPH3 Gefitinib
confocal laser scan microscopy
4′, 6-diamidino-2-phenylindole dihydrochloride
Dulbecco’s Modified Eagle’s Medium
epidermal growth factor receptor
epidermal growth factor receptor variant typeIII
enhanced permeability and retention
fetal bovine serum
golgi phosphoprotein 3
low-density lipoprotein receptor-related protein-1
phosphate buffer saline
poly (D, L-lactic-co-glycolic acid)
quantitative real-time polymerase chain reaction
receptor mediated transcytosis
small interfering RNA
The Cancer Genome Atlas
transmission electron microscopy
tyrosine kinase inhibitors
This work was financially supported by National Natural Science Foundation of China (Grant No. 81772665), Jiangsu Province, Key Research & Development Plan of Jiangsu Province (No. BE2016646), Jiangsu provincial Commission of Health and Family Planning (Grant No. Q201608), Postgraduate Research & Practice Innovation Program of Jiangsu Province (Grant No. KYCX18_2197), and Six Talents Peak Foundation of Jiangsu Province (No. 2018-WSW-071).
Compliance with ethical standards
Ethics approval and consent to participate
This study was performed according to the guidelines for the Care and Use of Laboratory Animals and the animal experimental protocols were approved by Xuzhou Medical University of China Animal Care and Use Committee.
The authors declare that they have no competing interests.
- 13.Sirisoma N, Pervin A, Zhang H, Jiang S, Willardsen JA, Anderson MB, Mather G, Pleiman CM, Kasibhatla S, Tseng B, Drewe J, Cai SX (2009) Discovery of N-(4-Methoxyphenyl)-N,2-dimethylquinazolin-4-amine, a potent apoptosis inducer and efficacious anticancer agent with high blood brain barrier penetration. J Med Chem. 52(8):2341–2351PubMedGoogle Scholar
- 16.Zhou XP, Xie S, Wu SS, Qi YH, Wang ZH, Zhang H, Lu D, Wang X, Dong Y, Liu G, Yang D, Shi Q, Bian W, Yu R (2017) Golgi phosphoprotein 3 promotes glioma progression via inhibiting Rab5-mediated endocytosis and degradation of epidermal growth factor receptor. Neuro-Oncology. 19(12):1628–1639PubMedPubMedCentralGoogle Scholar
- 22.Ni D, Zhang J, Bu W, Xing H, Han F, Xiao Q, Yao Z, Chen F, He Q, Liu J, Zhang S, Fan W, Zhou L, Peng W, Shi J (2014) Dual-targeting upconversion nanoprobes across the blood-brain barrier for magnetic resonance/fluorescence imaging of intracranial glioblastoma. ACS Nano. 8(2):1231–1242PubMedGoogle Scholar
- 26.Xu R, Ji JX, Zhang X, Han MZ, Zhang C, Xu YY, Wei Y, Wang S, Huang B, Chen A, Zhang D, Zhang Q, Li W, Jiang Z, Wang J, Li X (2017) PDGFA/PDGFRα-regulated GOLM1 promotes human glioma progression through activation of AKT. Journal of Experimental & Clinical Cancer Research. 36(1):193Google Scholar
- 27.Brennan CW, Verhaak RGW, McKenna A, Campos B, Noushmehr H, Salama SR et al (2014) The somatic genomic landscape of glioblastoma. Cell. 155(2):462–477Google Scholar
- 28.Inda MD, Bonavia R, Mukasa A, Narita Y, Sah DWY, Vandenberg S, Brennan C, Johns TG, Bachoo R, Hadwiger P, Tan P, Depinho RA, Cavenee W, Furnari F (2010) Tumor heterogeneity is an active process maintained by a mutant EGFR-induced cytokine circuit in glioblastoma. Genes & Development. 24(16):1731–1745Google Scholar
- 29.De Vries NA, Buckle T, Zhao J, Beijnen JH, Schellens JHM, van Tellingen O (2012) Restricted brain penetration of the tyrosine kinase inhibitor erlotinib due to the drug transporters P-gp and BCRP. Invest New Drug. 30(2):443–449Google Scholar
- 31.Chakravarti A, Wang M, Robins HI, Lautenschlaeger T, Curran WJ, Brachman DG et al (2013) RTOG 0211: A phase 1/2 study of radiation therapy with concurrent gefitinib for newly diagnosed glioblastoma patients. Int J Radiat Oncol. 85(5):1206–1211Google Scholar
- 32.Clarke JL, Molinaro AM, Phillips JJ, Butowski NA, Chang SM, Perry A, Costello JF, DeSilva A, Rabbitt JE, Prados MD (2014) A single-institution phase II trial of radiation, temozolomide, erlotinib, and bevacizumab for initial treatment of glioblastoma. Neuro-Oncology. 16(7):984–990PubMedPubMedCentralGoogle Scholar
- 33.Reardon DA, Nabors LB, Mason WP, Perry JR, Shapiro W, Kavan P, Mathieu D, Phuphanich S, Cseh A, Fu Y, Cong J, Wind S, Eisenstat DD, BI 1200 36 Trial Group and the Canadian Brain Tumour Consortium. Phase I/randomized phase II study of afatinib, an irreversible ErbB family blocker, with or without protracted temozolomide in adults with recurrent glioblastoma. Neuro-Oncology. 2015; 17 (3): 430- 439.Google Scholar
- 34.Hegi ME, Diserens AC, Bady P, Kamoshima Y, Kouwenhoven MCM, Delorenzi M, Lambiv WL, Hamou MF, Matter MS, Koch A, Heppner FL, Yonekawa Y, Merlo A, Frei K, Mariani L, Hofer S (2011) Pathway analysis of glioblastoma tissue after preoperative treatment with the EGFR tyrosine kinase inhibitor Gefitinib-A phase II trial. Molecular Cancer Therapeutics. 10(6):1102–1112PubMedGoogle Scholar
- 36.Shi YJ, Su C, Cui WY, Li HD, Liu LW, Feng B et al (2014) Gefitinib loaded folate decorated bovine serum albumin conjugated carboxymethyl-beta-cyclodextrin nanoparticles enhance drug delivery and attenuate autophagy in folate receptor-positive cancer cells. Journal of Nanobiotechnology. 12:43PubMedPubMedCentralGoogle Scholar
- 43.Bertrand Y, Currie JC, Poirier J, Demeule M, Abulrob A, Fatehi D, Stanimirovic D, Sartelet H, Castaigne JP, Béliveau R (2011) Influence of glioma tumor microenvironment on the transport of ANG1005 via low-density lipoprotein receptor-related protein 1. Br J Cancer. 105(11):1697–1707PubMedPubMedCentralGoogle Scholar
- 50.Wick W, Gorlia T, Bady P, Platten M, van den Bent MJ, Taphoorn MJB, Steuve J, Brandes AA, Hamou MF, Wick A, Kosch M, Weller M, Stupp R, Roth P, Golfinopoulos V, Frenel JS, Campone M, Ricard D, Marosi C, Villa S, Weyerbrock A, Hopkins K, Homicsko K, Lhermitte B, Pesce G, Hegi ME (2016) Phase II study of radiotherapy and temsirolimus versus radiochemotherapy with temozolomide in patients with newly diagnosed glioblastoma without MGMT promoter hypermethylation (EORTC 26082). Clinical Cancer Research. 22(19):4797–4806PubMedGoogle Scholar