CRISPR/Cas9 nickase mediated targeting of urokinase receptor gene inhibits neuroblastoma cell proliferation

Abstract

// Karina D. Rysenkova 1 , Ekaterina V. Semina 1, 2 , Maxim N. Karagyaur 3 , Anna A. Shmakova 1 , Daniyar T. Dyikanov 1 , Petr A. Vasiluev 1 , Yury P. Rubtsov 1, 4 , Kseniya A. Rubina 1 and Vsevolod A. Tkachuk 1, 2 1 Lomonosov Moscow State University, Faculty of Medicine, Laboratory of Gene and Cell Technologies, 119991, Moscow, Russian Federation 2 Federal State Budgetary Organization National Cardiology Research Center Ministry of Health of the Russian Federation, Institute of Experimental Cardiology, 121552, Moscow, Russian Federation 3 Institute of Regenerative Medicine, Lomonosov Moscow State University, 119991, Moscow, Russian Federation 4 Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry of the Russian Academy of Sciences, 117997, Moscow, Russian Federation Correspondence to: Ekaterina V. Semina, email: e-semina@yandex.ru Keywords: CRISPR/Cas9; genome editing; urokinase receptor; cell proliferation; neuroblastoma Received: January 29, 2018      Accepted: June 05, 2018      Published: June 29, 2018 ABSTRACT Neuroblastoma is a tumor arising from pluripotent sympathoadrenal precursor cells of neural cell origin. Neuroblastoma is one of the most aggressive childhood tumors with highly invasive and metastatic potential. The increased expression of urokinase and its receptor is often associated with a negative prognosis in neuroblastoma patients. We have shown that targeting of the Plaur gene in mouse neuroblastoma Neuro 2A cells by CRISPR/Cas9n results in ~60% decrease in cell proliferation (p<0.05), reduction in the number of Ki-67 positive cells, caspase 3 activation and PARP-1 cleavage. Knockout of uPAR leads to downregulation of mRNA encoding full-length TrkC receptor, which is involved in p38 MAPK and Akt signalling pathways. This finding provides a rationale to study a role of uPAR in neuroblastoma progression, since uPAR could be considered a potential therapeutic target in neuroblastoma treatment.